Figure 4
Infectious and transgenic HIV expression increase the expression of ALOX5 and release of leukotrienes (a) HIV increases the expression of ALOX5 and (b) Cysteinyl leukotrienes (Cys-LT) in monocyte-derived macrophages (MDM). MDM were cultured with HIV-1ADA for 1 h and processed for detection of ALOX5 mRNA expression by quantitative real time PCR and CysLT by ELISA (n = 5 for each condition). (c) Increased expression of ALOX5 and (d) CysLT in cultured HPAEC cultured with HIV-MDM conditioned media for 24 h. HIV-MDM medium was diluted to clinically-relevant levels of p24 or 50 pg/ml98. (e) HIV-1 transgene increases ALOX5 expression in vivo. HIV transgenic rat lungs were evaluated for the expression of ALOX5 mRNA by quantitative real time PCR, compared to wild-type rats, using GAPDH as internal control. The relative expression of ALOX5 in rat lungs is expressed as percent of control. (f) The increased expression of ALOX5 was confirmed by Western blot, normalized to GAPDH. For all datasets: *p < 0.05 when compared to control. **p < 0.001 when compared to controls. (g) HIV antiretroviral therapy reverses HIV-induced increase in leukotriene levels in bronchoalveolar lavage fluid. We measured the levels of leukotriene LTB4 (n = 3–4) and cysteinyl leukotrienes (CysLT, n = 6–10) in bronchoalveolar lavage fluid (BALF) from uninfected (HIV-), HIV-1 positive (HIV+), and HIV+ patients receiving antiretroviral therapy (HIV + ART). *p < 0.01 when compared to BALF from control, uninfected subjects. **p < 0.001 when compared to HIV+ subjects.
