Functional expression of a cattle MHC class II DR-like antigen on mouse L cells
- 50 Downloads
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
Unable to display preview. Download preview PDF.
- Baldwin, C. L., Morrison, W. I., and Naessens, J. Differentiation antigens and functional characteristics expressed by bovine leukocytes.In M. Miyasaka and Z. Trnka, (eds.):Comparative Aspects of Differentiation Antigens in Lympho-haemopoietic Tissues, pp 455–498, Marcel Dekker, New York, 1987Google Scholar
- Davies, C. J., Joosten, I., Andersson, L. Arriens, M. A., Bernoco, D., Bissumbhar, B., Byrns, G., van Eijk, M. J. T., Kristensen, B., Lewin, H. A., Mikko, S., Morgan, A. L. G., Muggli-Cockett, N. E., Nilsson, Ph. R., Oliver, R. A., Park, C. A., van der Poel, J. J., Polli, M., Spooner, R. L., and Stewart, J. A. Polymorphism of bovine MHC class II genes. Joint report of the Fifth International Bovine Lymphocyte (BoLA) Workshop, Interlaken, Switzerland, 1 August, 1992.Eur J Immunogenet 21: 259–289, 1994PubMedGoogle Scholar
- Davis, W. C., Marusic, S., Lewin, H. A., Splitter, G. A., Perryman, L. E., McGuire, T. C., and Gorham, J. R. The development and analysis of species specific and cross reactive monoclonal antibodies to leukocyte differentiation antigens and antigens of the major histocompatibility complex for use in the study of the immune system in cattle and other species.Vet Immunol Immunopathol 15: 337–376, 1987PubMedCrossRefGoogle Scholar
- Genetics Computer Group. Program Manual for the GCG Package, version 7 (April), Madison, 1991Google Scholar
- Hopkins, J., Dutia, B. M., and McConnell, I. Monoclonal antibodies to sheep lymphocytes. I. Identification of MHC class II molecules on lymphoid tissue and changes in the level of class II expression on lymph-borne cells following antigen stimulationin vivo.Immunology 59: 433–438, 1986PubMedGoogle Scholar
- Mikka, S. and Andersson, L. Extensive MHC class II DRB3 diversity in African and European cattle.Immunogenetics 42: 408–413, 1995Google Scholar
- Trucco, M. and de Petris, S. Determination of equilibrium binding. Parameters of monoclonal antibodies specific for cell surface antigens.In I. Lefkovits, and B. Pernis (eds.).Immunological Methods II, pp. 1–26, Academic Press, New York, 1981Google Scholar
- Wilkinson, D., De Vries, R. R. P., Madrigal, J. A., Lock, C. B., Morgenstern, J. P., Trowsdale, J., and Altmann, D. M. Analysis of HLA-DR glycoproteins by DNA-mediated gene transfer. Definition of DR2β gene products and antigen presentation to T cell clones from leprosy patients.J Exp Med 167: 1442–1458, 1988PubMedCrossRefGoogle Scholar