Baseline Type 2 Biomarker Levels and Clinical Remission Predictors in Children with Asthma

Baseline Type 2 Biomarker Levels and Clinical Remission Predictors in Children with Asthma | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Baseline Type 2 Biomarker Levels and Clinical Remission Predictors in Children with Asthma Mengmeng Chen, Congcong Li, Qiuyan Yang, Huijie Zhang, Yanli Zhang, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4182358/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Background Little study investigates the association between baseline type 2 biomarker levels and clinical features in children diagnosed with asthma. Characterizing clinical remission in Th2-high asthma could offer valuable insights into asthma prognosis. Objectives The study aims to investigate the association of baseline type 2 biomarker levels and clinical features in children with asthma, and to identify predictors of clinical remission of asthma in children. Methods A total of 172 children with baseline age of 6.87 ± 3.04 (mean ± SD) years were enrolled in the study including 119 with physician-diagnosed asthma who regularly attended a pediatric asthma center and 53 control subjects with no respiratory symptoms. Clinical tests included lung function examination, Fraction of exhaled NO (FeNO), total IgE, blood eosinophil, and skin test. Serum Th2 biomarkers were examined by ELISA. The enrolled patients have readjusted into Th2-high asthma according to clinical eosinophil count and total IgE, and Th2-high asthma subjects were further classified into acute attack asthma, persistent asthma, and clinical remission according to the recent GINA guidelines and clinical evaluation. To verify our results, the concentration of TSLP levels was measured in BALF, serum, and lung tissue by ELISA in mouse models. Results Compared with asthma and control groups, eosinophil counts and blood eosinophils (%) were significant, whereas, no correlation was observed between asthma subjects and controls including Th2 biomarkers, gender, or ages. Positive correlations were observed between Th2 inflammatory biomarkers (TSLP, TRAC IL-5, IL-13, and Periostin) at baseline. Th2-high asthma (n = 110) was defined based on clinical measurement of IgE > 100 IU/ml and a blood eosinophil count ≥ 140 cells/µl. Among those Th2-high asthma subjects, there were 48 in acute exacerbation (43.6%), and 36 in clinical remission (32.7%), 26 were clinical asthma persistence (23.6%). Lung function and serum TSLP had marked significance among the three categories. Compared with clinical remission asthmatic subjects and controls, serum TSLP levels were significantly higher in subjects experiencing acute exacerbation and subjects defined as asthma persistence. Spearman’s correlation outlined that serum TSLP levels were related to Total IgE (IU/mL), FEV1/FVC ratio, and FEF25-75, pred %. Multivariate logistic regression analysis demonstrated that serum TSLP levels were associated with clinical remission in Th2-high asthma children (OR = 1.009; 95% CI, 1.0087–1.0086 P = 0.023<0.05). It is also revealed that serum TSLP levels may help evaluate clinical remission in Th2-high asthma when using ROC curves analysis (AUC = 0.5887, 95% CI: 0.5052 to 0.7038, P < 0.05). A cutoff value of 373.363 pg/mL was found with the highest clinical sensitivity and specificity. PPV, and NPV were 100%, 39.1%, and 59.4%, 100% in two groups, respectively. However, there were no positive results in the analysis of multivariable logistic regression in determining the predictors of persistence in Th2-high asthma. In BALF mouse, TSLP concentration had no statistically significant change in the acute and remission stages when compared to the control, but it did increase noticeably in the chronic stage ( P < 0.001). Asthma Th2 Biomarkers Thymic stromal lymphopoietin Clinical asthma remission Asthma persistence Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 1 Introduction Asthma is one of the most common inflammatory airway diseases of childhood associated with significant heterogeneity, which refers to different asthma phenotypes having unique clinical and biomarker profiles [ 1 , 2 ]. The 2022 Global Initiative for Asthma (GINA) guidelines recommend using clinical characteristics and biomarkers to define asthma inflammatory phenotypes that may help physicians move toward more personalized management of asthma and benefit patients [ 3 ]. And type-2-high asthma which relates to inflammatory type presents an example of current asthma phenotypes (type 2-high or type 2-low), characterized by increased secretion of interleukin IL-4, IL-5, and IL-13 by Type 2 innate lymphoid cells and T-helper 2 cells [ 4 ]. The majority of asthma patients have evidence of type 2 (T2) inflammation, which also features high blood eosinophil counts and fractional exhaled nitric oxide (FeNO) levels. The Type 2 biomarkers involved in underlying airway inflammation are well-researched in the area. However, there are limited data when it comes to using biomarkers to predict the clinical trajectory of childhood asthma. Therefore, the aim of the study was to investigate the association of Th2 biomarker levels with clinical features in different phases of asthma in children and try to identify biomarkers whose level is expected to differ among those with persistent asthma, acute exacerbation asthma, or clinical asthma remission in cohorts, which may provide insight into clinical asthma trajectory. So, we determined the association of Th2 biomarkers with baseline characteristics and found out one of the Th2 biomarkers TSLP may predict asthma remission in a cohort of Th2 high asthma subjects which were divided into acute exacerbation asthma, persistent asthma, and clinical remission asthma. As reported, Thymic stromal lymphopoietin (TSLP) is an epithelial cell-derived cytokine that is implicated in the initiation and persistence of airway inflammation in asthma [ 5 – 7 ]. Based on a cross-sectional baseline of characteristics obtained during a medical visit along with specific serum biomarkers, we may make a close step toward individualized prognostication. 2 Methods 2.1.1 Study Design and population We enrolled 172 children from a clinic pediatric asthma center, among them, 119 subjects were diagnosed with asthma by doctors on the basis of the updated version of the 2022 statement by the Global Initiative for Asthma (GINA), and 53 were age- and sex-matched control subjects without respiratory symptoms. Th2-high asthma was defined based on clinical measurement of IgE > 100 IU/ml and blood eosinophil counts ≥ 140 cells/µl [ 8 , 9 ]. Th2 high asthma subjects (n = 110) were allocated into three groups (acute attack asthma, persistent asthma, and clinical remission). Those who presented with worsening wheezing, shortness of breath, cough, and chest tightness, and were diagnosed as acute asthma attacks by physicians were defined as asthma cases in acute exacerbation(n = 48) [ 3 , 10 ]. Asthma cases in clinical remission(n = 36) are characterized by an absence of asthma symptoms and medicine use for at least ≥ 1 year additionally without acute asthma attack within a year [ 11 , 12 ]. Persistent asthma(n = 26)was defined as those with the presence of asthma symptoms in the last year or use of long-term control medications daily to achieve and maintain control in the last year [ 13 , 14 ]. This trial was conducted under the ethical principles of the Declaration of Helsinki, the International Council for Harmonisation guidelines for good clinical practice, and applicable regulatory requirements. The protocol (2021-108-01) was initially approved by the ethics committee of the Third Affiliated Hospital of Zhengzhou University. All participants and their parents granted their written informed consent and all experiments were performed per the approved guidelines and regulations. 2.1.2 MICE BALB/c mouse, female, SPF grade, weight 18 + 2g, 6–8 weeks old, were purchased from Shanghai Jiesjie Laboratory Animal Co., LTD. They were fed in an SPF environment with free access to sterilized water and food, controlled with a 12-hour daytime to 12-hour night cycle, indoor temperature of 20–30 ℃, and humidity of 50%. After acclimation for 1 week, the mice were randomly divided into 4 groups (control, acute asthma, chronic asthma, and asthma remission),6 in each group. Mice in the control group were given 0.2mL normal saline intraperitoneal injection, and other asthma model groups were all intraperitoneally injected with 0.2mL suspension which is complexed 20µg of OVA (GradeV) with 2mg Aluminum hydroxide on days 0, 7, and 14 to induce asthma. To mimic different stagings of human asthma, OVA-sensitization mouse asthma models undergoing different durations of OVA-inhaled stimulation. From day 21, mice in the acute asthma group were challenged sensitization by ultrasonic atomizing inhalation of 3% OVA (Grade II) solution in the closed container for 30 minutes each day, lasting for 7 days. The same atomization condition applied for the chronic asthma model group was 3 times a week, continuing for 6 weeks. And the remission group was atomized once a month for 2 months. After the ceases of OVA inhalation exposure, all the experimental groups were determined for airway hyperresponsiveness (AHR) using different concentrations of methacholine. Type 2 diagnostic inflammation biomarkers were detected including eosinophils, specific-IgE, and Th2 cytokines (IL-4, IL5, and IL-13), both in blood and BALF. An overview of the results showed that the three murine models of asthma were successful. Compared with the control group, the other three groups were found airway methacholine hyperresponsiveness. Higher IgE and Th2 cytokines were detected in serum and BALF, and also more eosinophil infiltration in the lungs. All animal experiments are approved by the Laboratory Animal Ethics Committee (IACUC) of Fudan University and are conducted in strict accordance with the laboratory animal management regulations of Fudan University (2022JS Huashan hospital-497). 2.2 Study measurements 2.2.1 Clinical information collection All subjects' holistic history including signs and symptoms of asthma, medication use, and common triggers observed by parents are collected by well-trained clinicians. Height and weight were measured and body mass index (BMI) was calculated as an index of the physique. 2.2.2 Lung function tests Asthma-related pulmonary function was measured according to international standards. The forced expiratory volume in one second (FEV1) and forced vital capacity (FVC) were calculated using the formula of the American Thoracic Society/European Respiratory Society (ATS/ERS) guidelines [ 15 ]. FEF25-75 pred % were also recorded [ 16 ]. 2.3 Inflammatory parameters The fraction of exhaled nitric oxide was measured at a flow rate of 50 ml/s according to the recommended standards (SV-02, Wuxi Shangwo, China) [ 17 ]. Differential white blood cell counts were performed using the automatic blood cell count with a full automatic hemocyte analyzer (BC-5800, Mindary, Shenzhen, China). Serum total IgE was measured in an automatic biochemical analyzer (AU5400, Beckman Coulter Inc, Calif, USA) and related reagents were from Diasys diagnostic systems (Diasys, Catalog No: 17239). 2.4 Skin tests Skin prick testing (SPT) for common aeroallergens such as pteronyssinus and Dermatophagoides farina was measured among children. Weed and tree pollen mix was also included, as well as food allergens, molds, and animal dander. Histamine (10 mg/mL of histamine phosphate) and 0.9% saline were used as positive and negative controls, respectively. The positive reaction was a wheal observed 3 mm greater than the negative control [ 18 ]. 2.5 Serum concentrations of biomarkers and cytokines Venous blood samples were collected with a vacuum blood tube and centrifuged at 3000 g for 10 mins. Then the serum was isolated and stored in aliquots at − 80°C until use. Levels of serum IL-5, IL-13, IgE, IL-4, Periostin, TARC, and TSLP were measured by using specific human enzyme-linked immunosorbent assay kits (Shanghai Wellbio Technology Co., Ltd, No: EH6305M, EH6261M, EH6244M, EH6304M, EH6393M, EH6079M, EH6521M), according to the manufacturer’s instructions. Each sample was tested in duplicate. 2.6 Measurement of mouse TSLP by ELISA in serum, BALF, and lung tissue After the pulmonary function measurement, the mice were exsanguinated by eyeball enucleation. Their sera were separated and stored in aliquots at − 80°C until use. Lung lavage fluid was collected using a 1mL syringe that was connected to an indentation needle inserted into the trachea. Next, 0.2mL of pre-cooled PBS solution was injected into the trachea to irrigate the left lung and was quickly withdrawn. This process was repeated three times to ensure proper lavage of the lung. Afterward, the BALF was centrifuged at 4°C for 15 minutes, and the supernatant was collected and frozen in the − 80°C refrigerator for subsequent detection. Lung tissue was divided into different sections in need and stored in the − 80°C refrigerator for use. The concentration of mouse TSLP in serum, BALF, and lung tissue was tested by indirect ELISA, using a Mouse TSLP ELISA kit (Shanghai Wellbio Technology Co., Ltd, No: EM30546M). All experiment was performed according to the manufacturer’s instructions. 2.7 Statistical analysis Statistical analyses were performed using SPSS v 26.0 (IBM, Chicago, IL, USA) and GraphPad Prism 9 software (GraphPad Software Inc. La Jolla, CA, USA). Normally distributed continuous data were expressed as mean and standard deviation, and non-normally distributed continuous data as median and inter-quartile ranges (IQR). Basic between-group comparisons were made using Mann-Whitney tests, Student´s t-test, and multiple between-group comparisons were performed using Kruskal-Wallis tests with Dunn’s correction for multiple testing and ANOVA. For categorical variables, the chi-square test or Fisher's test was conducted. The correlation coefficients among Th2 biomarker levels and clinical characteristics were analyzed using Spearman's rank correlation coefficient. The association of clinical remission asthma classes with clinical variables was examined using regression models adjusted for potential confounders, including FEV1 (% predicted), FEV1/FVC ratio, FEF25-75, pred %, Total IgE (IU/mL), TSLP (pg/ml). The odds ratio (OR) and 95% confidence interval (CI) were reported. The diagnostic performance of serum TSLP levels to identify predicting biomarkers of clinical remission asthma in children was determined by receiver operating characteristic (ROC) curve analysis. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated for the selected cut-off point. A P level < 0.05 was considered statistically significant as shown in Fig. 1 . 3 Results 3.1 Baseline characteristics In total, 172 children were enrolled, among those 119 were diagnosed with asthma by the clinical physician or still experiencing acute asthma attacks, and 53 were control subjects with no respiratory disease. Baseline demographics, clinical characteristics, and levels of T2 biomarkers of the overall population are reported in Table 1 . There were no significant differences between the children with asthma and control groups regarding age and sex, while children with asthma had significantly higher blood eosinophil counts than controls ( P = 0.014). As for the Th2 biomarkers(IL-4 P = 0.142, IL-5 P = 0.4791, IL-13 P = 0.6885, Serum IgE P = 0.2534, TARC P = 0.2074, TSLP P = 0.6571), there are no significant statistical differences observed between asthma and control subjects. Table1: Demographic and laboratory characteristics of the study groups (n = 172). Asthma n=119 Control n=53 P value Sex(M/F) (70/49) (31/22) 0.967 Age（years） 6.7±3.1 7.2±2.9 0.2048 Eosinophils count/l 0.26（0.13-1.7） 0.16（0.1-0.7） 0.0413 Blood eosinophils (%) 2.1 (1.3-3.225) 3.35 (1.5-5.75) 0.014 IL-4（pg/ml) Mean±SD Median（min-max） 50.85±22.11 48.56（15.08-175.7） 54.26±17.94 53.85（17.06-107.1） 0.142 IL-5（pg/ml) Mean±SD Median（min-max） 246.3±114.9 238.8（52.94-890.5） 241.7±102.8 229.8（78.38-597.5） 0.4791 IL-13（pg/ml) Mean±SD Median（min-max） 174.3±70.92 178.8（50.74-335.8） 165.5±56.48 172.9±（39.75-263.9） 0.6885 Serum IgE（ng/ml） Mean±SD Median（min-max） 31.93±20.11 27.21（7.345-109.2） 29.32±20.42 22.15（7.832-104.3） 0.2534 Periostin（ng/ml） Mean±SD Median（min-max） 1.169±0.7573 1.002（0.2556-4.772） 1.275±0.7091 1.193（0.07387-3.106） 0.2074 TARC（pg/ml) Mean±SD Median（min-max） 251.7±93.31 247.8（54.47-533.5） 265.9±81.19 265.3（48.53-426.8） 0.2159 TSLP（pg/ml) Mean±SD Median（min-max） 374.2±285.7 297.2（72.38-1400） 356.6±285.1 250.2（86.81-1589） 0.6571 Table 1 : Data are presented as percentage, median (inter-quartile range), or mean ± standard deviation. Values in bold are statistically significant.Abbreviations: IgE, immunoglobulin E; IL, interleukin; SD, standard deviation; TARC, thymus, and activation-regulated chemokine; TSLP, thymic stromal lymphopoietin. 3.2 Baseline correlations between biomarkers At baseline, the level of serum TSLP has positive correlations with TRAC (r = 0.5), IL-4 (r = 0.2), IL-5 (r = 0.6), IL-13(r = 0.3), and Periostin (r = 0.3) ( P < 0 .005 in all) (Fig. 2 A–E). Positive correlations were also observed at baseline for IgE with IL-5 (r = 0.4), TRAC (r = 0.3), and IL-13 (r = 0.4) ( P < 0 .0001 in all) (Fig. 2 F–H). There were also positive correlations between TRAC and IL-5 (r = 0.5), and IL-13 (r = 0 .4), as well as between TRAC and Periostin (r = 0.5) ( P < 0 .0001 in all) (Fig. 2 I-K). Periostin had a positive correlation with IL-5 (r = 0.3) and IL-13 levels (r = 0.2) at baseline( P < 0 .05 in all) (Fig. 1 l-M). IL-13 and IL-5 (r = 0.7) levels were positively correlated at baseline (Fig. 2 N), with nominal P < 0.0001. Additionally, the levels of IL-13 were positively correlated with blood eosinophils (%) at baseline (r = 0.2) with nominal P < 0.05. No correlations were found between IgE, Periostin, IL-4, and Blood eosinophils (%) with any of the other biomarkers assessed. 3.3 Clinical features of Type-2-high patients Th2-high asthma criteria were defined based on blood eosinophil counts and allergen-specific serum IgE: IgE > 100 IU/ml and a blood eosinophil count ≥ 140 cells/µl. The enrolled subjects were recategorized into Th2 high asthma (n = 110) and controls(n = 53). The Th-2 high subjects were ulteriorly grouped into three categories (acute asthma, persistent asthma, and clinical remission), those who presented with worsening wheezing, shortness of breath, cough, and chest tightness, and were diagnosed as acute asthma attack by physicians were defined as asthma cases in acute exacerbation or acute asthma attack (n = 48). Asthma cases in clinical remission (n = 36) were characterized by an absence of asthma symptoms and medicine use for at least ≥ 1 year with no acute asthma attack within a year. Persistent asthma(n = 26)was defined as those with presence of asthma symptoms in the last year or the daily use of long-term control medications to achieve and maintain control in the last year. A comparison of clinical characteristics is presented in Table 2 . Obviously, gender or age was not a factor. No significant differences were found for allergic rhinitis, blood eosinophils (%), total IgE levels, aeroallergen sensitization, and FeNO (parts/billion) between groups. However, differences were statistically significant with regard to lung function measurements including FEV1 (% predicted) ( P = 0.015), FEV1/FVC, % ( P = 0.40), FEF25-75, pred % P = 0.0021). Table 2 characteristics of patients defined as acute attack asthma, persistent asthma, and clinical remission. Acute Attack Asthma n = 48 Persistent Asthma n = 26 Clinical Remmision n = 36 P value Sex(M/F) (33/21) (13/14) (24/14) 0.431 Age(years) 6.9 ± 2.7 6.5 ± 3.5 6.6 ± 3.1 0.816 Allergic rhinitis 48.1% 42.1% 51.9% 0.732 FEV1(% predicted) 80.15 ± 18.86 92.90 ± 14.23 103.0 ± 44.65 0.0015 FEV1/FVC, % 92.77 ± 14.48 99.62 ± 6.573 95.22 ± 22.45 0.040 FEF25-75, pred %, 56.53 ± 24.42 81.44 ± 25.66 71.74 ± 17.38 0.0021 FeNO(parts/billion) 20.72 ± 22.76 20.79 ± 24.52 21.57 ± 23.69 0.480 Blood eosinophils (%) 4.1(0.1–20.1) 2.9(0.1–12.4) 4.75(0.1–127) 0.267 Total IgE (IU/ml) 160.9(5.2–1814) 104(12.6–1346) 44.8(8-1905) 0.500 Skin tests Aeroallergen sensitization 60.9 47.4 50 0.452 House-dust mites 21.7 26.3 29.2 0.581 animal dander 8.7 21.1 8.3 0.674 food allergen sensitization 30.4 31.6 29.2 0.569 pollens 21.7 10.5 8.3 0.347 Mold 4.3 5.3 8.3 0.940 Table 2 : Data are mean ± SD, median (interquartile range), or percentage. Values in bold are statistically significant. Abbreviation: FEV1,forced expiratory volume during the first second; FVC, forced vital capacity; FEF25-75, pred %, forced expiratory flow between 25% and 75%. 3.4 Th2 biomarkers of clinical Th2 high asthma groups As shown in Fig. 3 , there was no obvious difference among the groups in serum concentration of Th2 biomarkers (Fig. 3 A-F) except for TSLP (Fig. 3 G). Notably, TSLP levels were significantly higher in subjects experiencing acute exacerbation and children defined as asthma persistence compared with non-asthmatic control subjects. There was no statistically significance between clinical remission and control subjects. 3.5 Correlation between serum TSLP level and clinical variables in Th2 high asthma Spearman's correlation coefficients were conducted to investigate whether serum TSLP level has associations with subjects’ clinical characteristics in Th2-high asthma and whether it can be used as a predictor to evaluate the ongoing eosinophilic inflammation remission. The results were shown in Table 3 . Serum TSLP level was found significantly correlated with total IgE (Spearman's rho [r] = 0.48, P < 0.0001), FEV1/FVC ratio (r = − 0.365, P < 0.05), and FEF25-75, pred % (r = − 0.06, P < 0.05), whereas no correlations were found with age, BMI, or allergic rhinitis, atopic dermatitis, blood eosinophil counts. Table 3 Spearman's correlation coefficients between serum TSLP level and other clinical characteristics in children with Th2 high asthma. Serum TSLP(pg/ml) r P value Age, y -0.03 0.770 BMI -0.081 0.455 Atopic dermatitis 0.372 0.155 Allergic rhinitis 0.098 0.119 Total IgE (IU/mL) 0.482 < .0001 Eosinophils (%) -0.072 0.502 FEV1, pred % -0.106 0.457 FEV1/FVC ratio -0.365 0.008 FEF25-75, pred % -0.306 0.031 Table 3 : Abbreviations: BMI, body mass index; FEF25-75, forced expiratory flow between 25% and 75%; FEV1, forced expiratory volume during the first second; FVC, forced vital capacity. 3.6 Multivariable regression and ROC analyses for predictors of clinical asthma remission and persistence in Th-2 high asthma To find out which predictors show relationships with clinical remission in Th2-high asthma, a multiple regression model was designed including the variables showing differences between readjusted Th2-high groups and variables associations with TSLP in Spearman’s analyses. The results of multivariable logistic regression analysis revealed that serum TSLP levels were associated with clinical remission in Th2-high asthma subjects (OR = 1.009; 95% CI:1.008–1.008, P = 0.023) (Table 4 ). Serum TSLP level correlated with clinical remission in Th2-high asthma in terms of ROC curves analysis (AUC = 0.5887, 95% CI: 0.5052 to 0.7038, P < 0 .05) (Fig. 4 ). Considering sensitivity, and specificity, for serum TSLP levels, a cutoff value of 373.363 pg/ml was found with the highest clinical sensitivity and specificity. PPV and NPV of 100%, 39.1%, and 59.4%, 100%, respectively. However, no positive result was found in the analysis of multivariable logistic regression in determining the predictors of Th2 high persistent asthma. Table 4 Multivariable logistic regression analysis for clinic remission in Th2 high asthma Variable OR (95% CI) P Value FEV1(% predicted) 0.963(0.959–0.966) 0.411 FEV1/FVC ration 1.085(1.096–1.074) 0.189 FEF25-75, pred % 0.974(0.972–0.976) 0.568 Total IgE (IU/mL) 1.014(1.014–1.013) 0.668 TSLP (pg/ml) 1.009(1.008–1.008) 0.023 3.7 Comparison of TSLP in serum, BALF, and lung tissue of different mouse model groups The hallmark of the Th2-high asthma was IL4, IL5, and IL13, and the mouse Elisa kit was used to screen the Th2 cytokines in BALF and serum of different stages mouse models first (data not shown). Measurements of TSLP levels was then examined in serum, BALF, and lung tissue of different stages of mouse asthma models to confirm and validate the observations from the study population. We aimed to inspect the expression of TSLP in different tissues of asthmatic mice and to find out the most indicative significance. When comparing the concentration of TSLP in serum, BALF, and lung tissue, it was found that BALF had the lowest level of TSLP concentration. There was no statistically significant change in the acute and remission stages when compared to the control, but TSLP did increase noticeably in the chronic stage (Fig. 5 A). There seems to be no significant difference in the levels of TSLP in mouse serum between control and asthma remission. However, it had been noted that during the acute and chronic stages, TSLP levels increased, as depicted in the figure. (Fig. 5 B) Furthermore, the lung tissue also demonstrated a comparable trend as serum, with significant statistical significance observed in this case (Fig. 5 C). 4 Discussion This cross-sectional study presents the positive association observed between Th2 biomarkers in circulating levels and demonstrated robust relationships between several biomarkers of T2 inflammation. In addition, it extended previous investigations regarding identifying the potential biomarkers predicting clinical remission in Th2-high asthma. We found that when subjects were simply partitioned into asthma and control, no significance was observed in Th2 biomarkers between these two groups, whereas clinical measurements such as blood eosinophils (%) and eosinophils count were statistically significant. When we defined Th2 high asthma based on eosinophils count (≥ 140 cells/µl ) and total IgE (IgE > 100 IU/ml), the serum TSLP levels were observed higher in subjects experiencing acute exacerbation and persistent asthma compared with no asthma control subjects. In addition, our results also found an association between serum TSLP levels and clinical remission independent of confounding factors such as lung function measurements or allergy sensitization. However, the role of serum TSLP in identifying clinical remission of Th2-high asthma still needs further holistic investigations to reconfirm its specificity and sensitivity to be used as a prognostic biomarker in asthma. 4.1 Th2 biomarkers at baseline Since asthma is a heterogeneous disease that can be divided into many subgroups, one of the widely accepted consensuses is that the research of it needs to evolve from phenotypes to endotypes [ 1 ]. Phenotypes are usually defined by clinical characteristics, allergen sensitization patterns, and physiologic lung function measurements which are readily observed or measured in clinical practice. Endotypes are a definition that is related to pathophysiological mechanisms underlying phenotype [ 19 ]. Thus, there is a growing urge to investigate asthma endotypes by connecting discernible and stable biomarkers with immunological mechanisms to make precision therapy decisions for asthma [ 20 ]. Th2-high asthma is a broad classification involving asthma endotypes based on type 2 (T2) inflammation pathobiology characterized by increased secretion of Th2 biomarkers such as IL-4, IL-5, IL-13, and IgE causing pathological changes, which can be summarized by a set of the cytokine-induced eosinophilic airway inflammation, epithelial dysfunction, and airway wall remodeling [ 21 , 22 ]. And it is believed that Th2-high asthma account for only half of the people diagnosed with asthma [ 1 , 23 ], which in a way supports our results that there was no considerable significance at the baseline levels of Th2 biomarkers between asthma and control when study population was not recruited specifically with Th2-high characteristics. When investigating the correlations between the biomarkers and clinical characteristics, there were no positive results observed between Th2 biomarkers and baseline blood eosinophil counts or total IgE except that IL-13 was positively correlated with blood eosinophils (%). Whereas, Corren, Pham, et al found that baseline levels of serum IL-5, IL-13, and Periostin correlated significantly with baseline blood eosinophil counts and FeNO levels in uncontrolled severe asthma subjects, which may account for the complex immune pathways in severe asthma [ 24 , 25 ]. After all, this is a relatively representative study measuring baseline circulating levels of Th2 biomarkers, and the positive correlations among specific biomarkers on one side further support the molecular mechanism of Th2 biomarkers, on the other side it may provide insight into combined immune biologic interventions in personalized therapy. 4.2 Asthma remission, persistence and predictive role of Th2 biomarkers Asthma remission is not emphasized in asthma treatment goals, unlike other chronic diseases, and it is widely accepted that clinical remission and complete remission have different criteria, and current research involving asthma remission is limited [ 26 , 27 ]. Wang, Datta [ 13 ] demonstrated that baseline lung function can be used as a clinical prognostic indicator of asthma remission. And other independent predictors of asthma remission include less BHR, no nasal polyps, younger age, better asthma control, lower doses of ICS, and lower levels of blood neutrophils [ 28 ]. Actually, when comes to asthma remission, clinical features are mostly considered [ 29 , 30 ]. However, with the prevalent use of biologics in asthma treatments, and the great development from phenotypes to endotypes in asthma pathophysiological mechanisms, identifying predicting biomarkers of asthma remission, especially in the Th2 high subtype may mean a step closer to clinical asthma remission evaluation. Thus, in this study, the study population was further divided into acute asthma attacks, persistent asthma, and clinical remission asthma among Th2-high subjects. The criteria of Th2-high and the definition of different phases were explained above. It is obvious that lung measurements including FEV1(% predicted), FEV1/FVC, %, FEF25-75, pred %, are statistically significant among groups which are in line with many research that lung measurements are sensitive clinical features evaluating asthma remission [ 13 , 30 , 31 ].When comparing circulating levels of Th2 biomarkers among groups, there was no marked significance except serum TSLP. However, Tan, Daniel J et al found that serum inflammatory cytokines including IL-4, and IL-5 have prognostic value in adults with spontaneous asthma remission. And there was solid evidence that Periostin was associated with asthma persistence, especially in severe uncontrolled asthma [ 32 , 33 ]. In addition,Christianson, Goplen [ 34 ] elucidated that IL-13 is a crucial factor in the feedback and feed-forward interactions that contribute to the persistence of asthma in mouse models. TARC(thymus activation-regulated chemokine) which is expressed in T helper (Th2) cells, also known as CCL17, belongs to the CC chemokine family. TARC is believed to have a pivotal role in the pathogenesis of allergic diseases such as atopic dermatitis, bronchial asthma, and allergic rhinitis [ 35 , 36 ]. What's more, the serial changes of plasma TARC concentrations in children with acute asthma attacks were monitored in a study which indicated that TARC is a potentially useful inflammatory maker to evaluate asthmatic exacerbation [ 37 ]. To sum up, these conflicts may due to the difference in study population and design, and it is crucial to emphasize the fact that the research on the levels of biomarkers associated with asthma remission and persistence is limited. Therefore, there is a pressing need to conduct more studies in this area to gain critical insights into this debilitating condition. 