重组小鼠热休克蛋白70对小鼠脾脏中NK细胞的影响(英文)

Effect of recombinant HSP70 on splenic NK cell population in mouse

目的观察皮下注射重组可溶性融合小鼠热休克蛋白70(rHSP70)后对小鼠脾脏中NK细胞在单核细胞中所占比例的影响。方法取C57/BL6小鼠18只,随机等分3组。第1组每只在颈部皮下注射20μgrHSP70(高剂量组),第2组每只注射10μg(低剂量组),第3组注射磷酸钠缓冲液(对照组),注射体积均为每只200μL。1周1次,共2次。第3周杀鼠,从脾分离其单个核细胞(MNBC),以藻红蛋白NK1.1单抗荧光标记后,在流式细胞仪上测定NK细胞及单个核细胞总数,计算NK细胞的比例。结果NK细胞数在高剂量组、低剂量组和对照组中的比例分别占(12.7±1.2)%、(34.3±11.3)%和(10.0±0.8)%,经方差分析,高剂量组与低剂量组间,以及低剂量组与对照组组间在统计学上差异均非常显著(P<0.01),其小鼠脾脏中NK细胞占MNBC之比例增加了2倍多。而3组的脾脏重量分别为(0.086±0.004)、(0.090±0.009)和(0.087±0.006)g,各组间差异均无统计学意义(P>0.05)。结论低剂量小鼠rHSP70可大幅提高小鼠脾脏NK细胞在MNBC中的比例,它有可能成为一种免疫治疗的潜在药物。每只20μg的高剂量反而不如10μg低剂量的促进效果明显,提示rHSP70对小鼠脾脏NK细胞的调剂存在双向模式。

【Objective】To construct the recombinant heat shock protein 70 （rHSP70） expression plasmid （pET-28b-HSP70）, express and purify the rHSP70, and finally investigate the effect of soluble murine rHSP70 on splenic natural killer （NK） cells in mice. 【Methods】Eighteen mice （C57BL/6） were randomly divided into three groups with six mice in each： High dose group （HDG）, low dose group （LDG） and control group （CG）. The mice in HDG, LDG and CG were injected with 20 μg, 10 μg of rHSP70 and phosphate buffered saline （PBS）, respectively. The injection was administered subcutaneously on the neck in a volume of 200 μL once a week for two weeks. Two weeks later, spleens were all weighed, mononuclear blood cells （MNBCs） were also isolated from spleen and directly stained with phycoerythrin （PE）-conjugated monoclonal anti-NK1.1 antibody. The NK cells and MNBCs were both assayed with flow cytometry. 【Results】The average proportions of splenic NK cells of HDG, LDG and CG were （12.7±1.2） %, （34.3±11.3） % and （10.0±0.8） %, respectively. There were significant differences between the low dose group LDG and each of the remaining two groups （both P 〈0.01）. The weights of spleens of three groups were （0.086±0.004）, （0.090±0.009） and （0.087±0.006）g, respectively,There were no differences between them （P 〉0.05）. 【Conclusion】 The results suggest that low dose of murine rHSP70 may increase the subpopulation of NK cells in murine spleen, which might be a potential biological regulation drug. The observation that the high dose of rHSP70 does not have so remarkable promoting effect on proportions of splenic NK cells in mice as the low dose suggests that there are bi-directional regulation effects of rHSP70 on splenic NK cells.