Induction of cell death in human immunodeficiency virus-infected macrophages and resting memory CD4 T cells by TRAIL/Apo2l - PubMed (original) (raw)

Induction of cell death in human immunodeficiency virus-infected macrophages and resting memory CD4 T cells by TRAIL/Apo2l

J J Lum et al. J Virol. 2001 Nov.

Abstract

Because the persistence of human immunodeficiency virus (HIV) in cellular reservoirs presents an obstacle to viral eradication, we evaluated whether tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/Apo2L) induces apoptosis in such reservoirs. Lymphocytes and monocyte-derived macrophages (MDM) from uninfected donors do not die following treatment with either leucine zipper human TRAIL (LZhuTRAIL) or agonistic anti-TRAIL receptor antibodies. By contrast, such treatment induces apoptosis of in vitro HIV-infected MDM as well as peripheral blood lymphocytes from HIV-infected patients, including CD4(+) CD45RO(+) HLA-DR(-) lymphocytes. In addition, LZhuTRAIL-treated cells produce less viral RNA and p24 antigen than untreated controls. Whereas untreated cultures produce large amounts of HIV RNA and p24 antigen, of seven treated CD4(+) CD45RO(+) HLA-DR(-) cell cultures, viral RNA production was undetectable in all, p24 antigen was undetectable in six, and proviral DNA was undetectable in four. These data demonstrate that TRAIL induces death of cells from HIV-infected patients, including cell types which harbor latent HIV reservoirs.

PubMed Disclaimer

Figures

FIG. 1

FIG. 1

(A) Analysis of TRAIL-R1, TRAIL-R2, TRAIL-R3, TRAIL-R4, and TRAIL mRNA expression by RT-PCR. (Top) HIV-infected Jurkat T cells show increased message for TRAIL-R2, TRAIL-R3, and TRAIL/Apo2L compared to mock-infected controls (normalized to β-actin). (Center) HIV-1Bal-infected MDM show increased message for all TRAIL receptors and TRAIL/Apo2L compared to mock-infected controls. (Bottom) PBL from four HIV-1-positive individuals show increased message for TRAIL-R2, -R3, and -R4 or TRAIL/Apo2L compared to PBL from four HIV-1-negative individuals. (B) Cell surface expression of TRAIL-R1, -R2, -R3, and -R4 in CD4+ T cells from HIV-infected patients and healthy controls. Open histograms with dotted lines, cells from HIV-infected patients stained with an isotype control MAb; shaded histograms, cells from HIV-1-infected patients stained with anti-TRAIL receptor MAbs; open histograms with solid lines, cells from healthy controls stained with anti-TRAIL receptor MAbs. (C) Jurkat T cells (top) or PBL (bottom) from HIV-negative patients were treated with gp120 or a control (BSA) as indicated and analyzed by flow cytometry for TRAIL receptor expression. Open histograms with dotted lines, isotype control staining; open histograms with solid lines, cells treated with BSA and stained with the indicated anti-TRAIL receptor antibody; shaded histograms, cells treated with gp120 and stained with the indicated anti-TRAIL receptor antibody.

FIG. 2

FIG. 2

Sensitivity of PBL from HIV-1-infected donors to titrated doses of LZhuTRAIL. Cells were isolated from HIV-1-infected donors and incubated with increasing concentrations of LZhuTRAIL as indicated. Cell death was measured by Hoechst staining. Data are representative of three independent experiments. Spontaneous levels of apoptosis are indicated by asterisks.

FIG. 3

FIG. 3

LZhuTRAIL induces apoptosis in cells from HIV-1-infected patients. PBL from 26 randomly selected HIV-1-infected patients or 5 uninfected controls were treated with 1 μg of LZhuTRAIL and analyzed for apoptosis by Hoechst staining, as were CD4+ T cells and CD8+ T cells.

FIG. 4

FIG. 4

Induction of apoptosis in macrophages by LZhuTRAIL. MDM from 11 HIV-1-negative donors were mock or HIV-1Bal infected with or without GM-CSF. Fourteen days following infection, cells were treated with LZhuTRAIL and analyzed by TUNEL to determine the levels of apoptosis.

FIG. 5

FIG. 5

LZhuTRAIL reduces viral gene expression in infected cells. CD4/DR− RO+ cells were isolated from seven HIV-1-infected patients, treated with LZhuTRAIL or agonistic TRAIL receptor antibodies (or isotype controls), and analyzed for p24 antigen production (A) and for the presence of integrated proviral DNA. (B) Viral RNA in culture supernatants. (C and D) Unfractionated cells from four HIV-1-infected patients with suppressed plasma viremia for more than 12 months were treated with or without LZhuTRAIL or agonistic MAbs to TRAIL-R2 (or isotype controls) and tested for p24 antigen (C) or viral RNA in culture supernatants (D).

Similar articles

Cited by

References

    1. Ashkenazi A, Pai R C, Fong S, Leung S, Lawrence D A, Marsters S A, Blackie C, Chang L, McMurtrey A E, Hebert A, DeForge L, Koumenis I L, Lewis D, Harris L, Bussiere J, Koeppen H, Shahrokh Z, Schwall R H. Safety and antitumor activity of recombinant soluble Apo2 ligand. J Clin Investig. 1999;104:155–162. - PMC - PubMed
    1. Badley A D, McElhinny J A, Leibson P J, Lynch D H, Alderson M R, Paya C V. Upregulation of Fas ligand expression by human immunodeficiency virus in human macrophages mediates apoptosis of uninfected T lymphocytes. J Virol. 1996;70:199–206. - PMC - PubMed
    1. Badley A D, Pilon A A, Landay A, Lynch D H. Mechanisms of HIV associated lymphocyte apoptosis. Blood. 2000;96:2951–2964. - 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. Chinnaiyan A M, Prasad U, Shankar S, Hamstra D A, Shanaiah M, Chenevert T L, Ross B D, Rehemtulla A. Combined effect of tumor necrosis factor-related apoptosis-inducing ligand and ionizing radiation in breast cancer therapy. Proc Natl Acad Sci USA. 2000;97:1754–1759. - PMC - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources