Folgueira L, McElhinny J A, Bren G D, MacMorran W S, Diaz-Meco M T, Moscat J, Paya C V
Department of Immunology, Mayo Clinic, Rochester, Minnesota 55905, USA.
J Virol. 1996 Jan;70(1):223-31. doi: 10.1128/JVI.70.1.223-231.1996.
The molecular mechanisms regulating human immunodeficiency virus (HIV) persistence in a major cell reservoir such as the macrophage remain unknown. NF-kappa B is a transcription factor involved in the regulation of the HIV long terminal repeat and is selectively activated following HIV infection of human macrophages. Although little information as to what signal transduction pathways mediate NF-kappa B activation in monocytes-macrophages is available, our previous work indicated that classical protein kinase C (PKC) isoenzymes were not involved in the HIV-mediated NF-kappa B activation. In this study, we have focused on atypical PKC isoenzymes. PKC-zeta belongs to this family and is known to be an important step in NF-kappa B activation in other cell systems. Immunoblotting experiments with U937 cells demonstrate that PKC-zeta is present in these cells, and its expression can be downmodulated by antisense oligonucleotides (AO). The HIV-mediated NF-kappa B activation is selectively reduced by AO to PKC-zeta. In addition, cotransfection of a negative dominant molecule of PKC-zeta (PKC-zeta mut) with NF-kappa B-dependent reporter genes selectively inhibits the HIV- but not phorbol myristate acetate- or lipopolysaccharide-mediated activation of NF-kappa B. That PKC-zeta is specific in regulating NF-kappa B is concluded from the inability of PKC-zeta(mut) to interfere with the basal or phorbol myristate acetate-inducible CREB- or AP1-dependent transcriptional activity. Lastly, we demonstrate a selective inhibition of p24 production by HIV-infected human macrophages when treated with AO to PKC-zeta. Altogether, these results suggest that atypical PKC isoenzymes, including PKC-zeta, participate in the signal transduction pathways by which HIV infection results in the activation of NF-kappa B in human monocytic cells and macrophages.
调节人类免疫缺陷病毒(HIV)在诸如巨噬细胞等主要细胞储存库中持续存在的分子机制仍不清楚。核因子κB(NF-κB)是一种参与HIV长末端重复序列调节的转录因子,在人类巨噬细胞受到HIV感染后被选择性激活。尽管关于哪些信号转导途径介导单核细胞-巨噬细胞中NF-κB激活的信息很少,但我们之前的研究表明,经典蛋白激酶C(PKC)同工酶不参与HIV介导的NF-κB激活。在本研究中,我们重点关注非典型PKC同工酶。PKC-ζ属于这个家族,已知在其他细胞系统中是NF-κB激活的重要步骤。对U937细胞进行的免疫印迹实验表明,PKC-ζ存在于这些细胞中,其表达可被反义寡核苷酸(AO)下调。AO作用于PKC-ζ可选择性降低HIV介导的NF-κB激活。此外,将PKC-ζ的负显性分子(PKC-ζ mut)与NF-κB依赖性报告基因共转染,可选择性抑制HIV介导的NF-κB激活,但不抑制佛波酯肉豆蔻酸酯乙酸盐或脂多糖介导的NF-κB激活。从PKC-ζ(mut)无法干扰基础或佛波酯肉豆蔻酸酯乙酸盐诱导的CREB或AP1依赖性转录活性可得出结论,PKC-ζ在调节NF-κB方面具有特异性。最后,我们证明,用AO处理PKC-ζ后,HIV感染的人类巨噬细胞中p24的产生受到选择性抑制。总之,这些结果表明,包括PKC-ζ在内的非典型PKC同工酶参与了HIV感染导致人类单核细胞和巨噬细胞中NF-κB激活的信号转导途径。