中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2011年
3期
225-228
,共4页
艾滋病病毒%疫苗%T细胞%调节/辅助蛋白%融合抗原
艾滋病病毒%疫苗%T細胞%調節/輔助蛋白%融閤抗原
애자병병독%역묘%T세포%조절/보조단백%융합항원
HIV/AIDS%Vaccine%T cell%Regulatory/accessory protein%Fusion antigen
目的 构建表达HIV-1 AE2f gp145-tat-rev-nef融合基因的DNA疫苗,并比较gp145-tat-rev-nef(AE-Gp145TRN)和tat-rev-integrase(c-half)(AE-TRIVN)两种不同基因融合方式对Tat、Rev和Nef蛋白免疫原性的影响.方法 按人源密码子使用频率对HIV-1 AE2f的gp145、tat、rev和nef基因序列进行优化并构建DNA疫苗.经Western blot进行体外表达检测后,利用小鼠模型比较AE-Gp145TRN与AE-TRIVN的免疫原性.结果 限制性酶切及DNA测序结果表明AE-Gp145TRN融合基因表达重组质粒构建正确;Western blot结果显示其能够在体外表达相应的融合蛋白.小鼠免疫后IFN-γ ELISPOT检测结果显示:AE-TRIVN所活化的针对Tat、Rev和Nef蛋白的总T细胞反应强度[(148±91)SFCs/106脾细胞]显著高于Gp145TRN[(55±28)SFCs/106脾细胞,P=0.017].T细胞反应分布情况显示:AE-TRIVN所活化的T细胞反应主要集中在Rev蛋白上;而Gp145TRN则可以显著提高对Nef蛋白的免疫识别.结论 AE-TRIVN与Gp145TRN在小鼠体内所活化的T细胞反应特征有明显区别,提示不同的抗原融合方式可能影响融合基因DNA疫苗的免疫结果.
目的 構建錶達HIV-1 AE2f gp145-tat-rev-nef融閤基因的DNA疫苗,併比較gp145-tat-rev-nef(AE-Gp145TRN)和tat-rev-integrase(c-half)(AE-TRIVN)兩種不同基因融閤方式對Tat、Rev和Nef蛋白免疫原性的影響.方法 按人源密碼子使用頻率對HIV-1 AE2f的gp145、tat、rev和nef基因序列進行優化併構建DNA疫苗.經Western blot進行體外錶達檢測後,利用小鼠模型比較AE-Gp145TRN與AE-TRIVN的免疫原性.結果 限製性酶切及DNA測序結果錶明AE-Gp145TRN融閤基因錶達重組質粒構建正確;Western blot結果顯示其能夠在體外錶達相應的融閤蛋白.小鼠免疫後IFN-γ ELISPOT檢測結果顯示:AE-TRIVN所活化的針對Tat、Rev和Nef蛋白的總T細胞反應彊度[(148±91)SFCs/106脾細胞]顯著高于Gp145TRN[(55±28)SFCs/106脾細胞,P=0.017].T細胞反應分佈情況顯示:AE-TRIVN所活化的T細胞反應主要集中在Rev蛋白上;而Gp145TRN則可以顯著提高對Nef蛋白的免疫識彆.結論 AE-TRIVN與Gp145TRN在小鼠體內所活化的T細胞反應特徵有明顯區彆,提示不同的抗原融閤方式可能影響融閤基因DNA疫苗的免疫結果.
목적 구건표체HIV-1 AE2f gp145-tat-rev-nef융합기인적DNA역묘,병비교gp145-tat-rev-nef(AE-Gp145TRN)화tat-rev-integrase(c-half)(AE-TRIVN)량충불동기인융합방식대Tat、Rev화Nef단백면역원성적영향.방법 안인원밀마자사용빈솔대HIV-1 AE2f적gp145、tat、rev화nef기인서렬진행우화병구건DNA역묘.경Western blot진행체외표체검측후,이용소서모형비교AE-Gp145TRN여AE-TRIVN적면역원성.결과 한제성매절급DNA측서결과표명AE-Gp145TRN융합기인표체중조질립구건정학;Western blot결과현시기능구재체외표체상응적융합단백.소서면역후IFN-γ ELISPOT검측결과현시:AE-TRIVN소활화적침대Tat、Rev화Nef단백적총T세포반응강도[(148±91)SFCs/106비세포]현저고우Gp145TRN[(55±28)SFCs/106비세포,P=0.017].T세포반응분포정황현시:AE-TRIVN소활화적T세포반응주요집중재Rev단백상;이Gp145TRN칙가이현저제고대Nef단백적면역식별.결론 AE-TRIVN여Gp145TRN재소서체내소활화적T세포반응특정유명현구별,제시불동적항원융합방식가능영향융합기인DNA역묘적면역결과.
Objective To construct DNA vaccine expressing HIV-1 AE2f gp145-tat-rev-nef fusion gene( AE-Gp145TRN) and to compare the immunogenicities of DNA vaccines expressing Tat, Rev and Nef in gene fusion formulations of tat-rev-integrase(c-half)-vif-nef( AE-TRIVN) and AE-Gpl45TRN. Methods DNA vaccine was constructed by inserting the codon optimized HIV-1 AE2( gp145-tat-rev-nef fusion gene into mammalian expression DNA vector. In vitro expression efficiency of the constructed DNA vaccine was determined by Western blot and the immunogenicities of AE-Gpl45TRN and AE-TRIVN were compared by immunizing female BALB/c mice. IFN-r ELISPOT assay was used to read out the specific T cell immunity. Results Western blot assay showed the constructed DNA vaccine could be expressed efficiently in vitro. After vaccination, AE-TRIVN mounted significantly higher T cell responses against Tat, Rev and Nef[(148±91)SFCs/106 splenocytes]than Gpl45TRN[(55±28) SFCs/106 splenocytes]. Specific T cell responses elicited by AE-TRIVN predominantly targeting Rev, whereas Gpl45TRN could significantly enhance T cell responses against Nef. Conclusion AE-TRIVN and Gpl45TRN induced distinct T cell response modalities, which implied different gene fusion formulations may affect the immunogenicity of specific DNA vaccines.
