Evaluation of aldrithiol-2-inactivated preparations of HIV type 1 subtypes A, B, and D as reagents to monitor T cell responses.
The development of HIV vaccines is an urgent priority and there is need to generate reagents representing multiple subtypes that can be used to screen HIV-1-specific responses. We used Aldrithiol-2 (AT-2), a mild oxidizing reagent, to eliminate the infectivity of HIV while maintaining its structure and ability to be processed for presentation to T cells. Inactivated subtype A, B, and D viruses were evaluated for their ability to stimulate T cell responses in PBMC samples from 18 U.S. subjects infected with HIV-1 subtype B and 32 Ugandan subjects infected with subtypes A and D or recombinants AC and AD. Five HIV-1-negative samples were also analyzed. T cell responses to AT-2-inactivated viral isolates were monitored by interferon-gamma (IFN-gamma) intracellular cytokine secretion (ICS) analysis; matched microvesicle preparations served as negative controls. Among the 18 subtype B infected subjects, 39% had CD3(+) CD4 (+) IFN-gamma responses and 67% had CD3(+) CD8(+) IFN-gamma responses. Of the 32 Ugandan subjects, 34% demonstrated CD3(+) CD4(+) IFN-gamma responses and 78% demonstrated CD3(+) CD8(+) IFN-gamma responses. Both subtype-specific and cross-reactive responses were observed. Responses to the AT-2 viruses tended to be lower in magnitude than those detected by a set of overlapping gag peptides. Robust lymphoproliferative responses to AT-2 viruses were seen in a subset of subjects. In conclusion, AT-2-inactivated HIV-1 virions stimulated both CD4 and CD8 HIV-1-specific responses and may provide an additional reagent for screening HIV-1-specific responses in HIV seropositives and vaccinees.
Rutebemberwa
A
A
U.S. Military HIV Research Program/Henry Jackson Foundation, 13 Taft Court, Rockville, MD 20850, USA.
Bess
J W
JW
Brown
B
B
Arroyo
M
M
Eller
M
M
Slike
B
B
Polonis
V
V
McCutchan
F
F
Currier
J R
JR
Birx
D
D
Robb
M
M
Marovich
M
M
Lifson
J D
JD
Cox
J H
JH
D015082:2,2'-Dipyridyl / Q000031:analogs & derivatives / Q000494:pharmacology; D016915:AIDS Vaccines; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000821:virology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology* / Q000821:virology; D002478:Cells, Cultured; D004220:Disulfides / Q000494:pharmacology; D015497:HIV-1 / Q000145:classification / Q000187:drug effects / Q000276:immunology*; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D016877:Oxidants / Q000494:pharmacology; D038301:Virus Inactivation
D016915:AIDS Vaccines; D004220:Disulfides; D016877:Oxidants; C001105:2,2'-dipyridyl disulfide; D015082:2,2'-Dipyridyl; D007371:Interferon-gamma