Allo-Antigen Stimulated CD8+ T-Cells Suppress NF-kappa B and Ets-1 DNA Binding Activity, and Inhibit Phosphorylated NF-kappa B p65 Nuclear Localization in CD4+ T-cells

作者：Nagashima Ryuichi; Kawakami Fumitaka; Takahashi Shinichiro; Obata Fumiya; Kubo Makoto^*

来源：Viral Immunology, 2014, 27(6): 305-315.

DOI：10.1089/vim.2013.0113

CD8+ T-cells of asymptomatic HIV-1 carriers (AC) suppress human immunodeficiency virus type 1 (HIV-1) replication in a class I major histocompatibility complex (MHC-I)-restricted and -unrestricted manner. In order to investigate the mechanism of MHC-I-unrestricted CD8+ T-cell-mediated HIV-1 suppression, we previously established allo-antigen stimulated CD8+ T-cells from HIV-1-uninfected donors. These allo-antigen stimulated CD8+ T-cells suppressed HIV-1 replication in acutely infected autologous CD4+ T-cells when directly co-cultured. To elucidate the mechanism of HIV-1 replication suppression, we analyzed DNA-binding activity and phosphorylation of transcriptional factors associated with HIV-1 replication by electrophoresis mobility shift assay and Western blotting. When CD4+ T-cells were cultured with allo-antigen stimulated CD8+ T-cells, the reduction of NF-kappa B and Ets-1 DNA-binding activity was observed. Nuclear localization of NF-kappa B p65 and Ets-1 was suppressed in CD4+ T-cells. Although NF-kappa B p65 and Ets-1 are known to be regulated by protein kinase A (PKA), no difference was observed in the expression and phosphorylation of the PKA catalytic subunit in CD4+ T-cells cultured with PHA-treated CD8+ T-cells or allo-antigen stimulated CD8+ T-cells. Cyclic AMP is also known to enter through gap junctions, but the suppression of HIV-1 replication mediated by allo-antigen stimulated CD8+ T-cells was not affected by the gap junction inhibitor. The nuclear transport of phosphorylated NF-kappa B p65 (Ser276) was inhibited only in CD4+ T-cells cultured with allo-antigen stimulated CD8+ T-cells. Our results indicate that allo-antigen stimulated CD8+ T-cells suppress the transcriptional activity of NF-kappa B p65 or Ets-1 in an antigen-nonspecific manner, and inhibit the nuclear transport of phosphorylated NF-kappa B p65 (Ser276).

• 出版日期2014-8