Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1989 Aug;84(2):613–618. doi: 10.1172/JCI114206

HLA gene amplification and hybridization analysis of polymorphism. HLA matching for bone marrow transplantation of a patient with HLA-deficient severe combined immunodeficiency syndrome.

L A Baxter-Lowe 1, J B Hunter 1, J T Casper 1, J Gorski 1
PMCID: PMC548923  PMID: 2668336

Abstract

The treatment of choice for certain immunodeficiency syndromes and hematological disorders is bone marrow transplantation (BMT). The success of BMT is influenced by the degree of HLA compatibility between recipient and donor. However, aberrant expression of HLA sometimes makes it difficult, if not impossible, to determine the patient's HLA type by standard serological and cellular techniques. We describe here the application of new molecular biological techniques to perform high resolution HLA typing independent of HLA expression. A patient with HLA-deficient severe combined deficiency was HLA typed using in vitro amplification of the HLA genes and sequence-specific oligonucleotide probe hybridization (SSOPH). Two major advances provided by this technology are:detection of HLA polymorphism at the level of single amino acid differences; and elimination of a requirement for HLA expression. Although the patient's lymphocytes lacked class II HLA proteins, polymorphism associated with DR7,w53;DQw2;DRw11a (a split of DR5), w52b (a split of DRw52);DQw7 were identified. The patient's class I expression was partially defective, and typing was accomplished by a combination of serological (HLA-A and -C) and SSOPH analysis (HLA-B). Complete patient haplotypes were predicted after typing of family members [A2;B35(w6); Cw4; DRw11a(w52b);DQw7 and A2;B13(w4); Cw6;DR7(w53); DQw2]. Potential unrelated donors were typed and a donor was selected for BMT.

Full text

PDF
613

Images in this article

615Image
on p.615

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Angelini G., de Preval C., Gorski J., Mach B. High-resolution analysis of the human HLA-DR polymorphism by hybridization with sequence-specific oligonucleotide probes. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4489–4493. doi: 10.1073/pnas.83.12.4489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bell J. I., Denney D., Jr, Foster L., Belt T., Todd J. A., McDevitt H. O. Allelic variation in the DR subregion of the human major histocompatibility complex. Proc Natl Acad Sci U S A. 1987 Sep;84(17):6234–6238. doi: 10.1073/pnas.84.17.6234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bidwell J. DNA-RFLP analysis and genotyping of HLA-DR and DQ antigens. Immunol Today. 1988 Jan;9(1):18–23. doi: 10.1016/0167-5699(88)91351-5. [DOI] [PubMed] [Google Scholar]
  4. Bjorkman P. J., Saper M. A., Samraoui B., Bennett W. S., Strominger J. L., Wiley D. C. Structure of the human class I histocompatibility antigen, HLA-A2. Nature. 1987 Oct 8;329(6139):506–512. doi: 10.1038/329506a0. [DOI] [PubMed] [Google Scholar]
  5. Boss J. M., Strominger J. L. Cloning and sequence analysis of the human major histocompatibility complex gene DC-3 beta. Proc Natl Acad Sci U S A. 1984 Aug;81(16):5199–5203. doi: 10.1073/pnas.81.16.5199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brown J. H., Jardetzky T., Saper M. A., Samraoui B., Bjorkman P. J., Wiley D. C. A hypothetical model of the foreign antigen binding site of class II histocompatibility molecules. Nature. 1988 Apr 28;332(6167):845–850. doi: 10.1038/332845a0. [DOI] [PubMed] [Google Scholar]
  7. Carlsson B., Wallin J., Böhme J., Möller E. HLA-DR-DQ haplotypes defined by restriction fragment analysis. Correlation to serology. Hum Immunol. 1987 Oct;20(2):95–113. doi: 10.1016/0198-8859(87)90025-5. [DOI] [PubMed] [Google Scholar]
  8. Gorski J., Mach B. Polymorphism of human Ia antigens: gene conversion between two DR beta loci results in a new HLA-D/DR specificity. Nature. 1986 Jul 3;322(6074):67–70. doi: 10.1038/322067a0. [DOI] [PubMed] [Google Scholar]
  9. Gregersen P. K., Moriuchi T., Karr R. W., Obata F., Moriuchi J., Maccari J., Goldberg D., Winchester R. J., Silver J. Polymorphism of HLA-DR beta chains in DR4, -7, and -9 haplotypes: implications for the mechanisms of allelic variation. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9149–9153. doi: 10.1073/pnas.83.23.9149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gregersen P. K., Shen M., Song Q. L., Merryman P., Degar S., Seki T., Maccari J., Goldberg D., Murphy H., Schwenzer J. Molecular diversity of HLA-DR4 haplotypes. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2642–2646. doi: 10.1073/pnas.83.8.2642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hows J. M. Histocompatible unrelated donors for bone marrow transplantation. Bone Marrow Transplant. 1987 Feb;1(3):259–263. [PubMed] [Google Scholar]
  12. Lee B. S., Rust N. A., McMichael A. J., McDevitt H. O. HLA-DR2 subtypes form an additional supertypic family of DR beta alleles. Proc Natl Acad Sci U S A. 1987 Jul;84(13):4591–4595. doi: 10.1073/pnas.84.13.4591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Marcadet A., Cohen D., Dausset J., Fischer A., Durandy A., Griscelli C. Genotyping with DNA probes in combined immunodeficiency syndrome with defective expression of HLA. N Engl J Med. 1985 May 16;312(20):1287–1292. doi: 10.1056/NEJM198505163122004. [DOI] [PubMed] [Google Scholar]
  14. Miller S. A., Dykes D. D., Polesky H. F. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 1988 Feb 11;16(3):1215–1215. doi: 10.1093/nar/16.3.1215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. doi: 10.1126/science.2448875. [DOI] [PubMed] [Google Scholar]
  16. Schwartz R. H. T-lymphocyte recognition of antigen in association with gene products of the major histocompatibility complex. Annu Rev Immunol. 1985;3:237–261. doi: 10.1146/annurev.iy.03.040185.001321. [DOI] [PubMed] [Google Scholar]
  17. Steimle V., Hinkkanen A., Schlesier M., Epplen J. T. A novel HLA-DR beta I sequence from the DRw11 haplotype. Immunogenetics. 1988;28(3):208–210. doi: 10.1007/BF00375861. [DOI] [PubMed] [Google Scholar]
  18. Tieber V. L., Abruzzini L. F., Didier D. K., Schwartz B. D., Rotwein P. Complete characterization and sequence of an HLA class II DR beta chain cDNA from the DR5 haplotype. J Biol Chem. 1986 Feb 25;261(6):2738–2742. [PubMed] [Google Scholar]
  19. Todd J. A., Bell J. I., McDevitt H. O. HLA-DQ beta gene contributes to susceptibility and resistance to insulin-dependent diabetes mellitus. Nature. 1987 Oct 15;329(6140):599–604. doi: 10.1038/329599a0. [DOI] [PubMed] [Google Scholar]
  20. Wood W. I., Gitschier J., Lasky L. A., Lawn R. M. Base composition-independent hybridization in tetramethylammonium chloride: a method for oligonucleotide screening of highly complex gene libraries. Proc Natl Acad Sci U S A. 1985 Mar;82(6):1585–1588. doi: 10.1073/pnas.82.6.1585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Wu S., Saunders T. L., Bach F. H. Polymorphism of human Ia antigens generated by reciprocal intergenic exchange between two DR beta loci. Nature. 1986 Dec 18;324(6098):676–679. doi: 10.1038/324676a0. [DOI] [PubMed] [Google Scholar]
  22. Zemmour J., Ennis P. D., Parham P., Dupont B. Comparison of the structure of HLA-Bw47 to HLA-B13 and its relationship to 21-hydroxylase deficiency. Immunogenetics. 1988;27(4):281–287. doi: 10.1007/BF00376123. [DOI] [PubMed] [Google Scholar]
  23. van Rood J. J., van Leeuwen A., Keuning J. J., van Oud Alblas A. B. The serological recognition of the human MLC determinants using a modified cytotoxicity technique. Tissue Antigens. 1975 Apr;5(2):73–79. doi: 10.1111/j.1399-0039.1975.tb00532.x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES