Eosinophils May Serve as CEA-Secreting Cells for ABPA Patients

Eosinophils May Serve as CEA-Secreting Cells for ABPA Patients | 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 Eosinophils May Serve as CEA-Secreting Cells for ABPA Patients Yang yanfei, Gao qiqi, Jin yangyi, Qi mengdie, Lu guohua, li Hequan This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-73116/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 Allergic bronchopulmonary aspergillosis (ABPA) is a condition characterized by an exaggerated response of the immune system to the fungus Aspergillus. This study aimed to assess the relationship between carcinoembryonic antigen (CEA) and eosinophils in ABPA patients. Methods: We describes a case of a 50-year-old patient who was diagnosed with ABPA presenting with high level of CEA and eosinophils. Besides,we used immunohistochemistry and immunofluorescence to identify eosinophils and CEA in sections which were obtained by Endobronchial ultrasound-guided transbronchial lung biopsy aspiration (EBUS-TBLB). The sections were then visualized using confocal microscopy. We also retrospectively analyzed a cohort of 37 ABPA patients between January 2013 and December 2019 in our hospital . Results: We found the patient whose serum CEA levels were consistent with eosinophils during the follow-up(r=0.929,P=0.022). The positive expression of CEA and abnormal expression of eosinophils was higher in the ABPA tissue compared to the normal lung tissue. The co-localization was represented as pixels containing both red and green color in the image (with various shades of orange and yellow) which signified that eosinophils were immunohistochemically positive for CEA. Patients with higher levels of eosinophils had higher levels of CEA in the serum (P<0.001). The results of Pearson correlation analysis showed that the levels of eosinophils were positively correlated with serum CEA levels(r=0.459 and r=0.506，P=0.004 and P=0.001). Conclusion: Serum CEA level is elevated in ABPA patients. The elevated serum CEA level was shown to be normalized after treatment. Increased CEA levels in ABPA patients may be positively correlated with eosinophil levels, and eosinophils may be served as CEA-secreting cells in patients with ABPA. Pulmonology Eosinophils ABPA Patients bronchopulmonary aspergillosis Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Background Allergic bronchopulmonary aspergillosis (ABPA) is characterized by an allergic inflammatory response to Aspergillus fumigatus colonized in the trachea(1). It is manifested as poorly controlled asthma and affected approximately 4 million patients worldwide(2). Some patients may even suffer from irreversible airway obstruction and pulmonary fibrosis(3). Besides, a significant eosinophil infiltration can be seen in the pathological specimens of ABPA patients(4). Carcinoembryonic antigen (CEA) was initially discovered as a tumor antigen(5). Serum CEA level is typically identified as a candidate biomarker for tumor progression(6). However, a patient with high levels of CEA was admitted to our hospital, while she had continuous cough and shortness of breath. Because of departed cognition of CEA, she was initially diagnosed with a malignant disease in another hospital. However, after careful clinical diagnosis, we could not exclude the diagnosis of ABPA. Thus, she received glucocorticoid therapy. After treatment, the patient’s symptoms were remarkably improved. Additionally, her serum CEA level and eosinophil levels were significantly decreased, indicating a subtle balance between the levels of eosinophils and CEA in the serum. Japanese scholars have reported 13 patients with ABPA, in which 7 of them had elevated levels of CEA in the peripheral blood. In case of pulmonary consolidation, the serum CEA levels were gradually returned to normal level(7). Regrettably, the study did not mention the precise source of CEA in ABPA patients. Recently, CEA has also been proposed as a clinical marker, reflecting the activity of acquired idiopathic generalized anhidrosis (AIGA)(8). Immunohistochemistry of 10 AIGA cases showed increased levels of CEA in the eccrine sweat glands(9). A previous study has pointed out that neutrophil extracellular traps (NETs) are associated with CEA(10). The above-mentioned studies indicated that the serum CEA level might be related to the presence of eosinophils. To prove this assumption, We tracked the clinical data of this ABPA patient and double-labeled the lung tissue samples with CEA and eosinophils. 37 cases with ABPA who were admitted to our hospital from 2013 to 2019 were reviewed. To the best of our knowledge, the presented study is the first to report the relationship between serum CEA level and eosinophils in patients with ABPA. Materials And Methods Materials The levels of aspergillus fumigatus-specific IgG and aspergillus fumigatus-specific IgE were detected by using a commercial enzyme-linked immunosorbent assay (ELISA) kit (Thermo Fisher Scientific, Waltham, MA, USA). Besides, interleukin-5 (IL-5) was detected by using human IL-5 platinum ELISA kit (eBioscience Inc., San Diego, CA, USA). Blood eosinophils were measured using a routine blood testing kit. Eosinophils were detected by myelin basic protein (MBP) antibody (Sangon Biotech Co., Ltd., Shanghai, China) and hematoxylin and eosin (H&E) staining. CEA was detceted by CEA monoclonal antibody(Thermo Fisher Scientific, Waltham, MA, USA).A secondary F(ab) polyclonal antibody, goat anti-rabbit IgG (H+L) highly cross-adsorbed secondary antibody, and goat anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody were used. Immunohistochemistry and immunofluorescence The present study was approved by the Ethics Committee of our hospital, and all patients signed written informed consent form prior to study initiation. All reagents used for immunohistochemistry were kindly provided by the Pathology Department of our hospital. The sections were de-waxed, rehydrated, and incubated with 3% hydrogen peroxide in methanol to block endogenous peroxidase activity at room temperature for 10 min. For antigen retrieval, deparaffinized and rehydrated sample tissue sections were pretreated by microwave irradiation for 20–30 min in 0.01 mol/l citrate-buffered saline (pH 6.0). Closed non-specific binding was performed, and normal goat serum was then added to the sections. Sections were then incubated for overnight at 4 °C with primary antibodies (mouse anti-human CEA and rabbit anti-human MBP antibodies) diluted in blocking solution. Membranes were incubated with a secondary antibody, and then goat anti-mouse/rabbit antibody, and diluted at an appropriate dilution in 1% bovine serum albumin (BSA) for 2 h at room temperature. Diaminobenzidine (DAB) color reactions were visualized under a microscope (5–10 min). After rinsing thricewith phosphate-buffered saline (PBS), staining was carried out by DAB before counterstaining with Hematoxylin. Double-immunofluorescence staining was performed as described previously(11). In brief, the sections were analyzed via antigen retrieval, blocked for endogenous peroxidase and non-specific epitopes, and incubated with primary antibody at 4°C overnight. Antibody detection was performed using a secondary F(ab) polyclonal antibody (goat anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody and goat anti-rabbit IgG (H+L) highly cross-adsorbed secondary antibody). The sections were counterstained with 4′,6-diamidino-2-phenylindole (DAPI), and visualized under a LSM880 Meta laser-scanning confocal microscope (Carl Zeiss). Inclusion and exclusion criteria The diagnosis of ABPA is currently done by combining clinical, radiological, and immunological findings based on the criteria proposed by The International Society for Human and Animal Mycology (ISHAM) in 2013(see Table 1).