Quantifying HIV-1-Specific CD8+ T-Cell Responses Using ELISPOT and Cytokine Flow Cytometry
Since the initial description and characterization of the agent that causes AIDS, human immunodeficiency virus (HIV-1), numerous research groups have characterized immune responses to this virus. Much effort has been directed towards identifying potential correlates of protection that may be useful for the development of vaccines and immunotherapies. In addition, several investigations have focused on comparing patients with rapid vs. slow disease progression profiles in an attempt to identify the characteristics of a “successful” immune response. Although many gaps remain in our understanding of the host–pathogen relationship, great progress has been made during the past 20 years in elucidating the adaptive, cell-mediated response to HIV-1. These investigations have benefited in recent years from the development of new approaches to the analysis of antigen-specific CD8+ T-cell function, notably the ELISPOT assay and cytokine flow cytometry. This chapter provides simple protocols for these two methods.
Key wordsCytokine ELISPOT flow cytometry T-cell
This work is supported by NIH/NIAID AI057020 and by the California Universitywide AIDS Research Program (UARP), grant CH05-D-606. The authors thank their colleagues in the Department of Medical Microbiology and Immunology, UC Davis, for assistance with protocol optimization. We also thank Drs. Douglas Nixon (University of California, San Francisco) and Michael Betts (University of Pennsylvania) for sharing protocols and technical information.
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