In vitro human immunodeficiency virus eradication by autologous CD8(+) T cells expanded with inactivated-virus-pulsed dendritic cells - PubMed (original) (raw)

In vitro human immunodeficiency virus eradication by autologous CD8(+) T cells expanded with inactivated-virus-pulsed dendritic cells

W Lu et al. J Virol. 2001 Oct.

Free PMC article

Abstract

Despite significant immune recovery with potent highly active antiretroviral therapy (HAART), eradication of human immunodeficiency virus (HIV) from the bodies of infected individuals represents a challenge. We hypothesized that an inadequate or inappropriate signal in virus-specific antigen presentation might contribute to the persistent failure to mount efficient anti-HIV immunity in most HIV-infected individuals. Here, we conducted an in vitro study with untreated (n = 10) and HAART-treated (n = 20) HIV type 1 (HIV-1) patients which showed that pulsing of monocyte-derived dendritic cells (DC) with aldrithiol-2-inactivated autologous virus resulted in the expansion of virus-specific CD8(+) T cells which were capable of killing HIV-1-infected cells and eradicating the virus from cultured patient peripheral blood mononuclear cells independently of the disease stages and HAART response statuses of the patients. This in vitro anti-HIV effect was further enhanced by the HIV protease inhibitor indinavir (at a nonantiviral concentration), which has been shown previously to be able to up-regulate directly patient T-cell proliferation following immune stimulation. However, following a 2-day treatment with culture supernatant derived from immune-activated T cells (which mimics an in vivo environment of HIV-disseminated and immune-activated lymphoid tissues), DC lost their capacity to present de novo inactivated-virus-derived antigens. These findings provide important information for understanding the establishment of chronic HIV infection and indicate a perspective for clinical use of DC-based therapeutic vaccines against HIV.

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Figures

FIG. 1

FIG. 1

Proliferation of patient T cells following stimulation with inactivated-virus-pulsed autologous DC in the absence or presence of HIV PI (indinavir, 10 nM). (A) Mean (± SD) [3H]thymidine incorporation in T cells from untreated patients in the absence (□) or presence (▪) of PI, in T cells from HAART-treated plasma viral load responders in the absence (▵) or presence (▴) of PI, and in T cells from plasma viral load nonresponders in the absence (○) or presence (●) of PI. (B) Mean (± SD) relative CD4/CD8 ratio in T cells from untreated patients (□), plasma viral load responders (▨), and plasma viral load nonresponders (▩). The baseline CD4/CD8 ratio in the absence of stimulation was normalized to 1. The baseline ranges of the CD3+ CD4+ phenotype in untreated patients, plasma viral load responders, and plasma viral load nonresponders were 15 to 32%, 24 to 53%, and 21 to 41%, respectively.

FIG. 2

FIG. 2

HIV-1 gag_-specific CTL activity in patient T cells expanded by inactivated-virus-pulsed autologous DC in the absence or presence of indinavir. (A) Mean (± SD) percent specific lysis (at an effector/target ratio of 10:1) of autologous B-LCL targets (infected with recombinant vaccinia virus containing a HIV-1_gag gene) by T cells stimulated with virus-pulsed DC with or without PI. T cells were from untreated patients (□), plasma viral load responders to HAART (▨), and plasma viral load nonresponders to HAART (▩). (B) Mean (± SD) percent specific lysis from all patients in the absence of antibodies (▤) or in the presence of blocking antibodies against CD4 (░⃞) or CD8 (▪). The background percent _gag_-specific lysis using unloaded DC-treated T cells was <10%.

FIG. 3

FIG. 3

Quantitative analysis of anti-HIV activity of patient T cells stimulated with inactivated-virus-pulsed autologous DC in the absence or presence of PI. Each result is the mean (± SD) number of proviral HIV DNA copies/106 cells (A and C) or the mean (± SD) number of supernatant HIV RNA copies per milliliter (B and D) in the coculture of autologous virus-pulsed-DC-stimulated T cells and superinfected T cells from untreated patients (□), plasma viral load responders to HAART (▨), and plasma viral load nonresponders to HAART (▩) or from all patients in the absence of antibodies (▤) or the presence of blocking antibodies against CD4 (░⃞) or CD8 (▪).

FIG. 4

FIG. 4

Functions of DC following treatment with activated-T-cell supernatant. (A) Mean (± SD) [3H]thymidine incorporation in patient T cells stimulated with virus-pulsed DC (○) or DC pretreated with activated-T-cell supernatant before (▴) or after (▪) pulsing with inactivated autologous virus. (B) Mean (± SD) percent HIV_gag_-specific lysis (at an effector/target ratio of 10:1) of autologous B-LCL targets by patient T cells expanded with virus-pulsed DC (□) or DC pretreated with activated-T-cell supernatant before (▨) or after (▩) pulsing with inactivated autologous virus. (C) Mean (± SD) number of proviral HIV DNA copies/106 cells or supernatant HIV RNA copies per milliliter in the coculture of superinfected T cells with autologous T cells expanded with virus-pulsed DC (□) or DC pretreated with activated-T-cell supernatant before (▨) or after (▩) pulsing with inactivated autologous virus.

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References

    1. Bohler T, Debatin K M, Wintergerst U. T-cell apoptosis in HIV-1-infected individuals receiving highly active antiretroviral therapy. Blood. 2001;97:1898–1901. - PubMed
    1. Cavert W, Notermans D W, Staskus K, Wietgrefe S W, Zupancic M, Gebhard K, Henry K, Zhang Z Q, Mills R, McDade H, Schuwirth C M, Goudsmit J, Danner S A, Haase A T. Kinetics of response in lymphoid tissues to antiretroviral therapy of HIV-1 infection. Science. 1997;276:960–964. - PubMed
    1. Chun T W, Engel D, Mizell S B, Hallahan C W, Fischette M, Park S, Davey R T, Jr, Dybul M, Kovacs J A, Metcalf J A, Mican J M, Berrey M M, Corey L, Lane H C, Fauci A S. Effect of interleukin-2 on the pool of latently infected, resting CD4+ T cells in HIV-1-infected patients receiving highly active anti-retroviral therapy. Nat Med. 1999;5:651–655. - PubMed
    1. Chun T W, Stuyver L, Mizell S B, Ehler L A, Mican J A, Baseler M, Lloyd A L, Nowak M A, Fauci A S. Presence of an inducible HIV-1 latent reservoir during highly active antiretroviral therapy. Proc Natl Acad Sci USA. 1997;94:13193–13197. - PMC - PubMed
    1. Collier A C, Coombs R W, Schoenfeld D A, Bassett R L, Timpone J, Baruch A, Jones M, Facey K, Whitacre C, McAuliffe V J, Friedman H M, Merigan T C, Reichman R C, Hooper C, Corey L. Treatment of human immunodeficiency virus infection with saquinavir, zidovudine, and zalcitabine. AIDS Clinical Trials Group. N Engl J Med. 1996;334:1011–1017. - PubMed

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