Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1991 Aug 15;88(16):7121–7125. doi: 10.1073/pnas.88.16.7121

Selection of unrelated donors for bone marrow transplantation is improved by HLA class II genotyping with oligonucleotide hybridization.

J M Tiercy 1, C Morel 1, A C Freidel 1, F Zwahlen 1, L Gebuhrer 1, H Bétuel 1, M Jeannet 1, B Mach 1
PMCID: PMC52245  PMID: 1908088

Abstract

As the demand for donors for bone marrow transplantation increases, the use of HLA-matched, genetically unrelated donors represents a promising strategy. It is well documented that the clinical outcome of bone marrow transplantation is directly dependent on optimal matching for HLA class I and class II specificities. Molecular studies have revealed the existence of a much larger number of HLA class II alleles than was anticipated, many of which cannot be recognized by routine serological typing. Currently this "hidden" polymorphism represents a major limitation to the generalized use of unrelated donors for bone marrow transplantation. It has recently become possible, however, to identify HLA allelic polymorphism directly at the DNA level by hybridization with sequence-specific oligonucleotide probes ("HLA oligotyping") after amplification of DNA by polymerase chain reaction. In this study, we have investigated whether donor-recipient pairs that are fully matched for HLA by serology are truly HLA-DR, -DQ, and -DP identical and to what extent class II differences influence the primary mixed lymphocyte culture. We show that HLA oligotyping, performed on 50 pairs of HLA class I and II serologically matched individuals, can indeed reveal phenotypically relevant allelic differences at either DRB or DQB loci in 56% of these pairs and can therefore improve HLA class II typing and the choice of bone marrow donors quite significantly. Oligotyping for DRB/DQB/DPB polymorphism also allows prediction of a positive mixed lymphocyte culture, as established in 31 donor/recipient combinations, and even detection of polymorphic differences that were not revealed by this test. This approach is well suited for accurate HLA typing of large pools of bone marrow donors and was successfully applied to select fully matched donors for bone marrow transplantation.

Full text

PDF
7121

Images in this article

7122Image
on p.7122
7122Image
on p.7122
7122Image
on p.7122
7122Image
on p.7122
7122Image
on p.7122
7124Image
on p.7124
7124Image
on p.7124

