Fraser D C, Craigmile S, Campbell J D, Oliver R A, Brown D J, Russell G C, Spooner R L, Glass E J
Roslin Institute, Midlothian, EH25 9PS, UK.
Immunogenetics. 1996;43(5):296-303. doi: 10.1007/BF02440997.
Cattle DRA and DRB genes, cloned by reverse-transcription polymerase chain reaction, were transfected into mouse L cells. The cattle DR-expressing L-cell transfectant generated was analyzed serologically, biochemically, and functionally. Sequence analysis of the transfected DRB gene clearly showed showed that it was DRB3 allele DRB3(*)0101 , which corresponds to the 1D-IEF-determined allele DRBF3. 1D-IEF analysis of the transfectant confirmed that the expressed DR product was DRBF3. Functional integrity of the transfected gene products was demonstrated by the ability of the transfectant cell line to present two antigens (the foot-and-mouth disease virus-derived peptide FMDV15, and ovalbumin) to antigen-specific CD4(+) T cells from both the original animal used to obtain the genes, and also from an unrelated DRBF3(+) heterozygous animal. Such transfectants will be invaluable tools, allowing us to dissect the precise contributions each locus product makes to the overall immune response in heterozygous animals, information essential for rational vaccine design.
通过逆转录聚合酶链反应克隆的牛DRA和DRB基因被转染到小鼠L细胞中。对产生的表达牛DR的L细胞转染子进行了血清学、生物化学和功能分析。对转染的DRB基因进行序列分析清楚地表明,它是DRB3等位基因DRB3(*)0101,这与一维等电聚焦确定的等位基因DRBF3相对应。对转染子进行的一维等电聚焦分析证实,表达的DR产物是DRBF3。转染基因产物的功能完整性通过转染细胞系向来自用于获取基因的原始动物以及无关的DRBF3(+)杂合动物的抗原特异性CD4(+)T细胞呈递两种抗原(口蹄疫病毒衍生肽FMDV15和卵清蛋白)的能力得以证明。此类转染子将是非常宝贵的工具,使我们能够剖析每个基因座产物对杂合动物整体免疫反应的确切贡献,而这些信息对于合理的疫苗设计至关重要。