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Critical contribution of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) to apoptosis of human CD4+ T cells in HIV-1-infected hu-PBL-NOD-SCID mice.

Miura Y, Misawa N, Maeda N, Inagaki Y, Tanaka Y, Ito M, Kayagaki N, Yamamoto N, Yagita H, Mizusawa H, Koyanagi Y - J. Exp. Med. (2001)

Bottom Line: Apoptosis is a key for CD4+ T cell destruction in HIV-1-infected patients.A combination of TUNEL and immunostaining for death-inducing tumor necrosis factor (TNF) family molecules indicated that the apoptotic cells were frequently found in conjugation with TNF-related apoptosis-inducing ligand (TRAIL)-expressing CD3+CD4+ human T cells.These results suggest that a large number of HIV-1-uninfected CD4+ T cells undergo TRAIL-mediated apoptosis in HIV-infected lymphoid organs.

View Article: PubMed Central - PubMed

Affiliation: Department of Virology, Tohoku University Graduate School of Medicine, Sendai 980-8575, Japan.

ABSTRACT
Apoptosis is a key for CD4+ T cell destruction in HIV-1-infected patients. In this study, human peripheral blood lymphocyte (PBL)-transplanted nonobese diabetic (NOD)-severe combined immunodeficient (SCID) (hu-PBL-NOD-SCID) mice were used to examine in vivo apoptosis after HIV-1 infection. As the hu-PBL-NOD-SCID mouse model allowed us to see extensive infection with HIV-1 and to analyze apoptosis in human cells in combination with immunohistological methods, we were able to quantify the number of apoptotic cells with HIV-1 infection. As demonstrated by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), massive apoptosis was predominantly observed in virus-uninfected CD4+ T cells in the spleens of HIV-1-infected mice. A combination of TUNEL and immunostaining for death-inducing tumor necrosis factor (TNF) family molecules indicated that the apoptotic cells were frequently found in conjugation with TNF-related apoptosis-inducing ligand (TRAIL)-expressing CD3+CD4+ human T cells. Administration of a neutralizing anti-TRAIL mAb in HIV-1-infected mice markedly inhibited the development of CD4+ T cell apoptosis. These results suggest that a large number of HIV-1-uninfected CD4+ T cells undergo TRAIL-mediated apoptosis in HIV-infected lymphoid organs.

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Apoptosis occurs predominantly in HIV-1–uninfected CD4+ T cells. (a) Triple immunofluorescent staining for HIV p24gag (FITC, green), human CD4 (Cy5, blue), and TUNEL (TRITC, red) of spleens from HIV-1–infected hu-PBL-NOD-SCID mice 2 wk after inoculation of human PBMCs (9 d after infection). Original magnification ×25 (left panel), ×100 (middle panel), and ×1,000 (right two panels). TUNEL (red) and CD4 (blue) double-positive cells are shown as pink in lower magnification (left panel). CD4 (blue) and HIV p24gag (green) double-positive cells are shown as light green. Arrow shows a small artery. Right upper panel indicates TUNEL+, CD4+, and p24gag+ cells, which represented <2% of both TUNEL+ and CD4+ cells. Right lower panel indicates TUNEL+, CD4+, and p24gag− cells, which accounted for >90% of both TUNEL+ and CD4+ cells. (b) Dual color detection for GFP-expressing HIV-1 (GFP, green) and TUNEL staining (TRITC, red) of spleens from GFP/HIV-1–infected hu-PBL-NOD-SCID mice 2 wk after human PBL transplantation (9 d after infection). The majority of TUNEL+ cells were GFP/HIV-1 negative. Original magnification ×100.
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Figure 1: Apoptosis occurs predominantly in HIV-1–uninfected CD4+ T cells. (a) Triple immunofluorescent staining for HIV p24gag (FITC, green), human CD4 (Cy5, blue), and TUNEL (TRITC, red) of spleens from HIV-1–infected hu-PBL-NOD-SCID mice 2 wk after inoculation of human PBMCs (9 d after infection). Original magnification ×25 (left panel), ×100 (middle panel), and ×1,000 (right two panels). TUNEL (red) and CD4 (blue) double-positive cells are shown as pink in lower magnification (left panel). CD4 (blue) and HIV p24gag (green) double-positive cells are shown as light green. Arrow shows a small artery. Right upper panel indicates TUNEL+, CD4+, and p24gag+ cells, which represented <2% of both TUNEL+ and CD4+ cells. Right lower panel indicates TUNEL+, CD4+, and p24gag− cells, which accounted for >90% of both TUNEL+ and CD4+ cells. (b) Dual color detection for GFP-expressing HIV-1 (GFP, green) and TUNEL staining (TRITC, red) of spleens from GFP/HIV-1–infected hu-PBL-NOD-SCID mice 2 wk after human PBL transplantation (9 d after infection). The majority of TUNEL+ cells were GFP/HIV-1 negative. Original magnification ×100.