(12) Statistical analysis All data were presented as M(P25, P75), and were analyzed using SPSS 25.0 software (IBM, Armonk, NY, USA) . P ≤ 0.05 was considered statistically significant, and P ≤ 0.01 as highly significant. Pearson correlation coefficient lower than 0.4 indicates poor reproducibility, which range from 0.4-0.75, denoting fair reproducibility, and equal to 0.75 indicates good reproducibility(13). Results A patient with high level of CEA was diagnosed as ABPA A 50-year-old female patient suffering from cough and dyspnea for 2 years was diagnosed with lung cancer because of high serum levels of CEA and lymphadenopathy before she was admitted to our hospital. But the result of positron emission tomography-computed tomography (PET-CT) examination and bone marrow biopsy showed no evidence of tumor. Initial laboratory tests after admission showed elevated CEA level (38.0 ng/mL, reference range < 5.2 ng/mL) and a high count of eosinophils (5700/μL). CT of the lung revealed bronchiectasis and infiltration of the right middle and right lower lobes (Figure 1a). EBUS-TBLB were employed to evaluate mediastinal lymph nodes and lung lesions, showing accumulation of eosinophils in the lungs and lymph nodes. According to the proposed diagnostic criteria for ABPA, the patient was diagnosed with ABPA based on the history of bronchial asthma, as well as the elevated levels of aspergillus fumigatus-specific IgG,serum total IgE and aspergillus fumigatus-specific IgE. Systemic corticosteroid therapy (prednisone 40 mg/day) was initiated, and the patient’s symptoms were dissipated. CT scan of the chest indicated disappearance of pulmonary infiltrates and mucoid impaction (Figure 1.a-d) after 6 months. Eosinophils and CEA were decreased simultaneously The decreased serum CEA levels were consistent with eosinophils during the follow-up (Figure 2). The correlation between the serum CEA levels and eosinophil levels in the 6-month follow-up duration was analyzed by using Pearson correlation analysis, wherein significant correlations were observed (Table 2, r=0.929, P=0.022). Activated CEA was partially co-localized on eosinophils Initially, H&E staining confirmed that a lot of eosinophils in the lung (Figure 3a). CEA was first detected by immunocytochemistry.The yellow-brown granules observed in the cells indicated a positive result for CEA. Additionally, a great number of CEA markers were found in the cells (Figure 3b). The results revealed that the eosinophils were immunohistochemically positive for CEA under high magnification (Figure 3c and d). Immunofluorescence analysis also unveiled that eosinophils ,as well as CEA,were identified in the lung . Moreover，Lung section with a MBP–positive cells were colabeled with CEA antigen which was localized on Cytoplasm(Figure 4 and Figure 5). Increased serum CEA levels were positively associated with eosinophil levels Clinical data of the patients are listed in Table 3. Twenty four male and 13 female patients were included in the current study. Before commencing the treatment, 25 (25/37) patients had serum CEA levels higher than normal. As shown in Table 4, the serum CEA levels were significantly increased, and the mean serum CEA level was markedly higher in ABPA patients than that in healthy subjects (5.05 ng/ml). Moreover, 37 patients were assigned to two groups based on peripheral blood eosinophil counts. Independent -sample t-test was employed to compare serum CEA levels between the two groups. The results showed that patients with higher eosinophil levels had higher serum levels of CEA (P<0.001, Figure 6). Besides, we noted that the eosinophil levels were positively correlated with serum CEA levels(r=0.459 and r=0.506，P=0.004 and p=0.001) by Pearson correlation coefficient (Figure 7). Discussion ABPA is the most significant manifestation of allergic aspergillosis that occurs worldwide, but much attention has not been paid by the scholars(14). Meanwhile, eosinophils promote inflammation, and are known to play a beneficial role in isolating and controlling a disease site(15). To our knowledge, CEA is the most commonly used serum marker in the management of breast cancer, and its expression showed correlation with clinicopathological characteristics of gastric carcinoma(16). CEA was first identified as colon cancer antigen in 1965. The serum CEA levels were higher in patients with colon cancer when compared to healthy individuals, and this led to its clinical application as a diagnostic biomarker for colorectal cancer(17). Recently, the serum levels of CEA and the trend of its changes in the treatment process have been previously validated(18). Thus, some patients with elevated CEA levels were easily misdiagnosed as lung cancer. In the current study, several ABPA patients had higher serum CEA levels than healthy controls. The comparative data demonstrated the necessity of assessment of relationships between the eosinophils and serum CEA level in ABPA patients. In the current study, the serum CEA level was shown to be decreased after treatment, indicating that glucocorticoid therapy might have inhibitory effects on CEA level in ABPA patients. The serum CEA level was found to be positively correlated with blood eosinophils. Previous studies have shown that elevated serum CEA levels in patients with ABPA are associated with consolidation of lungs, mucus plugs, and localized inflammation of lungs(19). Several scholars have pointed out that serum concentrations of CEA were significantly increased in asthmatic patients with mucoid impactions when compared to those patients without mucoid impactions or patients with bronchiectasis(20). However, in our research, the serum levels of CEA were found to be associated with eosinophil count. This is also the first report to present an ABPA case with the relationship between serum CEA level and eosinophils count, and our findings may be significant for clinicians. According to our study, patients with elevated serum CEA levels were diagnosed with benign diseases, (e.g., ABPA). According to the present research, CEA potentially serves as a clinical marker for ABPA patients. However, the present study has two important limitations. Firstly, this was a retrospective single-center study with a relatively small sample size. Secondly, we only included one patient who underwent biopsy, and it is more appropriate to study serum samples of ABPA patients labeled with both CEA and eosinophils. Therefore, further large-scale prospective study should be conducted to verify the preliminary results of the present study. Conclusion In summary, the serum CEA levels were shown to be elevated relatively in ABPA patients. However, the elevated serum CEA level can be normalized after treatment. Increased levels of CEA in ABPA patients might be positively correlated with eosinophil levels, and eosinophils can serve as CEA-secreting cells in patients with ABPA. Abbreviations ABPA Allergic bronchopulmonary aspergillosis CEA carcinoembryonic antigen EBUS-TBLB Endobronchial ultrasound-guided transbronchial lung biopsy aspiration AIGA acquired idiopathic generalized anhidrosis NETs neutrophil extracellular traps ELISA enzyme-linked immunosorbent assay IL-5 interleukin-5 MBP myelin basic protein H&E hematoxylin and eosin BSA bovine serum albumin PBS phosphate-buffered saline DAPI 4′,6-diamidino-2-phenylindole ISHAM The International Society for Human and Animal Mycology PET-CT positron emission tomography-computed tomography Declarations Acknowledgments Not applicable. Declarations Ethics approval and consent to participate The study was approved by Research Ethics Committee of The First Affiliated Hospital,College of Medicine, Zhejiang University(Reference Number:2019/196). Ethics approval and consent to participate have been uploaded. Consent for publication Not applicable. Competing interests The authors have no conflict of interest in relation to this work. Foundation: The authors have reported to respiratory medicine the following:Dr.Li Hequan is a doctor in The First Affiliated Hospital of ZheJiang University School of Medcine, and is supported by Natural Science Foundation of Zhejiang Province ( LY18H010002) National Natural Science Foundation of China (81970015). References Selvaggi TA, Walco JP, Parikh S, Walco GA. Hypersensitivity testing for Aspergillus fumigatus IgE is significantly more sensitive than testing for Aspergillus niger IgE. Am J Clin Pathol. 2012;137(2):203-6. Sasaki H, Miyata J, Maki Y, Kimizuka Y, Hayashi N, Fujikura Y, et al. Development of Allergic Bronchopulmonary Aspergillosis in a Patient with Crohn's Disease. Internal medicine (Tokyo, Japan). 2019;58(19):2835-8. Baxter CG, Dunn G, Jones AM, Webb K, Gore R, Richardson MD, et al. Novel immunologic classification of aspergillosis in adult cystic fibrosis. J Allergy Clin Immunol. 2013;132(3):560-6.e10. Ueki S, Hebisawa A, Kitani M, Asano K, Neves JS. Allergic Bronchopulmonary Aspergillosis-A Luminal Hypereosinophilic Disease With Extracellular Trap Cell Death. Front Immunol. 2018;9:2346. Tan K, Zelus BD, Meijers R, Liu JH, Bergelson JM, Duke N, et al. Crystal structure of murine sCEACAM1a[1,4]: a coronavirus receptor in the CEA family. The EMBO journal. 2002;21(9):2076-86. Rizeq B, Zakaria Z, Ouhtit A. Towards understanding the mechanisms of actions of carcinoembryonic antigen-related cell adhesion molecule 6 in cancer progression. Cancer science. 2018;109(1):33-42. Schupp JC, Tchaptchet S, Lützen N, Engelhard P, Müller-Quernheim J, Freudenberg MA, et al. Immune response to Propionibacterium acnes in patients with sarcoidosis--in vivo and in vitro. Bmc Pulm Med. 2015;15:75. Honma M, Iinuma S, Kanno K, Komatsu S, Minami-Hori M, Ishida-Yamamoto A. Correlation of disease activity and serum level of carcinoembryonic antigen in acquired idiopathic generalized anhidrosis: A case report. J Dermatol. 2015;42(9):900-2. Honma M, Iinuma S, Kanno K, Komatsu S, Minami-Hori M, Iizuka H, et al. Serum carcinoembryonic antigen (CEA) as a clinical marker in acquired idiopathic generalized anhidrosis: a close correlation between serum CEA level and disease activity. Journal of the European Academy of Dermatology and Venereology. 2016;30(8):1379-83. Honma M, Nozaki H, Nagahata H, Fujii M, Shibuya T, Kanno K, et al. Serum carcinoembryonic antigen specifically increases among various serum markers of adenocarcinoma in hypohidrosis or conditions related to hypohidrosis. J Dermatol. 2017;44(8):903-8. Mandile R, Maglio M, Pellino N, Russo M, Miele E, Spagnuolo MI, et al. Seronegative Villous Atrophy in Children: Clinical and Immunohistochemical Features. Journal of pediatric gastroenterology and nutrition. 2020. Agarwal R, Chakrabarti A, Shah A, Gupta D, Meis JF, Guleria R, et al. Allergic bronchopulmonary aspergillosis: review of literature and proposal of new diagnostic and classification criteria. Clin Exp Allergy. 2013;43(8):850-73. Rayes RF, Vourtzoumis P, Bou Rjeily M, Seth R, Bourdeau F, Giannias B, et al. Neutrophil Extracellular Trap-Associated CEACAM1 as a Putative Therapeutic Target to Prevent Metastatic Progression of Colon Carcinoma. J Immunol. 2020;204(8):2285-94. Rosenberg M, Patterson R, Mintzer R, Cooper BJ, Roberts M, Harris KE. Clinical and immunologic criteria for the diagnosis of allergic bronchopulmonary aspergillosis. Ann Intern Med. 1977;86(4):405-14. Bovo S, Mazzoni G, Bertolini F, Schiavo G, Galimberti G, Gallo M, et al. Genome-wide association studies for 30 haematological and blood clinical-biochemical traits in Large White pigs reveal genomic regions affecting intermediate phenotypes. Sci Rep. 2019;9(1):7003. Ivankovics IG, Fernandes LC, Saad SS, Matos D. Peripheral and mesenteric serum levels of CEA and cytokeratins, staging and histopathological variables in colorectal adenocarcinoma. World journal of gastroenterology. 2008;14(43):6699-703. Hao C, Zhang G, Zhang L. Serum CEA levels in 49 different types of cancer and noncancer diseases. Prog Mol Biol Transl Sci. 2019;162:213-27. Chen ZQ, Huang LS, Zhu B. Assessment of Seven Clinical Tumor Markers in Diagnosis of Non-Small-Cell Lung Cancer. Dis Markers. 2018;2018:9845123. Drent M, Cremers JP, Jansen TL, Baughman RP. Practical eminence and experience-based recommendations for use of TNF-α inhibitors in sarcoidosis. Sarcoidosis, vasculitis, and diffuse lung diseases : official journal of WASOG. 2014;31(2):91-107. Maeda Y, Hizawa N, Fukui Y, Nagai K, Kikuchi E, Takahashi D, et al. [Concentrations of carcinoembryonic antigen in serum and bronchoalveolar lavage fluid of asthmatic patients with mucoid impaction]. Nihon Kokyuki Gakkai Zasshi. 2004;42(12):988-93. Tables Table 1 .Newly proposed diagnostic criteria for allergic bronchopulmonary aspergillosis(12) Predisposing conditions Bronchial asthma, cystic fibrosis and Obligatory criteria (both should be present) （1）Type I Aspergillus skin test positive (immediate cutaneous hypersensitivity to Aspergillus antigen) or elevated IgE levels against Aspergillus fumigatus （2）Elevated total IgE levels (> 1000 IU/mL) a Other criteria (at least two of three) （1） Presence of precipitating or IgG antibodies against A . fumigatus in serum （2） Radiographic pulmonary opacities consistent with ABPA b （3） Total eosinophil count > 500 cells/μL in steroid naïve patients (may be historical) a If the patient meets all other criteria, an IgE value < 1000 IU/mL may be acceptable. b The chest radiographic features consistent with ABPA may be transient (i.e. consolidation, nodules, tram‐track opacities, toothpaste/finger‐in‐glove opacities, fleeting opacities) or permanent (i.e. parallel line and ring shadows, bronchiectasis and pleuropulmonary fibrosis). Table 2 ． The relationship between CEA and eosinophils in this person Basic situation Correlation coefficient P Eosinophils（%） 8.81（1.65，19.10） 0.929 0.022 CEA（ng/ml） 14.31（6.30，34.10） Table 3. Demographics and Clinical features of 37 patients with ABPA （ n=37 ） Clinical parameters Number (proportion) Age（years） 55.00±15.96 Gender（male/female） 24（64.86%）/13（35.14%） Symptom Cough 37（100.00%） Expectoration 29（78.38%） Hemoptysis 7（18.92%） Short of breath 20（54.05%） Heat 5（13.51%） Comorbidities Asthma 13（35.14%） COPD 9（24.32%） Interstitial lung disease 4（10.81%） Bronchiectasis 28（75.68%） Treatment Voriconazole 9 (24.32%) Voriconazole and glucocorticoid 14 (37.84%) Glucocorticoid 4（10.81%） Itraconazole and glucocorticoid 9（24.32%） Itraconazole 1 (2.70%) Ending Relief or (and) stability 21（56.76%） Relapse or (and) progression 16（43.24%） Table 4 . Laboratory Characteristics of ABPA patients Laboratory results Normal range people（n=37） Eosinophils（/uL） 20~500（/uL） 1080（540,1430） Eosinophils（%） 0.5~5.0 (%) 14.70（8.90,22.75） TIgE（KU/L） 0.0~100.0(KU/L) <5000 28（75.68%） ≥5000 9（14.32%） sIgE(kU/L) 0.00~0.35(KU/L) 5.69（2.83,12.02） ESR（mm/h） 14.00（7.00,33.00） CRP（mg/L） 6.76（2.48,13.83） CEA（ng/ml） 0.00~5.05(ng/ml) 7.20（3.65,12.70） CEA <5.00 12 (32.43%) 5≤CEA<10 14 (37.84%) ≥10.00 11(29.73%) Supplementary Files EthicsCommittee.pdf 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-73116","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research","associatedPublications":[],"authors":[{"id":2283228,"identity":"f62b8533-43d5-4e9f-b44f-95e6a0e717d0","order_by":0,"name":"Yang yanfei","email":"","orcid":"","institution":"First Hospital of Zhejiang Province: Zhejiang University School of Medicine First Affiliated Hospital","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yang","middleName":"","lastName":"yanfei","suffix":""},{"id":2283229,"identity":"996a4f88-86bb-4aab-bdf1-3689123f2425","order_by":1,"name":"Gao qiqi","email":"","orcid":"","institution":"First Hospital of Zhejiang Province: Zhejiang University School of Medicine First Affiliated Hospital","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Gao","middleName":"","lastName":"qiqi","suffix":""},{"id":2283230,"identity":"5b515a0f-cd46-4c85-8860-ce9bc25e2c36","order_by":2,"name":"Jin yangyi","email":"","orcid":"","institution":"First Hospital of Zhejiang Province: Zhejiang University School of Medicine First Affiliated Hospital","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jin","middleName":"","lastName":"yangyi","suffix":""},{"id":2283231,"identity":"45d5c53c-4688-4068-a8a1-f6616e9367b5","order_by":3,"name":"Qi mengdie","email":"","orcid":"","institution":"Tongde Hospital Of Zhejiang Province","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Qi","middleName":"","lastName":"mengdie","suffix":""},{"id":2283232,"identity":"516b35b4-a4b7-4164-9dc2-b3b54b38d325","order_by":4,"name":"Lu guohua","email":"","orcid":"","institution":"Zhejiang University School of Medicine First Affiliated Hospital","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Lu","middleName":"","lastName":"guohua","suffix":""},{"id":2283233,"identity":"035710fa-c4e3-4bcb-bbf0-0be978276ba6","order_by":5,"name":"li Hequan","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA1klEQVRIiWNgGAWjYBACPmYeCIOfgSEBSDET1sIG0yLZQLQWBqgWgwNgihgt7LwHH7xts8szPn/gmQRDhXViA/vZAwQcxpdsOLctudjswIE0CYYz6YkNPHkJBLTwmEnztjEnbjvYkCbB2HY4sUGCx4CQFvPfvG31iZubGYBa/hGnxYyZF2j4BjaQlgbitBhLzjl3PHHGGYZki4Rj6cZtPDn4tfDznzH88KasOrG//0zijQ811rL97GfwawEDSNTwJIAjk42wergW9gNEKR4Fo2AUjIKRBwCK1jvp8B9D3AAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0003-3882-9837","institution":"First Hospital of Zhejiang Province: Zhejiang University School of Medicine First Affiliated Hospital","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"li","middleName":"","lastName":"Hequan","suffix":""}],"badges":[],"createdAt":"2020-09-06 