Generation of CD4+ and CD8+ Antiinfluenza CTL and Assay of In Vitro Cytotoxicity
Mice that are primed in vivo at 6–12 weeks of age by immunizing intravenously with 100–300 hemagglutinating units (HAU) of virus in allantoic fluid are used as spleen donors three or more weeks afterwards. Immune spleen cells (100 × 106) are stimulated in vitro by co-culture with 20 × 106 virusinfected, irradiated (2000 rad) naive splenocytes in 75-cm2 flasks containing 40 ml of minimal essential medium (MEM) (GIBCO) plus 10% fetal bovine serum (FBS) and 5 × 10−5 m 2-mercaptoethanol (2-ME) (Braciale et al., 1982).
KeywordsMajor Histocompatibility Complex Allantoic Fluid Major Histocompatibility Complex Binding Immune Spleen Cell Peptide Major Histocompatibility Complex
- Braciale TJ, Braciale VL, Andrew ME (1982): Cloned continuous lines of H-2 restricted influenza virus-specific CTL: Probes of T lymphocyte specificity and heterogeneity. In: Isolation, Characterization and L tilination of T Lymphocytes, Fathman CG, Fitch FW, eds. New York: Academic PressGoogle Scholar
- Braciale TJ, Braciale VL, Winkler M, Stroynowski I, Hood L, Sambrook J, Gething M-J (1987): On the role of the transmembrane anchor sequence of influenza hemagglutinin in target cell recognition by class I MHC-restricted, hemagglutinin-specific cytolytic T lymphocytes. J Exy Med 166: 678–692CrossRefGoogle Scholar
- Morrison LA, Braciale VL, Braciale TJ (1988): Antigen form influences induction and frequency of influenza specific class I and class II MHC-restricted cytolytic T lymphocytes. J Immunol 141: 363–368Google Scholar