4.3 Association between clinical asthma remission and serum TSLP Thymic stromal lymphopoietin (TSLP), part of the IL-2 cytokine family, was discovered as a growth factor for lymphocyte progenitors and is now recognized as a protein primarily released from epithelial cells and has a sensitive response to irritating stimuli [ 38 , 39 ]. and research involving mice models to human subjects and the genome-wide association had demonstrated that TSLP is associated with asthma [ 40 – 42 ]. Indeed, TSLP regulates dendritic cells which are considered one of the central responders of Th2 immunity [ 43 ]. What’s more, with the approval of TSLP-targeted antibody for severe asthma, it is reported that TSLP plays a key role in allergen-induced airway responses and persistent airway inflammation in patients with allergic asthma [ 42 ]. However, whether it can be a predictor to evaluate the clinical remission of Th2-high asthma is still understudied. There was research based on sputum type 2 makers and found that the sputum TSLP levels were higher at baseline in the remission group compared with the non-remission group and TSLP was not listed as a potential predictor of remission by univariate regression analysis [ 44 ]. On the contrary, our results found that serum TSLP has a significant difference among acute asthma groups, asthma persistence groups, and clinical remission group, and multivariable regression also revealed that serum TSLP levels were associated with clinical remission in Th2 high asthma children. The fundamental role of TSLP in asthma pathogenesis was underscored by the fact that the latest clinical trials have reconfirmed that treatment with Tezepelumab, which blocks TSLP, can lead to sustained and significant reductions in asthma exacerbations in individuals with severe, uncontrolled asthma [ 45 ]. A multi-center prospective observational study suggests that TSLP may play a pathogenic role across different asthma phenotypes [ 46 ]. 4.4 The expression of TSLP in BALF, serum, and lung tissue of different stages of asthmatic mice. We further conducted an animal model of allergic asthma which shows the levels of TSLP vary in acute, chronic, and remission asthma mouse models. Our findings indicate that the TSLP levels are remarkably high in chronic asthma mouse models among the lung tissue, BALF, and serum. Furthermore, both in the serum and lung tissue samples, we have also observed that the decrease in TSLP levels may indicate the remission of asthma. Numerous murine research have demonstrated that TSLP plays an important role in the development of asthma [ 47 , 48 ]. The anti-TSLP method or TSLP-R deficiency can effectively attenuate AHR and TH2 airway inflammation [ 49 , 50 ]. Notably, an interesting research also revealed that mice received TSLP receptor-deficient CD4 + T cells had alleviated pathogenic TH2 cell responses, and asthmatic children showed an increase in the production of TH2 cells, promoted by TSLP and IL-4, which resulted in higher amounts of IL-5 and IL-13 in their peripheral blood, compared to healthy controls [ 51 ]. Although it is commonly believed that TSLP plays a crucial role in the pathogenesis of asthma, there is still a lack of research on whether TSLP can be utilized as a biomarker for indicating asthma remission or persistence. We believe that this possibility has not been discussed enough, and therefore we are interested in exploring this topic further. 4.5 Conclusion We report for the first time that serum TSLP may be used as one of the biomarkers to evaluate the remission of asthma along with other measurements such as lung function to improve the accuracy sensitivity and specificity and to testify our findings in acute chronic and remission asthma mouse models. However, despite the positive likelihood of evaluating remission shown in ROC curves analysis, its diagnostic role and prognostic role in identifying clinical asthma remission still need further exploration. We aim to expand our understanding of TSLP and its clinical significance while advancing our current research. 4.6 limitation and strength The limitations of this study lie in that subjects and the Th2 biomarkers should have been be detected in a larger cohort of multiple centers and the definition of asthma remission is still under-refined. Also, the steadiness of the circulating biomarkers should be validated in the longitudinal study. Furthermore, investigation related to other biomarkers of different pathways is expected to be conducted to form a more holistic understanding of asthma remission. It is the first research to investigate the role of circulating levels of Th2 biomarkers as predictors of remission in a cohort of asthma subjects though it is a preliminary and retrospective study. Declarations 4.7 Funding This work was supported by the National Natural Science Foundation of China (Grant No. 82205189, 82174170, 82205044), Shanghai science and technology commission Project (Grant No. 21S21902500), as Fund of Huashan Hospital affiliated to Fudan University (Grant No. 2021QD042). 4.8 Acknowledgments The authors thank the participants of the study, Qiuyan Yang collected the serum from the children, Mengmeng Chen, for her contributions to the analysis and critical review, and Na Wang, MS, Ph, for her contribution to the statistical analysis. Medical writing support was provided by Congcong Li, Huijie Zhang, and Zhiyong Liu. This study was co-sponsored by Yanli Wang,Na Wang and Jingcheng Dong. 4.9 Conflict of interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. 4.10 Ethical Approval and Consent to participate This trial was conducted under the ethical principles of the Declaration of Helsinki, the International Council for Harmonisation guidelines for good clinical practice, and applicable regulatory requirements. The protocol (2021-108-01) was initially approved by the ethics committee of the Third Affiliated Hospital of Zhengzhou University. All participants and their parents granted their written informed consent and all experiments were performed per the approved guidelines and regulations. All animal experiments are approved by the Laboratory Animal Ethics Committee (IACUC) of Fudan University and are conducted in strict accordance with the laboratory animal management regulations of Fudan University (2022JS Huashan hospital-497). 4.11 Availability of supporting data All data are available and the correspondent can be contacted if requested. References Wenzel SE. Asthma phenotypes: the evolution from clinical to molecular approaches. Nat Med. 2012;18(5):716–25. Pijnenburg MW et al. Childhood asthma- pathogenesis and phenotypes. Eur Respir J, 2021. Pijnenburg MW et al. -2022-FINAL -22-07-01-WMS.pdf> . Shute J. Biomarkers of asthma. Minerva Med; 2021. Allakhverdi Z, et al. Thymic stromal lymphopoietin is released by human epithelial cells in response to microbes, trauma, or inflammation and potently activates mast cells. J Exp Med. 2007;204(2):253–8. Soumelis V, et al. Human epithelial cells trigger dendritic cell mediated allergic inflammation by producing TSLP. Nat Immunol. 2002;3(7):673–80. Shikotra A et al. Increased expression of immunoreactive thymic stromal lymphopoietin in patients with severe asthma. J Allergy Clin Immunol, 2012. 129(1): p. 104 – 11.e1-9. Pavlidis S et al. T2-high in severe asthma related to blood eosinophil, exhaled nitric oxide and serum periostin. Eur Respir J, 2019. 53(1). Maison N et al. T2-high asthma phenotypes across lifespan. Eur Respir J, 2022. 60(3). Papi A, et al. Asthma Lancet. 2018;391(10122):783–800. Thomas D et al. Asthma remission: what is it and how can it be achieved? Eur Respir J, 2022. 60(5). Corbett ML, Oppenheimer JJ. Need for a consensus definition of remission in paediatric asthma. Lancet Child Adolesc Health. 2022;6(11):755–6. Wang AL, et al. Remission of persistent childhood asthma: Early predictors of adult outcomes. J Allergy Clin Immunol. 2019;143(5):1752–e17596. Sears MR. Predicting asthma outcomes. J Allergy Clin Immunol, 2015. 136(4): p. 829 – 36; quiz 837. Graham BL, et al. Standardization of Spirometry 2019 Update. An Official American Thoracic Society and European Respiratory Society Technical Statement. Am J Respir Crit Care Med. 2019;200(8):e70–88. Ibrialos P et al. Usefulness of FEF25-75% and FEF200-1200 for the graphic/computational interpretation of spirometry. Chest. 88(1): p. 157–8. Khatri SB, et al. Use of Fractional Exhaled Nitric Oxide to Guide the Treatment of Asthma: An Official American Thoracic Society Clinical Practice Guideline. Am J Respir Crit Care Med. 2021;204(10):e97–109. Bousquet J, et al. Practical guide to skin prick tests in allergy to aeroallergens. Allergy. 2012;67(1):18–24. Nelson RK, et al. Eosinophilic Asthma. J Allergy Clin Immunol Pract. 2020;8(2):465–73. Akar-Ghibril N, et al. Allergic Endotypes and Phenotypes of Asthma. J Allergy Clin Immunol Pract. 2020;8(2):429–40. Busse WW et al. Understanding the key issues in the treatment of uncontrolled persistent asthma with type 2 inflammation. Eur Respir J, 2021. 58(2). Hammad H, Lambrecht BN. The basic immunology of asthma. Cell. 2021;184(6):1469–85. Woodruff PG, et al. T-helper type 2-driven inflammation defines major subphenotypes of asthma. Am J Respir Crit Care Med. 2009;180(5):388–95. Corren J, et al. Baseline type 2 biomarker levels and response to tezepelumab in severe asthma. Allergy. 2022;77(6):1786–96. Wenzel SE. Severe Adult Asthmas: Integrating Clinical Features, Biology, and Therapeutics to Improve Outcomes. Am J Respir Crit Care Med. 2021;203(7):809–21. Upham JW, James AL. Remission of asthma: The next therapeutic frontier? Pharmacol Ther. 2011;130(1):38–45. Menzies-Gow A, et al. An expert consensus framework for asthma remission as a treatment goal. J Allergy Clin Immunol. 2020;145(3):757–65. Westerhof GA, et al. Clinical predictors of remission and persistence of adult-onset asthma. J Allergy Clin Immunol. 2018;141(1):104–e1093. Almqvist L et al. Remission of adult-onset asthma is rare: a 15-year follow-up study. ERJ Open Res, 2020. 6(4). Holm M, et al. Remission of asthma: a prospective longitudinal study from northern Europe (RHINE study). Eur Respir J. 2007;30(1):62–5. Ronmark E, et al. Outcome and severity of adult onset asthma–report from the obstructive lung disease in northern Sweden studies (OLIN). Respir Med. 2007;101(11):2370–7. Hanania NA, et al. Exploring the effects of omalizumab in allergic asthma: an analysis of biomarkers in the EXTRA study. Am J Respir Crit Care Med. 2013;187(8):804–11. Bobolea I, et al. Sputum periostin in patients with different severe asthma phenotypes. Allergy. 2015;70(5):540–6. Christianson CA, et al. Persistence of asthma requires multiple feedback circuits involving type 2 innate lymphoid cells and IL-33. J Allergy Clin Immunol. 2015;136(1):59–e6814. Sugawara N et al. TARC in allergic disease. Allergy. 57(2): p. 180–1. Radonjic-Hoesli S et al. Are blood cytokines reliable biomarkers of allergic disease diagnosis and treatment responses? J Allergy Clin Immunol 150(2): p. 251–8. Leung TF et al. Plasma TARC concentration may be a useful marker for asthmatic exacerb ation in children. Eur Respir J 21(4): p. 616–20. Mitchell PD, O'Byrne PM. Biologics and the lung: TSLP and other epithelial cell-derived cytokines in asthma. Pharmacol Ther. 2017;169:104–12. Dahlen SE. TSLP in asthma–a new kid on the block? N Engl J Med. 2014;370(22):2144–5. Harada M, et al. Thymic stromal lymphopoietin gene promoter polymorphisms are associated with susceptibility to bronchial asthma. Am J Respir Cell Mol Biol. 2011;44(6):787–93. An G, et al. Combined blockade of IL-25, IL-33 and TSLP mediates amplified inhibition of airway inflammation and remodelling in a murine model of asthma. Respirology. 2020;25(6):603–12. Gauvreau GM, et al. Effects of an anti-TSLP antibody on allergen-induced asthmatic responses. N Engl J Med. 2014;370(22):2102–10. Roan F, Obata-Ninomiya K, Ziegler SF. Epithelial cell–derived cytokines: more than just signaling the alarm. J Clin Invest. 2019;129(4):1441–51. Moermans C et al. Sputum Type 2 Markers Could Predict Remission in Severe Asthma Treated With Anti-IL-5. Chest, 2023. Menzies-Gow A et al. Long-term safety and efficacy of tezepelumab in people with severe, un controlled asthma (DESTINATION): a randomised, placebo-controlled exte nsion study. Lancet Respiratory Med. 11(5): p. 425–38. Ko HK, et al. Blood tryptase and thymic stromal lymphopoietin levels predict the risk of exacerbation in severe asthma. Sci Rep. 2021;11(1):8425. Al-Shami A, et al. A role for TSLP in the development of inflammation in an asthma model. J Exp Med. 2005;202(6):829–39. Kabata H, et al. Targeted deletion of the TSLP receptor reveals cellular mechanisms that promote type 2 airway inflammation. Mucosal Immunol. 2020;13(4):626–36. Hu Y, et al. TSLP signaling blocking alleviates E-cadherin dysfunction of airway epithelium in a HDM-induced asthma model. Cell Immunol. 2017;315:56–63. Toki S, et al. TSLP and IL-33 reciprocally promote each other's lung protein expression and ILC2 receptor expression to enhance innate type-2 airway inflammation. Allergy. 2020;75(7):1606–17. Rochman Y et al. TSLP signaling in CD4(+) T cells programs a pathogenic T helper 2 cell state. Sci Signal, 2018. 11(521). Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4182358","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":286260764,"identity":"beea5b70-9b28-40f0-a911-09d96ec052ca","order_by":0,"name":"Mengmeng Chen","email":"","orcid":"","institution":"Department of Integrative Medicine, Huashan Hospital, Fudan University, Shanghai, China","correspondingAuthor":false,"prefix":"","firstName":"Mengmeng","middleName":"","lastName":"Chen","suffix":""},{"id":286260765,"identity":"25e90361-9fe3-4707-a223-357b58cf3a81","order_by":1,"name":"Congcong Li","email":"","orcid":"","institution":"Department of Integrative Medicine, Huashan Hospital, Fudan University, Shanghai, China","correspondingAuthor":false,"prefix":"","firstName":"Congcong","middleName":"","lastName":"Li","suffix":""},{"id":286260766,"identity":"34e28201-4242-47a4-88fd-c1718c21758f","order_by":2,"name":"Qiuyan Yang","email":"","orcid":"","institution":"Department of Pediatrics, Third Affiliated Hospital of Zhengzhou University, Zhengzhou, China","correspondingAuthor":false,"prefix":"","firstName":"Qiuyan","middleName":"","lastName":"Yang","suffix":""},{"id":286260767,"identity":"068d386c-5e9b-47c6-8056-fd0c076bdb07","order_by":3,"name":"Huijie Zhang","email":"","orcid":"","institution":"Department of Integrative Medicine, Huashan Hospital, Fudan University, Shanghai, China","correspondingAuthor":false,"prefix":"","firstName":"Huijie","middleName":"","lastName":"Zhang","suffix":""},{"id":286260768,"identity":"295d4ca7-afc1-4f28-b04a-e43e747af609","order_by":4,"name":"Yanli Zhang","email":"","orcid":"","institution":"Department of Pediatrics, Third Affiliated Hospital of Zhengzhou University, Zhengzhou, China","correspondingAuthor":false,"prefix":"","firstName":"Yanli","middleName":"","lastName":"Zhang","suffix":""},{"id":286260769,"identity":"4f093fc3-0a54-462a-b319-c08162c82274","order_by":5,"name":"Na Wang","email":"","orcid":"","institution":"Children’s Hospital of Fudan University","correspondingAuthor":false,"prefix":"","firstName":"Na","middleName":"","lastName":"Wang","suffix":""},{"id":286260770,"identity":"8c852d63-798a-47cb-8a47-fd02ce109c7d","order_by":6,"name":"Jingcheng Dong","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA10lEQVRIiWNgGAWjYBACxgYGAyBlw8PGcPjAgQ8VxGtJk+NnPJZ4cMYZ4iwCaTlsLNl8xvgwbwsR6pnbD298XPCLOXHDsTMfDvA2MMjzix0g4LCetGLjmX1siRvOnN1wQHIHg+HM2QkEtMzgMZPm7eFJ3HADqMXwDEOCwW3itEgkbrj/5sGBxDZitfD8MDCWbDjDcOAgUVpAfuFtSJDjZzhmcLDhjARhvxiCQoznz39QVD7+/KfCRp5fmpCWBpBVbXC+BH7lICAPJv8QVjgKRsEoGAUjGAAAY7tOA2vJXL4AAAAASUVORK5CYII=","orcid":"","institution":"Department of Integrative Medicine, Huashan Hospital, Fudan University, Shanghai, China","correspondingAuthor":true,"prefix":"","firstName":"Jingcheng","middleName":"","lastName":"Dong","suffix":""}],"badges":[],"createdAt":"2024-03-28 12:46:03","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4182358/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4182358/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":53969559,"identity":"83c59a68-cd5f-4c4f-b2f8-84fdceecbc67","added_by":"auto","created_at":"2024-04-02 20:35:02","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":306732,"visible":true,"origin":"","legend":"\u003cp\u003eA total of 172 children were enrolled in the study among them, 119 were diagnosed with asthma and 53 were non-asthmatic control subjects. Clinical tests included measurement of lung function, Fraction of exhaled NO (FeNO), total IgE, and blood eosinophil. Serum Th2 biomarkers (TSLP, TRAC, IL-4 IL-5, IL-13, IgEand Periostin) were examined by ELISA. Based on clinical eosinophil count and total IgE, the enrolled patients were readjusted into Th2-high asthma and classified into acute attack asthma, persistent asthma, and clinical remission according to the recent GINA guidelines and clinical evaluation. Serum TSLP levels along with lung function measurements can be used as predictors to help evaluate clinical remission in Th2-high asthma. Abbreviations: FeNO, fractional exhaled nitric oxide; IgE, immunoglobulin E; IL, interleukin; TARC, thymus and activation-regulated chemokine; TSLP, thymic stromal lymphopoietin.\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-4182358/v1/e7f760953f2b8a01ff44617f.png"},{"id":53969556,"identity":"bab092c3-19ea-4d1d-9d00-d55b4e8e4209","added_by":"auto","created_at":"2024-04-02 20:35:02","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":626960,"visible":true,"origin":"","legend":"\u003cp\u003eBaseline correlations between serum levels of IL-5 and IL-13 and other biomarkers of inflammation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFIGURE 2:\u003c/strong\u003e Spearman correlation coefficients (r) are shown. Abbreviations: IL, interleukin; TARC, thymus and activation-regulated chemokine; TSLP, thymic stromal lymphopoietin; Eos%, blood eosinophils (%).\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-4182358/v1/68960e213e4f96d7ea0f9f80.png"},{"id":53970369,"identity":"6bb9c7d4-1633-47a0-badc-df4bccb1bf2d","added_by":"auto","created_at":"2024-04-02 20:43:02","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":522323,"visible":true,"origin":"","legend":"\u003cp\u003eTSLP levels among the groups of Th2 high asthma.\u003c/p\u003e","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-4182358/v1/3d69ba0c0bad46406dde7528.png"},{"id":53969557,"identity":"941102b5-e634-4afa-9a7b-2e8fe796d6ce","added_by":"auto","created_at":"2024-04-02 20:35:02","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":105059,"visible":true,"origin":"","legend":"\u003cp\u003eROC curve for clinical remission in Th2 high asthma.\u003c/p\u003e","description":"","filename":"Figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-4182358/v1/a7e1a696631d904a36244f51.png"},{"id":53969560,"identity":"f6efbbc3-720f-4918-ae7f-06b691244b0a","added_by":"auto","created_at":"2024-04-02 20:35:03","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":138948,"visible":true,"origin":"","legend":"\u003cp\u003eThe expression of TSLP in BALF, serum, and lung tissue of different stages of asthmatic mice.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFigure 5:\u003c/strong\u003eThe statistical analysis compared control group is represented as *\u003cem\u003eP\u003c/em\u003e\u0026lt;0.05; **\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.01;***\u003cem\u003eP\u003c/em\u003e\u0026lt;0.001 (n=6).\u003c/p\u003e","description":"","filename":"Figure5.png","url":"https://assets-eu.researchsquare.com/files/rs-4182358/v1/bfd992ef144bd2632f4f96df.png"},{"id":59585341,"identity":"03bba351-0858-4967-9646-8e0a4514c970","added_by":"auto","created_at":"2024-07-03 13:36:24","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2246323,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4182358/v1/061d5834-da94-4a6b-b50f-169ac6c3501a.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Baseline Type 2 Biomarker Levels and Clinical Remission Predictors in Children with Asthma","fulltext":[{"header":"1 Introduction","content":"\u003cp\u003eAsthma is one of the most common inflammatory airway diseases of childhood associated with significant heterogeneity, which refers to different asthma phenotypes having unique clinical and biomarker profiles [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. The 2022 Global Initiative for Asthma (GINA) guidelines recommend using clinical characteristics and biomarkers to define asthma inflammatory phenotypes that may help physicians move toward more personalized management of asthma and benefit patients [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. And type-2-high asthma which relates to inflammatory type presents an example of current asthma phenotypes (type 2-high or type 2-low), characterized by increased secretion of interleukin IL-4, IL-5, and IL-13 by Type 2 innate lymphoid cells and T-helper 2 cells [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. The majority of asthma patients have evidence of type 2 (T2) inflammation, which also features high blood eosinophil counts and fractional exhaled nitric oxide (FeNO) levels. The Type 2 biomarkers involved in underlying airway inflammation are well-researched in the area. However, there are limited data when it comes to using biomarkers to predict the clinical trajectory of childhood asthma. Therefore, the aim of the study was to investigate the association of Th2 biomarker levels with clinical features in different phases of asthma in children and try to identify biomarkers whose level is expected to differ among those with persistent asthma, acute exacerbation asthma, or clinical asthma remission in cohorts, which may provide insight into clinical asthma trajectory. So, we determined the association of Th2 biomarkers with baseline characteristics and found out one of the Th2 biomarkers TSLP may predict asthma remission in a cohort of Th2 high asthma subjects which were divided into acute exacerbation asthma, persistent asthma, and clinical remission asthma. As reported, Thymic stromal lymphopoietin (TSLP) is an epithelial cell-derived cytokine that is implicated in the initiation and persistence of airway inflammation in asthma [\u003cspan additionalcitationids=\"CR6\" citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Based on a cross-sectional baseline of characteristics obtained during a medical visit along with specific serum biomarkers, we may make a close step toward individualized prognostication.\u003c/p\u003e"},{"header":"2 Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1.1 Study Design and population\u003c/h2\u003e \u003cp\u003eWe enrolled 172 children from a clinic pediatric asthma center, among them, 119 subjects were diagnosed with asthma by doctors on the basis of the updated version of the 2022 statement by the Global Initiative for Asthma (GINA), and 53 were age- and sex-matched control subjects without respiratory symptoms. Th2-high asthma was defined based on clinical measurement of IgE\u0026thinsp;\u0026gt;\u0026thinsp;100 IU/ml and blood eosinophil counts\u0026thinsp;\u0026ge;\u0026thinsp;140 cells/\u0026micro;l [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Th2 high asthma subjects (n\u0026thinsp;=\u0026thinsp;110) were allocated into three groups (acute attack asthma, persistent asthma, and clinical remission). Those who presented with worsening wheezing, shortness of breath, cough, and chest tightness, and were diagnosed as acute asthma attacks by physicians were defined as asthma cases in acute exacerbation(n\u0026thinsp;=\u0026thinsp;48) [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Asthma cases in clinical remission(n\u0026thinsp;=\u0026thinsp;36) are characterized by an absence of asthma symptoms and medicine use for at least\u0026thinsp;\u0026ge;\u0026thinsp;1 year additionally without acute asthma attack within a year [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Persistent asthma(n\u0026thinsp;=\u0026thinsp;26)was defined as those with the presence of asthma symptoms in the last year or use of long-term control medications daily to achieve and maintain control in the last year [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e This trial was conducted under the ethical principles of the Declaration of Helsinki, the International Council for Harmonisation guidelines for good clinical practice, and applicable regulatory requirements. The protocol (2021-108-01) was initially approved by the ethics committee of the Third Affiliated Hospital of Zhengzhou University. All participants and their parents granted their written informed consent and all experiments were performed per the approved guidelines and regulations.\u003c/p\u003e \u003cdiv id=\"Sec4\" class=\"Section3\"\u003e \u003ch2\u003e2.1.2 MICE\u003c/h2\u003e \u003cp\u003eBALB/c mouse, female, SPF grade, weight 18\u0026thinsp;+\u0026thinsp;2g, 6\u0026ndash;8 weeks old, were purchased from Shanghai Jiesjie Laboratory Animal Co., LTD. They were fed in an SPF environment with free access to sterilized water and food, controlled with a 12-hour daytime to 12-hour night cycle, indoor temperature of 20\u0026ndash;30 ℃, and humidity of 50%. After acclimation for 1 week, the mice were randomly divided into 4 groups (control, acute asthma, chronic asthma, and asthma remission),6 in each group. Mice in the control group were given 0.2mL normal saline intraperitoneal injection, and other asthma model groups were all intraperitoneally injected with 0.2mL suspension which is complexed 20\u0026micro;g of OVA (GradeV) with 2mg Aluminum hydroxide on days 0, 7, and 14 to induce asthma. To mimic different stagings of human asthma, OVA-sensitization mouse asthma models undergoing different durations of OVA-inhaled stimulation. From day 21, mice in the acute asthma group were challenged sensitization by ultrasonic atomizing inhalation of 3% OVA (Grade II) solution in the closed container for 30 minutes each day, lasting for 7 days. The same atomization condition applied for the chronic asthma model group was 3 times a week, continuing for 6 weeks. And the remission group was atomized once a month for 2 months. After the ceases of OVA inhalation exposure, all the experimental groups were determined for airway hyperresponsiveness (AHR) using different concentrations of methacholine. Type 2 diagnostic inflammation biomarkers were detected including eosinophils, specific-IgE, and Th2 cytokines (IL-4, IL5, and IL-13), both in blood and BALF.\u003c/p\u003e \u003cp\u003eAn overview of the results showed that the three murine models of asthma were successful. Compared with the control group, the other three groups were found airway methacholine hyperresponsiveness. Higher IgE and Th2 cytokines were detected in serum and BALF, and also more eosinophil infiltration in the lungs.\u003c/p\u003e \u003cp\u003e All animal experiments are approved by the Laboratory Animal Ethics Committee (IACUC) of Fudan University and are conducted in strict accordance with the laboratory animal management regulations of Fudan University (2022JS Huashan hospital-497).\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.2 Study measurements\u003c/h2\u003e \u003cdiv id=\"Sec6\" class=\"Section3\"\u003e \u003ch2\u003e2.2.1 Clinical information collection\u003c/h2\u003e \u003cp\u003eAll subjects' holistic history including signs and symptoms of asthma, medication use, and common triggers observed by parents are collected by well-trained clinicians. Height and weight were measured and body mass index (BMI) was calculated as an index of the physique.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section3\"\u003e \u003ch2\u003e2.2.2 Lung function tests\u003c/h2\u003e \u003cp\u003eAsthma-related pulmonary function was measured according to international standards. The forced expiratory volume in one second (FEV1) and forced vital capacity (FVC) were calculated using the formula of the American Thoracic Society/European Respiratory Society (ATS/ERS) guidelines [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. FEF25-75 pred % were also recorded [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.3 Inflammatory parameters\u003c/h2\u003e \u003cp\u003eThe fraction of exhaled nitric oxide was measured at a flow rate of 50 ml/s according to the recommended standards (SV-02, Wuxi Shangwo, China) [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. Differential white blood cell counts were performed using the automatic blood cell count with a full automatic hemocyte analyzer (BC-5800, Mindary, Shenzhen, China). Serum total IgE was measured in an automatic biochemical analyzer (AU5400, Beckman Coulter Inc, Calif, USA) and related reagents were from Diasys diagnostic systems (Diasys, Catalog No: 17239).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.4 Skin tests\u003c/h2\u003e \u003cp\u003eSkin prick testing (SPT) for common aeroallergens such as pteronyssinus and Dermatophagoides farina was measured among children. Weed and tree pollen mix was also included, as well as food allergens, molds, and animal dander. Histamine (10 mg/mL of histamine phosphate) and 0.9% saline were used as positive and negative controls, respectively. The positive reaction was a wheal observed 3 mm greater than the negative control [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003e2.5 Serum concentrations of biomarkers and cytokines\u003c/h2\u003e \u003cp\u003eVenous blood samples were collected with a vacuum blood tube and centrifuged at 3000 g for 10 mins. Then the serum was isolated and stored in aliquots at \u0026minus;\u0026thinsp;80\u0026deg;C until use. Levels of serum IL-5, IL-13, IgE, IL-4, Periostin, TARC, and TSLP were measured by using specific human enzyme-linked immunosorbent assay kits (Shanghai Wellbio Technology Co., Ltd, No: EH6305M, EH6261M, EH6244M, EH6304M, EH6393M, EH6079M, EH6521M), according to the manufacturer\u0026rsquo;s instructions. Each sample was tested in duplicate.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003e2.6 Measurement of mouse TSLP by ELISA in serum, BALF, and lung tissue\u003c/h2\u003e \u003cp\u003eAfter the pulmonary function measurement, the mice were exsanguinated by eyeball enucleation. Their sera were separated and stored in aliquots at \u0026minus;\u0026thinsp;80\u0026deg;C until use. Lung lavage fluid was collected using a 1mL syringe that was connected to an indentation needle inserted into the trachea. Next, 0.2mL of pre-cooled PBS solution was injected into the trachea to irrigate the left lung and was quickly withdrawn. This process was repeated three times to ensure proper lavage of the lung. Afterward, the BALF was centrifuged at 4\u0026deg;C for 15 minutes, and the supernatant was collected and frozen in the \u0026minus;\u0026thinsp;80\u0026deg;C refrigerator for subsequent detection. Lung tissue was divided into different sections in need and stored in the \u0026minus;\u0026thinsp;80\u0026deg;C refrigerator for use. The concentration of mouse TSLP in serum, BALF, and lung tissue was tested by indirect ELISA, using a Mouse TSLP ELISA kit (Shanghai Wellbio Technology Co., Ltd, No: EM30546M). All experiment was performed according to the manufacturer\u0026rsquo;s instructions.