The lethal mechanism of lymphocyte-mediated cytotoxicity of tumor cells is unknown. Specific irreversible inhibitors, e.g., alpha-1-antichymotrypsin and haptenic active site reactants, were instrumental to our initial, indirect approaches that indicate that a serine proteinase with aromatic amino acid specificity must have a crucial role in human, natural killer (NK) lymphocyte-mediated cytotoxicity. We will use these inhibitors to tag NK-associated proteinases for subsequent antibody affinity isolation. The Mr, subunit structure, pI, and cellular localization of the isolated NK-associated proteinase(s) will be determined. NK cell lines and clones have been established. These and additional NK clones will be maintained as the source of material for these studies. Major emphasis will be placed on obtaining IL-2-independent killer clones. This will be done by creating hybridomas with HAT-sensitive cell lines by transformation and by mutation. The NK proteinase may function in NK-associated degranulation, in activation of a lethal substance, or in yet unrecognized processes. We will assess the importance of degranulation in NK and the potential proteinase control of degranulation. In addition, we will examine whether prolethal substances are cleaved prior to transfer of lethal materials to the tumor cells destined to die. Because of their experimental accessibility, the proteinases that control cytotoxicity could be the first components of the lymphocytes' lethal weaponry to be isolated. (LB)

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
3R01CA038942-03S1
Application #
3177448
Study Section
Allergy and Immunology Study Section (ALY)
Project Start
1984-05-11
Project End
1988-01-31
Budget Start
1986-05-01
Budget End
1988-01-31
Support Year
3
Fiscal Year
1987
Total Cost
Indirect Cost
Name
University of Nevada Reno
Department
Type
Schools of Medicine
DUNS #
146515460
City
Reno
State
NV
Country
United States
Zip Code
89557
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Alves, Bryce; Leong, Jeff; Tamang, David L et al. (2009) Hydrolysis of tumor cell lipids after CTL-mediated death. Int Immunol 21:543-53
Alves, Bryce N; Leong, Jeff; Tamang, David L et al. (2009) Pancreatic lipase-related protein 2 (PLRP2) induction by IL-4 in cytotoxic T lymphocytes (CTLs) and reevaluation of the negative effects of its gene ablation on cytotoxicity. J Leukoc Biol 86:701-12
Tamang, David L; Alves, Bryce N; Elliott, Viki et al. (2008) Low dose IL-15 induces snap arming of CD44(low) T lymphocytes in the absence of antigen. Cell Immunol 251:93-101
Tamang, David L; Redelman, Doug; Alves, Bryce N et al. (2006) Induction of granzyme B and T cell cytotoxic capacity by IL-2 or IL-15 without antigens: multiclonal responses that are extremely lytic if triggered and short-lived after cytokine withdrawal. Cytokine 36:148-59
Woolard, Matthew D; Hudig, Dorothy; Tabor, Leslie et al. (2005) NK cells in gamma-interferon-deficient mice suppress lung innate immunity against Mycoplasma spp. Infect Immun 73:6742-51
Kam, Chih-Min; Gotz, Marion G; Koot, Gretchen et al. (2004) Design and evaluation of inhibitors for dipeptidyl peptidase I (Cathepsin C). Arch Biochem Biophys 427:123-34
Barao, Isabel; Hudig, Dorothy; Ascensao, Joao L (2003) IL-15-mediated induction of LFA-1 is a late step required for cytotoxic differentiation of human NK cells from CD34+Lin- bone marrow cells. J Immunol 171:683-90
Tran, Tinh V; Ellis, Karen A; Kam, Chih Min et al. (2002) Dipeptidyl peptidase I: importance of progranzyme activation sequences, other dipeptide sequences, and the N-terminal amino group of synthetic substrates for enzyme activity. Arch Biochem Biophys 403:160-70

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