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Southern blot analysis of HLA-DP gene polymorphisms in Caucasoid rheumatoid arthritis (RA) patients and controls

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Abstract

Despite extensive analysis of the incidence ofHLA-DR andHLA-DQ allele frequencies in defined autoimmune disease groups, there is very little information available onHLA-DP allele frequencies. This is largely becauseHLA-DP typing has until recently been restricted to primed lymphocyte typing (PLT). However, allelic polymorphism of theHLA-DP subregion can now be studied by Southern blot analysis or genotyping withDPA1 andDPB1 probes. By direct counting of allele-specific DNA fragments, we have analyzed the frequencies of five majorDP genotypes (DPw1, DPw2, DPw3/6, DPw4, andDPw5), in a large number of Caucasoid rheumatoid arthritis (RA) patients (n=74), and controls (n=91). The predicted frequency ofDP alleles in both patient and control groups was comparable to PLT-determinedDP allele frequencies in normal Caucasoids. However, the gene frequency ofDPw4 was increased in the RA patients, with 51% of the patients studied scoring asDPw4, 4 homozygotes. With the exception of one possible combination (DPw5 andDRw6) in the controls, no significant linkage disequilibrium was detected betweenDP andDR alleles in either patient or control groups. Thus the prevalence ofDPw4 in the RA patients is independent of any disease association with theDR loci, and may represent a new class II association with RA.

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References

  • Auffray, C., Lillie, J. W., Arnot, D., Grossberger, D., Kappes, D., and Strominger, J. L.: Isotypic and allotypic variation of human class II histocompatibility antigen alpha-chain genes.Nature 308: 327–333, 1984

    Article  CAS  PubMed  Google Scholar 

  • Austin, P., Trowsdale, J., Rudd, C., Bodmer, W., Feldmann, M., and Lamb, J.: Functional expression ofHLA-DP genes transfected into mouse fibroblasts.Nature 313: 61–64, 1985

    Article  CAS  PubMed  Google Scholar 

  • Benaceraff, B.: Role of MHC gene products in immune regulation.Science 212: 1229–1238, 1981

    Google Scholar 

  • Bidwell, J., Bidwell, E. A., Laundy, G. J., Klouda, P. T., and Bradley, B. A.: Allogenotypes defined by short DQ alpha and DQ beta cDNA probes correlate with and define splits of HLA-DQ serological specificities.Mol Immunol 24: 513–522, 1987

    Article  CAS  PubMed  Google Scholar 

  • Bodmer, J., Bodmer, W., Heyes, J., So, A., Tonks, S., Trowsdale, J., and Young, A.: Identification of HLA-DP polymorphism with DP alpha and DP beta probes and monoclonal antibodies: Correlation with primed lymphocyte typing.Proc Natl Acad Sci USA 84: 4596–4600, 1987

    CAS  PubMed  Google Scholar 

  • Bugawan, T. L., Horn, G. T., Long, C. M., Mickelson, E., Hansen, J. A., Ferrara, G. B., Angelini, G., and Erlich, H. A.: Analysis of HLA-DP allelic polymorphism using the in vitro enzymatic DNA amplification ofDPA andDPB loci.J Immunol 141: 4024–4030, 1988

    CAS  PubMed  Google Scholar 

  • Carlsson, B., Wallin, J., Bohme, J., and Moller, E.:HLA-DR-DQ haplotypes defined by restriction fragment length analysis. Correlation to serology.Hum Immunol 20: 95–113, 1987

    Article  CAS  PubMed  Google Scholar 

  • Eckels, D., Lake, P., Lamb, J. R., Johnson, A. W., Shaw, S., Woody, J. N., and Hartzman, R. J.: SB-restricted presentation of influenza and herpes simplex virus antigens to human T-lymphocyte clones.Nature 301: 716–718, 1983

    Article  CAS  PubMed  Google Scholar 

  • Erlich, H., Lee, J. S., Peterson, J. W., Bugawan, T., and DeMars, R.: Molecular analysis of HLA class I and class II antigen loss mutants reveals a homozygous deletion of theDR, DQ, and part of theDP region: Implications for class II gene order.Hum Immunol 16: 205–219, 1986

    Article  CAS  PubMed  Google Scholar 

  • Gilles, R. C. and Capra, J. D.: Biochemistry ofMHC class II molecules.Tissue Antigens 25: 57–68, 1985

    Article  Google Scholar 

  • Gustafsson, K., Emmoth, E., Widmark, E., Bohme, J., Peterson, P. A., and Rask, L.: Isolation of a cDNA clone coding for an SB beta chain.Nature 309: 76–78, 1984

    Article  CAS  PubMed  Google Scholar 

  • Gustafsson, K., Widmark, E., Jonsson, A. K., Servenius, B., Sachs, D. H., Larhammar, D., Rask, L., and Peterson, P. A.: Class II genes of the major histocompatibility complex. Evolution of theDP region as deduced from nucleotide sequences of the four genes.J Biol Chem 262: 8778–8786, 1987

