Journal:

Article Title: Human and Simian Immunodeficiency Virus-Mediated Upregulation of the Apoptotic Factor TRAIL Occurs in Antigen-Presenting Cells from AIDS-Susceptible but Not from AIDS-Resistant Species ▿ †

doi: 10.1128/JVI.02616-06

Figure Lengend Snippet: Expression analysis of selected IFN-inducible genes in different nonhuman primate iDC. (a) RT activity in macrophage supernatants. (b) Tat RNA levels measured by real time RT-PCR. (c) Induction of selected IFN-inducible genes based on RNA levels from human and nonhuman primate iDC after HIVBal or SIVmac316 infection (day 14) and Tat expression (30 h). Results are reported as the average (n-fold) induction relative to that of the time zero mock-infected sample. Two samples from two independent donors are reported. A positive control for the real-time RT-PCR in RNAs from each species was provided by an RNA sample from CD14+ cells treated with IFN-α. All detection values are normalized according to the GAPDH internal control. (d) Cytokine detection in primate iDC supernatants after HIVBal or SIVmac316 infection (means ± standard errors of three independent samples).

Article Snippet: CD14 + cells, as determined by staining with an anti-CD14 antibody (Miltenyi Biotec, Auburn, CA) were seeded at a density of 10 6 cells/ml with interleukin 4 (100 ng/ml) and granulocyte-macrophage colony-stimulating factor (GM-CSF; 200 ng/ml) ( 3 ).

Techniques: Expressing, Activity Assay, Quantitative RT-PCR, Infection, Positive Control, Control

Journal:

Article Title: Human and Simian Immunodeficiency Virus-Mediated Upregulation of the Apoptotic Factor TRAIL Occurs in Antigen-Presenting Cells from AIDS-Susceptible but Not from AIDS-Resistant Species ▿ †

doi: 10.1128/JVI.02616-06

Figure Lengend Snippet: Apoptosis and TRAIL production in lentivirus-infected primate PBMC cultures. Levels of apoptosis in (a) HIV- and (b) SIV-infected PBMC from human and nonhuman primates. Panels in the left column display the percentage of CD4+ T cells in the PBMC cultures that stain for annexin V, a marker for apoptosis. Panels in the middle column report the flow cytometric analysis of CD4+ T-cell viability in PBMC. Values at each time point are expressed as the percentage of viable CD4+ T cells present in the mock-infected sample. Panels in the right column report virus production in infected primate PBMC measured by p24 or p27 ELISA assay. The data represent the mean percentages ± standard errors (SE) of five (human) and three (other species) experiments. PBMC subpopulation mean ± SE percentages on the day of infection were as follows: for human PBMC, CD3, 71 ± 4.5; CD4, 45 ± 2.8; CD8, 24 ± 3.6; CD14, 13 ± 3.7; CD11c, 1.04 ± 0.30; CD123, 0.70 ± 0.14; for chimpanzee PBMC, CD3, 73 ± 4.1; CD4, 41 ± 2.8; CD8, 21 ± 1.2; CD14, 9.2 ± 0.7; CD11c, 1.03 ± 0.19; CD123, 0.47 ± 0.12; for RM PBMC, CD3, 67 ± 4.2; CD4, 32 ± 3.9; CD8, 33 ± 2.1; CD14, 7.9 ± 0.9; CD11c, 1.87 ± 0.34; CD123, 0.53 ± 0.12; for AGM PBMC, CD3, 68 ± 5; CD4, 19 ± 3.6; CD8, 49 ± 6.9; CD14, 7.7 ± 2.5; CD11c, 0.77 ± 0.18; CD123, 0.59 ± 0.24; for SM PBMC, CD3, 72 ± 1.6; CD4, 23 ± 1.4; CD8, 50 ± 3; CD14, 14.6 ± 1.7; CD11c, 0.81 ± 0.30; CD123, 0.54 ± 0.13. HIV or SIV infection mean ± SE percentages (determined by p24 or p27 intracellular staining on day 7 postinfection in CD4+ T cells or in CD14+ cells) for HIV-infected human PBMC were CD4, 15 ± 2.9; CD14, 10.1 ± 2; for SEB/HIV-infected human PBMC were CD4, 24.8 ± 5.3; CD14, 15.2 ± 4.5; for HIV-infected chimpanzee PBMC were CD4, 9 ± 1.5; CD14, 5 ± 1.5; for SEB/HIV-infected chimp PBMC were CD4, 14.2 ± 1.8; CD14, 9 ± 2.1; for SIV-infected RM PBMC were CD4+, 12 ± 3; CD14, 5 ± 1.9; for SEB/SIV-infected RM PBMC were CD4, 18 ± 4; CD14, 8 ± 2.6; for SIV-infected AGM PBMC were CD4, 12.6 ± 2.2; CD14, 4.8 ± 1.7; for SEB/SIV-infected AGM PBMC were CD4, 16.9 ± 3.7; CD14, 7.3 ± 3.8; for SIV-infected SM PBMC were CD4, 8.9 ± 1.9; CD14, 5.5 ± 2.1; for SEB/SIV-infected SM PBMC were CD4, 18.1 ± 5.3; CD14, 8.7 ± 3.9. (c) Levels of IFN-α, IFN-γ, and soluble TRAIL in primate PBMC culture supernatants after infection with HIVBal or SIVmac239 during 10 days of culture (means ± SE from three experiments).

