Rapid whole blood analysis of virus-specific CD4 and CD8 T cell responses in persistent HIV infection.
OBJECTIVE
Upon HIV infection, strong antiviral cytotoxic and helper T cell responses are generated. They are considered to be an important component in the control of HIV viral load. A simple and rapid whole blood assay was established to quantify and simultaneously characterize HIV-reactive CD4 and CD8 cells. The assay was applied to evaluate the effect of antiretroviral therapy on HIV-specific T cell responses.


METHODS
Whole blood of 33 HIV-infected individuals was specifically stimulated by HIV-1 Pr55gag, and activation-induced intracellular cytokine expression in CD4 and CD8 T cells was analysed by flow cytometry.


RESULTS
HIV-1-specific CD8 and CD4 T cells can be quantified simultaneously. As specific antigen, HIV-1 Pr55gag virus-like particles were superior to soluble protein, especially for the activation of CD8 T cells. In untreated individuals, a high frequency of HIV-specific T cells was observed. The frequency of CD8 T cells was consistently higher than the respective CD4 T cell response, thus demonstrating a dominance in CD8 T cell expansion in persistent HIV infection. Patients on antiretroviral therapy showed a significant reduction in HIV-specific CD4 and, even more strikingly, CD8 T cells.


CONCLUSIONS
The whole blood assay provides a rapid estimate of the total antiviral T cell resources, and is highly suited for a clinical setting. It may thus have widespread applications for the evaluation of vaccination strategies and immunotherapy. Because antiretroviral therapy significantly reduces both HIV-specific cytotoxic and helper T cell responses, future therapeutic strategies should aim at improving cellular antiviral immunity.
Sester
M
M
Medical Department IV, Institute of Medical Microbiology and Hygiene, University of the Saarland, Homburg, Germany.
Sester
U
U
Köhler
H
H
Schneider
T
T
Deml
L
L
Wagner
R
R
Mueller-Lantzsch
N
N
Pees
H W
HW
Meyerhans
A
A
D000328:Adult; D019380:Anti-HIV Agents / Q000494:pharmacology / Q000627:therapeutic use; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D018414:CD8-Positive T-Lymphocytes / Q000187:drug effects / Q000276:immunology*; D002908:Chronic Disease; D005260:Female; D005434:Flow Cytometry; D015683:Gene Products, gag / Q000737:chemistry / Q000276:immunology; D015488:HIV Antigens / Q000737:chemistry / Q000276:immunology; D015658:HIV Infections / Q000097:blood* / Q000188:drug therapy / Q000276:immunology*; D015497:HIV-1 / Q000187:drug effects / Q000276:immunology*; D006801:Humans; D008213:Lymphocyte Activation / Q000187:drug effects; D018655:Lymphocyte Count; D008297:Male; D011498:Protein Precursors / Q000737:chemistry / Q000276:immunology; D012995:Solubility; D013997:Time Factors; D014771:Virion / Q000737:chemistry / Q000276:immunology
D019380:Anti-HIV Agents; D015683:Gene Products, gag; D015488:HIV Antigens; D011498:Protein Precursors; C059767:p55 gag precursor protein, Human immunodeficiency virus 1