Abstract
To investigate T cell involvement in antigen‐specific immune responses, it is important to detect accumulating T cells at a clonal level in vivo. However, thus far the clinical application of such analyses has been limited. Here we have established novel primers to anneal with T cell receptor (TCR) β genes of multiple Vβ families and applied them to reverse transcription‐polymerase chain reaction‐single strand conformation polymorphism (RT‐PCR‐SSCP) analysis to evaluate peripheral T cell clonality of autoimmune disease patients. As a result, the new Vβ primers could detect accumulating T cell clones in the periphery of healthy individuals and patients. It was revealed that patients with autoimmune diseases such as systemic lupus erythematosus (SLE) had a larger number of clonal accumulations of peripheral T cells compared with normal individuals. Thus, the RT‐PCR‐SSCP system using the new multifamily Vβ primers is the first such laboratory examination to detect T cell clonal expansion, and will provide a simple and sensitive tool to aid in the diagnosis and also in the investigation of the pathogenesis of autoimmune diseases. J. Clin. Lab. Anal. 12:162–167, 1998. © 1998 Wiley‐Liss, Inc.
Keywords: T cell clonality, T cell receptor, RT‐PCR‐SSCP, autoimmunity
References
- 1. Yamamoto K, Sakoda H, Nakajima T, et al.: Accumulation of multiple T cell clonotypes in the synovial lesions of patients with rheumatoid arthritis revealed by a novel clonality analysis. Int Immunol 4: 1219–1223, 1992. [DOI] [PubMed] [Google Scholar]
- 2. Masuko K, Kato T, Ikeda Y, et al.: Dynamic changes of accumulated T cell clonotypes during antigenic stimulation in vivo and in vitro. Int Immunol 6: 1959–1966, 1994. [DOI] [PubMed] [Google Scholar]
- 3. Masuko K, Kato S, Hagihara M, et al.: Stable clonal expansion of T cells induced by bone marrow transplantation. Blood 87: 789–799, 1996. [PubMed] [Google Scholar]
- 4. Ikeda Y, Masuko K, Nakai Y, et al.: High frequencies of identical T cell clonotypes in synovial tissues of rheumatoid arthritis patients suggest the occurrence of common antigen‐driven immune response. Arthritis Rheum 39: 446–453, 1996. [DOI] [PubMed] [Google Scholar]
- 5. Höger TA, Tokuyama M, Yonamine K, et al.: Time course analysis of α+β+ T cell clones during normal pregnancy. Eur J Immunol 26: 834–838, 1996. [DOI] [PubMed] [Google Scholar]
- 6. Kato T, Ikeda Y, Zong ZP, et al.: Characterization of T cell receptor beta chains of accumulating T cells in skin allografts in mice. Transplantation 62: 266–272, 1996. [DOI] [PubMed] [Google Scholar]
- 7. Komagata Y, Masuko K, Tashiro F, et al.: Clonal prevalence of T cells infiltrating into the pancreas of prediabetic NOD mice. Int Immunol 8: 807–814, 1996. [DOI] [PubMed] [Google Scholar]
- 8. Mitsuhashi M, Hosokawa T: Novel computerized method for designing nucleotide sequence used for DNA probes and PCR primers. Nippon Rinsho 52: 246–257, 1994. [PubMed] [Google Scholar]
- 9. Choi Y, Kotzin B, Herron L, Callahan J, Marrack P, Kappler J: Interaction of staphylococcus aureus toxin “superantigens” with human T cells. Proc Natl Acad Sci USA 86: 8941–8945, 1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 10. Tan EM, Cohen AS, Fries JF, et al.: The 1982 revised criteria for the classification of systemic lupus erythematosus. Arthritis Rheum 25: 1271–1277, 1982. [DOI] [PubMed] [Google Scholar]
- 11. Bohan A, Peter JB, Bowman RL, Pearson CM: A computer‐assisted analysis of 153 patients with polymyositis and dermatomyositis. Medicine 56: 255–286, 1977. [DOI] [PubMed] [Google Scholar]
- 12. Sharp GC, Irvin WS, Tan EM, Gould RG, Holman HR: Mixed connective tissue disease ‐ an apparently distinct rheumatic disease syndrome associated with a spcific antibody to an extractable nuclear antigen (ENA). Am J Med 52: 148–159, 1972. [DOI] [PubMed] [Google Scholar]
- 13. Arnett FC, Edworthy SM, Bloch DA, et al.: The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum 31: 315–324, 1988. [DOI] [PubMed] [Google Scholar]
- 14. Subcommittee for scleroderma criteria of the American Rheumatism Association diagnostic and therapeutic criteria committee: Preliminary criteria for the classification of systemic sclerosis (scleroderma). Arthritis Rheum 23: 581–590, 1980. [DOI] [PubMed] [Google Scholar]
- 15. Goronzy JJ, Bartz‐Bazzanella P, Hu W, Jendro MC, Walser‐Kuntz DR, Weyand CM: Dominant clonotypes in the repertoire of peripheral CD4+ T cells in rheumatoid arthritis. J Clin Invest 94: 2068–2076, 1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 16. Fitzgerald JE, Ricalton NS, Meyre AC, et al.: Analysis of clonal CD8+ T cell expansions in normal individuals and patients with rheumatoid arthritis. J Immunol 154: 3538–3547, 1995. [PubMed] [Google Scholar]
- 17. Hingaroni R, Monteiro J, Furie R, et al.: Oligoclonality of Vβ3 TCR chains in the CD8+ T cell population of rheumatoid arthritis patients. J Immunol 156: 852–858, 1996. [PubMed] [Google Scholar]
- 18. Waase I, Kayser C, Carlson PJ, Goronzy JJ, Weyand CM: Oligoclonal T cell proliferation in patients with rheumatoid arthritis and their unaffected siblings. Arthritis Rheum 39: 904–913, 1996. [DOI] [PubMed] [Google Scholar]
- 19. Mato T, Masuko K, Misaki Y, et al.: Correlation of clonal T cell expansion with disease activity in systemic lupus erythematosus. Int Immunol 9: 547–554, 1997. [DOI] [PubMed] [Google Scholar]
- 20. Yamamoto K, Masuko K, Takahashi S, et al.: Accumulation of distinct T cell clonotypes in human solid tumors. J Immunol 154: 1804–1809, 1995. [PubMed] [Google Scholar]