中国免疫学杂志2015，Vol.31Issue(12)：1648-1653,1658,7.DOI:10.3969/j.issn.1000-484X.2015.12.014
B群脑膜炎奈瑟菌外膜蛋白NMB0315核酸疫苗的免疫活性和免疫保护作用初步研究
Immunocompetence and Immunoprotection of a DNA vaccine NMB0315 against N.meningitidis serogroup B in mice
摘要
Abstract
Objective:To construct NMB0315 eukaryotic expression recombinant vector ,detect specific humoral and cellular immune response induced by the recombint DNA vaccine intramuscularly in female BALB /c mice,evaluate the immunocompetence and immunoprotection of the vaccine , so as to provide experimental basis for the development of a novel nucleic acid vaccine against N.meningitidis serogroup B .Methods: The whole NMB0315 gene was amplified by PCR from the standard strains MC 58 genomic DNA,cloned into a plasmid pcDNA3.1(+),identified by double digestion of the recombinant plasmid with restriction enzymes and se -quencing.The recombinant vector pcDNA 3.1 (+)/NMB0315 was transfected into eukaryotic COS-7 cells and RAW264.7 cells, the NMB0315 protein was detected by immunocytochemical method and Western blot respectively .The levels of specific humoral and cellular immune response were detected after inoculating in female BALB /c mice intramuscularly with the recombinant plasmid .The immune protective effect was investigated with the DNA vaccine and the bactericidal titer of the immune serum was deter mined by serum bactericidal assay ( SBA ) in vitro.Results: The recombinant pcDNA3.1 (+)/NMB0315 was effectively transcripted and expressed in eukaryotic cells and the specific humoral and cellular immune responses were induced in the inoculated mice .In the re-combinant pcDNA3.1(+)/NMB0315 group ,the levels of serum IgG,IgG1,IgG2a,IgG2b and IgG3 and genital tract sIgA were significantly higher than in controls ( P<0.001 ) .The stimulation index in the culture supernatant of the spleen lymphocytes of the vaccine group was higher than that of the control group (P<0.05).The ratios of serum IgG2a/IgG1 in the DNA vaccine group were less than 1.The bactericidal titer of the NMB 0315+CpG group reached 1:128 following three immunizations , the protection rate of the vaccine group was 70%against the N.meningitidis strain MC58.Conclusion:The NMB0315 nucleic acid vaccine could induce higher levels of humoral immunity and cellular immunity and showed effective protection against N .meningitidis serogroup B , the immune serum had strong bactericidal activity in vitro .关键词
B群脑膜炎奈瑟菌/B群脑膜炎奈瑟菌外膜蛋白0315/核酸疫苗/免疫活性/免疫保护Key words
Neisseria meningitides/Outer membrane protein NMB0315/DNA vaccine/Immunocompetence/Immunoprotection分类
医药卫生引用本文复制引用
李开铭,谢梅花,余敏君,唐双阳,张璐思,邓苏宏,胡四海..B群脑膜炎奈瑟菌外膜蛋白NMB0315核酸疫苗的免疫活性和免疫保护作用初步研究[J].中国免疫学杂志,2015,31(12):1648-1653,1658,7.基金项目
本文为国家自然科学基金(No.81172890)、特殊病原体防控湖南省重点实验室(湘科计字2014-5号 （No.81172890）
湘教通2012-312号)及湖南省重点学科建设项目资助. （）
