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目的通过检测肥胖小鼠的乙肝疫苗接种效果和免疫系统功能,探讨肥胖影响乙肝疫苗预防效果的免疫学机制。方法对WNIN/Ob肥胖小鼠(突变型)和正常体重小鼠(野生型)接种4μg乙肝疫苗(3,4月),第二针接种7d后,流式细胞检测脾脏细胞中总T细胞及T细胞亚群的比例,MTT法检测脾脏淋巴细胞对刀豆球蛋白A(ConA)、乙肝表面抗原(HBsAg)的增殖试验,酶联免疫吸附试验(ELISA)检测小鼠血清中抗-HBs浓度。脂多糖(LPS)刺激腹腔巨噬细胞,上清液中肿瘤坏死因子α(TNF-α)、一氧化氮(NO)浓度分别用ELISA和Griess法检测。结果与正常组相比,肥胖组小鼠脾脏中总T细胞和Th细胞比例下降(P<0.05)、血清抗-HBs降低(P<0.05)、脾脏淋巴细胞对ConA、HBsAg的增殖率都降低(P<0.05)。腹腔巨噬细胞上清液中NO、TNF-α浓度差异有统计学意义(P<0.05)。结论肥胖小鼠先天性免疫和特异性免疫功能均受损,导致对乙肝疫苗免疫应答降低。
OBJECTIVE: To investigate the immunological mechanism of obesity affecting the preventive effect of hepatitis B vaccine by detecting the effect of hepatitis B vaccination and immune system in obese mice. Methods WNIN / Ob obese mice (mutant) and normal weight mice (wild type) were inoculated with 4μg Hepatitis B vaccine (3,4 months). After the second injection for 7 days, the total T cells in spleen cells were detected by flow cytometry and T lymphocyte subsets, MTT assay was used to detect proliferation of ConA and HBsAg in splenic lymphocytes. Serum anti-HBs concentration was measured by enzyme-linked immunosorbent assay (ELISA) . Peritoneal macrophages were stimulated with lipopolysaccharide (LPS). The concentrations of tumor necrosis factor-alpha (TNF-α) and nitric oxide (NO) in the supernatant were detected by ELISA and Griess method respectively. Results Compared with the normal group, the percentage of total T cells and Th cells in obese mice decreased (P <0.05) and the serum anti-HBs decreased (P <0.05), and the proliferation rate of ConA and HBsAg in spleen lymphocytes decreased (P <0.05). The concentration of NO and TNF-αin the peritoneal macrophage supernatant had statistical significance (P <0.05). Conclusion The congenital immunity and specific immune function of obese mice are impaired, leading to the decrease of immune response to hepatitis B vaccine.