Figure 1
From: HIV infection and latency induce a unique metabolic signature in human macrophages
A small population of HIV infected macrophages survives infection and shows mitochondrial compromise. PBMCs were isolated from leukopacks provided by the NY blood center. Macrophages were isolated by adherence in the presence of M-CSF for 7 dpi. Cultures were exposed to 50 ng/ml of HIVADA for 24 h, washed, and maintained for 3, 7, 14, and 21 dpi for subsequent analysis. (A) HIV replication as determined by HIV-p24 ELISA in the supernatant of uninfected (control) and HIV infected macrophage cultures (early, mid, and late). HIV at the late stages becomes silent. HIV DNA staining and Alu-PCR confirmed these results. (B) Staining for HIV integrated DNA (nef), viral mRNA (nef) and HIV-p24 protein and subsequent analysis for microscopy. We quantified the % of positive cells at the early, mid, and late stage, indicating that at later time points, viral mRNA and protein expression is decreased in the cells. (C) However, quantification of the staining in B, indicates that integrated HIV DNA remains stable, viral mRNA decreased at the late stages, and HIV-p24 remain stable from the mid stages. (D) Percentage of cell death in uninfected (control conditions) and HIV infected conditions (HIV). HIV infection induces a large amount of cell death, but early on a population of HIV-p24 positive cells survive the infection. In parenthesis is the % of HIV infected cells. (E,F) correspond to representative pictures of transmission electron microscopy (TEM) of macrophages in control conditions (uninfected, UI) and HIV infected cells (HIV) to observe the distribution, numbers, and size of mitochondria. (G) Quantification of mitochondrial size (nm2) per area. *p = 0.0341, n = 7.
