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目的在我们先前的研究中发现,鼠疫疫苗免疫的猕猴脾组织中B细胞和T细胞数量明显增加。然而,是否这些细胞是由鼠疫疫苗引起的增殖性B细胞和T细胞还不得而知。方法为回答这个问题,本研究使用免疫组化双标记方法检测了猕猴脾组织中T细胞和B细胞的增殖。应用Ki67抗体以及T细胞和B细胞特异性单克隆抗体的免疫组化双标法,对脾组织T细胞和B细胞增殖进行检测。脾组织分别来自于亚单位疫苗SV1(20μg F1+10μg rV270)免疫的猕猴以及分别通过SV2(200μg F1+100μg rV270)、减毒活疫苗EV和铝佐剂免疫并分别攻毒的猕猴。结果与正常动物相比,受试动物脾组织生发中心有较多的B细胞增殖,边缘区有较多的静止B细胞,在动脉周围淋巴鞘区有较多的静止T细胞,提示原始T细胞可能在免疫早期发生增殖。经SV1、SV2或EV76免疫并攻毒的动物脾组织生发中心较仅用SV1免疫动物的生发中心扩大。此外,铝佐剂免疫并攻毒的猕猴脾组织生发中心不完整,这可能归因于强毒株鼠疫菌感染引起的病理损伤。B细胞增殖、静止B细胞增加、静止T细胞增多及生发中心扩大是诱导特异性体液免疫和保持免疫记忆反应的标志。结论这些结果与我们先前的发现的SV1、SV2或者EV76免疫动物激发较高的抗体和IL-4分泌相一致。
Aim In our previous study, we found that the number of B cells and T cells in macaque splenocytes immunized with plague vaccine was significantly increased. However, whether these cells are proliferative B cells and T cells caused by plague vaccine is unknown. Methods To answer this question, we used the immunohistochemical double-labeling method to detect the proliferation of T cells and B cells in the cynomolgus monkey spleen. Ki67 antibody and T cell and B cell specific monoclonal antibody immunohistochemical double labeling method was used to detect the spleen T cell and B cell proliferation. Spleen tissues were from macaques immunized with subunit vaccine SV1 (20 μg F1 + 10 μg rV270) and macaques immunized with SV2 (200 μg F1 + 100 μg rV270), live attenuated vaccine EV and aluminum adjuvant and challenged separately, respectively. Results Compared with normal animals, there were more B cell proliferation in the germinal centers of spleen tissues, more resting B cells in the marginal zone and more resting T cells in the perilymph peripheral area, suggesting that the original T cells Proliferation may occur early in the immunization. The spleen germinal centers of animals immunized and challenged with SV1, SV2 or EV76 were enlarged compared to the germinal centers of animals immunized with SV1 alone. In addition, the immunogenicity of aluminum adjuvant inoculated with rhesus macaque spleen hair germinal center is incomplete, which may be attributed to virulent strains of Y. infestans caused pathological damage. B cell proliferation, increased number of resting B cells, increased number of resting T cells and enlarged germinal centers are markers of specific humoral and immune memory responses. Conclusions These results are consistent with our previous findings that SV1, SV2 or EV76 immunized animals elicit higher antibodies and IL-4 secretion.