Abstract
Human HLA cDNA probes were used to analyze the restriction fragment length polymorphism (RFLP) of the SLA major histocompatibility complex in swine. Cellular genomic DNA from 19 SLA homozygous pigs representing 13 different haplotypes was digested with restriction endonucleases Eco RI, Hind III, or Bam H1, separated by electrophoresis, and transferred onto diazobenzyloxymethyl paper by the Southern blot technique. The blots were probed with 32P-labeled class I or beta-DR class II cDNA. Depending on the haplotypes and the endonucleases used, seven to ten restriction fragments hybridized with the class I probe, and five to seven with the beta-DR probe. Their sizes ranged from 3.4 to 22 kilobase-pairs. Few bands were common to all 13 haplotypes. With all but one haplotype, identical autoradiogram patterns were obtained from unrelated, but phenotypically SLA-identical pigs, suggesting that most of the RFLP revealed were controlled by the SLA region. Further polymorphism was found in a group of seven unrelated pigs which typed serologically as SLA A15 CI B18 homozygotes but could be divided into two subgroups, with five animals in one subgroup and two in the other, when the genomic DNA was hybridized with the class I probe. When the class 11 beta-DR probe was tested on the same seven pigs, another subdivision was seen, and this correlated with MLR data. These results demonstrate that HLA class I and class II probes can be used to identify certain well-established SLA haplotypes and to identify subclasses within at least one SLA haplotype.
Similar content being viewed by others
Abbreviations
- MHC:
-
major histocompatibility complex
- MLR:
-
mixed lymphocyte reaction
- kbp:
-
kilobase pair(s)
- RFLP:
-
restriction fragment length polymorphism
References
Alwine, J. C., Kemp, D. J., Parker, B. A., Reiser, J., Renart, J., Stark, G. R., and Wahl, G. M.: Detection of specific RNAs or specific fragments of DNA by fractionation in gels and transfer to diazobenzyloxymethyl paper. Methods Enzymol. 68: 220–242, 1979
Bohme, J., Owerbach, D., Denaro, M., Lernmark, A., Peterson, P. A., and Rask, L.: Human class II major histocompatibility antigen B-chains are derived from at least three loci. Nature 301: 82–84, 1983
Cami, B., Bregegere, F., Abastado, J. P., and Kourilsky, P.: Multiple sequences related to classical histocompatibility antigens in the mouse genome. Nature 291: 673–675, 1981
Cann, H. M., Ascanio, L., Paul, P., Marcadet, A., Dausset, J., and Cohen, D.: Polymorphic restriction endonuclease fragment segregates and correlates with the gene for HLA-B.8. Proc. Natl. Acad. Sci. U.S.A. 80: 1665–1668, 1983
Cohen, D., Paul, P., Font, M.-P., Cohen, O., Sayagh, B., Marcadet, A., Busson, M., Mahouy, G., Cann, H. M., and Dausset, J.: Analysis of HLA class I genes with restriction endonuclease fragments: Implications for polymorphism of the human major histocompatibility complex. Proc. Natl. Acad. Sci. U.S.A. 80: 6289–6292, 1983
Larhammar, D., Schenning, L., Gustafsson, K., Wiman, K., Claesson, L., Rask, L., and Peterson, P. A.: Complete amino acid sequence of an HLA-DR antigen-like beta-chain as predicted from the nucleotide sequence: Similarities with immunoglobulins and HLA-A, -B and -C antigens. Proc. Natl. Acad. Sci. U.S.A. 79: 3687–3691, 1982
Orr, H.T.: Use of Southern blotting to analyze the size and restriction fragment polymorphism of HLA class I DNA in the human population. Transplant. Proc. 15: 1900–1906, 1983
Pease, L. R., Nathenson, S. G., and Leinwand, L. A.: Mapping class I gene sequences in the major histocompatibility complex. Nature 298: 382–385, 1982
Renard, C,, Chardon, P., and Vaiman, M.: The pig histocompatibility system SLA: Serological study on a group of antigenic specificities. Animal Blood Groups Biochem. Genet. 13: 161–177, 1982
Rigby, P. W. J., Dieckmann, M., Rhodes, C., and Berg, P.: Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J. Mol. Biol. 113: 237–251, 1977
Singer, D. S., Otero, R. S. C., Satz, M. L., Osborne, B., Sachs, D., and Rudikoff, S.: Characterization of a porcine genomic clone encoding a major histocompatibility antigen: Expression in mouse L cells. Proc. Natl. Acad. Sci. U.S.A. 79: 1403–1407, 1982
Sood, A. K., Pereira, D., and Weissman, S. M.: Isolation and partial nucleotide sequence of a cDNA clone for human histocompatibility antigen HLA-B by use of an oligodeoxynucleotide primer. Proc. Natl. Acad. Sci. U.S.A. 78: 616–620, 1981
Southern, E. M.: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 98: 503–517, 1975
Steinmetz, M., Moore, K. W., Frelinger, J. G., Sher, B. T., Shen, F. W., Boyse, E. A., and Hood, L.: A pseudogene homologous to mouse transplantation antigens: Transplantation antigens are encoded by eight exons that correlated with protein domains. Cell 25: 683–692, 1981
Vaiman, M., Hauptmann, G.; and Mayer, S.: Influence of the major histocompatibility complex in the pig (SLA) on serum haemolytic complement levels. J. Immunogenet. 5: 59–65, 1978a
Vaiman, M., Metzger, J. J., Renard, C., and Vila, J. P.: Immune response gene(s) controlling the humoral anti-lysozyme response (Ir-Lys) linked to the major histocompatibility complex SL-A in the pig. Immunogenetics 7: 231–238, 1978b
Vaiman, M., Chardon, P., and Renard, C.: Genetic organization of the pig SLA complex. Studies on nine recombinants and biochemical and lysostrip analysis. Immunogenetics 9: 353–361, 1979
Weiss, E. H.. Golden, L., Fahrner, K., Mellor, A. L., Devlin, J. J., Bullman, H., Tiddens, H., and Flaveil, R. A.: Organization and evolution of the class I gene family in the major histocompatibility complex of the C57BL/10 mouse. Nature 310: 650–655, 1984
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Chardon, P., Vaiman, M., Kirszenbaum, M. et al. Restriction fragment length polymorphism of the major histocompatibility complex of the pig. Immunogenetics 21, 161–171 (1985). https://doi.org/10.1007/BF00364868
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00364868