The experiments in the present application are directed toward resolving: 1.) The mechanism of CTL-mediated lysis. The mechanism by which cytotoxic T lymphocytes kill specific target cells in still unknown. A study of lysis in lectin-mediated cytolysis (LDCC) led us to propose a new mechanism for CTL-mediated cytolysis, in which target cell death occurs as a result of distortion of target cell MHC proteins. Leakage at the interface between the transmembrane MHC proteins and surrounding lipids is postulated to lead to irreversible osmotic damage. Experiments are included in this proposal to test our hypothesis. Antibodies to MHC proteins, other transmembrane proteins, non-transmembrane proteins, and purely lipid antigens will be coupled to the surface of inert microspheres, microtiter wells, RBC or other nonlytic cell types. These will then be incubated with 51Cr-labeled target cells, and the rate of 51Cr release monitored. 2.) The relationship of lectin-mediated polyclonal activation of CTL function to LDCC and to antigen activation of CTL function. Certain plant lectins have been used both to activate (CTL function, and to mediate lysis in cases where MHC-restricted recognition is presumed not to occur. We have recently shown that target cell lysis in LDCC is in fact MHC-restricted, and we have obtained preliminary data indicating that lectin-mediated, polyclonal generation of CTL function is also controlled by MHC proteins involved in lectin presentation. We propose experiments to explore this point further, and to analyze its significance for CTL function generally. Using specific MHC antibodies, appropriate H-2 recombinant and congenic strains, and MHC positive and negative cell lines, we will probe the involvement of MHC proteins in activation of CTL function in primary and secondary reactions, in both syngeneic and allogeneic combination. We will also clone out individual CTL generated by lectin and compare their properties will CTL generated by specific antigen.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI014747-15
Application #
3125850
Study Section
Allergy and Immunology Study Section (ALY)
Project Start
1978-09-30
Project End
1988-08-31
Budget Start
1986-09-01
Budget End
1987-08-31
Support Year
15
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of California Los Angeles
Department
Type
Schools of Arts and Sciences
DUNS #
119132785
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Ratner, A; Clark, W R (1993) Role of TNF-alpha in CD8+ cytotoxic T lymphocyte-mediated lysis. J Immunol 150:4303-14
Ratner, A; Clark, W R (1991) Lack of target cell participation in cytotoxic T lymphocyte-mediated lysis. J Immunol 147:55-9
Ostergaard, H L; Clark, W R (1989) Evidence for multiple lytic pathways used by cytotoxic T lymphocytes. J Immunol 143:2120-6
Torbett, B E; Clark, W R (1988) The role of accessory molecules in T-helper activation induced by antigen, lectin, or CD3 antibodies. Cell Immunol 115:352-62
Gorman, K; Liu, C C; Blakely, A et al. (1988) Cloned cytotoxic T lymphocytes as target cells. II. Polarity of lysis revisited. J Immunol 141:2211-5
Ostergaard, H; Gorman, K; Clark, W R (1988) Cloned cytotoxic T lymphocyte target cells fail to induce early activation events in effector cytotoxic T lymphocytes. Cell Immunol 114:188-97
Gorman, K C; Kane, K P; Clark, W R (1987) Target cell recognition structures in LDCC and ODCC. J Immunol 138:1014-9
Blakely, A; Gorman, K; Ostergaard, H et al. (1987) Resistance of cloned cytotoxic T lymphocytes to cell-mediated cytotoxicity. J Exp Med 166:1070-83
Young, J D; Clark, W R; Liu, C C et al. (1987) A calcium- and perforin-independent pathway of killing mediated by murine cytolytic lymphocytes. J Exp Med 166:1894-9
Torbett, B E; Skidmore, B; Clark, W R (1986) Multiple pathways for antigen-independent activation of a T helper hybridoma. Eur J Immunol 16:933-8

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