Abstract
HLA class II molecules are surface heterodimers which are essential in the initiation of immune responses. The amount of polymorphism expressed by the different class II molecules is largely dependent on the polymorphic structure of their beta chains. Cross-hybridization between, class II beta genes frequently hampered restriction fragment length polymorphism analysis of donor genomic DNA. In this report we show that the cross-hybridization between human class II beta genes is mediated by a region of high homology, rich in C and G residues, between the first domain encoding sequences of DP, DQ, and DR genes. The removal of the DNA segment containing this region from the fragments used as labeled probes against the corresponding fragments of the genes at other loci or against endonuclease digested genomic DNA completely eliminated or drastically reduced the cross-hybridization. Also, the RFLP patterns generated with the shortened probes were more informative and much simpler to interpret than were these generated with probes made from the original genes.
Similar content being viewed by others
References
Bodmer, W.: The HLA system: 1984. In E. D. Albert, M. P. Baur, and W. R. Mayr (eds.): Histocompatibility Testing 1984, pp. 209–210, Springer-Verlag, Heidelberg, 1984
Cascino, I., Rosenshine, S., Turco, E., Marrari, M., Duquesnoy, R. J., and Trucco, M.: Relationship between DQ alpha and DQ beta RFLP and serologically defined class II HLA antigens, J. Immunogenet. 13: 387–400, 1986
Cohen, D., Gall, I., Marcadet, A., Font, M., Lalouel, J., and Dausset, J.: Clusters of HLA class II beta restriction fragments describe allelic series. Proc. Natl. Acad. Sci. U.S.A. 81: 7870–7874, 1984
Cohen, D., Paul, P., Le Gall, I., Marcadet, A., Font, M., Cohen-Haguenauer, O., Sayagh, B., Cann, H., Lalouel, J., and Dausset, J.: DNA polymorphisms of class I and class II regions. Immunol. Rev. 85: 87–122, 1985
Compagnone-Post, P., Turco, E., Robinson, C., and Trucco, M.: The beta chains of DP4 molecules from different haplotypes are encoded by the same gene. Genomics 2: 8–13, 1988
Klein, J.: Natural History of the Major Histocompatibility Complex, John Wiley & Sons, New York, 1986
Korman, A. J., Boss, J. M., Spies, T., Sorrentino, R., Okada, K., and Strominger, J. L.: Genetic complexity and expression of human class II histocompatibility antigens. Immunol. Rev. 85: 45–86, 1985
Long, E., Wake, C., Gorski, J., and Mach, B.: Complete sequence of an HLA-DR beta chain deduced from a cDNA clone and identification of multiple nonallelic DR beta chain genes. EMBO J. 2: 389–394, 1983
Long, E., Gorski, J., and Mach, B.: Structural relationship of the SB beta-chain gene to HLA-D-region genes and murine I-region gene. Nature 310: 233–235, 1984
Lotteau, V., Teyton, L., Burroughs, D., and Charron, D.: A novel HLA class II molecule (DR-alpha-DQ-beta) created by mismatched isotype pairing. Nature 329: 339–341, 1987
Maniatis, T., Fritsch, E., and Sambrook, J. (eds.): Molecular Cloning: A Laboratory Manual, pp. 280–281, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1982
Rask, L., Gustafsson, K., Larhammar, D., Ronne, H., and Peterson, P. A.: Generation of class II antigens polymorphism. Immunol. Rev. 85: 123–143, 1985
Rosenshine, S., Cascino, I., Zeevi, A., Duquesnoy, R. J., and Trucco, M.: DQ alpha and beta RFLP reveals the composition of the DQ molecule recognized by T-cell clones. Immunogenetics 23: 187–196, 1986
Schenning, L., Larhammar, D., Bill, P., Wiman, K., Jonsson, A. K., Rask, L., and Peterson, P. A.: Both alpha and beta chains of HLA-DC class II histocompatibility antigens display extensive polymorphism in their amino-terminal domains. EMBO J. 3: 447–452, 1984
Spielman, R., Lee, J., Bodmer, W., Bodmer, J., and Trowsdale, J.: Six HLA-D region alpha chain genes of human chromosome 6: polymorphisms and associations of DC alpha related sequences with DR types. Proc. Natl. Acad. Sci. U.S.A. 81: 3461–3465, 1984
Todd, J., Bell, J., and McDevitt, H.: HLA-DQ beta gene contributes to susceptibility and resistance to insulin dependent diabetes mellitus. Nature 329: 599–604, 1987
Trowsdale, J., Young, J. A., Kelly, A. P., Austin, P. J., Carson, S., Meunier, H., So, A., Ehrlich, H. A., Spielman, R. S., Bodmer, J., and Bodmer, W. F.: Structure, sequence and polymorphism in the HLA-D region. Immunol. Rev. 85: 1–44, 1985
Trucco, M. and Duquesnoy, R. J.: Polymorphisms of the HLA-DQ subregion. Immunol. Today 7: 297–300, 1986
Trucco, M., Rosenshine, S., Cascino, I., and Duquesnoy, R. J.: HLA-DQ polymorphism analyzed by sequential restriction endonuclease DNA digestion. Immunogenetics 24: 184–190, 1986
Turco, E., Care, A., Compagnone-Post, P., Robinson, C., Cascino, I., and Trucco, M.: Allelic forms of the alpha- and beta-chain genes encoding DQw1-positive heterodimers. Immunogenetics 26: 282–290, 1987
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Turco, E., Fritsch, R. & Trucco, M. First domain encoding sequence mediates human class II beta-chain gene cross-hybridization. Immunogenetics 28, 193–204 (1988). https://doi.org/10.1007/BF00375859
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00375859