A High-Throughput Assay for the Isolation of HIV-Specific Cytotoxic T Lymphocytes

Virus-specific Cytotoxic T Lymphocytes (CTLs) play an important role in restricting the replication of persistent viruses like the Human Immunodeficiency Virus (HIV). The restriction is, however, only transient for most individuals and fails to afford long-term protection. The reasons for the failure remain poorly understood. We present a high-throughput approach to monitor effector-mediated lysis of peptide-loaded targets by pairing both target cells and effector CTLs in arrays of microwells. Additionally, target specificity, kinetics of lysis and kinetics of interferon gamma/macrophage inflammation protein beta secretion of the effectors are determined in a quantitative manner. Finally, the identification of an HIV-1 specific CTL in a clinical sample is demonstrated. The assay should facilitate identification of the functional characteristics of an effective HIV-specific CTL response, by identifying and isolating CTLs from HIV-infected long-term non-progressors (LTNPs). Comparison of their functional profiles to the same types of cells isolated from progressors should highlight the characteristic responses required for effective T cell vaccines.