摘要
目的结核分枝杆菌(Mtb)耐药菌株的流行加剧了结核病(TB)的全球负担,凸显了对新的TB治疗策略的需求。方法表达环二腺苷酸(c-di-AMP)分解酶磷酸二酯酶B(CnpB)的重组腺病毒疫苗(rAd-CnpB)经黏膜免疫正常小鼠,单独或联合药物免疫治疗Mtb呼吸道感染小鼠。ELISA检测小鼠血清及肺泡灌洗液(BALF)抗体水平,实时定量PCR检测小鼠肺组织中细胞因子γ干扰素(IFN-γ)、白细胞介素10(IL-10)转录水平,流式细胞术检测肺、脾CD4^(+)、CD8^(+)T细胞亚群细胞比例,ELISA检测脾细胞抗原刺激后的细胞因子IFN-γ、IL-2、IL-10、炎症因子IL-6、肿瘤坏死因子α(TNF-α)分泌水平,平板计数法计数Mtb感染小鼠肺、脾的荷菌数。结果rAd-CnpB滴鼻免疫可诱导小鼠血清高滴度的IgG,以及BALF中IgG和IgA产生,肺、脾T淋巴细胞亚群改变。rAd-CnpB单独或联合药物治疗Mtb感染小鼠可诱导血清IgG水平、BALF中IgA和IgG水平显著增高,rAd-CnpB免疫促进Mtb感染小鼠的脾细胞CnpB抗原特异性的细胞因子和炎症因子分泌。rAd-CnpB单独或联合药物免疫治疗并不显著影响Mtb呼吸道感染小鼠肺、脾荷菌数。结论rAd-CnpB经黏膜免疫可诱导小鼠显著的黏膜、体液免疫反应,并显著增强Mtb感染小鼠中CnpB特异性细胞免疫应答。
Objective The prevalence of drug-resistant Mycobacterium tuberculosis(Mtb)strains is exacerbating the global burden of tuberculosis(TB),highlighting the urgent need for new treatment strategies for TB.Methods The recombinant adenovirus vaccine expressing cyclic di-adenosine monophosphate(c-di-AMP)phosphodiesterase B(CnpB)(rAd-CnpB),was administered to normal mice via mucosal immunization,either alone or in combination with drug therapy,to treat Mtb respiratory infections in mice.Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of antibodies in serum and bronchoalveolar lavage fluid(BALF).Real-time quantitative PCR was performed to assess the transcription levels of cytokines interferonγ(IFN-γ)and interleukin 10(IL-10)in mouse lungs.Flow cytometry was used to determine the proportions of CD4^(+)and CD8^(+)T cell subsets in the lungs and spleens.ELISA was employed to measure the levels of cytokines IFN-γ,IL-2,IL-10,inflammatory factors IL-6,and tumor necrosis factorα(TNF-α)secreted by spleen cells following antigen stimulation.The bacteria loads in the lungs and spleens of Mtb-infected mice were enumerated by plate counting methods.Resluts Intranasal immunization with rAd-CnpB induced high titers of IgG in mouse serum and the production of IgG and IgA in BALF,along with alterations in T lymphocyte subsets in the lungs and spleens.Administration of rAd-CnpB,either alone or in combination with drugs,to Mtb-infected mice significantly increased serum IgG levels as well as IgA and IgG levels in BALF.rAd-CnpB immunization promoted the secretion of CnpB-specific cytokines and inflammatory factors by splenocytes in Mtb-infected mice.However,rAd-CnpB immunotherapy,either alone or combined with drugs,did not significantly affect the bacterial loads in the lungs and spleens of mice with Mtb respiratory infections.Conclusion Mucosal immunization with rAd-CnpB induced significant mucosal,humoral and cellular immune responses in mice,and significantly enhanced CnpB-specific cellular immune responses in Mtb-infected mice.
作者
代婷
路延之
赵瑞华
宁唤唤
康健
郝乐冉
李嘉玲
常宇骁
柏银兰
DAI Ting;LU Yanzhi;ZHAO Ruihua;NING Huanhuan;KANG Jian;HAO Leran;LI Jialing;CHANG Yuxiao;BAI Yinlan(School of Life Sciences,Yan’an University,Yan’an 716000;Department of Microbiology and Pathogenic Biology,Air Force Military Medical University,Xi’an 710032;Military Medical Innovation Center,Air Force Military Medical University,Xi’an 710032;Department of Physiology and Neurobiology,School of Basic Medicine,Ningxia Medical University,Yinchuan 750001,China)
出处
《细胞与分子免疫学杂志》
北大核心
2025年第1期1-8,共8页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(82272343,81971560)
陕西省重点研发计划(2022ZDLSF01-07,2024SF-YBXM-153)。
