Anti-thrombospondin-1 autoantibody indicated increased disease activity and xerophthalmia incidence in Primary Sjögren's syndrome

Anti-thrombospondin-1 autoantibody indicated increased disease activity and xerophthalmia incidence in Primary Sjögren's syndrome | 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 Anti-thrombospondin-1 autoantibody indicated increased disease activity and xerophthalmia incidence in Primary Sjögren's syndrome Ruoxi Li, Yuanhao Li, Yixue Guo, Yaodong Zhao, Xuewu Zhang, Xiaolin Sun This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8660592/v1 This work is licensed under a CC BY 4.0 License Status: Under Revision Version 1 posted 9 You are reading this latest preprint version Abstract Objective. The relationship between anti- thrombospondin-1 antibody (anti-TSP-1) and the clinical manifestations of Primary Sjögren's syndrome（pSS) has not been fully clarified.We analyzed the clinical factors relevance of anti-TSP-1 in pSS. Methods. Serum ATSA was detected by ELISA in 70 pSS patients and 21 healthy controls enrolled. Results. Anti-TSP-1 antibody is significantly higher in pSS patients (10.63± 4.92 vs. 7.64 ± 4.70, t =2.403, P =0.019). Positivity of anti-TSP-1 antibody in pSS and HCs were respectively 51.4% (36/70) and 28.6%(6/21). The increase of anti-TSP-1 antibody was also significantly correlated with the increase of IgG ( r = 0.263, P =0.029) and RF ( r =0.421, P ＜0.001), and the decrease of C4 ( r =-0.253, P =0.035)in pSS patients. The increase of serum anti-TSP-1 antibody was associated with the increase of ESSDAI in pSS patients ( r =0.277, P =0.02). Further statistical analysis showed that γG% values of anti-TSP-1 positive group were significantly higher than those of anti-TSP-1 negative group ( t =2.554, P =0.014). γG% values of anti-TSP-1 positive group were significantly higher than those of anti-TSP-1 negative group ( t =2.554, P =0.014).What’s more,the incidence of xerophthalmia significantly increased in the anti-TSP-1 positive group ( c 2 =4.939, P =0.026). Conclusion. Anti-TSP-1antibody levels were associated with increased SS disease activity,and it may be an effective tool in screening and predicting prognosis in pSS patients Anti- thrombospodin-1 antibody Primary Sjögren's syndrome Figures Figure 1 Figure 2 1. Introduction TSP-1 is a multi-functional matricellular ECM and secreted protein. It was first discovered in platelets[ 1 , 2 ].Multiple types of cells can secret TSP-1,including endothelial cells,fibroblasts, adipocytes, smooth muscle cells, monocytes, macrophages, and malignant glioma cells[ 3 , 4 ]. Recently,more properties of TSP-1 are also gradually confirmed,including growth factor (EGF, VEGF, FGF) regulation, inhibition of angiogenesis and immune cell recruitment and activation[ 5 , 6 ].Moreover,targeting the TSP-1 pathway potentially represents a more selective approach to controlling activity in some diseases[ 7 ]. Primary Sjögren's syndrome (pSS) is a chronic autoimmune disease that mainly affects exocrine glands.Functional loss of these glands leads to clinical manifestations of dry mouth (xerostomia) and dry eye (xerophthalmia).However,a majority of pSS patients also experience multi-system impairment.Growing evidence indicates that TSP-derived peptide may attenuate Sjögren's syndrome-associated symptoms[ 8 ].However, there has been few research investigating the role of anti-TSP-1 antibodies in pSS patients.In this study,we determined the serum levels of anti-TSP-1 autoantibodies in pSS patients,and analyzed the clinical significance of serum anti-TSP-1 in patients with pSS.The results showed that TSP-1 autoantibody might serve as a new indicator of disease activity in pSS. 2. Materials and Methods 2.1 Patients and Controls. We retrospectively analyzed the data of 70 patients with primary Sjögren's syndrome (pSS) who had been admitted to The Fifth Affiliated Hospital of Zhengzhou University and Peking University People’s Hospital.All pSS patients were diagnosed based on the revised USEURO classification criteria for SS (2002)[ 9 ] or ACR classification criteria (2012)[ 10 ].Patients with other conditions such as RA,SLE and diabetes were excluded.21 healthy controls (HCs) were collected serum from age- and sex-matched disease-free subjects who underwent routine medical examinations at same hospital.All of them have no infection, autoimmune disease, tumor and other diseases.All subjects provided their written informed consents before the samples were obtained. The study was conducted in accordance with the Declaration of Helsinki.The study was approved by the Medical Ethical Committee of The Fifth Affiliated Hospital of Zhengzhou University and Peking University People’s Hospital. The study protocol was approved by the Ethical Committee of The Fifth Affiliated Hospital of Zhengzhou University and Peking University People’s Hospital (Y2021089、2016PHB168⁃01). 2.2 Serum samples. Venous blood was collected by vacuum coagulation vessel on an empty stomach. After mixing, the blood was centrifuged at 3500r/min within 2h, centrifuged at 4℃ for 5min. The serum samples of 70 pSS patients and 21 HCs were preserved at -80°C before tests. Their clinical and laboratory data were reviewed. 2.3 Methods. Serum levels of autoantibody to TSP-1 were determined by enzyme-linked immunosorbent assay (ELISA).Recombinant human TSP-1 (R&D Systems, Minneapolis,USA) of 4µg/L was dissolved in carbonate buffer to coat 96-well polysorp plates (Nunc,Denmark) at 4 ℃ overnight.Then,the well were washed for 4 times with phosphate-buffered saline containing 0.05% Tween-20 (PBS-T),and blocked with 3% albumin bovine V (BSA) for 3.5 hours at 37℃.After washing with PBS-T for four times,serum samples were diluted at 1:200 with PBS-T containing 1% BSA.After incubation for 1hour at 37℃,the wells were washed by PBS-T for four times.Then,100 µL of goat anti-human IgG (Solarbio,Beijing,China) conjugated to peroxidase,diluted at 1:10000, were added to each well and then incubated for 30 minutes at 37°C.100 µL of Tetramethylbenzidine (TMB) (Solarbio,Beijing ,China) was added after washing wells for four times,and fifteen minutes later,2 M sulfuric acid was added to stop the color reaction. Plates were read by a plate reader (Bio Tek) at an absorbance wavelength of 450 and 360nm (OD450 and OD360). Each serum sample was assayed in duplicate. A positive serum sample was included on each plate as a positive control and reference to correct for inter-assay variation. The values of OD of anti-TSP-1 antibody were transformed to arbitrary units (AU),calculated as follows: [OD peptide -OD non−specific background ] test serum / [OD peptide -OD non−specific background ] positive control serum ×100. In this study, we determined the cut-off values by mean + 2 × standard deviation of TSP-1 antibody in healthy controls. 2.4 Clinical and laboratory evaluation. Clinical and laboratory data of pSS patients were reviewed retrospectively from the medical records,including age, sex, gender,disease duration,clinical symptoms,white blood cell (WBC) count,red blood cell (RBC) count,hemoglobin (Hb),platelet count (PLT),rheumatoid factor (RF),erythrocyte sedimentation rate (ESR), immunoglobulin A (IgA), immunoglobulin G (IgG), immunoglobulin M (IgM), complement component 3 (C3), complement component 4 (C4),C-reactive protein (CRP),anti-nuclear antibodies (ANA),anti-ribosomal P protein (rRNP) antibody,anti-SSA antibody (SSA),anti-SSB antibody (SSB),anti α-fodrin antibody.The “EULAR Sjögren's syndrome disease activity index” (ESSDAI)[ 11 ]was used to assess disease activity. 2.5 Statistical analysis. SPSS25.0 for windows and GraphPad Prism 8.0 were used to analyze all data.The data of normal distribution were expressed as mean± standard deviation.Data of non-normal distribution is represented by median (range). Statistical significance between two groups was assessed with the Student t -test,the Mann-Whitney U test and χ 2 test.Collelation of data was examined by Pearson’s rank correlation coefficient and further verified by multiple linear regression analysis.P value less than 0.05 was considered to be statistically significant. 3. Results 3.1 Characteristics of the pSS patients and controls.In this study, 70 pSS patients and 21 healthy controls with matched age and gender were recruited.The characteristics of the pSS patients and healthy controls were enrolled in Table 1 . The average age of 70 pSS patients was 46.37 ± 9.63 years (range 30–67), and the average disease duration ranged from 3 to 312 months. Table 1 Characteristics of the pSS patients and controls General information pSS Controls t/u/x 2 P Sex(M/F) 19/51 2/19 1.920 0.166 Age (years) 46.37 ± 9.63 42.33 ± 7.53 2.012 0.051 Disease duration (months) 24 (3,312) - - - pSS: Primary Sjögren's syndrome; M: male;F: female; 3.2 Serum anti-TSP-1 antibodies were elevated in pSS patients.Anti-TSP-1 antibody is significantly higher in pSS patients (10.63 ± 4.92 v.s. 7.64 ± 4.70, t = 2.403,p = 0.019, Fig. 1 ).A cut-off value (9.79) for positivity was defined as the 95th percentile of the healthy sera.Positivity of anti-TSP-1 antibody in pSS and HCs were 51.4%(36/70) and 28.6% (6/21) respectively,which was not significantly different (χ 2 = 3.396,p = 0.065).The positive rates of TSP-1 antibody were 65.2% (15/23),62.5% (15/24) and 55.1% (27/49) in those patients with negative ANA,anti-SSA and anti-SSB,respectively. 