ovalbumin and Arenaviridae-Infections

The functional identification of antigen-specific CD8 T cell populations is critical to understanding host responses to infection by intracellular pathogens. Furthermore, assessing the properties of protective memory CD8 T cell populations generated by immunization is necessary in the rational design of vaccines. Recently, a classification scheme was proposed in which memory CD8 T cells were divided into one of two distinct subsets, based on CD62L expression, that have different functional properties and protective capacities. Intracellular cytokine staining functionally identifies antigen-specific CD8 T cell populations after short in vitro stimulation with cognate peptide. This short stimulation, however, results in the cleavage of CD62L from the cell surface of antigen-specific CD8 T cells and precludes distinguishing CD62L(hi)- and CD62L(lo)-expressing memory cell subsets within this population. Here, we describe a method of preserving CD62L expression by the antigen-specific CD8 T cell population during coculture with antigen. This methodology allows for the identification and functional assessment of antigen-specific memory CD8 T cell populations, while simultaneously characterizing the memory subset composition of that population. Using this method, we directly identify differences in IL-2 production capacity by CD62L(hi)- and CD62L(lo)-expressing antigen-specific memory CD8 T cell populations.

Topics: Adoptive Transfer; Animals; Arenaviridae Infections; CD8-Positive T-Lymphocytes; Cytokines; Flow Cytometry; Hydroxamic Acids; Immunologic Memory; Interferon-gamma; Interleukin-2; L-Selectin; Listeria monocytogenes; Listeriosis; Lymphocyte Activation; Lymphocytic choriomeningitis virus; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Transgenic; Oligopeptides; Ovalbumin; Receptors, Antigen, T-Cell; Spleen; T-Lymphocyte Subsets