10:59:06","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-73116/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-73116/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":2417880,"identity":"5c3fdaba-0ed7-4a89-9058-66bba120efbd","added_by":"auto","created_at":"2020-09-15 16:39:57","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":88784,"visible":true,"origin":"","legend":"Patient underwent follow-up CT\n（Figure 1 a）When this patient first came to our hospital, CT appeared as exudation with consolidation. （Figure 1 b-d）After treatment with glucocorticoids, the pulmonary consolidation significantly reduced.","description":"","filename":"Figure1.JPG","url":"https://assets-eu.researchsquare.com/files/rs-73116/v1/Figure1.JPG"},{"id":2417881,"identity":"80d0a816-b93e-4fe4-9938-ae1598918b6c","added_by":"auto","created_at":"2020-09-15 16:39:57","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":27297,"visible":true,"origin":"","legend":"The relationship between CEA and eosinophils in this patient","description":"","filename":"Figure2.JPG","url":"https://assets-eu.researchsquare.com/files/rs-73116/v1/Figure2.JPG"},{"id":2417882,"identity":"cb34617c-3609-49c4-bdb8-5f3561cd2516","added_by":"auto","created_at":"2020-09-15 16:39:57","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":62008,"visible":true,"origin":"","legend":"Lung tissue stained with H\u0026E and immunohistochemistry.\n (Figure 3 a)lung tissue stained with H\u0026E, Alveolar was infiltrated with eosinophils (×400 magnification). (Figure 3 b) There are CEA positive cells in lung tissue(×400 magnification). (Figure 3 c and d)Immunostaining for CEA showed that CEA was observed in the cells which were characteristic of eosinophils(×800 magnification).","description":"","filename":"Figure3.JPG","url":"https://assets-eu.researchsquare.com/files/rs-73116/v1/Figure3.JPG"},{"id":2417883,"identity":"07ba8666-6ae2-4568-9a34-8c952391c35e","added_by":"auto","created_at":"2020-09-15 16:39:57","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":61972,"visible":true,"origin":"","legend":"Lung section with a MBP–positive cell colabeled with CEA antigen localized on Cytoplasm, as indicated by the arrow. Scale bars = 20 μm.","description":"","filename":"Figure4.JPG","url":"https://assets-eu.researchsquare.com/files/rs-73116/v1/Figure4.JPG"},{"id":2417884,"identity":"1e12a586-d56c-4355-b9d0-847aae4701e8","added_by":"auto","created_at":"2020-09-15 16:39:57","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":52801,"visible":true,"origin":"","legend":"Lung section with a MBP–positive cell colabeled with CEA antigen localized on Cytoplasm, as indicated by the arrow. Scale bars = 10 μm. \nDual immunofluorescence was performed using the eosinophils marker MBP (green) and the CEA Monoclonal Antibody (RED). Nuclei stained with DAPI (blue).Both double-labeling immunofluorescence assays were performed in lung section because of their positive induced in eosinophils . DAPI, 4’,6’-diamino-2-phenylindole; MBP（also called Anti-PRG2 rabbit polyclonal antibody） eosinophil major basic protein ; CEA (CEA Monoclonal Antibody),Carcino Embryonic Antigen;","description":"","filename":"Figure5.JPG","url":"https://assets-eu.researchsquare.com/files/rs-73116/v1/Figure5.JPG"},{"id":2417885,"identity":"0502da38-9272-4346-b363-5d285d7ce618","added_by":"auto","created_at":"2020-09-15 16:39:57","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":31801,"visible":true,"origin":"","legend":"A higher eosinophil count is with the echo of higher levels of CEA","description":"","filename":"Figure6.JPG","url":"https://assets-eu.researchsquare.com/files/rs-73116/v1/Figure6.JPG"},{"id":2417886,"identity":"34547551-880a-4049-bc83-9642f75fa59d","added_by":"auto","created_at":"2020-09-15 16:39:57","extension":"jpg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":44653,"visible":true,"origin":"","legend":"Correlation of CEA and EOS in ABPA patients.\n(figure 7 a) Relationships among CEA and EOS%.( figure 7 b) Relationships among CEA and EOS counts.","description":"","filename":"Figure7.JPG","url":"https://assets-eu.researchsquare.com/files/rs-73116/v1/Figure7.JPG"},{"id":13594334,"identity":"9c2ddb21-01f9-4ed6-aadd-f91cc6ef6803","added_by":"auto","created_at":"2021-09-17 05:19:58","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":712224,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-73116/v1/f498e6e4-a464-43a7-93e0-80d9394a5c3f.pdf"},{"id":2417888,"identity":"afa8c7f2-3d9c-4936-848a-0579014a2281","added_by":"auto","created_at":"2020-09-15 16:39:58","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":210881,"visible":true,"origin":"","legend":"","description":"","filename":"EthicsCommittee.pdf","url":"https://assets-eu.researchsquare.com/files/rs-73116/v1/EthicsCommittee.pdf"}],"financialInterests":"","formattedTitle":"\u003cp\u003eEosinophils May Serve as CEA-Secreting Cells for ABPA Patients\u003c/p\u003e","fulltext":[{"header":"Background","content":"\u003cp\u003eAllergic bronchopulmonary aspergillosis (ABPA) is characterized by an allergic inflammatory response to Aspergillus fumigatus colonized in the trachea(1). It is manifested as poorly controlled asthma and affected approximately 4 million patients worldwide(2). Some patients may even suffer from irreversible airway obstruction and pulmonary fibrosis(3). Besides, a significant eosinophil infiltration can be seen in the pathological specimens of ABPA patients(4).\u003c/p\u003e\n\u003cp\u003eCarcinoembryonic antigen (CEA) was initially discovered as a tumor antigen(5). Serum CEA level is typically identified as a candidate biomarker for tumor progression(6).\u0026nbsp; However, a patient with high levels of CEA was admitted to our hospital, while she had continuous cough and shortness of breath. Because of departed cognition of CEA, she was initially diagnosed with a malignant disease in another hospital. However, after careful clinical diagnosis, we could not exclude the diagnosis of ABPA. Thus, she received glucocorticoid therapy. After treatment, the patient\u0026rsquo;s symptoms were remarkably improved. Additionally, her serum CEA level and eosinophil levels were significantly decreased, indicating a subtle balance between the levels of eosinophils and CEA in the serum.\u003c/p\u003e\n\u003cp\u003eJapanese scholars have reported 13 patients with ABPA, in which 7 of them had elevated levels of CEA in the peripheral blood. In case of pulmonary consolidation, the serum CEA levels were gradually returned to normal level(7). Regrettably, the study did not mention the precise source of CEA in ABPA patients. Recently, CEA has also been proposed as a clinical marker, reflecting the activity of acquired idiopathic generalized anhidrosis (AIGA)(8). Immunohistochemistry of 10 AIGA cases showed increased levels of CEA in the eccrine sweat glands(9). A previous study has pointed out that neutrophil extracellular traps (NETs) are associated with CEA(10). The above-mentioned studies indicated that the serum CEA level might be related to the presence of eosinophils. To prove this assumption, We tracked the clinical data of this ABPA patient and double-labeled the lung tissue samples with CEA and eosinophils. \u0026nbsp;37 cases with ABPA who were admitted to our hospital from 2013 to 2019 were reviewed. To the best of our knowledge, the presented study is the first to report the relationship between serum CEA level and eosinophils in patients with ABPA.\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003cp\u003e\u003cstrong\u003eMaterials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe levels of aspergillus fumigatus-specific IgG and aspergillus fumigatus-specific IgE were detected by using a commercial enzyme-linked immunosorbent assay (ELISA) kit (Thermo Fisher Scientific, Waltham, MA, USA). Besides, interleukin-5 (IL-5) was detected by using human IL-5 platinum ELISA kit (eBioscience Inc., San Diego, CA, USA).\u0026nbsp;Blood eosinophils were measured using a routine blood testing kit. Eosinophils were detected by myelin basic protein (MBP) antibody (Sangon Biotech Co., Ltd., Shanghai, China) and hematoxylin and eosin (H\u0026amp;E) staining. CEA was detceted by CEA monoclonal antibody(Thermo Fisher Scientific, Waltham, MA, USA).A secondary F(ab) polyclonal antibody, goat anti-rabbit IgG (H+L) highly cross-adsorbed secondary antibody, and goat anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody were used.