Mentions: A similar extent of splenomegaly was observed in HIV-1–infected mice 2 wk after human PBL transplantation. However, the splenomegaly in infected mice regressed more rapidly than that of uninfected mice at 4 wk (data not shown). A marked decrease in the number of human CD4+ T cells was observed in infected mice with immunostaining analysis (data not shown). HIV-1 p24gag–expressing cells were found in the human CD4+ T cell–rich region; however, the number of p24gag+ cells was far lower than the number of human CD4+ T cells. In contrast, the number and distribution of human CD8+ T cells, human CD20+ B cells, and human CD68+ macrophages was apparently unchanged. TUNEL staining showed a large number of apoptotic cells around the small arteries in the spleen 2 wk after HIV-1 infection (Fig. 1 a). In contrast, only a few TUNEL+ apoptotic cells were observed in the spleens of uninfected hu-PBL-NOD-SCID mice at the same time point (data not shown). Furthermore, a NOD-SCID mouse section without human PBL transplantation showed no staining. The specificity of the TUNEL staining was confirmed as described in Materials and Methods.


Critical contribution of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) to apoptosis of human CD4+ T cells in HIV-1-infected hu-PBL-NOD-SCID mice.

Miura Y, Misawa N, Maeda N, Inagaki Y, Tanaka Y, Ito M, Kayagaki N, Yamamoto N, Yagita H, Mizusawa H, Koyanagi Y - J. Exp. Med. (2001)

Apoptosis occurs predominantly in HIV-1–uninfected CD4+ T cells. (a) Triple immunofluorescent staining for HIV p24gag (FITC, green), human CD4 (Cy5, blue), and TUNEL (TRITC, red) of spleens from HIV-1–infected hu-PBL-NOD-SCID mice 2 wk after inoculation of human PBMCs (9 d after infection). Original magnification ×25 (left panel), ×100 (middle panel), and ×1,000 (right two panels). TUNEL (red) and CD4 (blue) double-positive cells are shown as pink in lower magnification (left panel). CD4 (blue) and HIV p24gag (green) double-positive cells are shown as light green. Arrow shows a small artery. Right upper panel indicates TUNEL+, CD4+, and p24gag+ cells, which represented <2% of both TUNEL+ and CD4+ cells. Right lower panel indicates TUNEL+, CD4+, and p24gag− cells, which accounted for >90% of both TUNEL+ and CD4+ cells. (b) Dual color detection for GFP-expressing HIV-1 (GFP, green) and TUNEL staining (TRITC, red) of spleens from GFP/HIV-1–infected hu-PBL-NOD-SCID mice 2 wk after human PBL transplantation (9 d after infection). The majority of TUNEL+ cells were GFP/HIV-1 negative. Original magnification ×100.
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Related In: Results  -  Collection