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003e2.7 Statistical analysis\u003c/h2\u003e \u003cp\u003eStatistical analyses were performed using SPSS v 26.0 (IBM, Chicago, IL, USA) and GraphPad Prism 9 software (GraphPad Software Inc. La Jolla, CA, USA). Normally distributed continuous data were expressed as mean and standard deviation, and non-normally distributed continuous data as median and inter-quartile ranges (IQR). Basic between-group comparisons were made using Mann-Whitney tests, Student\u0026acute;s t-test, and multiple between-group comparisons were performed using Kruskal-Wallis tests with Dunn\u0026rsquo;s correction for multiple testing and ANOVA. For categorical variables, the chi-square test or Fisher's test was conducted. The correlation coefficients among Th2 biomarker levels and clinical characteristics were analyzed using Spearman's rank correlation coefficient. The association of clinical remission asthma classes with clinical variables was examined using regression models adjusted for potential confounders, including FEV1 (% predicted), FEV1/FVC ratio, FEF25-75, pred %, Total IgE (IU/mL), TSLP (pg/ml). The odds ratio (OR) and 95% confidence interval (CI) were reported. The diagnostic performance of serum TSLP levels to identify predicting biomarkers of clinical remission asthma in children was determined by receiver operating characteristic (ROC) curve analysis. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated for the selected cut-off point. A \u003cem\u003eP\u003c/em\u003e level\u0026thinsp;\u0026lt;\u0026thinsp;0.05 was considered statistically significant as shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e "},{"header":"3 Results","content":"\u003cdiv id=\"Sec14\" class=\"Section2\"\u003e\n\u003ch2\u003e3.1 Baseline characteristics\u003c/h2\u003e\n\u003cp\u003eIn total, 172 children were enrolled, among those 119 were diagnosed with asthma by the clinical physician or still experiencing acute asthma attacks, and 53 were control subjects with no respiratory disease. Baseline demographics, clinical characteristics, and levels of T2 biomarkers of the overall population are reported in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e. There were no significant differences between the children with asthma and control groups regarding age and sex, while children with asthma had significantly higher blood eosinophil counts than controls (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.014). As for the Th2 biomarkers(IL-4 \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.142, IL-5 \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.4791, IL-13 \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.6885, Serum IgE \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.2534, TARC \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.2074, TSLP \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.6571), there are no significant statistical differences observed between asthma and control subjects.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\n\u003ch3\u003eTable1: Demographic and laboratory characteristics of the study groups (n = 172).\u003c/h3\u003e\n\u0026nbsp;\u003c/div\u003e\n\u003ctable style=\"width: 4.3e+2pt; border-collapse: collapse; border: none;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 106.2pt; border-top: solid windowtext 1.5pt; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: none; background: white; padding: 0in 5.4pt 0in 5.4pt; height: 30.6pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 106.45pt; border-top: solid windowtext 1.5pt; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: none; background: white; padding: 0in 5.4pt 0in 5.4pt; height: 30.6pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eAsthma\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003en=119\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 125.6pt; border-top: solid windowtext 1.5pt; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: none; background: white; padding: 0in 5.4pt 0in 5.4pt; height: 30.6pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eControl\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003en=53\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 95.7pt; border-top: solid windowtext 1.5pt; border-left: none; border-bottom: solid windowtext 1.0pt; border-right: none; background: white; padding: 0in 5.4pt 0in 5.4pt; height: 30.6pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eP value\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 106.2pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 14.6pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eSex(M/F)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 106.45pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 14.6pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e(70/49)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 125.6pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 14.6pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e(31/22)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 95.7pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 14.6pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e0.967\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 106.2pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 19.85pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eAge（years）\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 106.45pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 19.85pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e6.7\u0026plusmn;3.1\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 125.6pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 19.85pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e7.2\u0026plusmn;2.9\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 95.7pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 19.85pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e0.2048\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 106.2pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 29.2pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eEosinophils count/l\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 106.45pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 29.2pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e0.26（0.13-1.7）\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 125.6pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 29.2pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e0.16（0.1-0.7）\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 95.7pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 29.2pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e0.0413\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 106.2pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: .5in;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eBlood eosinophils (%)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 106.45pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: .5in;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e2.1 (1.3-3.225)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 125.6pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: .5in;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e3.35 (1.5-5.75)\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 95.7pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: .5in;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e0.014\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 106.2pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 58.35pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eIL-4（pg/ml)\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eMean\u0026plusmn;SD\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eMedian（min-max）\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 106.45pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 58.35pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e50.85\u0026plusmn;22.11\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e48.56（15.08-175.7）\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 125.6pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 58.35pt;\"\u003e\n\u003ctable style=\"float: right; width: 1.2e+2pt; border: none; border-collapse: collapse; margin-left: 6.75pt; margin-right: 6.75pt;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 123.7pt; padding: 0in 5.4pt 0in 5.4pt; height: 14.2pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e54.26\u0026plusmn;17.94\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 123.7pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e53.85（17.06-107.1）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 95.7pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 58.35pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e0.142\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 106.2pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 58.35pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eIL-5（pg/ml)\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eMean\u0026plusmn;SD\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eMedian（min-max）\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 106.45pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 58.35pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable style=\"width: 1.2e+2pt; border: none; border-collapse: collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e246.3\u0026plusmn;114.9\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e238.8（52.94-890.5）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 125.6pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 58.35pt;\"\u003e\n\u003ctable style=\"width: 1.4e+2pt; border: none; border-collapse: collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 145.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e241.7\u0026plusmn;102.8\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 145.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e229.8（78.38-597.5）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 95.7pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 58.35pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e0.4791\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 106.2pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 58.35pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eIL-13（pg/ml)\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eMean\u0026plusmn;SD\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eMedian（min-max）\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 106.45pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 58.35pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable style=\"width: 1.2e+2pt; border: none; border-collapse: collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e174.3\u0026plusmn;70.92\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e178.8（50.74-335.8）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 125.6pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 58.35pt;\"\u003e\n\u003ctable style=\"width: 1.4e+2pt; border: none; border-collapse: collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 145.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e165.5\u0026plusmn;56.48\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 145.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 20.4pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e172.9\u0026plusmn;（39.75-263.9）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 95.7pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 58.35pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e0.6885\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 106.2pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 72.95pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eSerum IgE（ng/ml）\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eMean\u0026plusmn;SD Median（min-max）\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 106.45pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 72.95pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable style=\"width: 1.2e+2pt; border: none; border-collapse: collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e31.93\u0026plusmn;20.11\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e27.21（7.345-109.2）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 125.6pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 72.95pt;\"\u003e\n\u003ctable style=\"width: 1.4e+2pt; border: none; border-collapse: collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 145.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e29.32\u0026plusmn;20.42\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 145.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 25.65pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e22.15（7.832-104.3）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 95.7pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 72.95pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e0.2534\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 106.2pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 72.95pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003ePeriostin（ng/ml）\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eMean\u0026plusmn;SD\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eMedian（min-max）\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 106.45pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 72.95pt;\"\u003e\n\u003ctable style=\"float: left; width: 1.2e+2pt; border: none; border-collapse: collapse; margin-left: 6.75pt; margin-right: 6.75pt;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 121.6pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e1.169\u0026plusmn;0.7573\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 121.6pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e1.002（0.2556-4.772）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 125.6pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 72.95pt;\"\u003e\n\u003ctable style=\"float: left; width: 1.4e+2pt; border: none; border-collapse: collapse; margin-left: 6.75pt; margin-right: 6.75pt;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 145.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e1.275\u0026plusmn;0.7091\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 145.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e1.193（0.07387-3.106）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 95.7pt; border: none; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 72.95pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e0.2074\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 106.2pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 72.95pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eTARC（pg/ml)\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eMean\u0026plusmn;SD Median（min-max）\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 106.45pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 72.95pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable style=\"width: 1.2e+2pt; border: none; border-collapse: collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e251.7\u0026plusmn;93.31\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e247.8（54.47-533.5）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 125.6pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 72.95pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable style=\"width: 1.4e+2pt; border: none; border-collapse: collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 145.