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为了筛选和确定用于检测表达HIV-1 B’/C亚型病毒6种抗原(gp160、gag、polr、evt、at和nef)的艾滋病疫苗免疫小鼠后H-2d限制的特异性T细胞表位,本研究使用表达上述6种抗原的复制型DNA疫苗和非复制型重组痘苗病毒疫苗联合免疫BALB/c小鼠,通过矩阵设计将HIV-1 B(C)亚型6种相应抗原全序列肽库分别混合成肽池,使用肽池对免疫小鼠进行IFN-γELISPOT检测,根据检测结果确定肽库中特异反应的优势表位肽。结果显示:筛选到七条针对Gag的特异表位肽,其中有5条与文献报道相同,另2条为新表位肽;筛选到3条针对Pol蛋白特异表位肽,其中一条为新表位肽;筛选到2条针对gp160特异表位肽,其中一条为新表位肽;在Nef肽库中筛选到一条新的表位肽;从Tat肽库中筛选到3条表位肽,这三条肽在肽库中是连续的序列,都包含(或部分包含)网上公布的表位序列;在Rev肽库中没有筛选到能够产生阳性反应的特异性表位肽。本研究使用IFN-γELISPOT方法筛选和确定了可用于检测表达HIV-1 B’/C亚型病毒6种抗原(gp160、gag、pol、revt、at和nef)的艾滋病疫苗免疫小鼠后H-2d限制的特异性T细胞表位。
To screen for and identify H-2d-restricted T-specific cells for immunization of mice immunized with an AIDS vaccine expressing the 6 antigens of the HIV-1 B ’/ C subtype virus (gp160, gag, polr, evt, at and nef) In this study, BALB / c mice were immunized with a replicative DNA vaccine expressing the above six antigens and a non-replicating recombinant vaccinia virus vaccine, and the six corresponding antigens of HIV-1 B (C) subtype The peptide libraries were mixed into peptide pools respectively, and IFN-γELISPOT was detected in the immunized mice using peptide pool. According to the results, the dominant epitope peptide in the peptide library was identified. The results showed that seven specific epitope peptides against Gag were screened out, of which five were the same as reported in the literature and the other two were neoepitope peptides. Three specific epitope peptides against Pol were selected and one of them was a new epitope Two epitopes specific to gp160 were selected and one of them was a new epitope peptide. A new epitope peptide was screened from the Nef peptide library. Three epitope peptides were screened from the Tat peptide library. The peptide is a contiguous sequence in the peptide library and contains (or partially contains) an epitope sequence published online. No specific epitope peptide capable of producing a positive reaction is screened in the Rev peptide library. In this study, the IFN-γELISPOT method was used to screen and identify the H-2 immunized mice that can be used to detect HIV-1 HIV-1 B ’/ C subtype 6 virus (gp160, gag, pol, revt, at and nef) 2d-restricted T-cell epitopes.