The objective of the proposed study is the identification and characterization of viral gene products recognized by influenza A virus specific cytotoxic T lymphocytes (CTL). This work is of both practical and general importance since: 1) it may aid production of influenza vaccines which maximally prime for a CTL response, and 2) it will help define the requirements for CTL recognition of non-self antigens. The study will focus on CTL recognition of non-glycosylated viral proteins, one of which, the nucleoprotein (NP), is clealy shown to serve as a CTL target antigen by preliminary results. CTL recognition of NP and other non-glycosylated viral proteins will be examined using eukaryotic expression vectors cntaining cloned viral genes. Monoclonal antibodies will be used to aid in the biochemical and antigenic characterization of the cell surface forms of NP and other non-glycosylated proteins which are found to serve as CTL target antigens. The basis for the expression of these predominantly cytoplasmic proteins on cell surfaces and their recognition by CTL will be further examined using target cells sensitized for CTL lysis by acid mediated fusion of viral and cellular membranes. Also a focus of this proposal is examination of CTL cross-reactive recognition of influenza A virus glycoproteins derived from different subtypes. This will be investigated using eukaryotic expression vectors containing genes encoding viral glycoproteins. If CTL are found to recognize glycoproteins in a cross-reactive manner, attempts will be made to produce monoclonal antibodies with similar specificities. These antibodies will be characterized with respect to their biological activities, and the degree of overlap between their epitopes and the epitopes defined by CTL will be determined.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI022114-01
Application #
3132815
Study Section
Experimental Virology Study Section (EVR)
Project Start
1985-09-01
Project End
1988-08-31
Budget Start
1985-09-01
Budget End
1986-08-31
Support Year
1
Fiscal Year
1985
Total Cost
Indirect Cost
Name
Wistar Institute
Department
Type
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Eager, K B; Hackett, C J; Gerhard, W U et al. (1989) Murine cell lines stably expressing the influenza virus hemagglutinin gene introduced by a recombinant retrovirus vector are constitutive targets for MHC class I- and class II-restricted T lymphocytes. J Immunol 143:2328-35
Yewdell, J W; Bennink, J R; Eager, K B et al. (1988) CTL recognition of adenovirus-transformed cells infected with influenza virus: lysis by anti-influenza CTL parallels adenovirus-12-induced suppression of class I MHC molecules. Virology 162:236-8
Yewdell, J; Bennink, J; Smith, G et al. (1988) Use of recombinant vaccinia viruses to examine cytotoxic T lymphocyte recognition of individual viral proteins. Adv Exp Med Biol 239:151-61
Yewdell, J W; Bennink, J R; Hosaka, Y (1988) Cells process exogenous proteins for recognition by cytotoxic T lymphocytes. Science 239:637-40
Bennink, J R; Yewdell, J W (1988) Murine cytotoxic T lymphocyte recognition of individual influenza virus proteins. High frequency of nonresponder MHC class I alleles. J Exp Med 168:1935-9
Hosaka, Y; Sasao, F; Yamanaka, K et al. (1988) Recognition of noninfectious influenza virus by class I-restricted murine cytotoxic T lymphocytes. J Immunol 140:606-10
Bennink, J R; Yewdell, J W; Smith, G L et al. (1987) Anti-influenza virus cytotoxic T lymphocytes recognize the three viral polymerases and a nonstructural protein: responsiveness to individual viral antigens is major histocompatibility complex controlled. J Virol 61:1098-102
Yewdell, J W; Bennink, J R; Mackett, M et al. (1986) Recognition of cloned vesicular stomatitis virus internal and external gene products by cytotoxic T lymphocytes. J Exp Med 163:1529-38