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e265.9\u0026plusmn;81.19\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 145.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e265.3（48.53-426.8）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 95.7pt; border: none; background: #E7E7E7; padding: 0in 5.4pt 0in 5.4pt; height: 72.95pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e0.2159\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 106.2pt; border: none; border-bottom: solid windowtext 1.5pt; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 59.75pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eTSLP（pg/ml)\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eMean\u0026plusmn;SD\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003eMedian（min-max）\u003c/span\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 106.45pt; border: none; border-bottom: solid windowtext 1.5pt; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 59.75pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003ctable style=\"width: 1.2e+2pt; border: none; border-collapse: collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e374.2\u0026plusmn;285.7\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 118.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 17.0pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e297.2（72.38-1400）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 125.6pt; border: none; border-bottom: solid windowtext 1.5pt; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 59.75pt;\"\u003e\n\u003ctable style=\"width: 1.4e+2pt; border: none; border-collapse: collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 145pt; padding: 0in 5.4pt; height: 17pt; vertical-align: bottom;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e356.6\u0026plusmn;285.1\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 145.0pt; padding: 0in 5.4pt 0in 5.4pt; height: 34.45pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e250.2（86.81-1589）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/td\u003e\n\u003ctd style=\"width: 95.7pt; border: none; border-bottom: solid windowtext 1.5pt; background: #CBCBCB; padding: 0in 5.4pt 0in 5.4pt; height: 59.75pt;\"\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u003cspan style=\"color: black;\"\u003e0.6571\u003c/span\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center; text-indent: -8.05pt; line-height: 150%; font-size: 16px; font-family: SimSun; margin: 0in 0in 6.0pt 8.05pt;\"\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eTable\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e: Data are presented as percentage, median (inter-quartile range), or mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation. Values in bold are statistically significant.Abbreviations: IgE, immunoglobulin E; IL, interleukin; SD, standard deviation; TARC, thymus, and activation-regulated chemokine; TSLP, thymic stromal lymphopoietin.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e\n\u003ch2\u003e3.2 Baseline correlations between biomarkers\u003c/h2\u003e\n\u003cp\u003eAt baseline, the level of serum TSLP has positive correlations with TRAC (r\u0026thinsp;=\u0026thinsp;0.5), IL-4 (r\u0026thinsp;=\u0026thinsp;0.2), IL-5 (r\u0026thinsp;=\u0026thinsp;0.6), IL-13(r\u0026thinsp;=\u0026thinsp;0.3), and Periostin (r\u0026thinsp;=\u0026thinsp;0.3) (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0 .005 in all) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eA\u0026ndash;E). Positive correlations were also observed at baseline for IgE with IL-5 (r\u0026thinsp;=\u0026thinsp;0.4), TRAC (r\u0026thinsp;=\u0026thinsp;0.3), and IL-13 (r\u0026thinsp;=\u0026thinsp;0.4) (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0 .0001 in all) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eF\u0026ndash;H). There were also positive correlations between TRAC and IL-5 (r\u0026thinsp;=\u0026thinsp;0.5), and IL-13 (r\u0026thinsp;=\u0026thinsp;0 .4), as well as between TRAC and Periostin (r\u0026thinsp;=\u0026thinsp;0.5) (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0 .0001 in all) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eI-K). Periostin had a positive correlation with IL-5 (r\u0026thinsp;=\u0026thinsp;0.3) and IL-13 levels (r\u0026thinsp;=\u0026thinsp;0.2) at baseline(\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0 .05 in all) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003el-M). IL-13 and IL-5 (r\u0026thinsp;=\u0026thinsp;0.7) levels were positively correlated at baseline (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003eN), with nominal \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001. Additionally, the levels of IL-13 were positively correlated with blood eosinophils (%) at baseline (r\u0026thinsp;=\u0026thinsp;0.2) with nominal \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05. No correlations were found between IgE, Periostin, IL-4, and Blood eosinophils (%) with any of the other biomarkers assessed.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec16\" class=\"Section2\"\u003e\n\u003ch2\u003e3.3 Clinical features of Type-2-high patients\u003c/h2\u003e\n\u003cp\u003eTh2-high asthma criteria were defined based on blood eosinophil counts and allergen-specific serum IgE: IgE\u0026thinsp;\u0026gt;\u0026thinsp;100 IU/ml and a blood eosinophil count\u0026thinsp;\u0026ge;\u0026thinsp;140 cells/\u0026micro;l. The enrolled subjects were recategorized into Th2 high asthma (n\u0026thinsp;=\u0026thinsp;110) and controls(n\u0026thinsp;=\u0026thinsp;53). The Th-2 high subjects were ulteriorly grouped into three categories (acute asthma, persistent asthma, and clinical remission), those who presented with worsening wheezing, shortness of breath, cough, and chest tightness, and were diagnosed as acute asthma attack by physicians were defined as asthma cases in acute exacerbation or acute asthma attack (n\u0026thinsp;=\u0026thinsp;48). Asthma cases in clinical remission (n\u0026thinsp;=\u0026thinsp;36) were characterized by an absence of asthma symptoms and medicine use for at least\u0026thinsp;\u0026ge;\u0026thinsp;1 year with no acute asthma attack within a year. Persistent asthma(n\u0026thinsp;=\u0026thinsp;26)was defined as those with presence of asthma symptoms in the last year or the daily use of long-term control medications to achieve and maintain control in the last year. A comparison of clinical characteristics is presented in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. Obviously, gender or age was not a factor. No significant differences were found for allergic rhinitis, blood eosinophils (%), total IgE levels, aeroallergen sensitization, and FeNO (parts/billion) between groups. However, differences were statistically significant with regard to lung function measurements including FEV1 (% predicted) (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.015), FEV1/FVC, % (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.40), FEF25-75, pred % \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.0021).\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab2\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003echaracteristics of patients defined as acute attack asthma, persistent asthma, and clinical remission.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eAcute Attack Asthma\u003c/p\u003e\n\u003cp\u003en\u0026thinsp;=\u0026thinsp;48\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003ePersistent\u003c/p\u003e\n\u003cp\u003eAsthma\u003c/p\u003e\n\u003cp\u003en\u0026thinsp;=\u0026thinsp;26\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eClinical Remmision\u003c/p\u003e\n\u003cp\u003en\u0026thinsp;=\u0026thinsp;36\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eP\u003c/em\u003e value\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSex(M/F)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(33/21)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(13/14)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e(24/14)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.431\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAge(years)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.9\u0026thinsp;\u0026plusmn;\u0026thinsp;2.7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.5\u0026thinsp;\u0026plusmn;\u0026thinsp;3.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6.6\u0026thinsp;\u0026plusmn;\u0026thinsp;3.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.816\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAllergic rhinitis\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e48.1%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e42.1%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e51.9%\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.732\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eFEV1(% predicted)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e80.15\u0026thinsp;\u0026plusmn;\u0026thinsp;18.86\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e92.90\u0026thinsp;\u0026plusmn;\u0026thinsp;14.23\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e103.0\u0026thinsp;\u0026plusmn;\u0026thinsp;44.65\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e\u003cstrong\u003e0.0015\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eFEV1/FVC, %\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e92.77\u0026thinsp;\u0026plusmn;\u0026thinsp;14.48\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e99.62\u0026thinsp;\u0026plusmn;\u0026thinsp;6.573\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e95.22\u0026thinsp;\u0026plusmn;\u0026thinsp;22.45\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e\u003cstrong\u003e0.040\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eFEF25-75, pred %,\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e56.53\u0026thinsp;\u0026plusmn;\u0026thinsp;24.42\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e81.44\u0026thinsp;\u0026plusmn;\u0026thinsp;25.66\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e71.74\u0026thinsp;\u0026plusmn;\u0026thinsp;17.38\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e\u003cstrong\u003e0.0021\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eFeNO(parts/billion)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e20.72\u0026thinsp;\u0026plusmn;\u0026thinsp;22.76\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e20.79\u0026thinsp;\u0026plusmn;\u0026thinsp;24.52\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e21.57\u0026thinsp;\u0026plusmn;\u0026thinsp;23.69\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.480\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eBlood eosinophils (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.1(0.1\u0026ndash;20.1)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e2.9(0.1\u0026ndash;12.4)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.75(0.1\u0026ndash;127)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.267\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTotal IgE (IU/ml)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e160.9(5.2\u0026ndash;1814)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e104(12.6\u0026ndash;1346)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e44.8(8-1905)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.500\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cstrong\u003eSkin tests\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAeroallergen sensitization\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e60.9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e47.4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e50\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.452\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eHouse-dust mites\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e21.7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e26.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e29.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.581\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eanimal dander\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e8.7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e21.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e8.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.674\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003efood allergen sensitization\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e30.4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e31.6\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e29.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.569\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003epollens\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e21.7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e8.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.347\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eMold\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e8.3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.940\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eTable\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e: Data are mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD, median (interquartile range), or percentage. Values in bold are statistically significant. Abbreviation: FEV1,forced expiratory volume during the first second; FVC, forced vital capacity; FEF25-75, pred %, forced expiratory flow between 25% and 75%.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec17\" class=\"Section2\"\u003e\n\u003ch2\u003e3.4 Th2 biomarkers of clinical Th2 high asthma groups\u003c/h2\u003e\n\u003cp\u003eAs shown in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e, there was no obvious difference among the groups in serum concentration of Th2 biomarkers (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eA-F) except for TSLP (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eG). Notably, TSLP levels were significantly higher in subjects experiencing acute exacerbation and children defined as asthma persistence compared with non-asthmatic control subjects. There was no statistically significance between clinical remission and control subjects.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec18\" class=\"Section2\"\u003e\n\u003ch2\u003e3.5 Correlation between serum TSLP level and clinical variables in Th2 high asthma\u003c/h2\u003e\n\u003cp\u003eSpearman's correlation coefficients were conducted to investigate whether serum TSLP level has associations with subjects\u0026rsquo; clinical characteristics in Th2-high asthma and whether it can be used as a predictor to evaluate the ongoing eosinophilic inflammation remission. The results were shown in Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e. Serum TSLP level was found significantly correlated with total IgE (Spearman's rho [r]\u0026thinsp;=\u0026thinsp;0.48, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001), FEV1/FVC ratio (r\u0026thinsp;=\u0026thinsp;\u0026minus;\u0026thinsp;0.365, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05), and FEF25-75, pred % (r\u0026thinsp;=\u0026thinsp;\u0026minus;\u0026thinsp;0.06, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05), whereas no correlations were found with age, BMI, or allergic rhinitis, atopic dermatitis, blood eosinophil counts.