重组嵌合腺病毒35型和重组痘苗病毒SIV疫苗在小鼠体内联合免疫的实验研究  

Study on rAd5F35-SIVenvT vaccine in combination with rMVA-SIVenvT vaccine in mice

作　　者：何小周[1] 杨靖 李红霞[1] 郝彦哲[1] 冯霞[1] He Xiaozhou;Yang Jing;Li Hongxia;Hao Yanzhe;Feng Xia(National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Bejing 102206,China)

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京102206

出　　处：《中华微生物学和免疫学杂志》2021年第6期455-459,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家科技重大专项(2018ZX10731101)。

摘　　要：目的研究嵌合腺病毒35型载体和痘苗病毒载体SIVenvT疫苗通过不同策略联合免疫小鼠所诱导的细胞和体液免疫应答。方法通过PCR和Western blot方法对表达SIV mac239株SIVenvT基因的重组嵌合腺病毒35型(rAd5F35-SIVenvT)和重组痘苗病毒安卡拉株(rMVA-SIVenvT)进行体外鉴定。采用两种不同的策略联合免疫BALB/c小鼠,即同源载体免疫和异源载体免疫。通过ELISPOT方法检测小鼠脾脏中分泌SIV Env特异性IFN-γ的淋巴细胞数量,ELISA方法检测血清SIV gp120抗体滴度,比较两种免疫策略诱导小鼠产生细胞和体液免疫应答的差异。结果rAd5F35-SIVenvT和rMVA-SIVenvT均能在HEK293细胞中有效表达SIVenvT蛋白。初次免疫后第4周,两组SIV Env特异性细胞免疫应答和SIV gp120抗体均达到峰值,随后呈波动下降趋势。异源免疫组诱导小鼠的应答水平高于同源组,其中第4周和第16周异源组细胞免疫应答强度显著高于同源组,第4周异源组SIV gp120抗体滴度显著高于同源组。结论rAd5F35-SIVenvT和rMVA-SIVenvT两种载体疫苗联合免疫策略能诱导小鼠产生较强细胞和体液免疫应答。Objective To analyze the immune response in mice after immunization with vaccine of rAd5F35-SIVenvT in combination with rMVA-SIVenvT to evaluate the efficacy of different immunization strategies.Methods Two recombinant viruses were identified in vitro by PCR and Western blot.The BALB/c mice were immunized with homologous and heterologous immune strategies.The numbers of splenic lymphocytes secreting IFN-γwere measured by ELISPOT assay,meanwhile SIV gp120 antibody titer were measured by ELISA assay.Results SIVenvT protein was expressed effectively by rAd5F35-SIVenvT and rMVA-SIVenvT in HEK293 cells.The specific immune response reached its peak at 4-week post first immunization,then decreased.SIV Env specific cellular immune response and SIV gp120 specific antibody could be detected at 4-16 weeks post first immunization.The specific cellular response was significant stronger in heterologous immunization group than homologous group at 4 week and 16 week.Furthermore,heterologous immunization induced significant higher titer of SIV gp120 antibody at 4 week than homologous group.Conclusions Specific immune response induced by rAd5F35-SIVenvT in combination with rMVA-SIVenvT was stronger than homologous vector immunization.The results provided references for further study in nonhuman primates.

关 键 词：重组嵌合腺病毒35型 重组痘苗病毒安卡拉株 细胞免疫 体液免疫 猴免疫缺陷病毒 

分 类 号：R392-33[医药卫生—免疫学]