Vaccinia assay for the rapid detection of functional HIV-specific CD8+ cytotoxic T lymphocytes.
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.
Xu
Jianqing
J
Research Institute for Genetic and Human Therapy (RIGHT), Washington, DC, USA.
Lori
Franco
F
Lisziewicz
Julianna
J
D000938:Antigen-Presenting Cells / Q000276:immunology; D001402:B-Lymphocytes / Q000821:virology; D002460:Cell Line; D018920:Coculture Techniques; D003601:Cytotoxicity Tests, Immunologic; D004274:DNA, Recombinant / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D015488:HIV Antigens / Q000276:immunology; D015497:HIV-1 / Q000276:immunology*; D007371:Interferon-gamma / Q000096:biosynthesis; D007963:Leukocytes, Mononuclear / Q000821:virology; D010455:Peptides / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D013997:Time Factors; D014615:Vaccinia / Q000235:genetics*
D004274:DNA, Recombinant; D015488:HIV Antigens; D010455:Peptides; D007371:Interferon-gamma