A vaccinia-based elispot assay for detection of CD8+ T cells from HIV-1 infected children.
HIV-specific CD8+ T lymphocytes participate in the control of viral replication in infected patients. These responses are of low intensity in young infants and are decreased by antiretroviral therapy. In the present study, we report on a recombinant Vaccinia virus (rVV)-based Elispot assay for the detection of HIV-specific CD8+ T cells immediately after isolation of peripheral blood mononuclear cells (PBMC). The rVV-based assay was highly sensitive; 48 out of 50 children had a positive response against the rVV encoding HIV Env-Gag-Pol antigen. Interferon-gamma was produced by CD8+ T cells, and CD14+/15+ cells were the main cell subset presenting antigens expressed by rVV. We observed that the cell input per well had a critical influence on the sensitivity of the assay. Results from the ex vivo Elispot assay correlated poorly with those of the 51Cr release assay performed after expansion of PBMC in vitro; thus, both assays gave information on different subsets and/or functions of the HIV-specific T cell response.
Buseyne
Florence
F
Unité Postulante d'Immunopathologie Virale, URA CNRS 1930, Institut Pasteur, Bat. Lwoff, 28 rue du Dr Roux, 75015 Paris, France. florence@pasteur.fr
Catteau
Adeline
A
Scott-Algara
Daniel
D
Corre
Béatrice
B
Porrot
Françoise
F
Rouzioux
Christine
C
Blanche
Stéphane
S
Rivière
Yves
Y
D000818:Animals; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002648:Child; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005822:Genetic Vectors; D015488:HIV Antigens / Q000276:immunology; D015658:HIV Infections / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000032:analysis / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014615:Vaccinia / Q000235:genetics / Q000276:immunology
D015488:HIV Antigens; D007371:Interferon-gamma