    CAS  PubMed  Google Scholar 

  • Heyes, J., Austin, P., Bodmer, J., Bodmer, W., Madrigal, A., Mazzili, M. C., and Trowsdale, J.: Monoclonal antibodies to HLA-DP transfected mouse L cells.Proc Natl Acad Sci USA 84: 4596–4600, 1986

    Google Scholar 

  • Howell, M. D., Austin, R. K., Kelleher, D., Nepom, G. T., and Kagnoff, M. F.: AnHLA-D region restriction fragment length polymorphism associated with coeliac disease.J Exp Med 164: 333–338, 1986

    Article  CAS  PubMed  Google Scholar 

  • Howell, M. D., Smith, J. R., Austin, R. K., Kelleher, D., Nepom, G. T., Volk, B., and Kagnoff, M. F.: An extendedHLA-D region haplotype associated with coeliac disease.Proc Natl Acad Sci USA 85: 222–226, 1988

    CAS  PubMed  Google Scholar 

  • Hyldig-Nielsen, J. J., Morling, N., Ødum, N., Ryder, L. P., Platz, P., Jakobsen, B., and Svejgaard, A.: Restriction fragment length polymorphism of theHLA-DP subregion and correlations toHLA-DP phenotypes.Proc Natl Acad Sci USA 84: 1644–1648, 1987

    CAS  PubMed  Google Scholar 

  • Johnson, A. H.: Joint report. American Blacks.In M. Aizawa, T. Natori, A. Wakisaka, and Y. Konoeda (eds.).Proceedings of the Third Asia-Oceania Histocompatibility Workshop Conference, pp. 320–323, Hokkaido University Press, Hokkaido, 1986

    Google Scholar 

  • Kappes, D. J., Arnot, D., Okada, K., and Strominger, J. L.: Structure and polymorphism of theHLA class IISB light chain genes.EMBO J 3: 2985–2993, 1984

    CAS  PubMed  Google Scholar 

  • Kappes, D. J. and Strominger, J. L.: Structure and evolution of theHLA class IISX beta gene.Immunogenetics 24: 1–7, 1986

    Article  CAS  PubMed  Google Scholar 

  • Kelly, A. and Trowsdale, J.: Complete nucleotide sequence of a functionalHLA-DP beta gene of the region betweenDPB1 andDPA1 genes; comparison of the 5′ ends ofHLA class II genes.Nucleic Acids Res 13: 1607–1621, 1985

    CAS  PubMed  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Lair, B., Alber, C., Yu, W-Y., Watts, R., Bahl, M., and Karr, R. W.: A newly characterizedHLA-DP beta-chain allele. Evidence forDP beta heterogeneity within theDPw4 specificity.J Immunol 141: 1353–1357, 1988

    CAS  PubMed  Google Scholar 

  • Maeda, M., Inoko, H., Ando, A., Uryu, N., Nagata, Y., and Tsuji, K.: HLA-DP typing by analysis of DNA restriction fragment length polymorphisms in theHLA-DP beta subregion.Hum Immunol 21: 239–248, 1988

    Article  CAS  PubMed  Google Scholar 

  • Mittal, K. K., Hasegawa, T., Ting, H., Mickey, M. R., and Terasaki, P. I.: Genetic variation in theHLA system between Ainus, Japanese, and Caucasians.In J. Dausset and J. Colombani (eds.),Histocompatibility Testing 1972, pp. 187–195, Munksgaard Copenhagen, 1973

    Google Scholar 

  • Mitsuishi, Y., Urlacher, A., Tongio, M.-T., and Mayer, S.: Restriction fragment length polymorphism ofDP genes defines three newDP alleles.Immunogenetics 26: 383–388, 1987

    Article  CAS  PubMed  Google Scholar 

  • Moen, T., Bratlie, A., Kiss, E., Bruserud, O., and Thorsby, E.: Identification of four SB antigens by cloned cells. Population studies of Norwegians.Tissue Antigens 22: 298–309, 1983

    Article  CAS  PubMed  Google Scholar 

  • Morling, N., Hartzman, R. J., Jakobsen, B. K., Oedum, N., Platz, P., Ryder, L. P., Shaw, S., and Svejgaard, A.: Typing for DP-like determinants.In E. Albert, M. Baur, and W. Mayr (eds.):Histocompatibility Testing 1984, pp. 469; Springer-Verlag, Berlin, 1984

    Google Scholar 

  • Ødum, N., Hartzman, R. J., Jakobsen, B. K., Morling, N., Platz, P., Robbins, F. M., Ryder, L. P., and Svejgaard, A.: The HLA DP polymorphism in Denmark investigated by local and international PLT reagents. Definition of two ‘new’ DP antigens.Tissue Antigens 28: 105–118, 1986

    Article  PubMed  Google Scholar 

  • Ødum, N., Hyldig-Nilsen, J. J., Morling, N., Sandberg-Wollheim, M., Platz, P., and Svejgaard, A.: HLA-DP antigens are involved in the susceptibility to multiple sclerosis.Tissue Antigens 31: 235–237, 1988