3.3 Serum anti-TSP-1 antibody levels were associated with increased pSS disease activity.As shown in Table 2 ,the increase of anti-TSP-1 antibody was significantly correlated with the increase of IgG (r = 0.263,P = 0.029,Table 2 and Fig. 2 a),RF (r = 0.421,P<0.001,Table 2 and Fig. 2 b),and the decrease of C4 (r=-0.253,P = 0.035,Table 2 and Fig. 2 c)in pSS patients.Meanwhile,the increase of serum anti-TSP-1 antibody was also associated with the increase of ESSDAI (r = 0.277,P = 0.02,Table 2 and Fig. 2 d). These results demonstrated that elevated serum anti-TSP-1 levels were associated with increased SS disease activity.Further statistical analysis showed that γG% values of anti-TSP-1 positive group were significantly higher than those of the anti-TSP-1 negative group ( t = 2.554,P = 0.014, Table 3 ).What’s more,the incidence of xerophthalmia significantly increased in the anti-TSP-1 positive group (χ 2 = 4.939,P = 0.026, Table 3 ),which implicated its pathogenic role in pSS development. Table 2 Correlations of anti-TSP-1 antibody with clinical parameters of pSS Clinical parameters Anti-TSP-1 antibody r P Age(years) -0.078 0.521 Disease duration (months) 0.046 0.708 WBC(10 9 /L) 0.087 0.480 Neutrophils (×10 9 ) 0.076 0.533 Lymphocyte (×10 9 ) 0.092 0.451 RBC(×10 12 /L) 0.003 0.979 Hb(g/L) -0.006 0.962 PLT(10 9 /L) -0.141 0.246 CRP(mg/L) -0.143 0.292 ESR(mm/h) 0.014 0.917 γG(%) 0.147 0.283 IgG(g/L) 0.263 0.029 IgA(g/L) -0.029 0.818 IgM(g/L) 0.101 0.414 C3(g/L) -0.04 0.745 C4(g/L) -0.253 0.035 RF(IU/mL) 0.421 <0.001 Anti-α-fodrin(U/ml) -0.083 0.537 ESSDAI 0.277 0.020 Table 2 . pSS: Primary Sjögren's syndrome; WBC: white blood cell; N: neutrophil;L:lymphocyte; RBC: erythrocyte;Hb: hemoglobin; PLT: platelet count; CRP: C-reaction protein; ESR: erythrocyte sedimentation rate; RF: rheumatoid factor; γG : γglobulin;IgG: immunoglobulin G; IgA: immunoglobulin A; IgM: immunoglobulin M;C3: complement component 3; C4: complement component 4; ESSDAI: EULAR Sjögren's syndrome disease activity index. Table 3 Comparison of clinical and laboratory parameters between anti-TSP-1 positive and negative pSS patients. Clinical and laboratory parameters Anti-TSP-1 positive (n = 36) Anti-TSP-1 negative (n = 34) t/Z/χ 2 P Xerophthalmia, n (%) 29(80.6%) 19(55.9%) 4.939 0.026 Xerostomia, n (%) 32(88.9%) 27(79.4%) 1.186 0.276 Lymphadenectasis, n(%) 7(19.4%) 3(8.8%) 1.611 0.204 Arthralgia,n(%) 18(50.0%) 15(44.1%) 0.243 0.622 Lung involvement,n(%) 19(52.8%) 11(32.4%) 2.979 0.084 renal lesion,n(%) 9(25.0%) 9(26.5%) 0.020 0.888 WBC(×10 9 ) 6.05 ± 2.96 5.52 ± 2.80 0.771 0.443 Neutrophils (×10 9 ) 3.77 ± 2.40 3.42 ± 2.61 0.591 0.557 Hb(g/L) 131.00 ± 21.94 130.81 ± 17.80 0.039 0.969 PLT(×10 9 ) 188.36 ± 95.58 173.55 ± 72.08 0.321 0.473 γG(%) 24.05 ± 10.79 19.07 ± 4.41 2.554 0.014 IgG(g/L) 17.35 ± 10.40 15.36 ± 3.71 1.039 0.302 IgA(g/L) 3.05 ± 1.06 3.43 ± 1.44 -1.268 0.209 IgM(g/L) 1.13 ± 0.72 1.22 ± 0.86 -0.467 0.642 C3(g/L) 0.92 ± 0.20 0.91 ± 0.21 0.205 0.838 C4(g/L) 0.19 ± 0.04 0.21 ± 0.07 -1.273 0.207 CRP(mg/L) 1.01(0.2,56.61) 1.35(0.2,200) -0.805 0.421 ESR(mm/h) 20.62 ± 21.51 19.28 ± 15.92 0.199 0.794 RF(IU/mL) 20(20,721) 21.3(20,379) -1.107 0.268 ANA positive 26(72.2%) 29(85.3%) 1.775 0.183 Anti-SSA positive 21(58.3%) 25(73.5%) 1.792 0.181 Anti-SSB possitive 9(25.0%) 11(32.4%) 0.463 0.496 Anti-α-fodrin (U/mL) 9.41(2.2,128.28) 17.08(0,65.7) -0.579 0.563 ESSDAI 4.06 ± 1.91 3.32 ± 1.61 1.728 0.088 Table 3 .pSS: Primary Sjögren's syndrome;WBC: white blood cell;N: neutrophil;L:lymphocyte; RBC:erythrocyte;Hb: hemoglobin;PLT:platelet count;CRP:C-reaction protein;ESR:erythrocyte sedimentation rate;RF:rheumatoid factor;γG:γglobulin;IgG:immunoglobulin G;IgA:immunoglobulin A;IgM:immunoglobulin M;C3:complement component 3;C4:complement component 4;ESSDAI:EULAR Sjögren's syndrome disease activity index. 4. Discussion According to the American-European classification criteria in 2016,diagnosis of the pSS is based on the clinical symptoms,salivary gland biopsy and detection of circulating autoantibodies, including anti-Sjögren's syndrome A (anti-SSA/Ro) or B (anti-SSB/La)[ 10 , 12 ].Although salivary gland biopsy can be used as the gold standard for the diagnosis of pSS,it is not the first choice of clinical examination for its trauma.Autoantibodies have been shown to be involved in the development of pSS.Thus,it is urgent to screen autoantibodies in pSS.However,some reports showed that anti-SSA antibody might emerge at relatively late stage of the disease.Therefore,these commonly used antibodies are less sensitive in the diagnosis of pSS[ 12 , 13 ].Our study found that TSP-1 autoantibody was significantly higher in serum from pSS patients than in healthy controls.Positivity of anti-TSP-1 antibody in pSS and HCs were respectively 51.4% and 28.6%.Therefore,TSP-1 autoantibodiesare also involve in pathogenesis of pSS. TSP-1 was first discovered in platelets[ 6 ].It is a matricellular protein and has multiple functions,including the modulation of NO signals[ 14 ],and immune cell recruitment and activation[ 1 , 2 , 6 ].TSP1 is widely upregulated in tissue injuries to accelerate tissue reparation[ 5 , 15 ].The pathological characteristics of pSS are lymphocytic infiltrate of the exocrine glands.In addition to lacrimal gland and salivary gland duct epithelial cells,it also includes alveolar epithelial cells,renal tubular epithelial cells and liver fibroblasts,which lead to multiple system dysfunction and destruction[ 16 ].In recent years, accumulating evidence suggests that the TSP-1 plays an important role in gland function and homeostasis[ 17 ].TSP-1 deficient antigen-presenting cells(APC) promotes Th17 cells mediated inflammatory response in the periphery and gland of SS murine models[ 17 , 18 , 19 ].In our study,the levels of anti-TSP-1 antibody in pSS patients were significantly associated with inflammation and disease severity parameters,such as hypocomplementemia, immunoglobulin G and RF. The level of γG in anti-TSP-1-positive pSS patients is significantly higher than that in anti-TSP-1-negitive pSS patients (P<0.05, Supplementary Fig. 2 and Tables 2 and 3 ).Our study further validated that the ESSDAI score was significantly correlated with the anti-TSP-1 serum concentration.These results suggest that anti-TSP-1 is might promote the development of pSS. Our study did not find a significant association of anti-TSP-1 antibody with other autoantibodies in pSS patients.Thus,anti-TSP-1 antibody is independent.But as an autoantigen,TSP-1 is localized in various cell types,including those in ocular tissues and exocrine glands[ 20 , 21 ].Interestingly,in our study,the group of anti-TSP-1-positive pSS patients had significantly higher prevalence of xerophthalmia.Bruce Turpie et al.[ 17 ] reported that mice deficient of TSP-1 eventually developed a chronic ocular surface disease.The pathological process is driven by apoptosis and deterioration of lacrimal gland cells,resulting in dysfunction and inflammatory infiltration,and abnormal tears.We hypothesized that TSP-1 antibody might be associated with SS ocular surface inflammation. During the occurrence and development of pSS,TSP-1 and other autoantigens might trigger and stimulate the abnormal activation of autoimmune T cell and B cell to produce more autoantibodies,which in turn damage the gland tissues and other organs[ 22 ]. A cohort research of 921 Spanish patients with pSS showed that the most frequently involved organ was the lung[ 23 ],and a proportion of patients might develop pulmonary interstitial fibrosis in the later stage of disease.As a major activator of TGF-β,TSP-1 plays an important role in fibrotic diseases.Joanne E et al.