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eImmunohistochemistry and immunofluorescence\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe present study was approved by the Ethics Committee of our hospital, and all patients signed written informed consent form prior to study initiation. All reagents used for immunohistochemistry were kindly provided by the Pathology Department of our hospital.\u003c/p\u003e\n\u003cp\u003eThe sections were de-waxed, rehydrated, and incubated with 3% hydrogen peroxide in methanol to block endogenous peroxidase activity at room temperature for 10\u0026thinsp;min. For antigen retrieval, deparaffinized and rehydrated sample tissue sections were pretreated by microwave irradiation for 20\u0026ndash;30 min in 0.01 mol/l citrate-buffered saline (pH 6.0).\u003c/p\u003e\n\u003cp\u003eClosed non-specific binding was performed, and normal goat serum was then added to the sections. Sections were then incubated for overnight at 4 \u0026deg;C with primary antibodies (mouse anti-human CEA and rabbit anti-human MBP antibodies) diluted in blocking solution. Membranes were incubated with a secondary antibody, and then goat anti-mouse/rabbit antibody, and diluted at an appropriate dilution in 1% bovine serum albumin (BSA) for 2 h at room temperature. Diaminobenzidine (DAB) color reactions were visualized under a microscope (5\u0026ndash;10 min). After rinsing thricewith phosphate-buffered saline (PBS), staining was carried out by DAB before counterstaining with Hematoxylin.\u003c/p\u003e\n\u003cp\u003eDouble-immunofluorescence staining was performed as described previously(11). In brief, the sections were analyzed via antigen retrieval, blocked for endogenous peroxidase and non-specific epitopes, and incubated with primary antibody at 4\u0026deg;C overnight. Antibody detection was performed using a secondary F(ab) polyclonal antibody (goat anti-mouse IgG (H+L) highly cross-adsorbed secondary antibody and goat anti-rabbit IgG (H+L) highly cross-adsorbed secondary antibody). The sections were counterstained with 4\u0026prime;,6-diamidino-2-phenylindole (DAPI), and visualized under a LSM880 Meta laser-scanning confocal microscope (Carl Zeiss).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInclusion and exclusion criteria\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe diagnosis of ABPA is currently done by combining clinical, radiological, and immunological findings based on the criteria proposed by The International Society for Human and Animal Mycology (ISHAM) in 2013(see Table 1).(12)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data were presented as M(P25, P75), and were analyzed using SPSS 25.0 software (IBM, Armonk, NY, USA)\u003cem\u003e.\u003c/em\u003e P \u0026le; 0.05 was considered statistically significant, and P \u0026le; 0.01 as highly significant. Pearson correlation coefficient lower than 0.4 indicates poor reproducibility, which range from 0.4-0.75, denoting fair reproducibility, and equal to 0.75 indicates good reproducibility(13).\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e\u003cstrong\u003eA \u0026nbsp;patient with high level of CEA was diagnosed as ABPA\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA 50-year-old female patient suffering from cough and dyspnea for 2 years was diagnosed with lung cancer because of high serum levels of CEA and lymphadenopathy before she was admitted to our hospital. But the result of positron emission tomography-computed tomography (PET-CT) examination \u0026nbsp;and bone marrow biopsy showed no evidence of tumor. Initial laboratory tests after admission showed elevated CEA level (38.0 ng/mL, reference range \u0026lt; 5.2 ng/mL) and a high count of eosinophils (5700/\u0026mu;L). CT of the lung revealed bronchiectasis and infiltration of the right middle and right lower lobes (Figure 1a). EBUS-TBLB were employed to evaluate mediastinal lymph nodes and lung lesions, showing accumulation of eosinophils in the lungs and lymph nodes. According to the proposed diagnostic criteria for ABPA, the patient was diagnosed with ABPA based on the history of bronchial asthma, as well as the elevated levels of aspergillus fumigatus-specific IgG,serum total IgE and aspergillus fumigatus-specific IgE. Systemic corticosteroid therapy (prednisone 40 mg/day) was initiated, and the patient\u0026rsquo;s symptoms were dissipated. CT scan of the chest indicated disappearance of pulmonary infiltrates\u0026nbsp; and mucoid impaction (Figure 1.a-d) after 6 months.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEosinophils and CEA were decreased simultaneously\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe decreased serum CEA levels were consistent with eosinophils during the follow-up (Figure 2). The correlation between the serum CEA levels and eosinophil levels in the 6-month follow-up duration was analyzed by using Pearson correlation analysis, wherein significant correlations were observed (Table 2, r=0.929, P=0.022).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eActivated CEA was partially co-localized on eosinophils\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eInitially, H\u0026amp;E staining confirmed that a lot of eosinophils in the lung (Figure 3a). CEA was first detected by immunocytochemistry.The yellow-brown granules observed in the cells indicated a positive result for CEA. Additionally, a great number of CEA markers were found in the cells (Figure 3b). The results revealed that the eosinophils were immunohistochemically positive for CEA under high magnification (Figure 3c and d). Immunofluorescence analysis also unveiled that eosinophils ,as well as CEA,were identified in the lung . Moreover，Lung section with a MBP\u0026ndash;positive cells were colabeled with CEA antigen which was localized on Cytoplasm(Figure 4 and Figure 5).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIncreased serum CEA levels were positively associated with eosinophil levels\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eClinical data of the patients are listed in Table 3. Twenty four male and 13 female patients were included in the current study. Before commencing the treatment, 25 (25/37) patients had serum CEA levels higher than normal. As shown in Table 4, the serum CEA levels were significantly increased, and the mean serum CEA level was markedly higher in ABPA patients than that in healthy subjects (5.05 ng/ml). Moreover, 37 patients were assigned to two groups based on peripheral blood eosinophil counts. Independent -sample t-test was employed to compare serum CEA levels between the two groups. The results\u0026nbsp;showed\u0026nbsp;that\u0026nbsp;patients\u0026nbsp;with higher eosinophil levels had higher serum levels of CEA (P\u0026lt;0.001, Figure 6). Besides, we noted that the eosinophil levels were positively correlated with serum CEA levels(r=0.459 and r=0.506，P=0.004 and p=0.001) by Pearson correlation coefficient (Figure 7).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eABPA is the most significant manifestation of allergic aspergillosis that occurs worldwide, but much attention has not been paid by the scholars(14). Meanwhile, eosinophils promote inflammation, and are known to play a beneficial role in isolating and controlling a disease site(15). To our knowledge, CEA is the most commonly used serum marker in the management of breast cancer, and its expression showed correlation with clinicopathological characteristics of gastric carcinoma(16).\u0026nbsp;\u0026nbsp;CEA was first identified as colon cancer antigen in 1965. The serum CEA levels were higher in patients with colon cancer when compared to healthy individuals, and this led to its clinical application as a diagnostic biomarker for colorectal cancer(17). Recently, the serum levels of CEA and the trend of its changes in the treatment process have been previously validated(18).\u0026nbsp; Thus, some patients with elevated CEA levels were easily misdiagnosed as lung cancer. In the current study, several ABPA patients had higher serum CEA levels than healthy controls. The comparative data demonstrated the necessity of assessment of relationships between the eosinophils and serum CEA level in ABPA patients. In the current study, the serum CEA level was shown to be decreased after treatment, indicating that glucocorticoid therapy might have inhibitory effects on CEA level in ABPA patients. The serum CEA level was found to be positively correlated with blood eosinophils. Previous studies have shown that elevated serum CEA levels in patients with ABPA are associated with consolidation of lungs, mucus plugs, and localized inflammation of lungs(19). Several scholars have pointed out that serum concentrations of CEA were significantly increased in asthmatic patients with mucoid impactions when compared to those patients without mucoid impactions or patients with bronchiectasis(20). However, in our research, the serum levels of CEA were found to be associated with eosinophil count.