IL-12对HIV痘病毒载体疫苗毒性和免疫原性的影响  

Pathogenicity and immunogenicity of recombinant HIV Tiantan vaccinia virus expressing IL12

作　　者：田美娟[1] 刘颖[1] 马燕[1] 刘铮[1] 凌大伟[1] 段丹丽[1] 邬超[1] 彭虹[1] 邵一鸣 

机构地区：[1]中国疾病预防控制中心性病艾滋病预防控制中心,北京102206

出　　处：《中国热带医学》2010年第6期653-655,683,共4页China Tropical Medicine

基　　金：十一五"艾滋病和病毒性肝炎等重大传染病防治"科技重大专项"创新性复制非复制载体艾滋病疫苗研究"(2008ZX10001-010);国家自然科学基金"复制型痘苗病毒天坛株载体的改造与免疫学研究"(30700742)

摘　　要：目的探讨提高HIV痘病毒天坛株载体疫苗(vTKgpe)的免疫原性及细胞免疫反应。方法用同源重组方法在HIV痘病毒天坛株载体疫苗基因组中插入鼠白介素12(IL12)基因,获得重组病毒vTKgpe-IL12。首先经鼻腔免疫检测其毒力,然后以酶联免疫斑点实验(ELISPOT)和胞内因子染色(ICS)检测vTKgpe-IL12的细胞免疫反应,以酶联免疫吸附试验(ELISA)检测体液免疫应答。结果重组病毒vTKgpe-IL12能正确表达mIL12和HIV抗原。mIL12对vTKgpe毒性和体液免疫应答无影响。相对vTKgpe而言,vTKgpe-IL12在末次免疫后10d,对细胞免疫应答的强度没有影响;在末次免疫后30d,针对Env的细胞免疫应答有所提高,针对gag-pol的细胞免疫应答略有下降。vTKgpe-IL12诱导的细胞免疫反应30d时比10d显著下降,尤其是针对gag-pol的细胞免疫应答显著降低。结论应用DNA初免-痘病毒加强免疫策略时,在痘病毒重组疫苗中加入IL12增强了对env的细胞免疫反应,而降低了对gag-pol的细胞免疫反应。Aim To improve the immunogenicity of HIV candidate vaccine （vTKgpe）,especially to improve the cellmediated immunity. Methods The recombinant vaecinia virus Tiantan expressing IL12 （vTKgpe-IL12） was constructed by homologous recombination .Then the virulence was decided with the weight loss of mice inoculated intranasally,cellmediated immunity was detected by enzyme-linked immunosorbent spot test （ELISPOT） and flow cytometry （ICS） and then humoral immune response was assayed by enzyme lnked immunosorbent assay （ELISA）. Results The ILl2 and antigens from HIV were expressed correctly. No difference was detected between the virulence of vTKgpe-IL12 and vTKgpe. vTKgpe-IL12 elicited the same humoral immune response as vTKgpe did. 10 days after the final inoculation ,vTKgpe-ILl2 elicited the same cell-mediated immune response and humoral immune response as vTKgpe did. 30 days after the final inoculation ,more env-specific IFN-vpreducing cells and less gag-,pol-specific IFN-γ producing cells were detected in splenecytes from mice inoculated by vTKgpe-IL12. The cell-mediated immune response elicited by vTKgpe was longer than by vTKgpe-IL12. Conclusions The results were helpful for preparation of HIV vaccine in the future.

关 键 词：重组痘病毒 HIV载体疫苗 鼠白介素12 

分 类 号：R512.9[医药卫生—内科学]