Functional expression of a cattle MHC class II DR-like antigen on mouse L cells
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CattleDRA andDRB 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 transfectedDRB gene clearly showed showed that it wasDRB3 alleleDRB3 *0101, which corresponds to the 1D-IEF-determined alleleDRBF3. 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 antigenspecific 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.
KeywordsMajor Histocompatibility Complex Major Histocompatibility Complex Class Invariant Chain Peripheral Blood Mononuclear Heterozygous Animal
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