\u0026nbsp;This is also the first report to present an ABPA case with the relationship between serum CEA level and eosinophils count, and our findings may be significant for clinicians. According to our study, patients with elevated serum CEA levels were diagnosed with benign diseases, (e.g., ABPA). According to the present research, CEA potentially serves as a clinical marker for ABPA patients.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; However, the present study has two important limitations. Firstly, this was a retrospective single-center study with a relatively small sample size. Secondly, we only included one patient who underwent biopsy, and it is more appropriate to study serum samples of ABPA patients labeled with both CEA and eosinophils. Therefore, further large-scale prospective study should be conducted to verify the preliminary results of the present study.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn summary, the serum CEA levels were shown to be elevated relatively in ABPA patients. However, the elevated serum CEA level can be normalized after treatment. Increased levels of CEA in ABPA patients might be positively correlated with eosinophil levels, and eosinophils can serve as CEA-secreting cells in patients with ABPA.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003ctable width=\"595\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003eABPA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"491\"\u003e\n\u003cp\u003eAllergic bronchopulmonary aspergillosis\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003eCEA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"491\"\u003e\n\u003cp\u003ecarcinoembryonic antigen\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003eEBUS-TBLB\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"491\"\u003e\n\u003cp\u003eEndobronchial ultrasound-guided transbronchial lung biopsy aspiration\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003eAIGA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"491\"\u003e\n\u003cp\u003eacquired idiopathic generalized anhidrosis\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003eNETs\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"491\"\u003e\n\u003cp\u003eneutrophil extracellular traps\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003eELISA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"491\"\u003e\n\u003cp\u003eenzyme-linked immunosorbent assay\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003eIL-5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"491\"\u003e\n\u003cp\u003einterleukin-5\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003eMBP\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"491\"\u003e\n\u003cp\u003emyelin basic protein\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003eH\u0026amp;E\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"491\"\u003e\n\u003cp\u003ehematoxylin and eosin\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003eBSA\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"491\"\u003e\n\u003cp\u003ebovine serum albumin\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003ePBS\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"491\"\u003e\n\u003cp\u003ephosphate-buffered saline\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003eDAPI\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"491\"\u003e\n\u003cp\u003e4\u0026prime;,6-diamidino-2-phenylindole\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003eISHAM\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"491\"\u003e\n\u003cp\u003eThe International Society for Human and Animal Mycology\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"104\"\u003e\n\u003cp\u003ePET-CT\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"491\"\u003e\n\u003cp\u003epositron emission tomography-computed tomography\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeclarations\u003c/strong\u003e\u003c/p\u003e\n\u003ch3\u003eEthics approval and consent to participate\u003c/h3\u003e\n\u003cp\u003eThe study was approved by Research Ethics Committee of The First Affiliated Hospital,College of Medicine, Zhejiang University(Reference Number:2019/196). Ethics approval and consent to participate have been uploaded.\u003c/p\u003e\n\u003ch3\u003eConsent for publication\u003c/h3\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003ch3\u003eCompeting interests\u003c/h3\u003e\n\u003cp\u003eThe authors have no conflict of interest in relation to this work.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFoundation:\u003c/strong\u003e\u0026nbsp;The authors have reported to\u0026nbsp;\u003cem\u003erespiratory medicine\u003c/em\u003e\u0026nbsp;the following:Dr.Li Hequan is a doctor in The First Affiliated Hospital of ZheJiang University School of Medcine, and is supported by \u003cstrong\u003eNatural Science Foundation of Zhejiang Province\u003c/strong\u003e ( LY18H010002) \u003cstrong\u003eNational Natural Science Foundation of China\u003c/strong\u003e (81970015).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eSelvaggi TA, Walco JP, Parikh S, Walco GA. Hypersensitivity testing for Aspergillus fumigatus IgE is significantly more sensitive than testing for Aspergillus niger IgE. Am J Clin Pathol. 2012;137(2):203-6.\u003c/li\u003e\n\u003cli\u003eSasaki H, Miyata J, Maki Y, Kimizuka Y, Hayashi N, Fujikura Y, et al. Development of Allergic Bronchopulmonary Aspergillosis in a Patient with Crohn's Disease. Internal medicine (Tokyo, Japan). 2019;58(19):2835-8.\u003c/li\u003e\n\u003cli\u003eBaxter CG, Dunn G, Jones AM, Webb K, Gore R, Richardson MD, et al. Novel immunologic classification of aspergillosis in adult cystic fibrosis. J Allergy Clin Immunol. 2013;132(3):560-6.e10.\u003c/li\u003e\n\u003cli\u003eUeki S, Hebisawa A, Kitani M, Asano K, Neves JS. Allergic Bronchopulmonary Aspergillosis-A Luminal Hypereosinophilic Disease With Extracellular Trap Cell Death. Front Immunol. 2018;9:2346.\u003c/li\u003e\n\u003cli\u003eTan K, Zelus BD, Meijers R, Liu JH, Bergelson JM, Duke N, et al. Crystal structure of murine sCEACAM1a[1,4]: a coronavirus receptor in the CEA family. The EMBO journal. 2002;21(9):2076-86.\u003c/li\u003e\n\u003cli\u003eRizeq B, Zakaria Z, Ouhtit A. Towards understanding the mechanisms of actions of carcinoembryonic antigen-related cell adhesion molecule 6 in cancer progression. Cancer science. 2018;109(1):33-42.\u003c/li\u003e\n\u003cli\u003eSchupp JC, Tchaptchet S, L\u0026uuml;tzen N, Engelhard P, M\u0026uuml;ller-Quernheim J, Freudenberg MA, et al. Immune response to Propionibacterium acnes in patients with sarcoidosis--in vivo and in vitro. Bmc Pulm Med. 2015;15:75.\u003c/li\u003e\n\u003cli\u003eHonma M, Iinuma S, Kanno K, Komatsu S, Minami-Hori M, Ishida-Yamamoto A. Correlation of disease activity and serum level of carcinoembryonic antigen in acquired idiopathic generalized anhidrosis: A case report. J Dermatol. 2015;42(9):900-2.\u003c/li\u003e\n\u003cli\u003eHonma M, Iinuma S, Kanno K, Komatsu S, Minami-Hori M, Iizuka H, et al. Serum carcinoembryonic antigen (CEA) as a clinical marker in acquired idiopathic generalized anhidrosis: a close correlation between serum CEA level and disease activity. Journal of the European Academy of Dermatology and Venereology. 2016;30(8):1379-83.\u003c/li\u003e\n\u003cli\u003eHonma M, Nozaki H, Nagahata H, Fujii M, Shibuya T, Kanno K, et al. Serum carcinoembryonic antigen specifically increases among various serum markers of adenocarcinoma in hypohidrosis or conditions related to hypohidrosis. J Dermatol. 2017;44(8):903-8.\u003c/li\u003e\n\u003cli\u003eMandile R, Maglio M, Pellino N, Russo M, Miele E, Spagnuolo MI, et al. Seronegative Villous Atrophy in Children: Clinical and Immunohistochemical Features. Journal of pediatric gastroenterology and nutrition. 2020.