Immunohistochemical localization of granzyme B antigen in cytotoxic cells in human tissues.

Human granzyme B antigen is expressed in cytoplasmic granules of activated cytotoxic T lymphocytes and natural killer cells. Recombinant granzyme B was generated using a prokaryotic expression vector under the control of T7 transcription and translation signals. The 25-kd recombinant protein (granzyme B) was used to develop a rabbit polyclonal antiserum. Purified anti-granzyme B antibodies were used to detect the antigen expression in cytotoxic cells in human tissues. Using the avidin-biotin-complex peroxidase technique, formalin-fixed, paraffin-embedded tissue sections from patients with acute mild or moderate allograft cardiac rejection were stained. A constant cytoplasmic staining of the lymphocytic allograft infiltrate was observed. These results provide a basis for using the anti-granzyme B antibodies for detection of cytotoxic cells in human tissues. The detection and quantitative analysis of the granzyme-associated cytotoxic cells may help to evaluate the significance of these functionally distinct cytotoxic cells in human tissues associated with increased expression of cytoplasmic granule effector molecules.

[1]  J. Tschopp,et al.  Inhibition of lymphocyte mediated cytotoxicity by perforin antisense oligonucleotides. , 1990, The EMBO journal.

[2]  H. Macdonald,et al.  Cell specificity of granzyme gene expression. , 1990, Journal of immunology.

[3]  L. Young,et al.  Perforin-mediated myocardial damage in acute myocarditis , 1990, The Lancet.

[4]  M. Clément,et al.  Granzyme B-gene expression: a marker of human lymphocytes "activated" in vitro or in renal allografts. , 1990, Human immunology.

[5]  P. Anderson,et al.  A monoclonal antibody reactive with a 15-kDa cytoplasmic granule-associated protein defines a subpopulation of CD8+ T lymphocytes. , 1990, Journal of immunology.

[6]  S. Burrows,et al.  Interleukin-2 receptors in infectious mononucleosis. , 1989, Immunology letters.

[7]  P. Henkart,et al.  Induction of target cell DNA release by the cytotoxic T lymphocyte granule protease granzyme A , 1989, The Journal of experimental medicine.

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

[9]  J. Tschopp,et al.  Granzymes, a Family of Serine Proteases Released from Granules of Cytolytic T Lymphocytes upon T Cell Receptor Stimulation , 1988, Immunological reviews.

[10]  I. Weissman,et al.  A high proportion of T lymphocytes that infiltrate H-2-incompatible heart allografts in vivo express genes encoding cytotoxic cell-specific serine proteases, but do not express the MEL-14-defined lymph node homing receptor , 1988, The Journal of experimental medicine.

[11]  J. Manning,et al.  Effect of Trypsinization on the Immunostaining of Formalin‐Fixed, Paraffin‐Embedded Tissues , 1988, The American journal of surgical pathology.

[12]  I. Weissman,et al.  Cloning and chromosomal assignment of a human cDNA encoding a T cell- and natural killer cell-specific trypsin-like serine protease. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[13]  C. Verret,et al.  Serine esterase and hemolytic activity in human cloned cytotoxic T lymphocytes , 1988, The Journal of experimental medicine.

[14]  C. Weissmann,et al.  Induction of mRNA for a serine protease and a beta-thromboglobulin-like protein in mitogen-stimulated human leukocytes. , 1987, Journal of immunology.

[15]  R. MacKie,et al.  An assessment of the diagnostic value of the monoclonal antibodies Leu 8, OKT9, OKT10 and Ki67 in cutaneous lymphocytic infiltrates , 1986, The British journal of dermatology.

[16]  T. Waldmann.  The structure, function, and expression of interleukin-2 receptors on normal and malignant lymphocytes. , 1986, Science.

[17]  F. Studier,et al.  Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. , 1986, Journal of molecular biology.

[18]  David A. Amato,et al.  Specific release of proteoglycans from human natural killer cells during target lysis , 1985, Nature.

[19]  G. Pinkus,et al.  Leu-M1--a marker for Reed-Sternberg cells in Hodgkin's disease. An immunoperoxidase study of paraffin-embedded tissues. , 1985, The American journal of pathology.

[20]  D. Redelman,et al.  The mechanism of cell-mediated cytotoxicity. III. Protease-specific inhibitors preferentially block later events in cytotoxic T lymphocyte-mediated lysis than do inhibitors of methylation or thiol-reactive agents. , 1983, Cellular immunology.

[21]  V. Diehl,et al.  Production of a monoclonal antibody specific for Hodgkin and Sternberg–Reed cells of Hodgkin's disease and a subset of normal lymphoid cells , 1982, Nature.

[22]  D. Redelman,et al.  The mechanism of cell-mediated cytotoxicity. I. Killing by murine cytotoxic T lymphocytes requires cell surface thiols and activated proteases. , 1980, Journal of Immunology.

[23]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[24]  M. Kramer,et al.  A novel serine proteinase (HuTSP) isolated from a cloned human CD8+ cytolytic T cell line is expressed and secreted by activated CD4+ and CD8+ lymphocytes , 1987, European journal of immunology.

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