[ 7 ] found that antagonists of the TSP-1/TGF-β pathway might be suitable drug candidates for fibrotic diseases,including diabetic nephropathy,liver fibrosis,pulmonary fibrosis and scleroderma[ 7 , 24 , 25 ].Besides,our study also found that the group of anti-TSP-1-positive pSS patients had increased trend of prevalence of lung involvement. However,the number of serum samples we collected is limited.Therefore,our future experiments need to expand the sample size to explore the role of TSP-1 autoantibodies in pSS.In addition,the relationship between TSP-1 antibody and disease activity and clinical phenotype should be verified again according to the severity of the disease. In conclusion, this study revealed the clinical relevance of TSP-1 autoantibody in pSS.It showed that TSP-1 autoantibody might be an effective tool in screening and evaluating disease activity in pSS patients.However, the pathogenic mechanism of TSP-1 autoantibodies in pSS demands further study. Declarations Funding This work was supported by the National Natural Science Foundation of China (81671602, 81801619); and the Key Technology Research and Development Program of Henan Province Department of Education (19A320016). Data Availability The data used to support the findings of the current study are available from the corresponding authors upon reasonable request. Conflicts of Interest The authors declare that they have no conflict of interests. Authors’ Contributions Ruoxi Li performed most of the experiments and drafted the manuscript.Ruoxi Li,Yuanhao Li,Yixue Guo, Yaodong Zhao and Yang Meng worked on data collection.Xuewu Zhang,Wang Yan and Xiaolin Sun conceived the study and participated in its design and coordination and interpreted the data.All authors read and approved the final manuscript. References Baenziger NL, Brodie GN, Majerus PW. Isolation and properties of a thrombin-sensitive protein of human platelets. Biol Chem. 1972;247(9):2723–31. Lawler JW, Slayter HS, Coligan JE. Isolation and characterization of a high molecular weight glycoprotein from human blood platelets. Biol Chem. 1978;253(23):8609–16. Wight TN, Raugi GJ, Mumby SM, Bornstein P. Light microscopic immunolocation of thrombospondin in human tissues. Histochem Cytochem. 1985;33(4):295–302. Naganuma H, Satoh E, Asahara T, et al. Quantification of thrombospondin-1 secretion and expression of alphavbeta3 and alpha3beta1 integrins and syndecan-1 as cell-surface receptors for thrombospondin-1 in malignant glioma cells. Neurooncol. 2004;70(3):309–17. Adams JC, Lawler J. The thrombospondins. Cold Spring Harb Perspect Biol. 2011;3(10):a009712. Grenier C, Caillon A, Munier M, et al. Dual Role of Thrombospondin-1 in Flow-Induced Remodeling. Int J Mol Sci. 2021;22(21):12086. Published 2021 Nov 8. Murphy-Ullrich JE, Suto MJ. Thrombospondin-1 regulation of latent TGF-β activation: A therapeutic target for fibrotic disease. Matrix Biol. 2018;68–69:28–43. Masli S, Dartt DA. Mouse Models of Sjögren's Syndrome with Ocular Surface Disease. Int J Mol Sci. 2020;21(23):9112. Vitali C, Bombardieri S, Jonsson R, et al. Classification criteria for Sjögren's syndrome: a revised version of the European criteria proposed by the American-European Consensus Group. Ann Rheum Dis. 2002;61(6):554–8. Shiboski CH, Shiboski SC, Seror R, et al. 2016 American College of Rheumatology/European League Against Rheumatism Classification Criteria for Primary Sjögren's Syndrome: A Consensus and Data-Driven Methodology Involving Three International Patient Cohorts. Arthritis Rheumatol. 2017;69(1):35–45. Seror R, Ravaud P, Bowman SJ, et al. EULAR Sjogren's syndrome disease activity index: development of a consensus systemic disease activity index for primary Sjogren's syndrome. Ann Rheum Dis. 2010;69(6):1103–9. Hu Q, Wang D, Chen W. The accuracy of the anti-α-fodrin antibody test for diagnosis of Sjögren's syndrome: a meta-analysis. Clin Biochem. 2013;46(15):1372–6. Jin Y, Li J, Chen J, et al. Tissue-Specific Autoantibodies Improve Diagnosis of Primary Sjögren's Syndrome in the Early Stage and Indicate Localized Salivary Injury. Immunol Res. 2019;2019:3642937. Published 2019 May 7. Isenberg JS, Martin-Manso G, Maxhimer JB, Roberts DD. Regulation of nitric oxide signalling by thrombospondin 1: implications for anti-angiogenic therapies. Nat Rev Cancer. 2009;9(3):182–94. Murphy-Ullrich JE, Sage EH. Revisiting the matricellular concept. Matrix Biol. 2014;37:1–14. Thorne I, Sutcliffe N. Sjögren's syndrome. Br J Hosp Med (Lond). 2017;78(8):438–42. Turpie B, Yoshimura T, Gulati A, Rios JD, Dartt DA, Masli S. Sjögren's syndrome-like ocular surface disease in thrombospondin-1 deficient mice. Am J Pathol. 2009;175(3):1136–47. Ríos JD, Horikawa Y, Chen LL, et al. Age-dependent alterations in mouse exorbital lacrimal gland structure, innervation and secretory response. Exp Eye Res. 2005;80(4):477–91. Rashid S, Jin Y, Ecoiffier T, Barabino S, Schaumberg DA, Dana MR. Topical omega-3 and omega-6 fatty acids for treatment of dry eye. Arch Ophthalmol. 2008;126(2):219–25. Masli S, Sheibani N, Cursiefen C, Zieske J. Matricellular protein thrombospondins: influence on ocular angiogenesis, wound healing and immuneregulation. Curr Eye Res. 2014;39(8):759–74. Gromova A, Voronov DA, Yoshida M, et al. Lacrimal Gland Repair Using Progenitor Cells. Stem Cells Transl Med. 2017;6(1):88–98. Jin Y, Li J, Chen J, et al. Tissue-Specific Autoantibodies Improve Diagnosis of Primary Sjögren's Syndrome in the Early Stage and Indicate Localized Salivary Injury. Immunol Res. 2019;2019:3642937. Published 2019 May 7. Ramos-Casals M, Brito-Zerón P, Solans R, et al. Systemic involvement in primary Sjogren's syndrome evaluated by the EULAR-SS disease activity index: analysis of 921 Spanish patients (GEAS-SS Registry). Rheumatology (Oxford). 2014;53(2):321–31. Chang Y, Lau WL, Jo H, et al. Pharmacologic Blockade of αvβ1 Integrin Ameliorates Renal Failure and Fibrosis In Vivo. Am Soc Nephrol. 2017;28(7):1998–2005. Munger JS, Huang X, Kawakatsu H, et al. The integrin alpha v beta 6 binds and activates latent TGF beta 1: a mechanism for regulating pulmonary inflammation and fibrosis. Cell. 1999;96(3):319–28. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Revision Version 1 posted Editorial decision: Revision requested 14 May, 2026 Reviews received at journal 04 May, 2026 Reviewers agreed at journal 26 Apr, 2026 Reviews received at journal 15 Feb, 2026 Reviewers agreed at journal 02 Feb, 2026 Reviewers invited by journal 28 Jan, 2026 Editor assigned by journal 27 Jan, 2026 Submission checks completed at journal 27 Jan, 2026 First submitted to journal 21 Jan, 2026 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. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-8660592","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":582568292,"identity":"6c15a5a0-642c-4b6e-a851-9bbd1325af88","order_by":0,"name":"Ruoxi Li","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAxElEQVRIiWNgGAWjYBACPmYwJSHHz9584MCHH0RoYYNqMZbsOZZ4cGYPMVqgdOKGGz7GhznY8CqGamHnfSbxsc0CqIXnw2EGHgZ5frEDhBzGbiY544yE8czbvRsOF1gwGM6cnUBICxubNE+FhGzfnbMbDs/gYUgwuE2UFgMJxoYbOQ8O87ARraVCQnHCjRwGorUwW4L8AgxkA2AgSxD2Cz//McYbH9vqQFH5+MOHHzby/NIEtKADCdKUj4JRMApGwSjADgC40z2hr3Zx0AAAAABJRU5ErkJggg==","orcid":"","institution":"Fifth Affiliated Hospital of Zhengzhou University","correspondingAuthor":true,"prefix":"","firstName":"Ruoxi","middleName":"","lastName":"Li","suffix":""},{"id":582568294,"identity":"cd7b681b-ab30-4765-8329-782d6e313ae5","order_by":1,"name":"Yuanhao Li","email":"","orcid":"","institution":"Henan Provincial Chest Hospital","correspondingAuthor":false,"prefix":"","firstName":"Yuanhao","middleName":"","lastName":"Li","suffix":""},{"id":582568295,"identity":"154b8cd1-1119-459f-a917-4cc6c7c9bbb3","order_by":2,"name":"Yixue Guo","email":"","orcid":"","institution":"Peking University People's Hospital","correspondingAuthor":false,"prefix":"","firstName":"Yixue","middleName":"","lastName":"Guo","suffix":""},{"id":582568296,"identity":"c8f71475-9045-4316-a611-db7e182e131f","order_by":3,"name":"Yaodong Zhao","email":"","orcid":"","institution":"Fifth Affiliated Hospital of Zhengzhou University","correspondingAuthor":false,"prefix":"","firstName":"Yaodong","middleName":"","lastName":"Zhao","suffix":""},{"id":582568297,"identity":"660909cb-b167-42a0-b89a-a1107f0ec7ea","order_by":4,"name":"Xuewu Zhang","email":"","orcid":"","institution":"Peking University People's Hospital","correspondingAuthor":false,"prefix":"","firstName":"Xuewu","middleName":"","lastName":"Zhang","suffix":""},{"id":582568298,"identity":"370e6fd4-132f-4e30-82eb-fb9f72fb7ce3","order_by":5,"name":"Xiaolin Sun","email":"","orcid":"","institution":"Peking University People's Hospital","correspondingAuthor":false,"prefix":"","firstName":"Xiaolin","middleName":"","lastName":"Sun","suffix":""}],"badges":[],"createdAt":"2026-01-21 13:54:13","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8660592/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8660592/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":101631139,"identity":"0578db9e-e003-4562-ab49-a129a20df955","added_by":"auto","created_at":"2026-02-02 05:28:58","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":11643,"visible":true,"origin":"","legend":"\u003cp\u003eSerum levels of anti-TSP-1 antibodies in SS patients and HCs.