OX40L通过激活树突状细胞促进重组狂犬病病毒免疫早期抗体的产生  

OX40L promotes early antibody production after recombinant rabies virus immunization by activating dendritic cells

作　　者：王玉芳 高婷 邢潇 王勇 字绍美 刘亚萍 胡杨 李康 李莹莹 WANG Yufang;GAO Ting;XING Xiao;WANG Yong;ZI Shaomei;LIU Yaping;HU Yang;LI Kang;LI Yingying(School of Basic Medical Sciences,Dali University,Dali 671000,China;Dali Nursing Vocational College,Dali 671000,China;School of Clinical Medicine,Dali University,Dali 671000,China)

机构地区：[1]大理大学基础医学院,671000 [2]大理护理职业学院,671000 [3]大理大学临床医学院,671000

出　　处：《免疫学杂志》2024年第5期425-432,共8页Immunological Journal

基　　金：国家自然科学基金(32360881);云南省科技厅基础研究专项-面上项目(202301AT070024);云南省教育厅科学研究基金(2023Y0954)。

摘　　要：目的明确重组狂犬病病毒(LBNSE-OX40L)过表达的OX40L能否通过激活树突状细胞增强先天性免疫反应进而激活早期抗体的产生。方法体外研究:从Balb/c小鼠股骨提取骨髓源树突状细胞(bone marrow dendritic cells,BMDCs)培养6 d,用感染复数(MOI)=1的亲本病毒LBNSE和重组病毒LBNSE-OX40L感染培养好的BMDCs,通过流式细胞术分析各病毒对BMDCs成熟情况的影响和ELISA方法检测感染BMDCs上清中与先天性免疫相关细胞因子α干扰素(interferon-α,IFN-α)和白细胞介素12p40(interleukin-12p40,IL-12p40)的表达情况。体内研究:Balb/c雌鼠双后肢肌肉注射106 FFU的亲本病毒LBNSE和重组病毒LBNSE-OX40L,免疫后第6天收集小鼠腹股沟淋巴结,经流式细胞术检测成熟DCs比例,在免疫后第6天收集小鼠血清,通过抗病毒中和抗体滴定法检测病毒中和抗体(virus neutralizing antibody,VNA)含量,通过ELISA检测小鼠血清中IgG抗体;在免疫后第2、4、6天通过ELISA检测IgM抗体亚类含量。结果与亲本病毒LBNSE相比,重组病毒LBNSE-OX40L能够在体外激活显著多的BMDCs,并产生显著高水平的IFN-α和IL-12p40,且重组病毒LBNSE-OX40L能刺激体内DCs成熟分化,迅速产生高水平的VNA和RABV特异性IgG和IgM抗体亚类。结论LBNSE-OX40L能激活树突状细胞促进机体先天性免疫反应,进而增强早期抗体的产生,可以作为早期有效的狂犬病疫苗候选。This study performed to determine whether OX40L overexpressed by recombinant rabies virus(LBNSEOX40L)can enhance the innate immune response through activation of dendritic cells and thus activate early antibody production.Bone marrow dendritic cells(BMDCs)were extracted from the femur of Balb/c mice and cultured for 6 days,and the cultured BMDCs were infected with the parental virus LBNSE and the recombinant virus LBNSE-OX40L with the multiplicity of infections(MOI)=1.The effect of each virus on the maturation of BMDCs was analyzed by flow cytometry;ELISA was used to detect the expression of innate immunity-related cytokines such as interferon-α(IFN-α)and interleukin-12p40(IL-12p40)in the supernatants of the infected BMDCs.For in vivo study,Balb/c female mice were injected intramuscularly with 106 FFU of parental virus LBNSE and recombinant virus LBNSE-OX40L in both hind limbs,and the inguinal lymph nodes of mice were collected on day 6 after immunization,and the proportion of mature DCs was detected by flow cytometry.The serum was collected on day 6 after immunization,and the content of virus-neutralizing antibody(VNA)was detected by antiviral neutralizing antibody titration.Mouse serum was collected on day 6 after immunization,and virus neutralizing antibody content was measured by titration of antiviral neutralizing antibody,while IgG antibody in mouse serum was detected by ELISA.IgM antibody subclasses were detected by ELISA on days 2,4,and 6 after immunization.Compared with the parental virus LBNSE,the recombinant virus LBNSE-OX40L was able to activate more BMDCs in vitro and produce significantly higher levels of IFN-αand IL-12p40.Furthermore,the recombinant virus LBNSE-OX40L stimulated the maturation and differentiation of the DCs in vivo,which led to the rapid production of high levels of VNA and RABV-specific IgG and IgM antibodies.Taken together,LBNSE-OX40L activates dendritic cells to promote the body's innate immune response,and in turn enhances early antibody production,thus can be an early effect

关 键 词：重组狂犬病病毒 OX40L 树突状细胞 先天性免疫反应 抗体免疫 

分 类 号：R392.11[医药卫生—免疫学]