Yin Ling-fan, Fan Yan-ying, Li Li, Wu Chang-you
Department of Immunology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2007 Jul;23(7):623-6.
To investigate the effect of the TLR ligand (R-848) and IL-12 on the production of IFN-gamma by human NK cell subsets.
PBMC or purified NK cells were isolated from normal human peripheral blood and cultured with R-848, IL-12 , R-848+ IL-12. The level of IFN-gamma in the culture supernatants was measured by ELISA. The cell subsets which produced IFN-gamma were detected and analyzed by flow cytometry(FCM).
PBMC cultured with different concentrations of TLR ligands (R-848, LPS and CpG) could induce IFN-gamma production in a dose dependent manner. Among these three TLR ligands, R-848 was the best one in induction of IFN-gamma production by PBMC. FCM analysis indicated that R-848 could induce IFN-gamma expression by CD56(+) cells, but not CD4(+) or CD8(+) T cells. Similarly, IL-12 could stimulate NK cells to produce IFN-gamma. In addition, R-848 and IL-12 had synergistic effect on the production of IFN-gamma by PBMC and purified NK cells including CD56(bright) and CD56(dim) subsets.
Engagement of TLR7/8 by R-848 stimulation with IL-12 could induce IFN-gamma production by CD56(bright) NK cells which indicated that TLRs and cytokines played an important role in the regulation of biologic function of NK cells.
研究Toll样受体（TLR）配体（R-848）和白细胞介素-12（IL-12）对人自然杀伤（NK）细胞亚群产生γ干扰素（IFN-γ）的影响。
从正常人外周血中分离出外周血单个核细胞（PBMC）或纯化的NK细胞，并用R-848、IL-12、R-848 + IL-12进行培养。通过酶联免疫吸附测定（ELISA）法检测培养上清液中IFN-γ的水平。采用流式细胞术（FCM）检测并分析产生IFN-γ的细胞亚群。
用不同浓度的TLR配体（R-848、脂多糖（LPS）和CpG）培养PBMC可呈剂量依赖性地诱导IFN-γ的产生。在这三种TLR配体中，R-848在诱导PBMC产生IFN-γ方面效果最佳。FCM分析表明，R-848可诱导CD56(+)细胞表达IFN-γ，但不能诱导CD4(+)或CD8(+) T细胞表达。同样，IL-12可刺激NK细胞产生IFN-γ。此外，R-848和IL-12对PBMC和纯化的NK细胞（包括CD56(bright)和CD56(dim)亚群）产生IFN-γ具有协同作用。
R-848刺激结合IL-12激活TLR7/8可诱导CD56(bright) NK细胞产生IFN-γ，这表明TLR和细胞因子在NK细胞生物学功能的调节中起重要作用。
