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目的:观测双价痢疾工程菌苗滴鼻免疫小鼠后,不同时间、不同部位淋巴组织细胞表型的变化,探讨痢疾菌苗滴鼻免疫对黏膜和系统免疫应答的影响。方法:BALB/c小鼠随机分为3组,每组30只,分别以FSM-2117和FS-5416痢疾菌苗经滴鼻途径免疫小鼠4次,菌量依次为5×106、1×107、4×107和4×107CFU/只,对照组给予PBS,间隔2 wk。4次免疫后7、30和90 d活杀,分离NALT、鼻通道、脾、小肠PP结淋巴细胞,采用流式细胞术检测其淋巴细胞表型的变化。结果:4次免疫后7 d,小鼠鼻相关淋巴组织(NALT)、鼻通道(NP)和Peyer’s结(PP)淋巴细胞中,CD3+T细胞数均显著增加,其中以CD4+T细胞增加为主。FSM-2117免疫组的脾细胞中B220+细胞显著增加;而FS+5416免疫组的脾细胞中CD3+T细胞显著增加。4次免疫后30 d,NALT、NP和脾淋巴细胞仍出现上述变化;而90 d,仅NALT和NP淋巴细胞出现上述同样变化。结论:两株双价痢疾菌苗滴鼻免疫小鼠后,能有效地诱导黏膜和系统免疫应答,且持续时间较长,但该免疫应答的减弱是从距免疫部位较远的部位而开始的。
OBJECTIVE: To observe the changes of lymphocyte phenotypes in mice immunized with bivalent dysentery bacterin after intranasal immunization, and to investigate the effect of dysentery vaccine on nasal mucosal and systemic immune responses. Methods: BALB / c mice were randomly divided into 3 groups with 30 rats in each group. Mice were immunized with FSM-2117 and FS-5416 dysentery vaccine by intranasal route respectively. The bacterial counts were 5 × 106 and 1 × 107,4 × 107 and 4 × 107CFU / only, control group given PBS, interval 2 wk. After 4, 4 and 7 d, 30 and 90 d, the lymphocytes were separated from NALT, nasal passages, spleen and small intestine. The lymphocyte phenotypes were detected by flow cytometry. Results: The number of CD3 + T cells in nasal lymphoid tissue (NALT), nasal passage (NP) and Peyer’s knot (PP) lymphocytes increased significantly on day 7 after four immunizations, in which CD4 + T cells increased Mainly. The number of B220 + cells in splenocytes of FSM-2117 immunized group increased significantly, while the number of CD3 + T cells in splenocytes of FS + 5416 immunized group increased significantly. The changes of NALT, NP and splenic lymphocytes still appeared on the 30th day after the four immunizations. On the 90th day, the same changes occurred in only NALT and NP lymphocytes. CONCLUSIONS: Two bivalent dysentery vaccines can instil mucosal and systemic immune responses effectively after intranasal immunization in mice, and they last longer. However, the immune response is attenuated from the site far away from the immune site .