Munis J R, Richman D D, Kornbluth R S
Department of Medicine, University of California, San Diego.
J Clin Invest. 1990 Feb;85(2):591-6. doi: 10.1172/JCI114478.
The synthesis of tumor necrosis factor (TNF)/cachectin was assessed in primary monocyte-derived macrophage (MDM) cultures after in vitro infection with a macrophage-tropic strain of HIV-1 (HTLV-IIIBa-L/85). Productive and cytopathic infections in MDM cultures were established using a high multiplicity of infection (m.o.i. = 3) under conditions that minimized endotoxin contamination. Culture supernatants were tested for TNF/cachectin activity by L929 cell cytotoxicity assay, and TNF/cachectin mRNA was assessed by a sensitive PCR amplification technique that could detect between 1 and 10 cells fully activated for TNF/cachectin expression. Unstimulated MDM cultures produced no detectable levels of TNF/cachectin activity or mRNA, consistent with previous demonstrations that production of this cytokine by macrophages is an inducible and not a constitutive event. HIV-1 infection failed to induce detectable TNF/cachectin activity or mRNA in these unstimulated cultures. In addition, the responsiveness of macrophages to lipopolysaccharide (LPS) induction of TNF/cachectin production was assessed in dose-response and kinetic experiments. No differences between infected and uninfected cultures were discernable. These results demonstrate that productive and cytopathic infection with a macrophage-tropic strain of HIV-1 does not alter the regulation of TNF/cachectin expression in macrophages.
在用巨噬细胞嗜性HIV-1株(HTLV-IIIBa-L/85)进行体外感染后,在原代单核细胞衍生巨噬细胞(MDM)培养物中评估肿瘤坏死因子(TNF)/恶病质素的合成。在将内毒素污染降至最低的条件下,使用高感染复数(感染复数=3)在MDM培养物中建立有生产性和细胞病变性的感染。通过L929细胞细胞毒性试验检测培养上清液中的TNF/恶病质素活性,并通过一种敏感的PCR扩增技术评估TNF/恶病质素mRNA,该技术能够检测1至10个被完全激活以表达TNF/恶病质素的细胞。未受刺激的MDM培养物未产生可检测水平的TNF/恶病质素活性或mRNA,这与之前的证明一致,即巨噬细胞产生这种细胞因子是一个诱导性而非组成性事件。在这些未受刺激的培养物中,HIV-1感染未能诱导可检测到的TNF/恶病质素活性或mRNA。此外,在剂量反应和动力学实验中评估了巨噬细胞对脂多糖(LPS)诱导TNF/恶病质素产生的反应性。未感染和感染的培养物之间没有明显差异。这些结果表明,用巨噬细胞嗜性HIV-1株进行有生产性和细胞病变性的感染不会改变巨噬细胞中TNF/恶病质素表达的调节。