\u003c/p\u003e","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-8660592/v1/7d7c5dfd8ecc93146f35123b.png"},{"id":101753668,"identity":"ee2af1ba-e5f2-470d-8f1b-c397300c4042","added_by":"auto","created_at":"2026-02-03 10:40:30","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":87244,"visible":true,"origin":"","legend":"\u003cp\u003e(\u003cstrong\u003ea\u003c/strong\u003e) Anti-TSP-1 antibody positively correlated with IgG in pSS patients;(\u003cstrong\u003eb\u003c/strong\u003e) Anti-TSP-1 antibody positively correlated with RF in pSS patients; (\u003cstrong\u003ec\u003c/strong\u003e) Anti-TSP-1 antibody negatively correlated with C4 in pSS patients; (\u003cstrong\u003ed\u003c/strong\u003e) Anti-TSP-1 antibody positively correlated with ESSDAI in pSS patients.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-8660592/v1/f6c6722741da80e6809b1ade.png"},{"id":101943038,"identity":"a4358731-57e7-4fa9-a82e-4380e41b9f52","added_by":"auto","created_at":"2026-02-05 09:39:50","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":773921,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8660592/v1/4391034c-d926-46a2-9414-d7830c9e407d.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Anti-thrombospondin-1 autoantibody indicated increased disease activity and xerophthalmia incidence in Primary Sjögren's syndrome","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eTSP-1 is a multi-functional matricellular ECM and secreted protein. It was first discovered in platelets[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e].Multiple types of cells can secret TSP-1,including endothelial cells,fibroblasts, adipocytes, smooth muscle cells, monocytes, macrophages, and malignant glioma cells[\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Recently,more properties of TSP-1 are also gradually confirmed,including growth factor (EGF, VEGF, FGF) regulation, inhibition of angiogenesis and immune cell recruitment and activation[\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].Moreover,targeting the TSP-1 pathway potentially represents a more selective approach to controlling activity in some diseases[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e].\u003c/p\u003e \u003cp\u003ePrimary Sj\u0026ouml;gren's syndrome (pSS) is a chronic autoimmune disease that mainly affects exocrine glands.Functional loss of these glands leads to clinical manifestations of dry mouth (xerostomia) and dry eye (xerophthalmia).However,a majority of pSS patients also experience multi-system impairment.Growing evidence indicates that TSP-derived peptide may attenuate Sj\u0026ouml;gren's syndrome-associated symptoms[\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e].However, there has been few research investigating the role of anti-TSP-1 antibodies in pSS patients.In this study,we determined the serum levels of anti-TSP-1 autoantibodies in pSS patients,and analyzed the clinical significance of serum anti-TSP-1 in patients with pSS.The results showed that TSP-1 autoantibody might serve as a new indicator of disease activity in pSS.\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cdiv id=\"Sec3\"\u003e\n \u003ch2\u003e2.1 Patients and Controls. We retrospectively analyzed the data of 70 patients with primary Sj\u0026ouml;gren\u0026apos;s\u003c/h2\u003e\n \u003cp\u003esyndrome (pSS) who had been admitted to The Fifth Affiliated Hospital of Zhengzhou University\u003c/p\u003e\n \u003cp\u003eand Peking University People\u0026rsquo;s Hospital.All pSS patients were diagnosed based on the revised USEURO classification criteria for SS (2002)[\u003cspan\u003e9\u003c/span\u003e] or ACR classification criteria (2012)[\u003cspan\u003e10\u003c/span\u003e].Patients with other conditions such as RA,SLE and diabetes were excluded.21 healthy controls (HCs) were\u003c/p\u003e\n \u003cp\u003ecollected serum from age- and sex-matched disease-free subjects who underwent routine medical\u003c/p\u003e\n \u003cp\u003eexaminations at same hospital.All of them have no infection, autoimmune disease, tumor and other\u003c/p\u003e\n \u003cp\u003ediseases.All subjects provided their written informed consents before the samples were obtained. The study was conducted in accordance with the Declaration of Helsinki.The study was approved by the Medical Ethical Committee of The Fifth Affiliated Hospital of Zhengzhou University and Peking University People\u0026rsquo;s Hospital. The study protocol was approved by the Ethical Committee of The Fifth Affiliated Hospital of Zhengzhou University and Peking University People\u0026rsquo;s Hospital (Y2021089、2016PHB168⁃01).\u003c/p\u003e\u003cspan\u003e\n \u003cp\u003e2.2 Serum samples. Venous blood was collected by vacuum coagulation vessel on an empty stomach. After mixing, the blood was centrifuged at 3500r/min within 2h, centrifuged at 4℃ for 5min. The serum samples of 70 pSS patients and 21 HCs were preserved at -80\u0026deg;C before tests. Their clinical and laboratory data were reviewed.\u003c/p\u003e\n \u003c/span\u003e \u003cspan\u003e\n \u003cp\u003e2.3 Methods. Serum levels of autoantibody to TSP-1 were determined by enzyme-linked immunosorbent assay (ELISA).Recombinant human TSP-1 (R\u0026amp;D Systems, Minneapolis,USA) of 4\u0026micro;g/L was dissolved in carbonate buffer to coat 96-well polysorp plates (Nunc,Denmark) at 4 ℃ overnight.Then,the well were washed for 4 times with phosphate-buffered saline containing 0.05% Tween-20 (PBS-T),and blocked with 3% albumin bovine V (BSA) for 3.5 hours at 37℃.After washing with PBS-T for four times,serum samples were diluted at 1:200 with PBS-T containing 1% BSA.After incubation for 1hour at 37℃,the wells were washed by PBS-T for four times.Then,100 \u0026micro;L of goat anti-human IgG (Solarbio,Beijing,China) conjugated to peroxidase,diluted at 1:10000, were added to each well and then incubated for 30 minutes at 37\u0026deg;C.100 \u0026micro;L of Tetramethylbenzidine (TMB) (Solarbio,Beijing ,China) was added after washing wells for four times,and fifteen minutes later,2 M sulfuric acid was added to stop the color reaction. Plates were read by a plate reader (Bio Tek) at an absorbance wavelength of 450 and 360nm (OD450 and OD360). Each serum sample was assayed in duplicate. A positive serum sample was included on each plate as a positive control and reference to correct for inter-assay variation. The values of OD of anti-TSP-1 antibody were transformed to arbitrary units (AU),calculated as follows: [OD\u003csub\u003epeptide\u003c/sub\u003e-OD\u003csub\u003enon\u0026minus;specific background\u003c/sub\u003e]\u003csub\u003etest serum\u003c/sub\u003e/ [OD\u003csub\u003epeptide\u003c/sub\u003e-OD\u003csub\u003enon\u0026minus;specific background\u003c/sub\u003e]\u003csub\u003epositive control serum\u003c/sub\u003e\u0026times;100. In this study, we determined the cut-off values by mean\u0026thinsp;+\u0026thinsp;2 \u0026times; standard deviation of TSP-1 antibody in healthy controls.\u003c/p\u003e\n \u003c/span\u003e \u003cspan\u003e\n \u003cp\u003e2.4 Clinical and laboratory evaluation. Clinical and laboratory data of pSS patients were reviewed retrospectively from the medical records,including age, sex, gender,disease duration,clinical symptoms,white blood cell (WBC) count,red blood cell (RBC) count,hemoglobin (Hb),platelet count (PLT),rheumatoid factor (RF),erythrocyte sedimentation rate (ESR), immunoglobulin A (IgA), immunoglobulin G (IgG), immunoglobulin M (IgM), complement component 3 (C3), complement component 4 (C4),C-reactive protein (CRP),anti-nuclear antibodies (ANA),anti-ribosomal P protein (rRNP) antibody,anti-SSA antibody (SSA),anti-SSB antibody (SSB),anti \u0026alpha;-fodrin antibody.The \u0026ldquo;EULAR Sj\u0026ouml;gren\u0026apos;s syndrome disease activity index\u0026rdquo; (ESSDAI)[\u003cspan\u003e11\u003c/span\u003e]was used to assess disease activity.\u003c/p\u003e\n \u003c/span\u003e \u003cspan\u003e\n \u003cp\u003e2.5 Statistical analysis. SPSS25.0 for windows and GraphPad Prism 8.0 were used to analyze all data.The data of normal distribution were expressed as mean\u0026plusmn; standard deviation.Data of non-normal distribution is represented by median (range). Statistical significance between two groups was assessed with the Student \u003cem\u003et\u003c/em\u003e-test,the Mann-Whitney \u003cem\u003eU\u003c/em\u003e test and \u0026chi;\u003csup\u003e2\u003c/sup\u003e test.Collelation of data was examined by Pearson\u0026rsquo;s rank correlation coefficient and further verified by multiple linear regression analysis.P value less than 0.05 was considered to be statistically significant.\u003c/p\u003e\n \u003c/span\u003e\n\u003c/div\u003e"},{"header":"3. Results","content":"\u003cp\u003e3.1 Characteristics of the pSS patients and controls.In this study, 70 pSS patients and 21 healthy controls with matched age and gender were recruited.The characteristics of the pSS patients and healthy controls were enrolled in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The average age of 70 pSS patients was 46.37\u0026thinsp;\u0026plusmn;\u0026thinsp;9.63 years (range 30\u0026ndash;67), and the average disease duration ranged from 3 to 312 months.