Institute of Nuclear Medicine and Allied Sciences, Delhi, India.
Cell Immunol. 2011;267(1):67-75. doi: 10.1016/j.cellimm.2010.11.005. Epub 2010 Nov 19.
In the present study, a semiquinone glucoside derivative (SQGD) isolated from a radioresistant bacterium Bacillus sp. INM-1 was evaluated for its immunostimulatory activities. Human peripheral blood mononuclear cells (PBMCs) were stimulated by different doses (30-90 microg/ml) of SQGD for different time (3-12h) intervals at 37°C, and IL-12p40, IL-23p19, IL-10, RelA and c-Jun gene expression analysis was carried out by qRT-PCR method. SQGD dose dependent cytokines protein expression kinetic analysis was carried out using western blotting. As the results of SQGD (30μg/ml) stimulation for 3h at 37°C, significant induction in IL-12p40, IL-23p19 and RelA gene expression was observed in PBMCs compared to unstimulated control cells. However, no such induction in IL-10 and c-Jun gene expression was observed. Time dependent protein expression study indicated significant increase in IL-12p40, IL-12p35, IL-23p19 and RelA protein expression at 3-6h, which was found decrease at 12h upon SQGD treatment. In contrast, IL-10 protein expression was found to enhance significantly at 12h after SQGD treatment to the PBMCs. SQGD dose dependent study showed approximately similar level of induction in IL-12p40, IL-12p35, IL-23p19 and RelA proteins expression at all tested concentration (30-90 microg/ml) compared to control. However, no significant change in the IL-10 and c-Jun protein expression was observed at any SQGD concentration. SQGD treatment (0.25mg/kgbwt.) was also found to enhance anti-keyhole Limpet Hemocynin (KLH) IgM antibodies significantly in the mice immunized by KLH. Thus, SQGD fraction stimulates cellular immunity by inducing immunostimulatory cytokines and humoral immunity by enhancing IgM antibodies and could be a promising immunostimulant. Further studies related to molecular mechanisms offering immunostimulation is underway, will certainly helpful to unravel its mode of action in the biological system.
在本研究中，从耐辐射细菌芽孢杆菌 INM-1 中分离出一种半醌糖苷衍生物（SQGD），评估其免疫刺激活性。将人外周血单核细胞（PBMC）在 37°C 下用不同剂量（30-90μg/ml）的 SQGD 刺激不同时间（3-12h），并用 qRT-PCR 方法进行 IL-12p40、IL-23p19、IL-10、RelA 和 c-Jun 基因表达分析。用 Western 印迹法进行 SQGD 剂量依赖性细胞因子蛋白表达动力学分析。结果表明，在 37°C 下用 SQGD（30μg/ml）刺激 3h 后，与未刺激对照细胞相比，PBMC 中观察到 IL-12p40、IL-23p19 和 RelA 基因表达的显著诱导。然而，在 IL-10 和 c-Jun 基因表达中没有观察到这种诱导。时间依赖性蛋白表达研究表明，在 SQGD 处理后 3-6h，IL-12p40、IL-12p35、IL-23p19 和 RelA 蛋白表达显著增加，12h 时减少。相反，在 SQGD 处理后 12h，发现 IL-10 蛋白表达显著增强。SQGD 剂量依赖性研究表明，与对照相比，在所有测试浓度（30-90μg/ml）下，IL-12p40、IL-12p35、IL-23p19 和 RelA 蛋白表达的诱导水平大致相似。然而，在任何 SQGD 浓度下，IL-10 和 c-Jun 蛋白表达均无明显变化。还发现 SQGD 处理（0.25mg/kgbwt.）可显著增强 KLH 免疫小鼠的抗 KLH IgM 抗体。因此，SQGD 级分通过诱导免疫刺激性细胞因子刺激细胞免疫，通过增强 IgM 抗体刺激体液免疫，并且可以是有前途的免疫刺激剂。正在进行与提供免疫刺激的分子机制相关的进一步研究，这将有助于揭示其在生物系统中的作用模式。
