Figure 1.
Figure 1. Activation status, signaling response to phagocytic triggers, and bacterial clearance in HIV-1–infected macrophages. (A–D) Primary human macrophages were noninfected or infected with HIV-1ADAWT for 8 d. Total lysates were subjected to Western blotting with anti–phospho-ERK1/2 (A), anti–phospho-p38 (B), anti–phospho-SAPK/JNK (C), and anti–phospho-p65/RelA (D). The chemiluminescent signal was quantified and expressed as related to the noninfected condition, showing basal activation by HIV infection. (E) Macrophages infected for 8 d were incubated for different times with IgG-SRBCs at 37°C and then analyzed by Western blotting with anti–phospho ERK1/2 and anti–ERK1/2. (F) Results are expressed as a fold increase related to the basal condition for noninfected or HIV-1–infected cells. Means ± SEM of three different experiments are plotted. (G) Primary human macrophages were noninfected, infected with HIV-1ADAWT, or treated with lipopolysaccharide (LPS) or polyinosinic:polycytidylic acid (Poly:IC) for 8 d. The cells were then incubated for different times with IgG-SRBCs at 37°C and then analyzed by Western blotting with anti–phospho-ERK1/2, anti-ERK1/2, or anti-clathrin as a loading control. One of three representative experiments is presented. (H) Human macrophages were infected with HIV-1ADA WT or mock infected for 8 d. They were incubated or not (basal) with IgG-SRBCs (FcR), with complement-SRBCs (CR3), or with invasive S. typhimurium for 6 h at 37°C. Supernatants were collected and analyzed on human cytokine antibody arrays. Semiquantitative analysis was performed and the results are presented as a table, with white indicating no differential expression compared with noninfected conditions, light blue indicating down-regulation, and dark blue indicating higher down-regulation. n.d., not detected in control as well as HIV-infected conditions. Three independent experiments were performed with similar results. (I) The number of intracellular S. typhimurium at 24 h was divided by the number of bacteria at 1 h in HIV-1–infected or noninfected macrophages and results are expressed as related to noninfected cells. The mean ± SEM of four independent experiments is presented. *, P < 0.05.

Activation status, signaling response to phagocytic triggers, and bacterial clearance in HIV-1–infected macrophages. (A–D) Primary human macrophages were noninfected or infected with HIV-1ADAWT for 8 d. Total lysates were subjected to Western blotting with anti–phospho-ERK1/2 (A), anti–phospho-p38 (B), anti–phospho-SAPK/JNK (C), and anti–phospho-p65/RelA (D). The chemiluminescent signal was quantified and expressed as related to the noninfected condition, showing basal activation by HIV infection. (E) Macrophages infected for 8 d were incubated for different times with IgG-SRBCs at 37°C and then analyzed by Western blotting with anti–phospho ERK1/2 and anti–ERK1/2. (F) Results are expressed as a fold increase related to the basal condition for noninfected or HIV-1–infected cells. Means ± SEM of three different experiments are plotted. (G) Primary human macrophages were noninfected, infected with HIV-1ADAWT, or treated with lipopolysaccharide (LPS) or polyinosinic:polycytidylic acid (Poly:IC) for 8 d. The cells were then incubated for different times with IgG-SRBCs at 37°C and then analyzed by Western blotting with anti–phospho-ERK1/2, anti-ERK1/2, or anti-clathrin as a loading control. One of three representative experiments is presented. (H) Human macrophages were infected with HIV-1ADA WT or mock infected for 8 d. They were incubated or not (basal) with IgG-SRBCs (FcR), with complement-SRBCs (CR3), or with invasive S. typhimurium for 6 h at 37°C. Supernatants were collected and analyzed on human cytokine antibody arrays. Semiquantitative analysis was performed and the results are presented as a table, with white indicating no differential expression compared with noninfected conditions, light blue indicating down-regulation, and dark blue indicating higher down-regulation. n.d., not detected in control as well as HIV-infected conditions. Three independent experiments were performed with similar results. (I) The number of intracellular S. typhimurium at 24 h was divided by the number of bacteria at 1 h in HIV-1–infected or noninfected macrophages and results are expressed as related to noninfected cells. The mean ± SEM of four independent experiments is presented. *, P < 0.05.

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