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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 observe the effect of subcutaneous injection of recombinant soluble fusion mouse heat shock protein 70 (rHSP70) on the proportion of NK cells in monocytes in the spleen of mice. Methods Eighteen C57 / BL6 mice were randomly divided into three groups. Group 1 received subcutaneous injection of 20 μg rHSP70 (high dose group) in the neck, group 2 received 10 μg (low dose), group 3 received sodium phosphate buffer (control group), and each injection volume was 200 μL . 1 week 1 time, 2 times. The rats were sacrificed at week 3, and their mononuclear cells (MNBCs) were isolated from the spleen. Fluorescently labeled with phycoerythrin NK1.1 monoclonal antibody, the total number of NK cells and mononuclear cells was determined by flow cytometry, and the proportion of NK cells . Results The proportion of NK cells in high-dose group, low-dose group and control group were (12.7 ± 1.2)%, (34.3 ± 11.3)% and (10.0 ± 0.8)%, respectively. The difference between the dose group and the low dose group and the control group was statistically significant (P <0.01), and the ratio of NK cells to MNBC in the spleen of mice increased more than 2 folds. The spleen weights of the three groups were (0.086 ± 0.004), (0.090 ± 0.009) and (0.087 ± 0.006) g, respectively. There was no significant difference among the three groups (P> 0.05). Conclusion Low dose mice rHSP70 can significantly increase the proportion of spleen NK cells in MNBC, which may become a potential drug for immunotherapy. Each 20μg of high-dose but not as good as 10μg of low-dose promotion effect is obvious, suggesting that rHSP70 mouse spleen NK cells in two-way mode of adjustment.