\u003c/li\u003e\n\u003cli\u003eAgarwal R, Chakrabarti A, Shah A, Gupta D, Meis JF, Guleria R, et al. Allergic bronchopulmonary aspergillosis: review of literature and proposal of new diagnostic and classification criteria. Clin Exp Allergy. 2013;43(8):850-73.\u003c/li\u003e\n\u003cli\u003eRayes RF, Vourtzoumis P, Bou Rjeily M, Seth R, Bourdeau F, Giannias B, et al. Neutrophil Extracellular Trap-Associated CEACAM1 as a Putative Therapeutic Target to Prevent Metastatic Progression of Colon Carcinoma. J Immunol. 2020;204(8):2285-94.\u003c/li\u003e\n\u003cli\u003eRosenberg M, Patterson R, Mintzer R, Cooper BJ, Roberts M, Harris KE. Clinical and immunologic criteria for the diagnosis of allergic bronchopulmonary aspergillosis. Ann Intern Med. 1977;86(4):405-14.\u003c/li\u003e\n\u003cli\u003eBovo S, Mazzoni G, Bertolini F, Schiavo G, Galimberti G, Gallo M, et al. Genome-wide association studies for 30 haematological and blood clinical-biochemical traits in Large White pigs reveal genomic regions affecting intermediate phenotypes. Sci Rep. 2019;9(1):7003.\u003c/li\u003e\n\u003cli\u003eIvankovics IG, Fernandes LC, Saad SS, Matos D. Peripheral and mesenteric serum levels of CEA and cytokeratins, staging and histopathological variables in colorectal adenocarcinoma. World journal of gastroenterology. 2008;14(43):6699-703.\u003c/li\u003e\n\u003cli\u003eHao C, Zhang G, Zhang L. Serum CEA levels in 49 different types of cancer and noncancer diseases. Prog Mol Biol Transl Sci. 2019;162:213-27.\u003c/li\u003e\n\u003cli\u003eChen ZQ, Huang LS, Zhu B. Assessment of Seven Clinical Tumor Markers in Diagnosis of Non-Small-Cell Lung Cancer. Dis Markers. 2018;2018:9845123.\u003c/li\u003e\n\u003cli\u003eDrent M, Cremers JP, Jansen TL, Baughman RP. Practical eminence and experience-based recommendations for use of TNF-\u0026alpha; inhibitors in sarcoidosis. Sarcoidosis, vasculitis, and diffuse lung diseases : official journal of WASOG. 2014;31(2):91-107.\u003c/li\u003e\n\u003cli\u003eMaeda Y, Hizawa N, Fukui Y, Nagai K, Kikuchi E, Takahashi D, et al. [Concentrations of carcinoembryonic antigen in serum and bronchoalveolar lavage fluid of asthmatic patients with mucoid impaction]. Nihon Kokyuki Gakkai Zasshi. 2004;42(12):988-93.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003eTable 1 .Newly proposed diagnostic criteria for allergic bronchopulmonary aspergillosis(12)\u003c/p\u003e\n\u003ctable border=\"1\" width=\"586\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"208\"\u003e\n\u003cp\u003e\u003cstrong\u003ePredisposing conditions\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"378\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"208\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd width=\"378\"\u003e\n\u003cp\u003eBronchial asthma, cystic fibrosis and\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"208\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd width=\"378\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"208\"\u003e\n\u003cp\u003e\u003cstrong\u003eObligatory criteria\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e(both should be present)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"378\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"208\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd width=\"378\"\u003e\n\u003cp\u003e（1）Type I\u0026nbsp;\u003cem\u003eAspergillus\u0026nbsp;\u003c/em\u003eskin test positive (immediate cutaneous hypersensitivity to\u0026nbsp;\u003cem\u003eAspergillus\u0026nbsp;\u003c/em\u003eantigen) or elevated IgE levels against\u0026nbsp;\u003cem\u003eAspergillus fumigatus\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"208\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd width=\"378\"\u003e\n\u003cp\u003e（2）Elevated total IgE levels (\u0026gt;\u0026nbsp;1000\u0026nbsp;IU/mL)\u003cem\u003e\u003csup\u003ea\u003c/sup\u003e\u003c/em\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"208\"\u003e\n\u003cp\u003e\u003cstrong\u003eOther criteria\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e(at least two of three)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"378\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"208\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd width=\"378\"\u003e\n\u003cp\u003e（1）\u0026nbsp;\u0026nbsp;\u0026nbsp; Presence of precipitating or IgG antibodies against\u0026nbsp;\u003cem\u003eA\u0026nbsp;\u003c/em\u003e.\u0026nbsp;\u003cem\u003efumigatus\u0026nbsp;\u003c/em\u003ein serum\u003c/p\u003e\n\u003cp\u003e（2）\u0026nbsp;\u0026nbsp;\u0026nbsp; Radiographic pulmonary opacities consistent with ABPA\u003cem\u003e\u003csup\u003eb\u003c/sup\u003e\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e（3）\u0026nbsp;\u0026nbsp;\u0026nbsp; Total eosinophil count \u0026gt;\u0026nbsp;500 cells/\u0026mu;L in steroid na\u0026iuml;ve patients (may be historical)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cul\u003e\n\u003cli\u003e\u003cem\u003e\u003csup\u003ea\u003c/sup\u003e\u003c/em\u003e\u0026nbsp;If the patient meets all other criteria, an IgE value \u0026lt;\u0026nbsp;1000\u0026nbsp;IU/mL may be acceptable.\u003c/li\u003e\n\u003cli\u003e\u003cem\u003e\u003csup\u003eb\u003c/sup\u003e\u003c/em\u003e\u0026nbsp;The chest radiographic features consistent with ABPA may be transient (i.e. consolidation, nodules, tram‐track opacities, toothpaste/finger‐in‐glove opacities, fleeting opacities) or permanent (i.e. parallel line and ring shadows, bronchiectasis and pleuropulmonary fibrosis).\u003c/li\u003e\n\u003c/ul\u003e\n\u003cp\u003e\u003cstrong\u003e\u003cbr /\u003e\u003c/strong\u003e\u003cstrong\u003eTable 2\u003c/strong\u003e\u003cstrong\u003e．\u003c/strong\u003e\u003cstrong\u003eThe relationship between CEA and eosinophils in this person\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" width=\"558\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"161\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd width=\"161\"\u003e\n\u003cp\u003eBasic situation\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"151\"\u003e\n\u003cp\u003e\u0026nbsp; Correlation coefficient\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003eP\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"161\"\u003e\n\u003cp\u003eEosinophils（%）\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"198\"\u003e\n\u003cp\u003e8.81（1.65，19.10）\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\n\u003cp\u003e0.929\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\n\u003cp\u003e\u003cstrong\u003e0.022\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"161\"\u003e\n\u003cp\u003eCEA（ng/ml）\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd colspan=\"2\" width=\"198\"\u003e\n\u003cp\u003e14.31（6.30，34.10）\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"113\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd width=\"85\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3. Demographics and Clinical features of 37 patients with ABPA\u003c/strong\u003e\u003cstrong\u003e（\u003c/strong\u003e\u003cstrong\u003en=37\u003c/strong\u003e\u003cstrong\u003e）\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" width=\"548\"\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003e\u003cstrong\u003eClinical parameters\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e\u003cstrong\u003eNumber (proportion)\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eAge（years）\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e55.00\u0026plusmn;15.96\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eGender（male/female）\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e24（64.86%）/13（35.14%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003e\u003cstrong\u003eSymptom\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eCough\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e37（100.00%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eExpectoration\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e29（78.38%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eHemoptysis\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e7（18.92%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eShort of breath\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e20（54.05%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eHeat\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e5（13.51%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003e\u003cstrong\u003eComorbidities\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eAsthma\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e13（35.14%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eCOPD\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e9（24.32%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eInterstitial