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003e Characteristics of the pSS patients and controls\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGeneral information\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003epSS\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eControls\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003et/u/x\u003c/em\u003e\u003csup\u003e\u003cem\u003e2\u003c/em\u003e\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eP\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSex(M/F)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e19/51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2/19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.920\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.166\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAge (years)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e46.37\u0026thinsp;\u0026plusmn;\u0026thinsp;9.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e42.33\u0026thinsp;\u0026plusmn;\u0026thinsp;7.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.012\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.051\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDisease duration (months)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e24 (3,312)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003epSS: Primary Sj\u0026ouml;gren's syndrome; M: male;F: female;\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003e3.2\u003c/b\u003e Serum anti-TSP-1 antibodies were elevated in pSS patients.Anti-TSP-1 antibody is significantly higher in pSS patients (10.63\u0026thinsp;\u0026plusmn;\u0026thinsp;4.92 \u003cem\u003ev.s.\u003c/em\u003e 7.64\u0026thinsp;\u0026plusmn;\u0026thinsp;4.70, t\u0026thinsp;=\u0026thinsp;2.403,p\u0026thinsp;=\u0026thinsp;0.019, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).A cut-off value (9.79) for positivity was defined as the 95th percentile of the healthy sera.Positivity of anti-TSP-1 antibody in pSS and HCs were 51.4%(36/70) and 28.6% (6/21) respectively,which was not significantly different (χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;3.396,p\u0026thinsp;=\u0026thinsp;0.065).The positive rates of TSP-1 antibody were 65.2% (15/23),62.5% (15/24) and 55.1% (27/49) in those patients with negative ANA,anti-SSA and anti-SSB,respectively.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003e3.3\u003c/b\u003eSerum anti-TSP-1 antibody levels were associated with increased pSS disease activity.As shown in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e,the increase of anti-TSP-1 antibody was significantly correlated with the increase of IgG (r\u0026thinsp;=\u0026thinsp;0.263,P\u0026thinsp;=\u0026thinsp;0.029,Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ea),RF (r\u0026thinsp;=\u0026thinsp;0.421,P\u0026lt;0.001,Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eb),and the decrease of C4 (r=-0.253,P\u0026thinsp;=\u0026thinsp;0.035,Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ec)in pSS patients.Meanwhile,the increase of serum anti-TSP-1 antibody was also associated with the increase of ESSDAI (r\u0026thinsp;=\u0026thinsp;0.277,P\u0026thinsp;=\u0026thinsp;0.02,Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003ed). These results demonstrated that elevated serum anti-TSP-1 levels were associated with increased SS disease activity.Further statistical analysis showed that γG% values of anti-TSP-1 positive group were significantly higher than those of the anti-TSP-1 negative group (\u003cem\u003et\u003c/em\u003e\u0026thinsp;=\u0026thinsp;2.554,P\u0026thinsp;=\u0026thinsp;0.014, Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).What\u0026rsquo;s more,the incidence of xerophthalmia significantly increased in the anti-TSP-1 positive group (χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;4.939,P\u0026thinsp;=\u0026thinsp;0.026, Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e),which implicated its pathogenic role in pSS development.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eCorrelations of anti-TSP-1 antibody with clinical parameters of pSS\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eClinical parameters\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eAnti-TSP-1 antibody\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003er\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eP\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAge(years)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e-0.078\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.521\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDisease duration (months)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.046\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.708\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWBC(10\u003csup\u003e9\u003c/sup\u003e/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.087\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.480\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNeutrophils (\u0026times;10\u003csup\u003e9\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.076\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.533\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLymphocyte (\u0026times;10\u003csup\u003e9\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.092\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.451\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRBC(\u0026times;10\u003csup\u003e12\u003c/sup\u003e/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.003\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.979\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHb(g/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e-0.006\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.962\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePLT(10\u003csup\u003e9\u003c/sup\u003e/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e-0.141\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.246\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCRP(mg/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e-0.143\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.292\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eESR(mm/h)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.014\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.917\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eγG(%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.147\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.283\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIgG(g/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e0.263\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e0.029\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIgA(g/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e-0.029\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.818\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIgM(g/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.101\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.414\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC3(g/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e-0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.745\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC4(g/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e-0.253\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e0.035\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRF(IU/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e0.421\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e\u0026lt;0.001\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAnti-α-fodrin(U/ml)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e-0.083\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.537\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eESSDAI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e0.277\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003e0.020\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. pSS: Primary Sj\u0026ouml;gren's syndrome; WBC: white blood cell; N: neutrophil;L:lymphocyte; RBC: erythrocyte;Hb: hemoglobin; PLT: platelet count; CRP: C-reaction protein; ESR: erythrocyte sedimentation rate; RF: rheumatoid factor; γG : γglobulin;IgG: immunoglobulin G; IgA: immunoglobulin A; IgM: immunoglobulin M;C3: complement component 3; C4: complement component 4; ESSDAI: EULAR Sj\u0026ouml;gren's syndrome disease activity index.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eComparison of clinical and laboratory parameters between anti-TSP-1 positive and negative pSS patients.