Article Snippet: CD14 + cells, as determined by staining with an anti-CD14 antibody (Miltenyi Biotec, Auburn, CA) were seeded at a density of 10 6 cells/ml with interleukin 4 (100 ng/ml) and granulocyte-macrophage colony-stimulating factor (GM-CSF; 200 ng/ml) ( 3 ).

Techniques: Infection, Staining, Marker, Virus, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Human and Simian Immunodeficiency Virus-Mediated Upregulation of the Apoptotic Factor TRAIL Occurs in Antigen-Presenting Cells from AIDS-Susceptible but Not from AIDS-Resistant Species ▿ †

doi: 10.1128/JVI.02616-06

Figure Lengend Snippet: TRAIL and TRAIL death receptor (DR4 and DR5) expression in HIV- or SIV-exposed PBMC. (a) Percentage of mTRAIL+ PBMC in PBMC from human (closed symbols) and chimpanzee (open symbols) PBMC during 10 days of culture. The data represent the means ± standard errors (SE) of five experiments. (b) Percentage of human CD4+ T cells, monocytes (CD14+), and iDC (CD11c/HLA-DR+) expressing mTRAIL after HIV infection within each population in the total PBMC cultures. The data represent the means ± SE of three experiments. (c) Analysis of TRAIL receptors DR4 and DR5 in human and chimpanzee PBMC infected with HIV. Mean percentages ± SE of cells expressing the receptor are reported for total PBMC and for CD4+ T cells present in the PBMC culture (data derived from three independent donors). (d) Effect of antibodies against TRAIL and TRAIL receptors on HIV-induced PBMC apoptosis, showing the percentage of apoptotic human CD4+ T cells present on day 7 of culture under different conditions. Five experiments were carried out, and the average percentages of apoptosis observed for the mock-treated samples were subtracted from the other experimental values. (e) Effect of antibodies against CD95, CD95L, IFN-α, and IFN-αR on HIV-induced CD4+ T-cell apoptosis (mean percentages of three independent samples). Rates of HIV or SIV infection shown by ICS on day 7 in all the virus-infected cultures were comparable.

Article Snippet: CD14 + cells, as determined by staining with an anti-CD14 antibody (Miltenyi Biotec, Auburn, CA) were seeded at a density of 10 6 cells/ml with interleukin 4 (100 ng/ml) and granulocyte-macrophage colony-stimulating factor (GM-CSF; 200 ng/ml) ( 3 ).

Techniques: Expressing, Infection, Derivative Assay, Virus

Journal:

Article Title: Human and Simian Immunodeficiency Virus-Mediated Upregulation of the Apoptotic Factor TRAIL Occurs in Antigen-Presenting Cells from AIDS-Susceptible but Not from AIDS-Resistant Species ▿ †

doi: 10.1128/JVI.02616-06

Figure Lengend Snippet: In vivo TRAIL, DR4, and DR5 expression analysis. (a) Detection of soluble TRAIL in supernatants of nonhuman primate CD14+ cells by a human TRAIL ELISA, showing means ± standard errors of supernatant levels after rIFN-α stimulation of three PBMC-derived CD14+monocyte samples. (b) sTRAIL serum levels in SIV-infected and noninfected RM and SM samples. These samples were obtained from 20 macaques sampled multiple times before and after infection with SIVmac251 and from 74 SIV-positive and 35 SIV-negative SM. The SIV-positive SM were naturally infected, with the exception of two animals experimentally infected with SIVmac239. (c) Percentages of mTRAIL+ cells in PBMC and CD4+ T cells obtained from SIV-infected and noninfected RM and SM. (d) Percentages of DR4 and DR5+ cells in CD4+ T cells obtained from SIV-infected and noninfected RM and SM. (e) Correlation between SIV RNA viral loads in RM and serum TRAIL levels.

Article Snippet: CD14 + cells, as determined by staining with an anti-CD14 antibody (Miltenyi Biotec, Auburn, CA) were seeded at a density of 10 6 cells/ml with interleukin 4 (100 ng/ml) and granulocyte-macrophage colony-stimulating factor (GM-CSF; 200 ng/ml) ( 3 ).

Techniques: In Vivo, Expressing, Enzyme-linked Immunosorbent Assay, Derivative Assay, Infection