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Figure 1: Apoptosis occurs predominantly in HIV-1–uninfected CD4+ T cells. (a) Triple immunofluorescent staining for HIV p24gag (FITC, green), human CD4 (Cy5, blue), and TUNEL (TRITC, red) of spleens from HIV-1–infected hu-PBL-NOD-SCID mice 2 wk after inoculation of human PBMCs (9 d after infection). Original magnification ×25 (left panel), ×100 (middle panel), and ×1,000 (right two panels). TUNEL (red) and CD4 (blue) double-positive cells are shown as pink in lower magnification (left panel). CD4 (blue) and HIV p24gag (green) double-positive cells are shown as light green. Arrow shows a small artery. Right upper panel indicates TUNEL+, CD4+, and p24gag+ cells, which represented <2% of both TUNEL+ and CD4+ cells. Right lower panel indicates TUNEL+, CD4+, and p24gag− cells, which accounted for >90% of both TUNEL+ and CD4+ cells. (b) Dual color detection for GFP-expressing HIV-1 (GFP, green) and TUNEL staining (TRITC, red) of spleens from GFP/HIV-1–infected hu-PBL-NOD-SCID mice 2 wk after human PBL transplantation (9 d after infection). The majority of TUNEL+ cells were GFP/HIV-1 negative. Original magnification ×100.
Mentions: A similar extent of splenomegaly was observed in HIV-1–infected mice 2 wk after human PBL transplantation. However, the splenomegaly in infected mice regressed more rapidly than that of uninfected mice at 4 wk (data not shown). A marked decrease in the number of human CD4+ T cells was observed in infected mice with immunostaining analysis (data not shown). HIV-1 p24gag–expressing cells were found in the human CD4+ T cell–rich region; however, the number of p24gag+ cells was far lower than the number of human CD4+ T cells. In contrast, the number and distribution of human CD8+ T cells, human CD20+ B cells, and human CD68+ macrophages was apparently unchanged. TUNEL staining showed a large number of apoptotic cells around the small arteries in the spleen 2 wk after HIV-1 infection (Fig. 1 a). In contrast, only a few TUNEL+ apoptotic cells were observed in the spleens of uninfected hu-PBL-NOD-SCID mice at the same time point (data not shown). Furthermore, a NOD-SCID mouse section without human PBL transplantation showed no staining. The specificity of the TUNEL staining was confirmed as described in Materials and Methods.

Bottom Line: Apoptosis is a key for CD4+ T cell destruction in HIV-1-infected patients.A combination of TUNEL and immunostaining for death-inducing tumor necrosis factor (TNF) family molecules indicated that the apoptotic cells were frequently found in conjugation with TNF-related apoptosis-inducing ligand (TRAIL)-expressing CD3+CD4+ human T cells.These results suggest that a large number of HIV-1-uninfected CD4+ T cells undergo TRAIL-mediated apoptosis in HIV-infected lymphoid organs.

View Article: PubMed Central - PubMed

Affiliation: Department of Virology, Tohoku University Graduate School of Medicine, Sendai 980-8575, Japan.

ABSTRACT
Apoptosis is a key for CD4+ T cell destruction in HIV-1-infected patients. In this study, human peripheral blood lymphocyte (PBL)-transplanted nonobese diabetic (NOD)-severe combined immunodeficient (SCID) (hu-PBL-NOD-SCID) mice were used to examine in vivo apoptosis after HIV-1 infection. As the hu-PBL-NOD-SCID mouse model allowed us to see extensive infection with HIV-1 and to analyze apoptosis in human cells in combination with immunohistological methods, we were able to quantify the number of apoptotic cells with HIV-1 infection. As demonstrated by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), massive apoptosis was predominantly observed in virus-uninfected CD4+ T cells in the spleens of HIV-1-infected mice. A combination of TUNEL and immunostaining for death-inducing tumor necrosis factor (TNF) family molecules indicated that the apoptotic cells were frequently found in conjugation with TNF-related apoptosis-inducing ligand (TRAIL)-expressing CD3+CD4+ human T cells. Administration of a neutralizing anti-TRAIL mAb in HIV-1-infected mice markedly inhibited the development of CD4+ T cell apoptosis. These results suggest that a large number of HIV-1-uninfected CD4+ T cells undergo TRAIL-mediated apoptosis in HIV-infected lymphoid organs.

Show MeSH
Related in: MedlinePlus