    Article  PubMed  Google Scholar 

  • Nose, Y., Nakatsuji, T., and Reinsmoen, N. L.: A new HLA-DP (SB) specificity (Cp63) defined in Japanese and Caucasian cell populations by cloned PLT cells.Hum Immunol 13: 145–149, 1985

    Article  CAS  PubMed  Google Scholar 

  • Pawelec, G., Shaw, S., Schneider, M., Blaurock, M., Frauer, M., and Wernet, P.: Population studies of HLA-linked SB antigens and their relative importance in primary MLC typing. Analysis of HLA-D homozygous typing cells and normal heterozygous populations.Hum Immunol 5: 215–223, 1982

    Article  CAS  PubMed  Google Scholar 

  • Roux-Dosetto, M., Auffray, C., Lillie, J. W., Boss, J. M., Cohen, D., De Mars, R., Mawas, C., Seidman, J. G., and Strominger, J. L.: Genetic mapping of a human class II antigen beta-chain cDNA clone to theSB region of theHLA complex.Proc Natl Acad Sci USA 80: 6036–6040, 1983

    Google Scholar 

  • Servenius, B., Gustafsson, K., Widmark, E., Emmoth, E., Andersson, G., Larhammar, D., Rask, L., and Peterson, P. A.: Molecular map of the humanHLA-SB (HLA-DP) region and sequence of anSB alpha (DP alpha) pseudogene.EMBO J 3: 3209–3214, 1984

    CAS  PubMed  Google Scholar 

  • Shaw, S., Pollack, M. S., Payne, S. M., and Johnson, A. H.: HLA-linked B cell antigens of a new segregant series: Population and family studies of the SB antigens.Hum Immunol 1: 177–185, 1980

    Article  CAS  PubMed  Google Scholar 

  • Shaw, S., Duquesnoy, R. J., and Smith, P. L.: Population studies of the HLA-linked SB antigens.Immunogenetics 14: 153–162, 1981

    Article  CAS  PubMed  Google Scholar 

  • Southern, E. M.: Detection of specific sequences among DNA fragments separated by gel electrophoresis.J Mol Biol 98: 503–517, 1975

    CAS  PubMed  Google Scholar 

  • Stephens, H. A. F., Sakkas, L. I., Vaughan, R. W., Teitsson, I., Welsh, K. I., and Panayi, G. S.: HLA-DQw7 is a disease severity marker in patients with rheumatoid arthritis.Immunogenetics, in press, 1989

  • Teyton, L., Lotteau, V., Turmel, P., Arezana-Seiosdedos, F., Virelizier, J. L., Pujol, J. P., Loyau, G., Piater-Tonneau, D., Auffray, C., and Charron, D. J.: HLA DR, DQ, and DP antigen expression in rheumatoid synovial cells: A biochemical and quantitative study.J Immunol 138: 1730–1738, 1987

    CAS  PubMed  Google Scholar 

  • Todd, J., Acha-Orbea, H., Bell, J. I., Chao, N., Fronek, Z., Jacob, C. O., McDermott, M., Sinha, A. A., Timmerman, L., Steinman, L., and McDevitt, H. O.: A molecular basis for MHC class II-associated autoimmunity.Science 240: 1003–1009, 1988

    CAS  PubMed  Google Scholar 

  • Trowsdale, J., Kelly, A., Lee, J., Carson, S., Austin, P., and Travers, P.: Linkage map of twoHLA-SB beta and twoHLA-SB alpha related genes: An intron in one of theSB beta genes contains a processed pseudogene.Cell 38: 241–249, 1984

    Article  CAS  PubMed  Google Scholar 

  • Trowsdale, J., Young, J. A. T., Kelly, A. P., Austin, P. J., Carson, S., Meunier, H., So, A., Erlich, H. A., Spielman, R. S., Bodmer, J., and Bodmer, W. F.: Structure, sequence, and polymorphism in theHLA-D region.Immunol Rev 85: 5–43, 1985

    Article  CAS  PubMed  Google Scholar 

  • Wank, R. and Schendel, D. J.: Genetic analysis of HLA-D region products defined by PLT.In E. Albert, M. Baur, and W. Mayr (eds.):Histocompatibility Testing 1984, pp. 289–299, Springer-Verlag, Berlin, 1984

    Google Scholar 

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Authors and Affiliations

  1. Rheumatology Units, Division of Medicine, United Medical and Dental Schools, Guy's Hospital, SE1 9RT, London, UK

    Henry A. F. Stephens, Robert W. Vaughan, Lazaros I. Sakkas, Ken I. Welsh & Gabriel S. Panayi

  2. Molecular Immunogenetics Units, Division of Medicine, United Medical and Dental Schools, Guy's Hospital, SE1 9RT, London, UK

    Henry A. F. Stephens, Robert W. Vaughan, Lazaros I. Sakkas, Ken I. Welsh & Gabriel S. Panayi

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  1. Henry A. F. Stephens
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Stephens, H.A.F., Vaughan, R.W., Sakkas, L.I. et al. Southern blot analysis of HLA-DP gene polymorphisms in Caucasoid rheumatoid arthritis (RA) patients and controls. Immunogenetics 30, 149–155 (1989). https://doi.org/10.1007/BF02421199

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