摘要
背景结核病是世界范围内单一致病菌感染导致死亡率最高的慢性感染性疾病,目前缺乏安全有效的疫苗预防成人结核病。而预防接种主要是利用机体具有免疫记忆的特征以阻止病原体的再次入侵,因此,研究T淋巴细胞的免疫记忆及增殖能力对新型结核病疫苗的设计至关重要。目的探讨新型结核病疫苗菌株(简称B/R菌株)对T淋巴细胞免疫记忆及增殖能力的影响。方法2019年1月,选取6~8周龄、SPF级雌性C57BL/6小鼠48只,随机分为磷酸盐缓冲液(PBS)组、卡介苗(BCG)组、结核分枝杆菌国际标准毒株(H37Ra)组和新型结核病疫苗(B/R)菌株组,各12只。PBS组、BCG组、H37Ra组和B/R菌株组分别皮下接种PBS、BCG、H37Ra和B/R菌株。4组分别于免疫后8周、12周、16周各取3只小鼠提取脾淋巴细胞体外刺激培养,利用流式细胞技术检测中央型记忆性淋巴细胞(TCM)及效应型记忆性淋巴细胞(TEM),酶联免疫吸附试验(ELISA)检测脾淋巴细胞分泌的抗原特异性Th1/Th2型细胞因子[白介素2(IL-2)、干扰素γ(IFN-γ)和白介素4(IL-4)]水平;羟基荧光素二醋酸盐琥珀酰亚胺酯(CFSE)标记法检测CD4+T淋巴细胞增殖率、CD8+T淋巴细胞增殖率。结果免疫后8、12、16周BCG组、H37Ra组、B/R菌株组CD4+的TCM和TEM高于PBS组(P<0.05);免疫后8周B/R菌株组CD4+的TEM低于H37Ra组(P<0.05);免疫后16周B/R菌株组CD4+的TCM高于H37Ra组和BCG组(P<0.05);免疫后16周B/R菌株组CD4+的TEM高于BCG组(P<0.05)。免疫后8、12、16周BCG组、H37Ra组、B/R菌株组CD8+的TCM和TEM高于PBS对照组(P<0.05);免疫后12周B/R菌株组CD8+的TEM高于H37Ra组和BCG组(P<0.05);免疫后16周,B/R菌株组CD8+的TCM和TEM高于BCG组(P<0.05)。免疫后8、12、16周PBS组IL-2和IFN-γ低于BCG组、H37Ra组、B/R菌株组(P<0.05);免疫后12周,B/R菌株组IFN-γ高于BCG组(P<0.05);免疫后16周,B/R菌株组IFN-γ和IL-2高于BCG组和H37Ra组(P<0.05)。BCG组、H37Ra组和B/R菌株组CD4+T淋巴细胞增殖率、CD8+T淋巴细胞增殖率高于PBS组(P<0.05);B/R菌株组的CD4+T淋巴细胞增殖率、CD8+T淋巴细胞增殖率均高于BCG组和H37Ra组(P<0.05)。结论B/R菌株免疫小鼠后,可促进机体产生更高水平的免疫保护性记忆细胞,以Th1型细胞免疫应答为主,并可增强T淋巴细胞增殖能力。
Background Tuberculosis(TB)is a global chronic infectious disease with the highest mortality caused by an infective agent.But there is a lack of safe and effective vaccines to prevent adult tuberculosis.Vaccination mainly uses body’s immune memory to prevent the re-invasion of pathogens.Therefore,studying the immune memory and proliferation capacity of T cells is crucial for the development of new tuberculosis vaccines.Objective To investigate the effects of a new tuberculosis vaccine strain(B/R strain)on T cell immune memory and proliferation capacity.Methods In January 2019,48 SPF female C57 BL/6 mice aged 6-8 weeks were randomly divided into PBS group,BCG group,H37 Ra group and B/R strain group,with 12 mice in each group,and were subcutaneously inoculated,respectively.At 8,12,and 16 weeks after immunization,the splenic lymphocytes of three mice in each group were extracted and cultured in vitro,and the levels of central memory lymphocytes(TCM)and effector memory lymphocytes(TEM)were detected by flow cytometry.ELISA was used to detect antigen-specific Th1/Th2 cytokines(IL-2,IFN-γand IL-4)levels secreted by splenic lymphocytes.CFSE labeling method was used to detect the proliferation of CD4+and CD8+T lymphocytes.Results At 8,12,and 16 weeks after immunization,the TCM and TEM levels of CD4+in BCG,H37 Ra,and B/R strain groups were higher than those of PBS group(P<0.05).At 8 weeks post-inoculation,the level of CD4+TEM in B/R group was lower than that in H37 Ra group(P<0.05).At 16 weeks post-inoculation,B/R strain group showed higher level of TCM of CD4+than H37 Ra and BCG groups(P<0.05),and higher level of TEM of CD4+than BCG group(P<0.05).At 8,12,and 16 weeks after immunization,the TCM and TEM levels of CD8+in BCG,H37 Ra,and B/R strain groups were higher than those of PBS group(P<0.05).At 12 weeks post-inoculation,the level of TEM of CD8+in B/R strain group was higher than that in H37 Ra and BCG groups(P<0.05).At 16 weeks post-inoculation,the TCM and TEM levels of CD8+in B/R strain group were higher than those of BCG group(P<0.05).At 8,12,and 16 weeks post-inoculation,IL-2 and IFN-γlevels in PBS group were lower than those of other three groups(P<0.05).At 12 weeks post-inoculation,the level of IFN-γin B/R strain group was higher than that of BCG group(P<0.05).At 16 weeks post-inoculation,IFN-γand IL-2 levels in B/R strain group were higher than those of BCG and H37 Ra groups(P<0.05).The proliferation rates of CD4+and CD8+T lymphocytes in BCG,H37 Ra and B/R strain groups were higher than those of PBS group(P<0.05).The proliferation rates of CD4+and CD8+T lymphocytes in B/R strain group were higher than those of BCG and H37 Ra groups(P<0.05).Conclusion The immunizing with the B/R strain in mice could awake up the body to produce higher levels of protective immune memory cells,mainly Th1 type immune responses,and enhance the proliferation capacity of T cells.
作者
邵萌
吴芳
张杰
董江涛
柳小玲
吴江东
章乐
赵海军
张万江
SHAO Meng;WU Fang;ZHANG Jie;DONG Jiangtao;LIU Xiaoling;WU Jiangdong;ZHANG Le;ZHAO Haijun;ZHANG Wanjiang(Shihezi University School of Medicine/Xinjiang High Incidence of Local and Ethnic Diseases,Key Laboratory of the Ministry of Education,Shihezi 832002,China;First Affiliated Hospital,School of Medicine,Shihezi University,Shihezi 832002,China)
出处
《中国全科医学》
CAS
北大核心
2021年第11期1376-1382,共7页
Chinese General Practice
基金
NSFC-新疆联合基金项目(U1803127)
兵团重点领域科技攻关计划项目(2018AB019)。
关键词
结核
疫苗
结核菌苗
卡介苗
T淋巴细胞
免疫
细胞
Tuberculosis
Vaccines
Tuberculosis vaccines
BCG Vaccine
T-Lymphocytes
Immunity,cellular
