CAJ | 학술논문

目的比较我国不同艾滋病(AIDS)候选疫苗小鼠细胞免疫应答水平.方法将我国6种艾滋病候选疫苗按其各自免疫程序,免疫BALB/c小鼠,分离脾细胞,利用酶联免疫斑点法(ELISPOT)与胞内因子染色法(ICS)检测其对HIV特异多肽的细胞免疫应答水平.结果 6种艾滋病候选疫苗利用潜在T细胞表位(PTE)Gag、Env和Pol肽库和ELISPOT法检测IFN-γ的分泌情况,结果各疫苗的小鼠细胞免疫阳转率为70%～100%,各疫苗引起特异细胞免疫应答的反应肽库不同,强度亦不相同.1#和2#疫苗利用ELISPOT检测分泌Th1类细胞因子IFN-γ和IL-2的小鼠脾细胞数量,两疫苗的IFN-γ和IL-2检测结果均呈中度相关性(r1=0.62,P1＜0.01;r2=0.79,P2＜0.01).ELISPOT和ICS检测1#疫苗免疫后分泌IFN-γ的小鼠脾细胞数,结果两种方法的小鼠阳转率均为66.7%(10/15),ELISPOT阳性细胞数与ICS阳性细胞比例呈中度相关(r=0.55,P＜0.05).结论我国不同AIDS候选疫苗引起的小鼠细胞免疫应答在广度和强度上存在差别.ELISPOT或ICS检测IFN-γ等Th1类细胞因子可作为疫苗诱导小鼠细胞免疫应答的评价指标.
목적 비교아국불동애자병(AIDS)후선역묘소서세포면역응답수평.방법 장아국6충애자병후선역묘안기각자면역정서,면역BALB/c소서,분리비세포,이용매련면역반점법(ELISPOT)여포내인자염색법(ICS)검측기대HIV특이다태적세포면역응답수평.결과 6충애자병후선역묘이용잠재T세포표위(PTE)Gag、Env화Pol태고화ELISPOT법검측IFN-γ적분비정황,결과각역묘적소서세포면역양전솔위70%～100%,각역묘인기특이세포면역응답적반응태고불동,강도역불상동.1#화2#역묘이용ELISPOT검측분비Th1류세포인자IFN-γ화IL-2적소서비세포수량,량역묘적IFN-γ화IL-2검측결과균정중도상관성(r1=0.62,P1＜0.01;r2=0.79,P2＜0.01).ELISPOT화ICS검측1#역묘면역후분비IFN-γ적소서비세포수,결과량충방법적소서양전솔균위66.7%(10/15),ELISPOT양성세포수여ICS양성세포비례정중도상관(r=0.55,P＜0.05).결론 아국불동AIDS후선역묘인기적소서세포면역응답재엄도화강도상존재차별.ELISPOT혹ICS검측IFN-γ등Th1류세포인자가작위역묘유도소서세포면역응답적평개지표.
Objective To compare cellular immune responses in mice elicited by Chinese different AIDS candidate vaccines.Methods According to their different immunization procedures,BALB/c mice were immunized with 6 AIDS candidate vaccines,separately.Spleen cells were isolated for the detection of cellular immune response to HIV-specific peptides using enzyme-linked immunosorbent spot(ELISPOT)assay and intracellular cytokine staining(ICS)method.Results AIDS vaccines were evaluated by using potential T-cell epitopes(PTE)Gag,Env and Pol peptides pool and ELISPOT.The positive conversion rates for cellular immune response of 1#-6# vaccines fluctuated from 70% to 100%.The vaccine-induced cellular immune responses to specific peptides pool are different not only in magnitude but also in breadth.The Th1type cytokines,IFN-γand IL-2,were detected with ELISPOT in 1# and 2# vaccines.The productions of IFN-γand IL-2 induced by both of the two vaccines showed a moderate correlation(r1 =0.62,P1 ＜0.01 ;r2=0.79,P2 ＜ 0.01).The positive conversion rate of IFN-γ secreting cells of 1 # vaccine was 66.7%(10/15)mice detected with both ELISPOT and ICS.And the results tested by ELISPOT and ICS showed moderate correlation(r = 0.55,P ＜ 0.05).Conclusion The magnitude and breadth of cellular immune responses induced by different AIDS candidate vaccines are different.Being induced by different AIDS candidate vaccines,the IFN-γand other Th1 type cytokines detected by ELISPOT or ICS could be used to evaluate the cellular immune responses in mice.