Fig 7.

1,25D₃ rescue of M. tuberculosis-mediated TNF release in human alveolar macrophages from HIV⁺ persons is dependent on CD14. (A) AMs were incubated for 24 h in the presence or absence of 1,25D₃ pretreatment and then stained with PE-labeled anti-CD14 or isotype control antibody. Surface expression was measured by flow cytometry. Left panels show isotype control (gray lines)- and receptor (black lines)-labeled cells; right panels show receptor-labeled cells (gray lines) and 1,25D₃-treated receptor-labeled cells (black lines) (n = 3). (B) HIV⁺ AMs were treated with M. tuberculosis (MOI of 0.25:1), BCG (MOI of 10:1), or LPS (1 μg/ml) in the presence or absence of 1,25D₃ pretreatment (24 h) and the indicated antibodies. TNF in cell culture supernatants was measured by ELISA (n = 2). (C) 25D₃ levels were measured in the cell-free BALF of healthy and HIV-infected Indian patients with and without active M. tuberculosis infection (for HIV⁻ M. tuberculosis⁻ individuals, n = 38; for HIV⁻ M. tuberculosis⁺ individuals, n = 35; for HIV⁺ M. tuberculosis⁻ individuals, n = 12; and for HIV⁺ M. tuberculosis⁺ individuals, n = 17) by ELISA. Quantitative data represent means ± SEM. *, P < 0.05.