The laboratory has been focused during the last year on immune-mediated tumor cell destruction. Natural killer (NK) cells and cytotoxic T cells (CTL) lyse virally-infected and tumor cells by 2 main mechanisms:(1) the exocytosis of lytic granules and (2) the expression of cytotoxic molecules like tumor necrosis factor (TNF) or related molecules like Fas-ligand. We have investigated the importance of these mechanisms in cytotoxic immune responses to a murine renal cancer Renca. We successfully isolated CTL specific for the Renca tumor. These CTL exclusively used FasL or perforin/granzymes to lyse Renca cells in vitro. Furthermore,the CTL showed some therapeutic efficacy when transfered to tumor-bearing mice in vivo. Interestingly CTL derived from perforin """"""""knockout"""""""" mice could efficiently destroy pulmonary metastases of Renca. This suggests that the perforin pathway was not essential for antitumor effects of the CTL, at least in this metastasis model. We are currently investigating mechanisms underlying how metastases of this tumor in the lungs and various other anatomical sites (such as the peritoneum or liver) are eliminated by CTL. The cytolytic granules of cytotoxic lymphocytes contain a variety of unique proteins, including a pore-forming protein (perforin) and a family of serine protease enzymes called granzymes. We have further studied the expression of granzyme M (met-ase), a novel granzyme that we had previously cloned. Using a monoclonal antibody specific for granzyme M we have detected this granzyme in freshly isolated NK cells, NKT cells and T gamma delta cells by western blotting and immunohistochemistry. T alpha beta cells do not seem to express this granzyme even after activation. The expression pattern of Granzyme M contrasts with that of the other granzymes and perforin, which can be easily detected in activated T alpha beta cells. This suggests that Granzyme M may be involved in innate rather than adaptive immunity. We are trying to define the biological role of this enzyme, and have obtained mice where granzyme M has been ablated by gene targeting.

Agency
National Institute of Health (NIH)
Institute
Division of Basic Sciences - NCI (NCI)
Type
Intramural Research (Z01)
Project #
1Z01BC010012-06
Application #
6559089
Study Section
(LEI)
Project Start
Project End
Budget Start
Budget End
Support Year
6
Fiscal Year
2001
Total Cost
Indirect Cost
Name
Basic Sciences
Department
Type
DUNS #
City
State
Country
United States
Zip Code
Sun, Kai; Li, Minghui; Sayers, Thomas J et al. (2008) Differential effects of donor T-cell cytokines on outcome with continuous bortezomib administration after allogeneic bone marrow transplantation. Blood 112:1522-9
Cretney, Erika; Shanker, Anil; Yagita, Hideo et al. (2006) TNF-related apoptosis-inducing ligand as a therapeutic agent in autoimmunity and cancer. Immunol Cell Biol 84:87-98
Sayers, Thomas J; Murphy, William J (2006) Combining proteasome inhibition with TNF-related apoptosis-inducing ligand (Apo2L/TRAIL) for cancer therapy. Cancer Immunol Immunother 55:76-84
Khan, Tahira; Stauffer, Jimmy K; Williams, Rebecca et al. (2006) Proteasome inhibition to maximize the apoptotic potential of cytokine therapy for murine neuroblastoma tumors. J Immunol 176:6302-12
Sun, Kai; Wilkins, Danice E C; Anver, Miriam R et al. (2005) Differential effects of proteasome inhibition by bortezomib on murine acute graft-versus-host disease (GVHD): delayed administration of bortezomib results in increased GVHD-dependent gastrointestinal toxicity. Blood 106:3293-9
Brooks, Alan D; Sayers, Thomas J (2005) Reduction of the antiapoptotic protein cFLIP enhances the susceptibility of human renal cancer cells to TRAIL apoptosis. Cancer Immunol Immunother 54:499-505
Smyth, Mark J; Swann, Jeremy; Kelly, Janice M et al. (2004) NKG2D recognition and perforin effector function mediate effective cytokine immunotherapy of cancer. J Exp Med 200:1325-35
Shafer-Weaver, Kimberly; Sayers, Thomas; Kuhns, Douglas et al. (2004) Evaluating the cytotoxicity of innate immune effector cells using the GrB ELISPOT assay. J Transl Med 2:31
Sedelies, Karin A; Sayers, Thomas J; Edwards, Kirsten M et al. (2004) Discordant regulation of granzyme H and granzyme B expression in human lymphocytes. J Biol Chem 279:26581-7
Sun, Kai; Welniak, Lisbeth A; Panoskaltsis-Mortari, Angela et al. (2004) Inhibition of acute graft-versus-host disease with retention of graft-versus-tumor effects by the proteasome inhibitor bortezomib. Proc Natl Acad Sci U S A 101:8120-5

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