\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"char\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"char\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab3\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eSpearman's correlation coefficients between serum TSLP level and other clinical characteristics in children with Th2 high asthma.\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eSerum TSLP(pg/ml)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\u0026nbsp;\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003er\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eP\u003c/em\u003e value\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAge, y\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-0.03\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.770\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eBMI\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-0.081\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.455\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAtopic dermatitis\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.372\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.155\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAllergic rhinitis\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.098\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.119\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTotal IgE (IU/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.482\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e\u003cstrong\u003e\u0026lt;\u0026thinsp;.0001\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eEosinophils (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-0.072\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.502\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eFEV1, pred %\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-0.106\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.457\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eFEV1/FVC ratio\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-0.365\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e\u003cstrong\u003e0.008\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eFEF25-75, pred %\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e-0.306\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e\u003cstrong\u003e0.031\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eTable\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e: Abbreviations: BMI, body mass index; FEF25-75, forced expiratory flow between 25% and 75%; FEV1, forced expiratory volume during the first second; FVC, forced vital capacity.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.6 Multivariable regression and ROC analyses for predictors of clinical asthma remission and persistence in Th-2 high asthma\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTo find out which predictors show relationships with clinical remission in Th2-high asthma, a multiple regression model was designed including the variables showing differences between readjusted Th2-high groups and variables associations with TSLP in Spearman\u0026rsquo;s analyses. The results of multivariable logistic regression analysis revealed that serum TSLP levels were associated with clinical remission in Th2-high asthma subjects (OR\u0026thinsp;=\u0026thinsp;1.009; 95% CI:1.008\u0026ndash;1.008, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.023) (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). Serum TSLP level correlated with clinical remission in Th2-high asthma in terms of ROC curves analysis (AUC\u0026thinsp;=\u0026thinsp;0.5887, 95% CI: 0.5052 to 0.7038, P\u0026thinsp;\u0026lt;\u0026thinsp;0 .05) (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). Considering sensitivity, and specificity, for serum TSLP levels, a cutoff value of 373.363 pg/ml was found with the highest clinical sensitivity and specificity. PPV and NPV of 100%, 39.1%, and 59.4%, 100%, respectively. However, no positive result was found in the analysis of multivariable logistic regression in determining the predictors of Th2 high persistent asthma.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cdiv class=\"gridtable\"\u003e\n\u003cdiv class=\"colspec\" align=\"left\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"char\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003cdiv class=\"colspec\" align=\"char\"\u003e\u0026nbsp;\u003c/div\u003e\n\u003ctable id=\"Tab4\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eMultivariable logistic regression analysis for clinic remission in Th2 high asthma\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eVariable\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eOR (95% CI)\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eP\u003c/em\u003e Value\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eFEV1(% predicted)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.963(0.959\u0026ndash;0.966)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.411\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eFEV1/FVC ration\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.085(1.096\u0026ndash;1.074)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.189\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eFEF25-75, pred %\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.974(0.972\u0026ndash;0.976)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.568\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTotal IgE (IU/mL)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.014(1.014\u0026ndash;1.013)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.668\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTSLP (pg/ml)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e1.009(1.008\u0026ndash;1.008)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"char\" char=\".\"\u003e\n\u003cp\u003e0.023\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003c/div\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec19\" class=\"Section2\"\u003e\n\u003ch2\u003e3.7 Comparison of TSLP in serum, BALF, and lung tissue of different mouse model groups\u003c/h2\u003e\n\u003cp\u003eThe hallmark of the Th2-high asthma was IL4, IL5, and IL13, and the mouse Elisa kit was used to screen the Th2 cytokines in BALF and serum of different stages mouse models first (data not shown). Measurements of TSLP levels was then examined in serum, BALF, and lung tissue of different stages of mouse asthma models to confirm and validate the observations from the study population. We aimed to inspect the expression of TSLP in different tissues of asthmatic mice and to find out the most indicative significance.\u003c/p\u003e\n\u003cp\u003eWhen comparing the concentration of TSLP in serum, BALF, and lung tissue, it was found that BALF had the lowest level of TSLP concentration. There was no statistically significant change in the acute and remission stages when compared to the control, but TSLP did increase noticeably in the chronic stage (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eA).\u003c/p\u003e\n\u003cp\u003eThere seems to be no significant difference in the levels of TSLP in mouse serum between control and asthma remission. However, it had been noted that during the acute and chronic stages, TSLP levels increased, as depicted in the figure. (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eB) Furthermore, the lung tissue also demonstrated a comparable trend as serum, with significant statistical significance observed in this case (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003eC).\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4 Discussion","content":"\u003cp\u003eThis cross-sectional study presents the positive association observed between Th2 biomarkers in circulating levels and demonstrated robust relationships between several biomarkers of T2 inflammation. In addition, it extended previous investigations regarding identifying the potential biomarkers predicting clinical remission in Th2-high asthma.\u003c/p\u003e \u003cp\u003eWe found that when subjects were simply partitioned into asthma and control, no significance was observed in Th2 biomarkers between these two groups, whereas clinical measurements such as blood eosinophils (%) and eosinophils count were statistically significant. When we defined Th2 high asthma based on eosinophils count (\u0026ge;\u0026thinsp;140 cells/\u0026micro;l ) and total IgE (IgE\u0026thinsp;\u0026gt;\u0026thinsp;100 IU/ml), the serum TSLP levels were observed higher in subjects experiencing acute exacerbation and persistent asthma compared with no asthma control subjects. In addition, our results also found an association between serum TSLP levels and clinical remission independent of confounding factors such as lung function measurements or allergy sensitization. However, the role of serum TSLP in identifying clinical remission of Th2-high asthma still needs further holistic investigations to reconfirm its specificity and sensitivity to be used as a prognostic biomarker in asthma.\u003c/p\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003e4.1 Th2 biomarkers at baseline\u003c/h2\u003e \u003cp\u003eSince asthma is a heterogeneous disease that can be divided into many subgroups, one of the widely accepted consensuses is that the research of it needs to evolve from phenotypes to endotypes [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Phenotypes are usually defined by clinical characteristics, allergen sensitization patterns, and physiologic lung function measurements which are readily observed or measured in clinical practice. Endotypes are a definition that is related to pathophysiological mechanisms underlying phenotype [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. Thus, there is a growing urge to investigate asthma endotypes by connecting discernible and stable biomarkers with immunological mechanisms to make precision therapy decisions for asthma [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. Th2-high asthma is a broad classification involving asthma endotypes based on type 2 (T2) inflammation pathobiology characterized by increased secretion of Th2 biomarkers such as IL-4, IL-5, IL-13, and IgE causing pathological changes, which can be summarized by a set of the cytokine-induced eosinophilic airway inflammation, epithelial dysfunction, and airway wall remodeling [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]. And it is believed that Th2-high asthma account for only half of the people diagnosed with asthma [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e], which in a way supports our results that there was no considerable significance at the baseline levels of Th2 biomarkers between asthma and control when study population was not recruited specifically with Th2-high characteristics. When investigating the correlations between the biomarkers and clinical characteristics, there were no positive results observed between Th2 biomarkers and baseline blood eosinophil counts or total IgE except that IL-13 was positively correlated with blood eosinophils (%). Whereas, Corren, Pham, et al found that baseline levels of serum IL-5, IL-13, and Periostin correlated significantly with baseline blood eosinophil counts and FeNO levels in uncontrolled severe asthma subjects, which may account for the complex immune pathways in severe asthma [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. After all, this is a relatively representative study measuring baseline circulating levels of Th2 biomarkers, and the positive correlations among specific biomarkers on one side further support the molecular mechanism of Th2 biomarkers, on the other side it may provide insight into combined immune biologic interventions in personalized therapy.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003e4.2 Asthma remission, persistence and predictive role of Th2 biomarkers\u003c/h2\u003e \u003cp\u003eAsthma remission is not emphasized in asthma treatment goals, unlike other chronic diseases, and it is widely accepted that clinical remission and complete remission have different criteria, and current research involving asthma remission is limited [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eWang, Datta [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e] demonstrated that baseline lung function can be used as a clinical prognostic indicator of asthma remission. And other independent predictors of asthma remission include less BHR, no nasal polyps, younger age, better asthma control, lower doses of ICS, and lower levels of blood neutrophils [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Actually, when comes to asthma remission, clinical features are mostly considered [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. However, with the prevalent use of biologics in asthma treatments, and the great development from phenotypes to endotypes in asthma pathophysiological mechanisms, identifying predicting biomarkers of asthma remission, especially in the Th2 high subtype may mean a step closer to clinical asthma remission evaluation. Thus, in this study, the study population was further divided into acute asthma attacks, persistent asthma, and clinical remission asthma among Th2-high subjects. The criteria of Th2-high and the definition of different phases were explained above. It is obvious that lung measurements including FEV1(% predicted), FEV1/FVC, %, FEF25-75, pred %, are statistically significant among groups which are in line with many research that lung measurements are sensitive clinical features evaluating asthma remission [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e].When comparing circulating levels of Th2 biomarkers among groups, there was no marked significance except serum TSLP. However, Tan, Daniel J et al found that serum inflammatory cytokines including IL-4, and IL-5 have prognostic value in adults with spontaneous asthma remission. And there was solid evidence that Periostin was associated with asthma persistence, especially in severe uncontrolled asthma [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]. In addition,Christianson, Goplen [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e] elucidated that IL-13 is a crucial factor in the feedback and feed-forward interactions that contribute to the persistence of asthma in mouse models. TARC(thymus activation-regulated chemokine) which is expressed in T helper (Th2) cells, also known as CCL17, belongs to the CC chemokine family. TARC is believed to have a pivotal role in the pathogenesis of allergic diseases such as atopic dermatitis, bronchial asthma, and allergic rhinitis [\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]. What's more, the serial changes of plasma TARC concentrations in children with acute asthma attacks were monitored in a study which indicated that TARC is a potentially useful inflammatory maker to evaluate asthmatic exacerbation [\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e]. To sum up, these conflicts may due to the difference in study population and design, and it is crucial to emphasize the fact that the research on the levels of biomarkers associated with asthma remission and persistence is limited. Therefore, there is a pressing need to conduct more studies in this area to gain critical insights into this debilitating condition.