摘要
目的研究登革病毒(Dengue virus,DENV)共有序列候选DNA疫苗pVAX1-cE80在小鼠体内的免疫原性及其保护作用。方法首先利用pVAX1-cE80 DNA免疫BALB/c小鼠,通过ELISA和噬斑减少中和试验(plaque reduction neutralization test,PRNT)分别检测免疫后小鼠血清中抗登革病毒的4种血清型(dengue virus serotype 1-4,DENV1-4)IgG抗体和中和抗体(neutralizing antibody,nAb)效价;利用酶联免疫斑点试验(enzyme-linked immunospot assay,ELISPOT)检测小鼠脾细胞分泌DENV1-4特异性Th1/Th2型细胞因子的能力;通过攻毒试验评价疫苗对DENV1-4感染的保护作用。结果接种疫苗的小鼠可产生针对DENV1-4的四价抗体,其中抗DENV2抗体水平最高,IgG抗体和nAb效价分别为1∶1086和1∶120;免疫后小鼠脾细胞在DENV1-4抗原刺激下均可分泌较高水平的IFN-γ和IL-4;疫苗免疫可为小鼠提供针对DENV1-4的部分保护作用。结论单一共有序列登革病毒候选DNA疫苗可诱导小鼠产生四价体液免疫反应和细胞免疫反应,并为小鼠提供部分保护作用,为登革病毒共有序列疫苗的研发奠定了基础。
Objective To study the immunogenicity and protective efficacy in BALB/c mice induced by a dengue DNA vaccine candidate pVAX1-cE80.Methods After immunized with DNA of pVAX1-cE80,titers of IgG antibody and neutralizing antibody(nAb)against dengue virus serotypes 1-4(DENV1-4)were detected by ELISA and PRNT(plaque reduction neutralization test),respectively;the secretions of Th1/Th2-type cytokines in splenocytes upon stimulation with DENV1-4 antigens were determined by an ELISPOT assay(enzyme-linked immunospot assay);challenge experiments were used for evaluating the protective efficacy of the vaccine against DENV1-4 in mice.Results Tetravalent antibodies against DENV1-4 were induced by the vaccination,among which the titers of antibodies against DENV2 were the highest,IgG antibody and nAb titers were 1∶1086 and 1∶120,respectively;splenocytes from immunized mice could secret high levels of IFN-γand IL-4 upon stimulations with DENV1-4 antigens;the vaccination conferred partial protection against DENV1-4 for mice.Conclusion A single consensus-sequence-based dengue DNA vaccine candidate was used in this study and it could induce tetravalent humoral and cellular immune responses and confer partial protection from DENV challenge for vaccinated mice.This study provides a useful data for development of consensus-based dengue vaccine.
作者
杨馥嘉
王然
范东瀛
陈辉
安静
YANG Fu-jia;WANG Ran;FAN Dong-ying;CHEN Hui;AN Jing(Department of Medical Microbiology,School of Basic Medical Sciences,Capital Medical University,Beijing 100069,China)
出处
《微生物学免疫学进展》
2020年第1期1-7,共7页
Progress In Microbiology and Immunology
基金
国家自然科学基金项目(81772172、81671971、U1602223)
北京自然科学基金B类项目(KZ201810025035).
关键词
登革病毒
共有序列
包膜蛋白
DNA疫苗
保护作用
Dengue virus(DENV)
Consensus sequence
Envelope protein
DNA vaccine
Protection
