CD4+ T cells predominate in salivary gland (SG) inflammatory lesions in Sjögren's syndrome (SS). However, their antigen specificity, degree of clonal expansion, and relationship to clinical disease features remain unknown. We used multiplex reverse-transcriptase PCR to amplify paired T cell receptor α (TCRα) and β transcripts of single CD4+CD45RA- T cells from SG and peripheral blood (PB) of 10 individuals with primary SS, 9 of whom shared the HLA DR3/DQ2 risk haplotype. TCRα and β sequences were obtained from a median of 91 SG and 107 PB cells per subject. The degree of clonal expansion and frequency of cells expressing two productively rearranged α genes were increased in SG versus PB. Expanded clones from SG exhibited complementary-determining region 3 (CDR3) sequence similarity both within and among subjects, suggesting antigenic selection and shared antigen recognition. CDR3 similarities were shared among expanded clones from individuals discordant for canonical Ro and La autoantibodies, suggesting recognition of alternative SG antigen(s). The extent of SG clonal expansion correlated with reduced saliva production and increased SG fibrosis, linking expanded SG T cells with glandular dysfunction. Knowledge of paired TCRα and β sequences enables further work toward identification of target antigens and development of novel therapies.

Arthritis and Clinical Immunology Program Oklahoma Medical Research Foundation Oklahoma City Oklahoma USA

Arthritis and Clinical Immunology Program Oklahoma Medical Research Foundation Oklahoma City Oklahoma USA; Section of Endocrinology and Diabetes College of Medicine OUHSC Oklahoma City Oklahoma USA

Department of Ophthalmology College of Medicine OUHSC Oklahoma City Oklahoma USA

Department of Oral and Maxillofacial Pathology College of Dentistry OUHSC Oklahoma City Oklahoma USA

Department of Oral Diagnosis and Radiology College of Dentistry OUHSC Oklahoma City Oklahoma USA

Shemyakin Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences Moscow Russia; Pirogov Russian National Research Medical University Moscow Russia; Central European Institute of Technology Masaryk University Brno Czech Republic

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Jonsson R, Vogelsang P, Volchenkov R, Espinosa A, Wahren-Herlenius M, Appel S. The complexity of Sjogren’s syndrome: novel aspects on pathogenesis. Immunol Lett. 2011;141(1):1–9. doi: 10.1016/j.imlet.2011.06.007. PubMed DOI

Mavragani CP, Moutsopoulos HM. Sjogren’s syndrome. Annu Rev Pathol. 2014;9:273–285. doi: 10.1146/annurev-pathol-012513-104728. PubMed DOI

Vitali C, et al. Classification criteria for Sjogren’s syndrome: a revised version of the European criteria proposed by the American-European Consensus Group. Ann Rheum Dis. 2002;61(6):554–558. doi: 10.1136/ard.61.6.554. PubMed DOI PMC

Daniels TE. Labial salivary gland biopsy in Sjogren’s syndrome. Assessment as a diagnostic criterion in 362 suspected cases. Arthritis Rheum. 1984;27(2):147–156. doi: 10.1002/art.1780270205. PubMed DOI

Fox RI, Carstens SA, Fong S, Robinson CA, Howell F, Vaughan JH. Use of monoclonal antibodies to analyze peripheral blood and salivary gland lymphocyte subsets in Sjogren’s syndrome. Arthritis Rheum. 1982;25(4):419–426. doi: 10.1002/art.1780250410. PubMed DOI

Adamson TC, 3rd, Fox RI, Frisman DM, Howell FV. Immunohistologic analysis of lymphoid infiltrates in primary Sjogren’s syndrome using monoclonal antibodies. J Immunol. 1983;130(1):203–208. PubMed

Talal N, Sylvester RA, Daniels TE, Greenspan JS, Williams RC., Jr T and B lymphocytes in peripheral blood and tissue lesions in Sjogren’s syndrome. J Clin Invest. 1974;53(1):180–189. doi: 10.1172/JCI107536. PubMed DOI PMC

Moutsopoulos HM, Hooks JJ, Chan CC, Dalavanga YA, Skopouli FN, Detrick B. HLA-DR expression by labial minor salivary gland tissues in Sjogren’s syndrome. Ann Rheum Dis. 1986;45(8):677–683. doi: 10.1136/ard.45.8.677. PubMed DOI PMC

Zumla A, Mathur M, Stewart J, Wilkinson L, Isenberg D. T cell receptor expression in Sjogren’s syndrome. Ann Rheum Dis. 1991;50(10):691–693. doi: 10.1136/ard.50.10.691. PubMed DOI PMC

Ohyama Y, et al. T-cell receptor Vα and Vβ gene use by infiltrating T cells in labial glands of patients with Sjogren’s syndrome. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 1995;79(6):730–737. doi: 10.1016/S1079-2104(05)80308-7. PubMed DOI

Matthews JB, et al. Primed and naive helper T cells in labial glands from patients with Sjogren’s syndrome. Virchows Arch A Pathol Anat Histopathol. 1991;419(3):191–197. doi: 10.1007/BF01626347. PubMed DOI

Christodoulou MI, Kapsogeorgou EK, Moutsopoulos HM. Characteristics of the minor salivary gland infiltrates in Sjogren’s syndrome. J Autoimmun. 2010;34(4):400–407. doi: 10.1016/j.jaut.2009.10.004. PubMed DOI

Petrie HT, Livak F, Schatz DG, Strasser A, Crispe IN, Shortman K. Multiple rearrangements in T cell receptor alpha chain genes maximize the production of useful thymocytes. J Exp Med. 1993;178(2):615–622. doi: 10.1084/jem.178.2.615. PubMed DOI PMC

Padovan E, Casorati G, Dellabona P, Meyer S, Brockhaus M, Lanzavecchia A. Expression of two T cell receptor alpha chains: dual receptor T cells. Science. 1993;262(5132):422–424. doi: 10.1126/science.8211163. PubMed DOI

Smith MD, et al. Selective expression of V beta families by T cells in the blood and salivary gland infiltrate of patients with primary Sjogren’s syndrome. J Rheumatol. 1994;21(10):1832–1837. PubMed

Kay RA, et al. An abnormal T cell repertoire in hypergammaglobulinaemic primary Sjogren’s syndrome. Clin Exp Immunol. 1991;85(2):262–264. PubMed PMC

Matsumoto I, et al. Common T cell receptor clonotype in lacrimal glands and labial salivary glands from patients with Sjogren’s syndrome. J Clin Invest. 1996;97(8):1969–1977. doi: 10.1172/JCI118629. PubMed DOI PMC

Sasaki M, et al. Accumulation of common T cell clonotypes in the salivary glands of patients with human T lymphotropic virus type I-associated and idiopathic Sjogren’s syndrome. J Immunol. 2000;164(5):2823–2831. doi: 10.4049/jimmunol.164.5.2823. PubMed DOI

Sumida T, et al. T cell receptor repertoire of infiltrating T cells in lips of Sjogren’s syndrome patients. J Clin Invest. 1992;89(2):681–685. doi: 10.1172/JCI115635. PubMed DOI PMC

Ajjan RA, et al. Analysis of the T-cell receptor Valpha repertoire and cytokine gene expression in Sjogren’s syndrome. Br J Rheumatol. 1998;37(2):179–185. doi: 10.1093/rheumatology/37.2.179. PubMed DOI

Sumida T, et al. TCR in Fas-sensitive T cells from labial salivary glands of patients with Sjogren’s syndrome. J Immunol. 1997;158(2):1020–1025. PubMed

Matsumoto I, et al. Single cell analysis of T cells infiltrating labial salivary glands from patients with Sjogren’s syndrome. Int J Mol Med. 1999;4(5):519–527. PubMed

Sumida T, Kita Y, Yonaha F, Maeda T, Iwamoto I, Yoshida S. T cell receptor Vα repertoire of infiltrating T cells in labial salivary glands from patients with Sjogren’s syndrome. J Rheumatol. 1994;21(9):1655–1661. PubMed

Krischer JP, et al. Screening strategies for the identification of multiple antibody-positive relatives of individuals with type 1 diabetes. J Clin Endocrinol Metab. 2003;88(1):103–108. doi: 10.1210/jc.2002-020760. PubMed DOI

Weiner ES, et al. Prognostic significance of anticentromere antibodies and anti-topoisomerase I antibodies in Raynaud’s disease. A prospective study. Arthritis Rheum. 1991;34(1):68–77. doi: 10.1002/art.1780340111. PubMed DOI

van Venrooij WJ, van Beers JJ, Pruijn GJ. Anti-CCP antibodies: the past, the present and the future. Nat Rev Rheumatol. 2011;7(7):391–398. doi: 10.1038/nrrheum.2011.76. PubMed DOI

Michels AW, von Herrath M. 2011 Update: antigen-specific therapy in type 1 diabetes. Curr Opin Endocrinol Diabetes Obes. 2011;18(4):235–240. doi: 10.1097/MED.0b013e32834803ae. PubMed DOI PMC

Steinman L. Inverse vaccination, the opposite of Jenner’s concept, for therapy of autoimmunity. J Intern Med. 2010;267(5):441–451. doi: 10.1111/j.1365-2796.2010.02224.x. PubMed DOI

Walczak A, Siger M, Ciach A, Szczepanik M, Selmaj K. Transdermal application of myelin peptides in multiple sclerosis treatment. JAMA Neurol. 2013;70(9):1105–1109. doi: 10.1001/jamaneurol.2013.3022. PubMed DOI

Turner SJ, Doherty PC, McCluskey J, Rossjohn J. Structural determinants of T-cell receptor bias in immunity. Nat Rev. 2006;6(12):883–894. PubMed

Lessard CJ, et al. Variants at multiple loci implicated in both innate and adaptive immune responses are associated with Sjogren’s syndrome. Nat Genet. 2013;45(11):1284–1292. doi: 10.1038/ng.2792. PubMed DOI PMC

Zal T, Weiss S, Mellor A, Stockinger B. Expression of a second receptor rescues self-specific T cells from thymic deletion and allows activation of autoreactive effector function. Proc Natl Acad Sci U S A. 1996;93(17):9102–9107. doi: 10.1073/pnas.93.17.9102. PubMed DOI PMC

Sarukhan A, Garcia C, Lanoue A, von Boehmer H. Allelic inclusion of T cell receptor alpha genes poses an autoimmune hazard due to low-level expression of autospecific receptors. Immunity. 1998;8(5):563–570. doi: 10.1016/S1074-7613(00)80561-0. PubMed DOI

Ni PP, Solomon B, Hsieh CS, Allen PM, Morris GP. The ability to rearrange dual TCRs enhances positive selection, leading to increased Allo- and Autoreactive T cell repertoires. J Immunol. 2014;193(4):1778–1786. doi: 10.4049/jimmunol.1400532. PubMed DOI PMC

Morris GP, Uy GL, Donermeyer D, Dipersio JF, Allen PM. Dual receptor T cells mediate pathologic alloreactivity in patients with acute graft-versus-host disease. Sci Transl Med. 2013;5(188):e85609. PubMed PMC

Robins HS, et al. Comprehensive assessment of T-cell receptor beta-chain diversity in alphabeta T cells. Blood. 2009;114(19):4099–4107. doi: 10.1182/blood-2009-04-217604. PubMed DOI PMC

Venturi V, Kedzierska K, Turner SJ, Doherty PC, Davenport MP. Methods for comparing the diversity of samples of the T cell receptor repertoire. J Immunol Methods. 2007;321(1-2):182–195. doi: 10.1016/j.jim.2007.01.019. PubMed DOI

Spreafico R, et al. A circulating reservoir of pathogenic-like CD4+ T cells shares a genetic phenotypic signature with the inflamed synovial micro-environment. Ann Rheum Dis. 2016;75(2):459–465. doi: 10.1136/annrheumdis-2014-206226. PubMed DOI PMC

Venturi V, et al. Sharing of T cell receptors in antigen-specific responses is driven by convergent recombination. Proc Natl Acad Sci U S A. 2006;103(49):18691–18696. doi: 10.1073/pnas.0608907103. PubMed DOI PMC

Britanova OV, et al. Age-related decrease in TCR repertoire diversity measured with deep and normalized sequence profiling. J Immunol. 2014;192(6):2689–2698. doi: 10.4049/jimmunol.1302064. PubMed DOI

Egorov ES, et al. Quantitative profiling of immune repertoires for minor lymphocyte counts using unique molecular identifiers. J Immunol. 2015;194(12):6155–6163. doi: 10.4049/jimmunol.1500215. PubMed DOI

Shugay M, et al. VDJtools: unifying post-analysis of T cell receptor repertoires. PLoS Comput Biol. 2015;11(11):e85609. doi: 10.1371/journal.pcbi.1004503. PubMed DOI PMC

Leehan KM, et al. Precisely quantified fibrosis in labial salivary glands predicts Sjögren’s syndrome classification in a multiple regression model. Arthritis Rheumatol. 2014;66:e85609

Ji Q, Perchellet A, Goverman JM. Viral infection triggers central nervous system autoimmunity via activation of CD8+ T cells expressing dual TCRs. Nat Immunol. 2010;11(7):628–634. doi: 10.1038/ni.1888. PubMed DOI PMC

He X, et al. Dual receptor T cells extend the immune repertoire for foreign antigens. Nat Immunol. 2002;3(2):127–134. doi: 10.1038/ni751. PubMed DOI

Sumida T, Matsumoto I, Maeda T, Nishioka K. T-cell receptor in Sjogren’s syndrome. Br J Rheumatol. 1997;36(6):622–629. doi: 10.1093/rheumatology/36.6.622. PubMed DOI

Ishigaki K, et al. Quantitative and qualitative characterization of expanded CD4+ T cell clones in rheumatoid arthritis patients. Sci Rep. 2015;5:e85609. PubMed PMC

Jonsson MV, Skarstein K, Jonsson R, Brun JG. Serological implications of germinal center-like structures in primary Sjogren’s syndrome. J Rheumatol. 2007;34(10):2044–2049. PubMed

Maier-Moore JS, et al. Antibody-secreting cell specificity in labial salivary glands reflects the clinical presentation and serology in patients with Sjogren’s syndrome. Arthritis Rheumatol. 2014;66(12):3445–3456. doi: 10.1002/art.38872. PubMed DOI PMC

Tengner P, Halse AK, Haga HJ, Jonsson R, Wahren-Herlenius M. Detection of anti-Ro/SSA and anti-La/SSB autoantibody-producing cells in salivary glands from patients with Sjogren’s syndrome. Arthritis Rheum. 1998;41(12):2238–2248. PubMed

Theander E, et al. Lymphoid organisation in labial salivary gland biopsies is a possible predictor for the development of malignant lymphoma in primary Sjogren’s syndrome. Ann Rheum Dis. 2011;70(8):1363–1368. doi: 10.1136/ard.2010.144782. PubMed DOI PMC

Vissink A, Bootsma H, Spijkervet FK, Hu S, Wong DT, Kallenberg CG. Current and future challenges in primary Sjogren’s syndrome. Curr Pharm Biotechnol. 2012;13(10):2026–2045. doi: 10.2174/138920112802273254. PubMed DOI

Daniels TE, et al. Associations between salivary gland histopathologic diagnoses and phenotypic features of Sjogren’s syndrome among 1,726 registry participants. Arthritis Rheum. 2011;63(7):2021–2030. doi: 10.1002/art.30381. PubMed DOI PMC

Malladi AS, et al. Primary Sjogren’s syndrome as a systemic disease: a study of participants enrolled in an international Sjogren’s syndrome registry. Arthritis Care Res (Hoboken) 2012;64(6):911–918. doi: 10.1002/acr.21610. PubMed DOI PMC

Jensen SB, Vissink A. Salivary gland dysfunction and xerostomia in Sjogren’s syndrome. Oral Maxillofac Surg Clin North Am. 2014;26(1):35–53. doi: 10.1016/j.coms.2013.09.003. PubMed DOI

Bookman AA, et al. Whole stimulated salivary flow: correlation with the pathology of inflammation and damage in minor salivary gland biopsy specimens from patients with primary Sjogren’s syndrome but not patients with sicca. Arthritis Rheum. 2011;63(7):2014–2020. doi: 10.1002/art.30295. PubMed DOI

Blank U, Boitel B, Mege D, Ermonval M, Acuto O. Analysis of tetanus toxin peptide/DR recognition by human T cell receptors reconstituted into a murine T cell hybridoma. Eur J Immunol. 1993;23(12):3057–3065. doi: 10.1002/eji.1830231203. PubMed DOI

Delong T, et al. Pathogenic CD4 T cells in type 1 diabetes recognize epitopes formed by peptide fusion. Science. 2016;351(6274):711–714. doi: 10.1126/science.aad2791. PubMed DOI PMC

Shiboski SC, et al. American College of Rheumatology classification criteria for Sjogren’s syndrome: a data-driven, expert consensus approach in the Sjogren’s International Collaborative Clinical Alliance cohort. Arthritis Care Res (Hoboken) 2012;64(4):475–487. doi: 10.1002/acr.21591. PubMed DOI PMC

Rasmussen A, et al. Comparison of the American-European Consensus Group Sjogren’s syndrome classification criteria to newly proposed American College of Rheumatology criteria in a large, carefully characterised sicca cohort. Ann Rheum Dis. 2014;73(1):31–38. doi: 10.1136/annrheumdis-2013-203845. PubMed DOI PMC

Bunce M, et al. Phototyping: comprehensive DNA typing for HLA-A, B, C, DRB1, DRB3, DRB4, DRB5 & DQB1 by PCR with 144 primer mixes utilizing sequence-specific primers (PCR-SSP) Tissue Antigens. 1995;46(5):355–367. doi: 10.1111/j.1399-0039.1995.tb03127.x. PubMed DOI

Zheng X, et al. HIBAG--HLA genotype imputation with attribute bagging. Pharmacogenomics J. 2014;14(2):192–200. doi: 10.1038/tpj.2013.18. PubMed DOI PMC

Wang GC, Dash P, McCullers JA, Doherty PC, Thomas PG. T cell receptor αβ diversity inversely correlates with pathogen-specific antibody levels in human cytomegalovirus infection. Sci Transl Med. 2012;4(128):e85609. PubMed PMC

Lefranc MP, et al. IMGT, the international ImMunoGeneTics information system. Nucleic Acids Res. 2009;37(Database issue):D1006–D1012. PubMed PMC

Brochet X, Lefranc MP, Giudicelli V. IMGT/V-QUEST: the highly customized and integrated system for IG and TR standardized V-J and V-D-J sequence analysis. Nucleic Acids Res. 2008;36(Web Server issue):W503–W508. PubMed PMC

Caporaso JG, et al. QIIME allows analysis of high-throughput community sequencing data. Nat Methods. 2010;7(5):335–336. doi: 10.1038/nmeth.f.303. PubMed DOI PMC

Edgar RC. Search and clustering orders of magnitude faster than BLAST. Bioinformatics. 2010;26(19):2460–2461. doi: 10.1093/bioinformatics/btq461. PubMed DOI

Camacho C, et al. BLAST+: architecture and applications. BMC Bioinformatics. 2009;10:e85609. PubMed PMC

Stothard P. The sequence manipulation suite: JavaScript programs for analyzing and formatting protein and DNA sequences. Biotechniques. 2000;28(6):e85609. PubMed

Kivioja T, et al. Counting absolute numbers of molecules using unique molecular identifiers. Nat Methods. 2012;9(1):72–74. PubMed