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eClinical and laboratory parameters\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAnti-TSP-1 positive (n\u0026thinsp;=\u0026thinsp;36)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAnti-TSP-1 negative (n\u0026thinsp;=\u0026thinsp;34)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003et/Z/χ\u003csup\u003e2\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eP\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eXerophthalmia, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e29(80.6%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e19(55.9%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e4.939\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e0.026\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eXerostomia, n (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e32(88.9%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e27(79.4%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.186\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.276\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLymphadenectasis, n(%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e7(19.4%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3(8.8%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.611\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.204\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eArthralgia,n(%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e18(50.0%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e15(44.1%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.243\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.622\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLung involvement,n(%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e19(52.8%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e11(32.4%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2.979\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.084\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003erenal lesion,n(%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9(25.0%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9(26.5%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.020\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.888\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eWBC(\u0026times;10\u003csup\u003e9\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.05\u0026thinsp;\u0026plusmn;\u0026thinsp;2.96\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e5.52\u0026thinsp;\u0026plusmn;\u0026thinsp;2.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.771\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.443\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNeutrophils (\u0026times;10\u003csup\u003e9\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.77\u0026thinsp;\u0026plusmn;\u0026thinsp;2.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.42\u0026thinsp;\u0026plusmn;\u0026thinsp;2.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.591\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.557\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHb(g/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e131.00\u0026thinsp;\u0026plusmn;\u0026thinsp;21.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e130.81\u0026thinsp;\u0026plusmn;\u0026thinsp;17.80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.039\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.969\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePLT(\u0026times;10\u003csup\u003e9\u003c/sup\u003e)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e188.36\u0026thinsp;\u0026plusmn;\u0026thinsp;95.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e173.55\u0026thinsp;\u0026plusmn;\u0026thinsp;72.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.321\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.473\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eγG(%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e24.05\u0026thinsp;\u0026plusmn;\u0026thinsp;10.79\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e19.07\u0026thinsp;\u0026plusmn;\u0026thinsp;4.41\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003e2.554\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003e0.014\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIgG(g/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e17.35\u0026thinsp;\u0026plusmn;\u0026thinsp;10.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e15.36\u0026thinsp;\u0026plusmn;\u0026thinsp;3.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.039\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.302\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIgA(g/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3.05\u0026thinsp;\u0026plusmn;\u0026thinsp;1.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.43\u0026thinsp;\u0026plusmn;\u0026thinsp;1.44\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e-1.268\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.209\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIgM(g/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.22\u0026thinsp;\u0026plusmn;\u0026thinsp;0.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e-0.467\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.642\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC3(g/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.92\u0026thinsp;\u0026plusmn;\u0026thinsp;0.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.91\u0026thinsp;\u0026plusmn;\u0026thinsp;0.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.205\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.838\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC4(g/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.19\u0026thinsp;\u0026plusmn;\u0026thinsp;0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.21\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e-1.273\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.207\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCRP(mg/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.01(0.2,56.61)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.35(0.2,200)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e-0.805\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.421\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eESR(mm/h)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e20.62\u0026thinsp;\u0026plusmn;\u0026thinsp;21.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e19.28\u0026thinsp;\u0026plusmn;\u0026thinsp;15.92\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.199\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.794\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRF(IU/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e20(20,721)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e21.3(20,379)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e-1.107\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.268\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eANA positive\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e26(72.2%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e29(85.3%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.775\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAnti-SSA positive\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e21(58.3%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e25(73.5%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.792\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.181\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAnti-SSB possitive\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9(25.0%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e11(32.4%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.463\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.496\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAnti-α-fodrin (U/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e9.41(2.2,128.28)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e17.08(0,65.7)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e-0.579\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.563\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eESSDAI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4.06\u0026thinsp;\u0026plusmn;\u0026thinsp;1.91\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.32\u0026thinsp;\u0026plusmn;\u0026thinsp;1.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.728\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.088\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eTable\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e.pSS: Primary Sj\u0026ouml;gren's syndrome;WBC: white blood cell;N: neutrophil;L:lymphocyte; RBC:erythrocyte;Hb: hemoglobin;PLT:platelet count;CRP:C-reaction protein;ESR:erythrocyte sedimentation rate;RF:rheumatoid factor;γG:γglobulin;IgG:immunoglobulin G;IgA:immunoglobulin A;IgM:immunoglobulin M;C3:complement component 3;C4:complement component 4;ESSDAI:EULAR Sj\u0026ouml;gren's syndrome disease activity index.\u003c/p\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eAccording to the American-European classification criteria in 2016,diagnosis of the pSS is based on the clinical symptoms,salivary gland biopsy and detection of circulating autoantibodies, including anti-Sj\u0026ouml;gren's syndrome A (anti-SSA/Ro) or B (anti-SSB/La)[\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e].Although salivary gland biopsy can be used as the gold standard for the diagnosis of pSS,it is not the first choice of clinical examination for its trauma.Autoantibodies have been shown to be involved in the development of pSS.Thus,it is urgent to screen autoantibodies in pSS.However,some reports showed that anti-SSA antibody might emerge at relatively late stage of the disease.Therefore,these commonly used antibodies are less sensitive in the diagnosis of pSS[\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e].Our study found that TSP-1 autoantibody was significantly higher in serum from pSS patients than in healthy controls.Positivity of anti-TSP-1 antibody in pSS and HCs were respectively 51.4% and 28.6%.Therefore,TSP-1 autoantibodiesare also involve in pathogenesis of pSS.\u003c/p\u003e \u003cp\u003eTSP-1 was first discovered in platelets[\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].It is a matricellular protein and has multiple functions,including the modulation of NO signals[\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e],and immune cell recruitment and activation[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].TSP1 is widely upregulated in tissue injuries to accelerate tissue reparation[\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e].The pathological characteristics of pSS are lymphocytic infiltrate of the exocrine glands.In addition to lacrimal gland and salivary gland duct epithelial cells,it also includes alveolar epithelial cells,renal tubular epithelial cells and liver fibroblasts,which lead to multiple system dysfunction and destruction[\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e].In recent years, accumulating evidence suggests that the TSP-1 plays an important role in gland function and homeostasis[\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e].TSP-1 deficient antigen-presenting cells(APC) promotes Th17 cells mediated inflammatory response in the periphery and gland of SS murine models[\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e].In our study,the levels of anti-TSP-1 antibody in pSS patients were significantly associated with inflammation and disease severity parameters,such as hypocomplementemia, immunoglobulin G and RF. The level of γG in anti-TSP-1-positive pSS patients is significantly higher than that in anti-TSP-1-negitive pSS patients (P\u0026lt;0.05,\u003cb\u003eSupplementary Fig.\u0026nbsp;2 and\u003c/b\u003e Tables\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and \u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).Our study further validated that the ESSDAI score was significantly correlated with the anti-TSP-1 serum concentration.These results suggest that anti-TSP-1 is might promote the development of pSS.\u003c/p\u003e \u003cp\u003eOur study did not find a significant association of anti-TSP-1 antibody with other autoantibodies in pSS patients.Thus,anti-TSP-1 antibody is independent.But as an autoantigen,TSP-1 is localized in various cell types,including those in ocular tissues and exocrine glands[\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e].Interestingly,in our study,the group of anti-TSP-1-positive pSS patients had significantly higher prevalence of xerophthalmia.Bruce Turpie et al.[\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e] reported that mice deficient of TSP-1 eventually developed a chronic ocular surface disease.The pathological process is driven by apoptosis and deterioration of lacrimal gland cells,resulting in dysfunction and inflammatory infiltration,and abnormal tears.We hypothesized that TSP-1 antibody might be associated with SS ocular surface inflammation. During the occurrence and development of pSS,TSP-1 and other autoantigens might trigger and stimulate the abnormal activation of autoimmune T cell and B cell to produce more autoantibodies,which in turn damage the gland tissues and other organs[\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eA cohort research of 921 Spanish patients with pSS showed that the most frequently involved organ was the lung[\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e],and a proportion of patients might develop pulmonary interstitial fibrosis in the later stage of disease.As a major activator of TGF-β,TSP-1 plays an important role in fibrotic diseases.Joanne E et al.[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e] found that antagonists of the TSP-1/TGF-β pathway might be suitable drug candidates for fibrotic diseases,including diabetic nephropathy,liver fibrosis,pulmonary fibrosis and scleroderma[\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e].Besides,our study also found that the group of anti-TSP-1-positive pSS patients had increased trend of prevalence of lung involvement.\u003c/p\u003e \u003cp\u003eHowever,the number of serum samples we collected is limited.Therefore,our future experiments need to expand the sample size to explore the role of TSP-1 autoantibodies in pSS.In addition,the relationship between TSP-1 antibody and disease activity and clinical phenotype should be verified again according to the severity of the disease.\u003c/p\u003e \u003cp\u003eIn conclusion, this study revealed the clinical relevance of TSP-1 autoantibody in pSS.It showed that TSP-1 autoantibody might be an effective tool in screening and evaluating disease activity in pSS patients.However, the pathogenic mechanism of TSP-1 autoantibodies in pSS demands further study.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003eThis work was supported by the National Natural Science Foundation of China (81671602, 81801619); and the Key Technology Research and Development Program of Henan Province Department of Education (19A320016).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability\u0026nbsp;\u003c/strong\u003eThe data used to support the findings of the current study are available from the corresponding authors upon reasonable request.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of Interest\u0026nbsp;\u003c/strong\u003eThe authors declare that they have no conflict of interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026rsquo; Contributions\u0026nbsp;\u003c/strong\u003eRuoxi Li performed most of the experiments and drafted the manuscript.Ruoxi Li,Yuanhao Li,Yixue Guo, Yaodong Zhao and Yang Meng worked on data collection.Xuewu Zhang,Wang Yan and Xiaolin Sun conceived the study and participated in its design and coordination and interpreted the data.All authors read and approved the final manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eBaenziger NL, Brodie GN, Majerus PW. Isolation and properties of a thrombin-sensitive protein of human platelets. Biol Chem. 1972;247(9):2723\u0026ndash;31.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLawler JW, Slayter HS, Coligan JE. Isolation and characterization of a high molecular weight glycoprotein from human blood platelets. Biol Chem. 1978;253(23):8609\u0026ndash;16.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWight TN, Raugi GJ, Mumby SM, Bornstein P. Light microscopic immunolocation of thrombospondin in human tissues. Histochem Cytochem. 1985;33(4):295\u0026ndash;302.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNaganuma H, Satoh E, Asahara T, et al. Quantification of thrombospondin-1 secretion and expression of alphavbeta3 and alpha3beta1 integrins and syndecan-1 as cell-surface receptors for thrombospondin-1 in malignant glioma cells. Neurooncol. 2004;70(3):309\u0026ndash;17.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAdams JC, Lawler J. The thrombospondins. Cold Spring Harb Perspect Biol. 2011;3(10):a009712.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGrenier C, Caillon A, Munier M, et al. Dual Role of Thrombospondin-1 in Flow-Induced Remodeling. Int J Mol Sci. 2021;22(21):12086. Published 2021 Nov 8.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMurphy-Ullrich JE, Suto MJ. Thrombospondin-1 regulation of latent TGF-β activation: A therapeutic target for fibrotic disease. Matrix Biol. 2018;68\u0026ndash;69:28\u0026ndash;43.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMasli S, Dartt DA. Mouse Models of Sj\u0026ouml;gren's Syndrome with Ocular Surface Disease. Int J Mol Sci. 2020;21(23):9112.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVitali C, Bombardieri S, Jonsson R, et al. Classification criteria for Sj\u0026ouml;gren's syndrome: a revised version of the European criteria proposed by the American-European Consensus Group. Ann Rheum Dis. 2002;61(6):554\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eShiboski CH, Shiboski SC, Seror R, et al. 2016 American College of Rheumatology/European League Against Rheumatism Classification Criteria for Primary Sj\u0026ouml;gren's Syndrome: A Consensus and Data-Driven Methodology Involving Three International Patient Cohorts. Arthritis Rheumatol. 2017;69(1):35\u0026ndash;45.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSeror R, Ravaud P, Bowman SJ, et al. EULAR Sjogren's syndrome disease activity index: development of a consensus systemic disease activity index for primary Sjogren's syndrome. Ann Rheum Dis. 2010;69(6):1103\u0026ndash;9.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHu Q, Wang D, Chen W. The accuracy of the anti-α-fodrin antibody test for diagnosis of Sj\u0026ouml;gren's syndrome: a meta-analysis. Clin Biochem. 2013;46(15):1372\u0026ndash;6.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJin Y, Li J, Chen J, et al. Tissue-Specific Autoantibodies Improve Diagnosis of Primary Sj\u0026ouml;gren's Syndrome in the Early Stage and Indicate Localized Salivary Injury. Immunol Res. 2019;2019:3642937. Published 2019 May 7.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIsenberg JS, Martin-Manso G, Maxhimer JB, Roberts DD. Regulation of nitric oxide signalling by thrombospondin 1: implications for anti-angiogenic therapies. Nat Rev Cancer. 2009;9(3):182\u0026ndash;94.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMurphy-Ullrich JE, Sage EH. Revisiting the matricellular concept. Matrix Biol. 2014;37:1\u0026ndash;14.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eThorne I, Sutcliffe N. Sj\u0026ouml;gren's syndrome. Br J Hosp Med (Lond). 2017;78(8):438\u0026ndash;42.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTurpie B, Yoshimura T, Gulati A, Rios JD, Dartt DA, Masli S. Sj\u0026ouml;gren's syndrome-like ocular surface disease in thrombospondin-1 deficient mice. Am J Pathol. 2009;175(3):1136\u0026ndash;47.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eR\u0026iacute;os JD, Horikawa Y, Chen LL, et al. Age-dependent alterations in mouse exorbital lacrimal gland structure, innervation and secretory response. Exp Eye Res. 2005;80(4):477\u0026ndash;91.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRashid S, Jin Y, Ecoiffier T, Barabino S, Schaumberg DA, Dana MR. Topical omega-3 and omega-6 fatty acids for treatment of dry eye. Arch Ophthalmol. 2008;126(2):219\u0026ndash;25.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMasli S, Sheibani N, Cursiefen C, Zieske J. Matricellular protein thrombospondins: influence on ocular angiogenesis, wound healing and immuneregulation. Curr Eye Res. 2014;39(8):759\u0026ndash;74.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGromova A, Voronov DA, Yoshida M, et al. Lacrimal Gland Repair Using Progenitor Cells. Stem Cells Transl Med. 2017;6(1):88\u0026ndash;98.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eJin Y, Li J, Chen J, et al. Tissue-Specific Autoantibodies Improve Diagnosis of Primary Sj\u0026ouml;gren's Syndrome in the Early Stage and Indicate Localized Salivary Injury. Immunol Res. 2019;2019:3642937. Published 2019 May 7.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRamos-Casals M, Brito-Zer\u0026oacute;n P, Solans R, et al. Systemic involvement in primary Sjogren's syndrome evaluated by the EULAR-SS disease activity index: analysis of 921 Spanish patients (GEAS-SS Registry). Rheumatology (Oxford). 2014;53(2):321\u0026ndash;31.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChang Y, Lau WL, Jo H, et al. Pharmacologic Blockade of αvβ1 Integrin Ameliorates Renal Failure and Fibrosis In Vivo. Am Soc Nephrol. 2017;28(7):1998\u0026ndash;2005.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMunger JS, Huang X, Kawakatsu H, et al. The integrin alpha v beta 6 binds and activates latent TGF beta 1: a mechanism for regulating pulmonary inflammation and fibrosis. Cell. 1999;96(3):319\u0026ndash;28.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"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":"advances-in-rheumatology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"adrh","sideBox":"Learn more about [Advances in Rheumatology](https://advancesinrheumatology.biomedcentral.com/)","snPcode":"42358","submissionUrl":"https://submission.springernature.com/new-submission/42358/3","title":"Advances in Rheumatology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Anti- thrombospodin-1 antibody, Primary Sjögren's syndrome","lastPublishedDoi":"10.21203/rs.3.rs-8660592/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8660592/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eObjective. \u003c/strong\u003eThe relationship between anti- thrombospondin-1 antibody (anti-TSP-1) and the clinical manifestations of Primary Sjögren's syndrome（pSS) has not been fully clarified.We analyzed the clinical factors relevance of anti-TSP-1 in pSS.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods.\u003c/strong\u003e Serum ATSA was detected by ELISA in 70 pSS patients and 21 healthy controls enrolled.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults. \u003c/strong\u003eAnti-TSP-1 antibody is significantly higher in pSS patients (10.63± 4.92 \u003cem\u003evs.\u003c/em\u003e7.64 ± 4.70, \u003cem\u003et\u003c/em\u003e=2.403,\u003cem\u003eP\u003c/em\u003e=0.019). Positivity of anti-TSP-1 antibody in pSS and HCs were respectively 51.4% (36/70) and 28.6%(6/21). The increase of anti-TSP-1 antibody was also significantly correlated with the increase of IgG (\u003cem\u003er\u003c/em\u003e= 0.263, \u003cem\u003eP\u003c/em\u003e=0.029) and RF (\u003cem\u003er\u003c/em\u003e=0.421,\u003cem\u003eP\u003c/em\u003e＜0.001), and the decrease of C4 (\u003cem\u003er\u003c/em\u003e=-0.253,\u003cem\u003eP\u003c/em\u003e=0.035)in pSS patients. The increase of serum anti-TSP-1 antibody was associated with the increase of ESSDAI in pSS patients (\u003cem\u003er\u003c/em\u003e=0.277, \u003cem\u003eP\u003c/em\u003e=0.02). Further statistical analysis showed that γG% values of anti-TSP-1 positive group were significantly higher than those of anti-TSP-1 negative group (\u003cem\u003et\u003c/em\u003e=2.554,\u003cem\u003eP\u003c/em\u003e=0.014). γG% values of anti-TSP-1 positive group were significantly higher than those of anti-TSP-1 negative group (\u003cem\u003et\u003c/em\u003e=2.554,\u003cem\u003eP\u003c/em\u003e=0.014).What’s more,the incidence of xerophthalmia significantly increased in the anti-TSP-1 positive group (\u003cem\u003ec\u003c/em\u003e\u003csup\u003e2\u003c/sup\u003e=4.939,\u003cem\u003eP\u003c/em\u003e=0.026).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion. \u003c/strong\u003eAnti-TSP-1antibody levels were associated with increased SS disease activity,and it may be an effective tool in screening\u0026nbsp; and predicting prognosis in pSS patients\u003c/p\u003e","manuscriptTitle":"Anti-thrombospondin-1 autoantibody indicated increased disease activity and xerophthalmia incidence in Primary Sjögren's syndrome","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-02 05:28:54","doi":"10.21203/rs.3.rs-8660592/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2026-05-15T01:59:18+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-05-04T09:53:35+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"226685239551880896900411504428242177559","date":"2026-04-26T12:42:01+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-02-15T11:41:58+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"85639014894645415806508238513205523783","date":"2026-02-02T09:53:40+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2026-01-28T13:54:09+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2026-01-27T08:35:57+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2026-01-27T08:32:56+00:00","index":"","fulltext":""},{"type":"submitted","content":"Advances in Rheumatology","date":"2026-01-21T13:17:15+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"advances-in-rheumatology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"adrh","sideBox":"Learn more about [Advances in Rheumatology](https://advancesinrheumatology.biomedcentral.com/)","snPcode":"42358","submissionUrl":"https://submission.springernature.com/new-submission/42358/3","title":"Advances in Rheumatology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"e69f5585-b427-40cb-9b06-8544254ade93","owner":[],"postedDate":"February 2nd, 2026","published":true,"recentEditorialEvents":[{"type":"decision","content":"Revision requested","date":"2026-05-15T01:59:18+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2026-05-04T09:53:35+00:00","index":42,"fulltext":""}],"rejectedJournal":[],"revision":"","amendment":"","status":"in-revision","subjectAreas":[],"tags":[],"updatedAt":"2026-05-15T02:09:25+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-02 05:28:54","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8660592","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8660592","identity":"rs-8660592","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}