Vaccinia assay for the rapid detection of functional HIV-specific CD8+ cytotoxic T lymphocytes.

Abstract

Human immunodeficiency virus (HIV)-specific T cells play a critical role in anti-virus immunity. Therefore, during the clinical development of immune-based therapies, it is important to perform a diagnostic test that rapidly quantifies and characterizes cellular immune responses. For detection of functional HIV-specific CD8(+) cytotoxic T cells (CTL), we used a rapid vaccinia assay that employs recombinant vaccinia virus-infected peripheral blood mononuclear cells (PBMC). This assay was compared with the traditional 51Cr-release CTL assay and with the peptide pool-based enzyme-linked immunospot assay (ELISPOT). We demonstrated a close correlation between these assays by using identical antigens in parallel assays. However, the vaccinia assay was the least expensive and time- and labor-consuming test. Regarding sensitivity, the vaccinia assay was similar to both the peptide pool-based ELISPOT and CTL assays. We showed that human histocompatibility leukocyte antigen (HLA)-Dr(+) cells are responsible for presenting recombinant vaccinia antigens to T cells. The recombinant vaccinia-based assay is a rapid, sensitive and quantitative test and is thus suitable for the detection of functional antigen-specific CD8(+) CTL in large-scale clinical studies.

• Authors: Jianqing Xu, Franco Lori, Julianna Lisziewicz
• Journal: Journal of immunological methods (J Immunol Methods) Vol. 276 Issue 1-2 Pg. 45-57 (May 01 2003) ISSN: 0022-1759 [Print] Netherlands
• PMID: 12738358 (Publication Type: Comparative Study, Evaluation Study, Journal Article, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• DNA, Recombinant
• HIV Antigens
• Peptides
• Interferon-gamma

Topics

• Antigen-Presenting Cells (immunology)
• B-Lymphocytes (virology)
• Cell Line
• Coculture Techniques
• Cytotoxicity Tests, Immunologic
• DNA, Recombinant (genetics)
• Enzyme-Linked Immunosorbent Assay
• Flow Cytometry
• HIV Antigens (immunology)
• HIV-1 (immunology)
• Interferon-gamma (biosynthesis)
• Leukocytes, Mononuclear (virology)
• Peptides (immunology)
• T-Lymphocytes, Cytotoxic (immunology)
• Time Factors
• Vaccinia (genetics)