中国免疫学杂志2018，Vol.34Issue(4)：508-512,519,6.DOI:10.3969/j.issn.1000-484X.2018.04.006
梅毒螺旋体黏附素Tp0751核酸菌影的构建及免疫原性研究
Construction and immunocompetence of recombinant DNA E.coli ghosts expressing Treponema pallidum adhesin Tp0751
摘要
Abstract
Objective:To construct the recombinant DNA E.coli ghosts (EBGs) expressing Treponema pallidum adhesin Tp0751 (pcD/Tp0751-BG) and determine its immunocompetence in immunized mice,and provide a potential novel method for syphilis vaccine developing.Methods:The recombinant eukaryotic expression plasmid pcDNA3.1(+)/Tp0751 was constructed and loaded into empty EBGs to create pcD/Tp0751-BG.The loading rate was determined accordingly.Macrophages cell line RAW264.7 was transfected with pcD/Tp0751-BG,and the expression of recombinant Tp0751(rTp0751) protein was detected by Western blot(WB).For immuno-competence in mice,the female BALB/c mice were randomly divided into 6 groups,including three control groups,A (PBS),B (EBG),C (empty pcDNA),and experimental group D(naked pcD/Tp0751),E (pcD/Tp0751-BG) and F (pcD/Tp0751-BG+rTp0751).All the mice were immunized as indicated for three times by intramuscular injection at two weeks intervals.The levels of specific IgG in sera and SIgA in genital tract lavage fluid were measured by ELISA.Levels of lymphocyte proliferation and IFN-γ secretion in spleen cells were measured by CCK-8 Cell Counting Kit and ELISA as well,respectively.Results:The loading rate of pcD/Tp0751 to EBGs was 76.1%.WB showed that the target recombinant protein pcD/Tp0751 expressed in RAW264.7 cells was active with Tp-infected rabbit sera.The titers of specific IgG and SIgA in group D,E,F gradually increased to significantly higher level as compared to the control groups (P<0. 01),which reached its peak at wk 8 after last immunization(the titers of IgG and SIgA were 1 :102 400 and 1 :12 800 in group F,respectively). Higher levels of specific IgG and SIgA were observed in groups E and F as compared to group D after first boost (P<0. 01),with groups F higher than group E after last boost(P<0. 01). At wk 8 after the last boost,the stimulation index (SI) and levels of IFN-γ in group D,E,F were all significant higher than the control groups (P<0. 01), with group E and F higher than group D (P<0. 01),and group E higher than group F (P<0. 05). Conclusion: The recombinant DNA EBGs of T. pallidum adhesin Tp0751 (pcD/ Tp0751-BG) possesses the immunocompetence to induce not only strong mucosal and systemic humoral immune response but also systemic cellular immune response in BALB/ c mice. The heterologous boost can be more efficient than homologous boost during immunization process.关键词
梅毒螺旋体/Tp0751/核酸疫苗/菌影/免疫原性Key words
Treponema pallidum/Tp0751/Nucleic acid vaccine/Bacterial ghost/Immunocompetence分类
医药卫生引用本文复制引用
张佳俐,曹二龙,曹龙古,赵飞骏,余坚,唐一之,符波,段武,曾铁兵..梅毒螺旋体黏附素Tp0751核酸菌影的构建及免疫原性研究[J].中国免疫学杂志,2018,34(4):508-512,519,6.基金项目
本文受国家自然科学基金(81273322 （）
81373230)、湖南省分子靶标新药研究协同创新中心项目(湘教通[2014]405号)、特殊病原体防控湖南省重点实验室资助项目(湘科计字2014-5号)、湖南省教育厅开放创新平台基金(15K110)和湖南省教育厅资助科研项目(15C1256 （湘教通[2014]405号）
17B245)资助. （）