Selected References

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

  1. Anasetti C., Amos D., Beatty P. G., Appelbaum F. R., Bensinger W., Buckner C. D., Clift R., Doney K., Martin P. J., Mickelson E. Effect of HLA compatibility on engraftment of bone marrow transplants in patients with leukemia or lymphoma. N Engl J Med. 1989 Jan 26;320(4):197–204. doi: 10.1056/NEJM198901263200401. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Ash R. C., Casper J. T., Chitambar C. R., Hansen R., Bunin N., Truitt R. L., Lawton C., Murray K., Hunter J., Baxter-Lowe L. A. Successful allogeneic transplantation of T-cell-depleted bone marrow from closely HLA-matched unrelated donors. N Engl J Med. 1990 Feb 22;322(8):485–494. doi: 10.1056/NEJM199002223220801. [DOI] [PubMed] [Google Scholar]
  4. Awdeh Z. L., Alper C. A., Eynon E., Alosco S. M., Stein R., Yunis E. J. Unrelated individuals matched for MHC extended haplotypes and HLA-identical siblings show comparable responses in mixed lymphocyte culture. Lancet. 1985 Oct 19;2(8460):853–856. doi: 10.1016/s0140-6736(85)90123-0. [DOI] [PubMed] [Google Scholar]
  5. Baxter-Lowe L. A., Hunter J. B., Casper J. T., Gorski J. HLA gene amplification and hybridization analysis of polymorphism. HLA matching for bone marrow transplantation of a patient with HLA-deficient severe combined immunodeficiency syndrome. J Clin Invest. 1989 Aug;84(2):613–618. doi: 10.1172/JCI114206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Beatty P. G., Ash R., Hows J. M., McGlave P. B. The use of unrelated bone marrow donors in the treatment of patients with chronic myelogenous leukemia: experience of four marrow transplant centers. Bone Marrow Transplant. 1989 May;4(3):287–290. [PubMed] [Google Scholar]
  7. Beatty P. G., Clift R. A., Mickelson E. M., Nisperos B. B., Flournoy N., Martin P. J., Sanders J. E., Stewart P., Buckner C. D., Storb R. Marrow transplantation from related donors other than HLA-identical siblings. N Engl J Med. 1985 Sep 26;313(13):765–771. doi: 10.1056/NEJM198509263131301. [DOI] [PubMed] [Google Scholar]
  8. Beatty P. G., Dahlberg S., Mickelson E. M., Nisperos B., Opelz G., Martin P. J., Hansen J. A. Probability of finding HLA-matched unrelated marrow donors. Transplantation. 1988 Apr;45(4):714–718. doi: 10.1097/00007890-198804000-00010. [DOI] [PubMed] [Google Scholar]
  9. Bodmer J. G., Marsh S. G., Albert E. D., Bodmer W. F., Dupont B., Erlich H. A., Mach B., Mayr W. R., Parham P., Sasazuki T. Nomenclature for factors of the HLA system, 1990. Tissue Antigens. 1991 Mar;37(3):97–104. doi: 10.1111/j.1399-0039.1991.tb01853.x. [DOI] [PubMed] [Google Scholar]
  10. Bugawan T. L., Horn G. T., Long C. M., Mickelson E., Hansen J. A., Ferrara G. B., Angelini G., Erlich H. A. Analysis of HLA-DP allelic sequence polymorphism using the in vitro enzymatic DNA amplification of DP-alpha and DP-beta loci. J Immunol. 1988 Dec 1;141(11):4024–4030. [PubMed] [Google Scholar]
  11. Clay T. M., Jones H. P., Bidwell J. L., Darke C., Harvey J., Bradley B. A. A comparison of DNA-RFLP typing with serology and mixed lymphocyte reaction in the selection of matched unrelated bone marrow donors. Bone Marrow Transplant. 1989 Sep;4(5):493–497. [PubMed] [Google Scholar]
  12. Freidel A. C., Betuel H., Gebuhrer L., Farre A., Lambert J. Cellular and serological subtypes of HLA-DR2. Tissue Antigens. 1987 Mar;29(3):129–140. doi: 10.1111/j.1399-0039.1987.tb01565.x. [DOI] [PubMed] [Google Scholar]
  13. Hansen J. A., Clift R. A., Thomas E. D., Buckner C. D., Storb R., Giblett E. R. Transplantation of marrow from an unrelated donor to a patient with acute leukemia. N Engl J Med. 1980 Sep 4;303(10):565–567. doi: 10.1056/NEJM198009043031007. [DOI] [PubMed] [Google Scholar]
  14. Hows J. M., Yin J. L., Marsh J., Swirsky D., Jones L., Apperley J. F., James D. C., Smithers S., Batchelor J. R., Goldman J. M. Histocompatible unrelated volunteer donors compared with HLA nonidentical family donors in marrow transplantation for aplastic anemia and leukemia. Blood. 1986 Dec;68(6):1322–1328. [PubMed] [Google Scholar]
  15. Irlé C., Chapuis B., Jeannet M., Kaestli M., Montandon N., Speck B. Detection of anti-non-MHC-directed T cell reactivity following in vivo priming after HLA-identical marrow transplantation and following in vitro priming in limiting dilution cultures. Transplant Proc. 1987 Feb;19(1 Pt 3):2674–2677. [PubMed] [Google Scholar]
  16. Jeannet M., Speck B., Sartorius J. Donor selection for bone marrow transplantation. Predictive value of DR typing for mixed lymphocyte culture compatibility between unrelated individuals. Transplantation. 1978 Dec;26(6):448–449. [PubMed] [Google Scholar]
  17. Kaminski E., Hows J., Man S., Brookes P., Mackinnon S., Hughes T., Avakian O., Goldman J. M., Batchelor J. R. Prediction of graft versus host disease by frequency analysis of cytotoxic T cells after unrelated donor bone marrow transplantation. Transplantation. 1989 Oct;48(4):608–613. [PubMed] [Google Scholar]
  18. Mach B., Gorski J., Rollini P., Berte C., Amaldi I., Berdoz J., Ucla C. Polymorphism and regulation of HLA class II genes of the major histocompatibility complex. Cold Spring Harb Symp Quant Biol. 1986;51(Pt 1):67–74. doi: 10.1101/sqb.1986.051.01.009. [DOI] [PubMed] [Google Scholar]
  19. Mach B., Tiercy J. M. Genotypic typing of HLA class II: from the bench to the bedside. Hum Immunol. 1991 Apr;30(4):278–284. doi: 10.1016/0198-8859(91)90007-v. [DOI] [PubMed] [Google Scholar]
  20. Morel C., Zwahlen F., Jeannet M., Mach B., Tiercy J. M. Complete analysis of HLA-DQB1 polymorphism and DR-DQ linkage disequilibrium by oligonucleotide typing. Hum Immunol. 1990 Sep;29(1):64–77. doi: 10.1016/0198-8859(90)90070-6. [DOI] [PubMed] [Google Scholar]
  21. Mullis K. B., Faloona F. A. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987;155:335–350. doi: 10.1016/0076-6879(87)55023-6. [DOI] [PubMed] [Google Scholar]
  22. Odum N., Platz P., Jakobsen B. K., Petersen C. M., Jacobsen N., Møller J., Ryder L. P., Lamm L., Svejgaard A. HLA-DP and bone marrow transplantation: DP-incompatibility and severe acute graft versus host disease. Tissue Antigens. 1987 Nov;30(5):213–216. doi: 10.1111/j.1399-0039.1987.tb01624.x. [DOI] [PubMed] [Google Scholar]
  23. Olerup O., Möller E., Persson U. HLA-DP incompatibilities induce significant proliferation in primary mixed lymphocyte cultures in HLA-A, -B, -DR and -DQ compatible individuals: implications for allogeneic bone marrow transplantation. Tissue Antigens. 1990 Nov;36(5):194–202. doi: 10.1111/j.1399-0039.1990.tb01829.x. [DOI] [PubMed] [Google Scholar]
  24. Pollack M. S., Chin-Louie J., Callaway C., Blanco M. A., Slavin K., Sullivan K., O'Reilly R. J., Dupont B. Mixed lymphocyte reactions for individuals with phenotypic identity for specific HLA-B,DR determinants: the role of linkage disequilibrium and of specific DR and other class II determinants. J Clin Immunol. 1983 Oct;3(4):341–351. doi: 10.1007/BF00915795. [DOI] [PubMed] [Google Scholar]
  25. Saiki R. K., Bugawan T. L., Horn G. T., Mullis K. B., Erlich H. A. Analysis of enzymatically amplified beta-globin and HLA-DQ alpha DNA with allele-specific oligonucleotide probes. Nature. 1986 Nov 13;324(6093):163–166. doi: 10.1038/324163a0. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Sell T. W., Eckels D. D. Primary mixed lymphocyte responses to HLA-DP. Hum Immunol. 1990 Sep;29(1):23–30. doi: 10.1016/0198-8859(90)90066-x. [DOI] [PubMed] [Google Scholar]
  28. Thomas E. D. Karnofsky Memorial Lecture. Marrow transplantation for malignant diseases. J Clin Oncol. 1983 Sep;1(9):517–531. doi: 10.1200/JCO.1983.1.9.517. [DOI] [PubMed] [Google Scholar]
  29. Tiercy J. M., Gorski J., Jeannet M., Mach B. Identification and distribution of three serologically undetected alleles of HLA-DR by oligonucleotide.DNA typing analysis. Proc Natl Acad Sci U S A. 1988 Jan;85(1):198–202. doi: 10.1073/pnas.85.1.198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tiercy J. M., Goumaz C., Mach B., Jeannet M. Application of HLA-DR oligotyping to 110 kidney transplant patients with doubtful serological typing. Transplantation. 1991 May;51(5):1110–1114. doi: 10.1097/00007890-199105000-00034. [DOI] [PubMed] [Google Scholar]
  31. Tiercy J. M., Jeannet M., Mach B. A new HLA-DRB1 allele within the DRw52 supertypic specificity (DRw13-DwHAG): sequencing and direct identification by oligonucleotide typing. Eur J Immunol. 1990 Feb;20(2):237–241. doi: 10.1002/eji.1830200202. [DOI] [PubMed] [Google Scholar]
  32. Tiercy J. M., Jeannet M., Mach B. A new approach for the analysis of HLA class II polymorphism: 'HLA oligotyping'. Blood Rev. 1990 Mar;4(1):9–15. doi: 10.1016/0268-960x(90)90012-h. [DOI] [PubMed] [Google Scholar]
  33. Wake C. T., Long E. O., Mach B. Allelic polymorphism and complexity of the genes for HLA-DR beta-chains--direct analysis by DNA-DNA hybridization. Nature. 1982 Nov 25;300(5890):372–374. doi: 10.1038/300372a0. [DOI] [PubMed] [Google Scholar]
  34. de Koster H. S., Anderson D. C., Termijtelen A. T cells sensitized to synthetic HLA-DR3 peptide give evidence of continuous presentation of denatured HLA-DR3 molecules by HLA-DP. J Exp Med. 1989 Mar 1;169(3):1191–1196. doi: 10.1084/jem.169.3.1191. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES