Demonstration of granzyme A and perforin messenger RNA in the synovium of patients with rheumatoid arthritis.

OBJECTIVE To examine the gene expression of 2 highly specific markers of cytotoxic T lymphocyte (CTL) activation, the serine protease granzyme A and the pore-forming protein perforin, in synovial tissue of patients with rheumatoid arthritis (RA), and to compare the findings with those in osteoarthritis (OA) synovial tissue. METHODS Snap-frozen synovial tissue specimens from 9 patients with RA and 5 patients with OA were examined. The number of CTL that expressed granzyme A or perforin messenger RNA was determined by in situ hybridization using nonradioactive riboprobes for granzyme A and perforin, and by a novel in situ reverse transcriptase technique. The signals were visualized by an immunogold-silver immunohistochemistry technique and compared with immunohistochemical labeling of T and B cells. Additional double-labeling was achieved using anti-type IV collagen, anti-macrophage (anti-CD68), anti-T lymphocyte (anti-CD45RO), anti-B lymphocyte (anti-CD20), and anti-natural killer cell (anti-CD56) antibodies in an alkaline phosphatase-anti-alkaline phosphatase assay. RESULTS Granzyme A and perforin messenger RNA (mRNA) was observed in CTL in synovial specimens from all of the RA patients, whereas in specimens from OA patients only a few, single cells with a positive mRNA signal for these molecules could be detected. In the RA specimens, the number of lymphocytes showing a positive mRNA signal for granzyme A or perforin varied from 10% to 50%, reflecting the recent findings of other investigators studying synovial fluid. CONCLUSION Our results demonstrate that gene expression of at least 2 CTL products, granzyme A and perforin, is up-regulated in the synovium of patients with RA compared with that in the synovium of patients with OA. These molecules presumably play an important role not only in lymphocyte-mediated cytotoxicity, but also in facilitating the migration of blood-borne mononuclear cells through the vascular basement membrane into the rheumatoid synovium.

[1]  F. Breedveld,et al.  Granzyme-positive cytotoxic cells are specifically increased in early rheumatoid synovial tissue. , 1994, Arthritis and rheumatism.

[2]  J. Tschopp,et al.  Cytolytic T-cell cytotoxicity is mediated through perforin and Fas lytic pathways , 1994, Nature.

[3]  Hans Hengartner,et al.  Cytotoxicity mediated by T cells and natural killer cells is greatly impaired in perforin-deficient mice , 1994, Nature.

[4]  L. Moreland,et al.  Virus-like particles in synovial fluids from patients with rheumatoid arthritis. , 1993, British journal of rheumatology.

[5]  S. Fuggle,et al.  In situ cDNA polymerase chain reaction. A novel technique for detecting mRNA expression. , 1993, The American journal of pathology.

[6]  R. Gay,et al.  Molecular and cellular mechanisms of joint destruction in rheumatoid arthritis: two cellular mechanisms explain joint destruction? , 1993, Annals of the rheumatic diseases.

[7]  I. Weissman,et al.  Perforin and granzyme A expression identifying cytolytic lymphocytes in rheumatoid arthritis. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[8]  G. Griffiths,et al.  Expression of perforin and granzymes in vivo: potential diagnostic markers for activated cytotoxic cells. , 1991, Immunology today.

[9]  J. D. Young,et al.  Subcellular localization of perforin and serine esterase in lymphokine-activated killer cells and cytotoxic T cells by immunogold labeling. , 1991, Journal of immunology.

[10]  M. Kramer,et al.  Mouse T-cell associated serine proteinase 1 degrades collagen type IV: a structural basis for the migration of lymphocytes through vascular basement membranes. , 1991, Immunology.

[11]  D A Bloch,et al.  The American College of Rheumatology 1990 criteria for the classification of hypersensitivity vasculitis. , 2010, Arthritis and rheumatism.

[12]  P. Miossec,et al.  Functional studies of soluble low-affinity interleukin-2 receptors in rheumatoid synovial fluid. , 1990, Arthritis and rheumatism.

[13]  M. Lark,et al.  Discoordinate expression of stromelysin, collagenase, and tissue inhibitor of metalloproteinases-1 in rheumatoid human synovial fibroblasts. Synergistic effects of interleukin-1 and tumor necrosis factor-alpha on stromelysin expression. , 1990, The Journal of biological chemistry.

[14]  R. Zinkernagel,et al.  Detection of perforin and granzyme A mRNA in infiltrating cells during infection of mice with lymphocytic choriomeningitis virus , 1989, European journal of immunology.

[15]  J. D. Young,et al.  IN VIVO EXPRESSION OF PERFORIN BY CD8+ LYMPHOCYTES DURING AN ACUTE VIRAL INFECTION BY LUCY , 1989 .

[16]  P. Guerne,et al.  Synovium as a source of interleukin 6 in vitro. Contribution to local and systemic manifestations of arthritis. , 1989, The Journal of clinical investigation.

[17]  E. Podack,et al.  Structure and function of perforin. , 1989, Current topics in microbiology and immunology.

[18]  J. Tschopp,et al.  Granzymes: a family of serine proteases in granules of cytolytic T lymphocytes. , 1989, Current topics in microbiology and immunology.

[19]  A. Lanzavecchia,et al.  Characterization of granzymes A and B isolated from granules of cloned human cytotoxic T lymphocytes. , 1988, Journal of immunology.

[20]  G. Lemm,et al.  Interleukin-2 secretion by synovial fluid lymphocytes in rheumatoid arthritis. , 1988, British journal of rheumatology.

[21]  D. Heinegård,et al.  Detection of tumor necrosis factor alpha but not tumor necrosis factor beta in rheumatoid arthritis synovial fluid and serum. , 1988, Arthritis and rheumatism.

[22]  Chau‐Ching Liu,et al.  Perforin‐Dependent and ‐Independent Pathways of Cytotoxicity Mediated by Lymphocytes , 1988, Immunological reviews.

[23]  C. Frégeau,et al.  The Isolation and Characterization of a Family of Serine Protease Genes Expressed in Activated Cytotoxic T Lymphocytes , 1988, Immunological reviews.

[24]  J. Tschopp,et al.  Cytotoxic T lymphocyte mediated cytolysis. , 1988, Biochemistry.

[25]  M. Liang,et al.  The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. , 1988, Arthritis and rheumatism.

[26]  M. Kramer,et al.  A secretable serine proteinase with highly restricted specificity from cytolytic T lymphocytes inactivates retrovirus‐associated reverse transcriptase , 1987, FEBS letters.

[27]  M. Goto,et al.  T cytotoxic and helper cells are markedly increased, and T suppressor and inducer cells are markedly decreased, in rheumatoid synovial fluids. , 1987, Arthritis and rheumatism.

[28]  J. Tschopp,et al.  A family of serine esterases in lytic granules of cytolytic T lymphocytes , 1987, Cell.

[29]  S. Gay,et al.  Characterization and isolation of poly- and monoclonal antibodies against collagen for use in immunohistochemistry. , 1987, Methods in enzymology.

[30]  P. Russell,et al.  Widespread and selective induction of major histocompatibility complex- determined antigens in vivo by gamma interferon , 1985, The Journal of experimental medicine.

[31]  J. Tschopp,et al.  Isolation of a lytic, pore-forming protein (perforin) from cytolytic T-lymphocytes. , 1985, The Journal of biological chemistry.

[32]  N. Zvaifler,et al.  Characterization of the natural killer-like lymphocytes in rheumatoid synovial fluid. , 1985, Journal of immunology.

[33]  P. Henkart.  Mechanism of lymphocyte-mediated cytotoxicity. , 1985, Annual review of immunology.

[34]  N. Sabharwal,et al.  Synovial fluid lymphocytes differ from peripheral blood lymphocytes in patients with rheumatoid arthritis. , 1982, Journal of immunology.

[35]  A. Notkins,et al.  Immune interferon in the circulation of patients with autoimmune disease. , 1979, The New England journal of medicine.