lung disease\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e4（10.81%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eBronchiectasis\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e28（75.68%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003e\u003cstrong\u003eTreatment \u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eVoriconazole\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e9 (24.32%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eVoriconazole and glucocorticoid\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e14 (37.84%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eGlucocorticoid\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e4（10.81%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eItraconazole and\u0026nbsp; glucocorticoid\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e9（24.32%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eItraconazole\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e1 (2.70%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003e\u003cstrong\u003eEnding\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eRelief or (and) stability\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e21（56.76%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"293\"\u003e\n\u003cp\u003eRelapse or (and) progression\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"255\"\u003e\n\u003cp\u003e16（43.24%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTable 4\u003c/strong\u003e\u003cstrong\u003e. \u003c/strong\u003e\u003cstrong\u003eLaboratory Characteristics of ABPA patients\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" width=\"582\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"168\"\u003e\n\u003cp\u003eLaboratory results\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"217\"\u003e\n\u003cp\u003eNormal range\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"197\"\u003e\n\u003cp\u003epeople（n=37）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"168\"\u003e\n\u003cp\u003eEosinophils（/uL）\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"217\"\u003e\n\u003cp\u003e20~500（/uL）\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"197\"\u003e\n\u003cp\u003e1080（540,1430）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"168\"\u003e\n\u003cp\u003eEosinophils（%）\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"217\"\u003e\n\u003cp\u003e0.5~5.0 (%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"197\"\u003e\n\u003cp\u003e14.70（8.90,22.75）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"168\"\u003e\n\u003cp\u003eTIgE（KU/L）\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"217\"\u003e\n\u003cp\u003e0.0~100.0(KU/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"197\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"168\"\u003e\n\u003cp\u003e\u0026lt;5000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"217\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd width=\"197\"\u003e\n\u003cp\u003e28（75.68%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"168\"\u003e\n\u003cp\u003e\u0026ge;5000\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"217\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd width=\"197\"\u003e\n\u003cp\u003e9（14.32%）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"168\"\u003e\n\u003cp\u003esIgE(kU/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"217\"\u003e\n\u003cp\u003e0.00~0.35(KU/L)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"197\"\u003e\n\u003cp\u003e5.69（2.83,12.02）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"168\"\u003e\n\u003cp\u003eESR（mm/h）\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"217\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd width=\"197\"\u003e\n\u003cp\u003e14.00（7.00,33.00）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"168\"\u003e\n\u003cp\u003eCRP（mg/L）\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"217\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd width=\"197\"\u003e\n\u003cp\u003e6.76（2.48,13.83）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"168\"\u003e\n\u003cp\u003eCEA（ng/ml）\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"217\"\u003e\n\u003cp\u003e0.00~5.05(ng/ml)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"197\"\u003e\n\u003cp\u003e7.20（3.65,12.70）\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"168\"\u003e\n\u003cp\u003eCEA \u0026lt;5.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"217\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd width=\"197\"\u003e\n\u003cp\u003e12 (32.43%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"168\"\u003e\n\u003cp\u003e5\u0026le;CEA\u0026lt;10\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"217\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd width=\"197\"\u003e\n\u003cp\u003e14 (37.84%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"168\"\u003e\n\u003cp\u003e\u0026ge;10.00\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"217\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd width=\"197\"\u003e\n\u003cp\u003e11(29.73%)\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\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":"Eosinophils, ABPA Patients, bronchopulmonary aspergillosis","lastPublishedDoi":"10.21203/rs.3.rs-73116/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-73116/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground \u003c/strong\u003eAllergic bronchopulmonary aspergillosis (ABPA) is a condition characterized by an exaggerated response of the immune system to the fungus Aspergillus. This study aimed to assess the relationship between carcinoembryonic antigen (CEA) and eosinophils in ABPA patients.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eMethods: \u003c/strong\u003eWe\u0026nbsp;describes a case of a 50-year-old patient who was diagnosed with ABPA presenting with high level of CEA and eosinophils. Besides,we used \u0026nbsp;immunohistochemistry and immunofluorescence to identify eosinophils and CEA in sections which were obtained by Endobronchial ultrasound-guided transbronchial lung biopsy aspiration (EBUS-TBLB). The sections were then visualized using confocal microscopy. We also retrospectively analyzed a cohort of 37 ABPA patients between January 2013 and December 2019 in our hospital\u003cem\u003e.\u003c/em\u003e\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e We found the patient whose serum CEA levels were consistent with eosinophils during the follow-up(r=0.929,P=0.022). The positive expression of CEA and abnormal expression of eosinophils was higher in the ABPA tissue compared to the normal lung tissue. The co-localization was represented as pixels containing both red and green color in the image\u0026nbsp;(with various shades of orange and yellow) which signified that eosinophils were immunohistochemically positive for CEA. Patients\u0026nbsp;with higher levels of eosinophils had higher levels of CEA in the serum (P\u0026lt;0.001). The results of \u003cem\u003ePearson correlation analysis \u003c/em\u003eshowed that the levels of eosinophils were positively correlated with serum CEA levels(r=0.459 and r=0.506，P=0.004 and P=0.001). \u0026nbsp;\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConclusion: \u003c/strong\u003eSerum CEA level is elevated in ABPA patients. The elevated serum CEA level was shown to be normalized after treatment. Increased CEA levels in ABPA patients may be positively correlated with eosinophil levels, and eosinophils may be served as CEA-secreting cells in patients with ABPA.\u003c/p\u003e","manuscriptTitle":"Eosinophils May Serve as CEA-Secreting Cells for ABPA Patients","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2020-09-15 16:39:55","doi":"10.21203/rs.3.rs-73116/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":"9035e02e-b6e1-4828-ab5c-6274020730a2","owner":[],"postedDate":"September 15th, 2020","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":522330,"name":"Pulmonology"}],"tags":[],"updatedAt":"2020-09-23T20:39:08+00:00","versionOfRecord":[],"versionCreatedAt":"2020-09-15 16:39:55","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-73116","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-73116","identity":"rs-73116","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}