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec23\" class=\"Section2\"\u003e \u003ch2\u003e4.3 Association between clinical asthma remission and serum TSLP\u003c/h2\u003e \u003cp\u003eThymic stromal lymphopoietin (TSLP), part of the IL-2 cytokine family, was discovered as a growth factor for lymphocyte progenitors and is now recognized as a protein primarily released from epithelial cells and has a sensitive response to irritating stimuli [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. and research involving mice models to human subjects and the genome-wide association had demonstrated that TSLP is associated with asthma [\u003cspan additionalcitationids=\"CR41\" citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. Indeed, TSLP regulates dendritic cells which are considered one of the central responders of Th2 immunity [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. What\u0026rsquo;s more, with the approval of TSLP-targeted antibody for severe asthma, it is reported that TSLP plays a key role in allergen-induced airway responses and persistent airway inflammation in patients with allergic asthma [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. However, whether it can be a predictor to evaluate the clinical remission of Th2-high asthma is still understudied. There was research based on sputum type 2 makers and found that the sputum TSLP levels were higher at baseline in the remission group compared with the non-remission group and TSLP was not listed as a potential predictor of remission by univariate regression analysis [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. On the contrary, our results found that serum TSLP has a significant difference among acute asthma groups, asthma persistence groups, and clinical remission group, and multivariable regression also revealed that serum TSLP levels were associated with clinical remission in Th2 high asthma children. The fundamental role of TSLP in asthma pathogenesis was underscored by the fact that the latest clinical trials have reconfirmed that treatment with Tezepelumab, which blocks TSLP, can lead to sustained and significant reductions in asthma exacerbations in individuals with severe, uncontrolled asthma [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. A multi-center prospective observational study suggests that TSLP may play a pathogenic role across different asthma phenotypes [\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e].\u003c/p\u003e \u003cp\u003e \u003cb\u003e4.4 The expression of TSLP in BALF, serum, and lung tissue of different stages of asthmatic mice.\u003c/b\u003e \u003c/p\u003e \u003cp\u003eWe further conducted an animal model of allergic asthma which shows the levels of TSLP vary in acute, chronic, and remission asthma mouse models. Our findings indicate that the TSLP levels are remarkably high in chronic asthma mouse models among the lung tissue, BALF, and serum. Furthermore, both in the serum and lung tissue samples, we have also observed that the decrease in TSLP levels may indicate the remission of asthma. Numerous murine research have demonstrated that TSLP plays an important role in the development of asthma [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]. The anti-TSLP method or TSLP-R deficiency can effectively attenuate AHR and TH2 airway inflammation [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e, \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e]. Notably, an interesting research also revealed that mice received TSLP receptor-deficient CD4\u0026thinsp;+\u0026thinsp;T cells had alleviated pathogenic TH2 cell responses, and asthmatic children showed an increase in the production of TH2 cells, promoted by TSLP and IL-4, which resulted in higher amounts of IL-5 and IL-13 in their peripheral blood, compared to healthy controls [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. Although it is commonly believed that TSLP plays a crucial role in the pathogenesis of asthma, there is still a lack of research on whether TSLP can be utilized as a biomarker for indicating asthma remission or persistence. We believe that this possibility has not been discussed enough, and therefore we are interested in exploring this topic further.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec24\" class=\"Section2\"\u003e \u003ch2\u003e4.5 Conclusion\u003c/h2\u003e \u003cp\u003eWe report for the first time that serum TSLP may be used as one of the biomarkers to evaluate the remission of asthma along with other measurements such as lung function to improve the accuracy sensitivity and specificity and to testify our findings in acute chronic and remission asthma mouse models.\u003c/p\u003e \u003cp\u003eHowever, despite the positive likelihood of evaluating remission shown in ROC curves analysis, its diagnostic role and prognostic role in identifying clinical asthma remission still need further exploration.\u003c/p\u003e \u003cp\u003eWe aim to expand our understanding of TSLP and its clinical significance while advancing our current research.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec25\" class=\"Section2\"\u003e \u003ch2\u003e4.6 limitation and strength\u003c/h2\u003e \u003cp\u003eThe limitations of this study lie in that subjects and the Th2 biomarkers should have been be detected in a larger cohort of multiple centers and the definition of asthma remission is still under-refined. Also, the steadiness of the circulating biomarkers should be validated in the longitudinal study. Furthermore, investigation related to other biomarkers of different pathways is expected to be conducted to form a more holistic understanding of asthma remission. It is the first research to investigate the role of circulating levels of Th2 biomarkers as predictors of remission in a cohort of asthma subjects though it is a preliminary and retrospective study.\u003c/p\u003e \u003c/div\u003e"},{"header":"Declarations","content":"\n\u003cp\u003e4.7 Funding\u003c/p\u003e\n\u003cp\u003eThis work was supported by the National Natural Science Foundation of China (Grant No. 82205189, 82174170, 82205044), Shanghai science and technology commission Project (Grant No. 21S21902500), as Fund of Huashan Hospital affiliated to Fudan University (Grant No. 2021QD042).\u003c/p\u003e\n\u003ch3\u003e4.8 Acknowledgments\u003c/h3\u003e\n\u003cp\u003eThe authors thank the participants of the study, Qiuyan Yang collected the serum from the children, Mengmeng Chen, for her contributions to the analysis and critical review, and Na Wang, MS, Ph, for her contribution to the statistical analysis. Medical writing support was provided by Congcong Li, Huijie Zhang, and Zhiyong Liu. This study was co-sponsored by Yanli Wang,Na Wang and Jingcheng Dong.\u003c/p\u003e\n\u003cp\u003e4.9 Conflict of interest\u003c/p\u003e\n\u003cp\u003eThe authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e4.10 Ethical Approval and Consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis trial was conducted under the ethical principles of the Declaration of Helsinki, the International Council for Harmonisation guidelines for good clinical practice, and applicable regulatory requirements. The protocol (2021-108-01) was initially approved by the ethics committee of the Third Affiliated Hospital of Zhengzhou University. All participants and their parents granted their written informed consent and all experiments were performed per the approved guidelines and regulations.\u003c/p\u003e\n\u003cp\u003eAll animal experiments are approved by the Laboratory Animal Ethics Committee (IACUC) of Fudan University and are conducted in strict accordance with the laboratory animal management regulations of Fudan University (2022JS Huashan hospital-497).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e4.11 Availability of supporting data\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data are available and the correspondent can be contacted if requested.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eWenzel SE. Asthma phenotypes: the evolution from clinical to molecular approaches. Nat Med. 2012;18(5):716\u0026ndash;25.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePijnenburg MW et al. Childhood asthma- pathogenesis and phenotypes. 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ERJ Open Res, 2020. 6(4).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHolm M, et al. Remission of asthma: a prospective longitudinal study from northern Europe (RHINE study). Eur Respir J. 2007;30(1):62\u0026ndash;5.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRonmark E, et al. Outcome and severity of adult onset asthma\u0026ndash;report from the obstructive lung disease in northern Sweden studies (OLIN). Respir Med. 2007;101(11):2370\u0026ndash;7.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHanania NA, et al. Exploring the effects of omalizumab in allergic asthma: an analysis of biomarkers in the EXTRA study. Am J Respir Crit Care Med. 2013;187(8):804\u0026ndash;11.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBobolea I, et al. Sputum periostin in patients with different severe asthma phenotypes. 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Biologics and the lung: TSLP and other epithelial cell-derived cytokines in asthma. Pharmacol Ther. 2017;169:104\u0026ndash;12.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDahlen SE. TSLP in asthma\u0026ndash;a new kid on the block? N Engl J Med. 2014;370(22):2144\u0026ndash;5.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHarada M, et al. Thymic stromal lymphopoietin gene promoter polymorphisms are associated with susceptibility to bronchial asthma. Am J Respir Cell Mol Biol. 2011;44(6):787\u0026ndash;93.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAn G, et al. Combined blockade of IL-25, IL-33 and TSLP mediates amplified inhibition of airway inflammation and remodelling in a murine model of asthma. Respirology. 2020;25(6):603\u0026ndash;12.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGauvreau GM, et al. Effects of an anti-TSLP antibody on allergen-induced asthmatic responses. 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Sci Rep. 2021;11(1):8425.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAl-Shami A, et al. A role for TSLP in the development of inflammation in an asthma model. J Exp Med. 2005;202(6):829\u0026ndash;39.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKabata H, et al. Targeted deletion of the TSLP receptor reveals cellular mechanisms that promote type 2 airway inflammation. Mucosal Immunol. 2020;13(4):626\u0026ndash;36.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHu Y, et al. TSLP signaling blocking alleviates E-cadherin dysfunction of airway epithelium in a HDM-induced asthma model. Cell Immunol. 2017;315:56\u0026ndash;63.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eToki S, et al. TSLP and IL-33 reciprocally promote each other's lung protein expression and ILC2 receptor expression to enhance innate type-2 airway inflammation. Allergy. 2020;75(7):1606\u0026ndash;17.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRochman Y et al. \u003cem\u003eTSLP signaling in CD4(+) T cells programs a pathogenic T helper 2 cell state.\u003c/em\u003e Sci Signal, 2018. 11(521).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Asthma, Th2 Biomarkers, Thymic stromal lymphopoietin, Clinical asthma remission, Asthma persistence","lastPublishedDoi":"10.21203/rs.3.rs-4182358/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4182358/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003eBackground\u003c/h2\u003e \u003cp\u003eLittle study investigates the association between baseline type 2 biomarker levels and clinical features in children diagnosed with asthma. Characterizing clinical remission in Th2-high asthma could offer valuable insights into asthma prognosis.\u003c/p\u003e\u003ch2\u003eObjectives\u003c/h2\u003e \u003cp\u003eThe study aims to investigate the association of baseline type 2 biomarker levels and clinical features in children with asthma, and to identify predictors of clinical remission of asthma in children.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003eA total of 172 children with baseline age of 6.87\u0026thinsp;\u0026plusmn;\u0026thinsp;3.04 (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD) years were enrolled in the study including 119 with physician-diagnosed asthma who regularly attended a pediatric asthma center and 53 control subjects with no respiratory symptoms. Clinical tests included lung function examination, Fraction of exhaled NO (FeNO), total IgE, blood eosinophil, and skin test. Serum Th2 biomarkers were examined by ELISA. The enrolled patients have readjusted into Th2-high asthma according to clinical eosinophil count and total IgE, and Th2-high asthma subjects were further classified into acute attack asthma, persistent asthma, and clinical remission according to the recent GINA guidelines and clinical evaluation. To verify our results, the concentration of TSLP levels was measured in BALF, serum, and lung tissue by ELISA in mouse models.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eCompared with asthma and control groups, eosinophil counts and blood eosinophils (%) were significant, whereas, no correlation was observed between asthma subjects and controls including Th2 biomarkers, gender, or ages. Positive correlations were observed between Th2 inflammatory biomarkers (TSLP, TRAC IL-5, IL-13, and Periostin) at baseline. Th2-high asthma (n\u0026thinsp;=\u0026thinsp;110) was defined based on clinical measurement of IgE\u0026thinsp;\u0026gt;\u0026thinsp;100 IU/ml and a blood eosinophil count\u0026thinsp;\u0026ge;\u0026thinsp;140 cells/\u0026micro;l. Among those Th2-high asthma subjects, there were 48 in acute exacerbation (43.6%), and 36 in clinical remission (32.7%), 26 were clinical asthma persistence (23.6%). Lung function and serum TSLP had marked significance among the three categories. Compared with clinical remission asthmatic subjects and controls, serum TSLP levels were significantly higher in subjects experiencing acute exacerbation and subjects defined as asthma persistence. Spearman\u0026rsquo;s correlation outlined that serum TSLP levels were related to Total IgE (IU/mL), FEV1/FVC ratio, and FEF25-75, pred %. Multivariate logistic regression analysis demonstrated that serum TSLP levels were associated with clinical remission in Th2-high asthma children (OR\u0026thinsp;=\u0026thinsp;1.009; 95% CI, 1.0087\u0026ndash;1.0086 \u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.023\u0026lt;0.05). It is also revealed that serum TSLP levels may help evaluate clinical remission in Th2-high asthma when using ROC curves analysis (AUC\u0026thinsp;=\u0026thinsp;0.5887, 95% CI: 0.5052 to 0.7038, \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). A cutoff value of 373.363 pg/mL was found with the highest clinical sensitivity and specificity. PPV, and NPV were 100%, 39.1%, and 59.4%, 100% in two groups, respectively. However, there were no positive results in the analysis of multivariable logistic regression in determining the predictors of persistence in Th2-high asthma. In BALF mouse, TSLP concentration had no statistically significant change in the acute and remission stages when compared to the control, but it did increase noticeably in the chronic stage (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.001).\u003c/p\u003e","manuscriptTitle":"Baseline Type 2 Biomarker Levels and Clinical Remission Predictors in Children with Asthma","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-04-02 20:34:57","doi":"10.21203/rs.3.rs-4182358/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"f4cfbb70-f1a6-4ac1-81a6-8be1411de230","owner":[],"postedDate":"April 2nd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-07-03T13:28:15+00:00","versionOfRecord":[],"versionCreatedAt":"2024-04-02 20:34:57","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4182358","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4182358","identity":"rs-4182358","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}