HLA-A, -B, -DR en receptores de trasplante de médula ósea en Uruguay: Sistema nacional de registro, tipificación y búsqueda de donantes de médula ósea y progenitores hematopoyéticos (SINDOME)

Milka Bengochea; Inés Álvarez; Pedro C. Hidalgo; Adriana Cabrera; Olga Senatore; Roberto Toledo; Elena Carreto; Martha Sosa; Doris Abilleira; Ely Silva

Authors

Milka Bengochea Universidad de la República, Facultad de Medicina, Hospital de Clínicas, Banco Nacional de Órganos y Tejidos, Prof. Adjunta
Inés Álvarez Universidad de la República, Facultad de Medicina, Hospital de Clínicas, Banco Nacional de Órganos y Tejidos, Profesora
Pedro C. Hidalgo Universidad de la República, Facultad de Medicina, Hospital de Clínicas, Banco Nacional de Órganos y Tejidos, Genetista
Adriana Cabrera Universidad de la República, Facultad de Medicina, Hospital de Clínicas, Banco Nacional de Órganos y Tejidos, Laboratorio de Inmunogenética e Histocompatibilidad, Laboratorio Clínico, Licenciada
Olga Senatore Universidad de la República, Facultad de Medicina, Hospital de Clínicas, Banco Nacional de Órganos y Tejidos, Programa SINDOME, Asistente Social
Roberto Toledo Universidad de la República, Facultad de Medicina, Hospital de Clínicas, Banco Nacional de Órganos y Tejidos, Prof. Agregado
Elena Carreto Universidad de la República, Facultad de Medicina, Hospital de Clínicas, Banco Nacional de Órganos y Tejidos, Ex asistente
Martha Sosa Universidad de la República, Facultad de Medicina, Hospital de Clínicas, Banco Nacional de Órganos y Tejidos, Laboratorio de Inmunogenética e Histocompatibilidad, Técnico en Hemoterapia
Doris Abilleira Universidad de la República, Facultad de Medicina, Hospital de Clínicas, Banco Nacional de Órganos y Tejidos, Laboratorio de Inmunogenética e Histocompatibilidad, Laboratorio Clínico, Licenciada
Ely Silva Universidad de la República, Facultad de Medicina, Hospital de Clínicas, Banco Nacional de Órganos y Tejidos, Laboratorio de Inmunogenética e Histocompatibilidad, Laboratorio Clínico, Licenciada

Keywords:

HLA-A ANTIGENS, HLA-B ANTIGENS, HLA-DR ANTIGENS, HISTOCOMPATIBILITY, BONE MARROW TRANSPLANTATION, TRANSPLANTATION OF HEMATOPOIETIC STEM CELLS, URUGUAY

Abstract

Background and Objective: as starting point Uruguay has as starting pointa transplant solidarity system based upon the Fondo Nacional de Recursos and the Banco Nacional de Organos y Tejidos (BNOT). HLA compatibility is a biological boundary in allogenic bone marrow transplantation (BMT). HLA allele and haplotype frequencies are used to determine the probability to find a donor with a particular HLA phenotype and to predict the degree of HLA mismatching in a bone marrow donation scheme. In Uruguay, all potential donors are typified in In the Immunogenetic and Histocompatibility Laboratory of the BNOT are typified all potentials bone marrow recipients and donors from all the country. In this context a program was developed for the register, to register, evaluate, and procure the non-related evaluation and procurement of non-related potential bone marrow donors at national level known as SINDOME. Our aim was to analyzed bone marrow donors and recipients for allogenic transplantations registered in our register from January 1997 to May 2002. We also determined the genetic constitution of HLA system for 298 bone marrow recipients.
Method: 346 recipients and 1083 donors were analyzed. Allele and haplotype frequencies were determined in a sample of 298 recipients. HLA class I alleles were typing by standard complement-dependent microcytotoxic assays. HLA class II alleles were was determined by polymerase chain reactionsequence specific oligonucleotide (PCR-SSO) technique, with medium-high resolution.
We analyzed allele and haplotype frequencies in a sample of 298 recipients.
Results: among the 346 bone marrow recipient subjects recorded in the SINDOME, 58% were younger than 30 years. Donor/recipient rate was 3.13 but only 45% of potential receptors matched with a compatible donor. HLA-A, -B, -DR polymorphism analysis of the sample showed that prevalent alleles were A2 (28.97%), B35 (12.49%) and DR04 (15.24%). Only the HLA-DRB1 loci locus showed a highly significant deviation from Hardy-Weinberg equilibrium. Most common haplotypes were A2-B51 and A2-B7 for HLA-A, -B, A2-DR11 and A2-DR04 for HLA-A -DRB1, and haplotype B8-DR03 for HLA-B-DR. Highest frequencies were observed in HLA-A1-B8-DR03 and HLA-A2-B51-DR13 for haplotypes HLA-A-B-DRB1.
Conclusion: our findings showed that HLA system is heterogeneous, with a wide range of polymorphisms and haplotype distribution. The possibility to find compatible donors for a group of patients with malignant diseases without sibling donors is low in our country. In these cases, searching in international donor records might be an alternative to find potential suitable marrow donors for allogenic transplantations.

References

1) Thomas E, Storb R, Clift RA, Fefer A, Johnson FL, Neiman PE, et al. Bone-marrow transplantation. N Engl J Med 1975; 292: 832-43.
2) Remberger M, Ringdén O, Blau I, Ottinger H, Kremens B, Kiehl M, et al. No difference in graft-versus-host disease, relapse, and survival comparing peripheral stem cells to bone marrow using unrelated donors. Blood 2001; 98: 1739-45.
3) Kurtzberg J, Laughlin M, Graham L, Smith C, Olson J, Halperin E, et al. Placental Blood as a source of hematopoietic stem cells for transplantation into unrelated recipients. N Eng J Med 1996; 335: 157-66.
4) Rocha V, Cornish J, Sievers E, Filipovich A, Locatelli F, Peters C, et al. Comparison of outcomes of unrelated bone marrow and umbilical cord blood transplants in children with acute leukemia. Blood 2001; 97: 2962-71.
5) Molina A, Popplewell L, Kashyap A, Nademanee A. Hematopoietic Stem cell transplantation in the new Millenium: Report from City of Hope National Medical Center. In: Cecka JN, Terasaki PI, eds. Clinical Transplants 1999. Los Angeles: UCLA Immunogenetics Center, 1999: 317-42.
6) International Bone Marrow Transplant Registry (IBMTR) and the Autologous Blood and Marrow Transplant Registry. 2001 IBMTR/ABMTR statistical center report of survival statistics for blood and marrow transplants. http://www.ibmtr.org/infoserv/info_rep.html (visto el 17/9/2003).
7) Fondo Nacional de Recursos. Medicina Altamente Especializada. Informe: Programa de Seguimiento Trasplante de Médula Ósea. Montevideo: FNR, 2002.
8) National Marrow Donor Program. 1999 Data & Scientific Report of the National Donor Program. U.S. Department of Health and Human Services. Health Resources and Services Administration Office of Special Programs. Division of Transplantation. Miniapolis: NMDP, 2000.
9) Bishop M, Flowers ME, Foss F, Couriel D. Emerging Strategies in the tr eatment of Chronic Graft-versus-Host Disease. Blood Marrow Transplant Rev 2002; 12: 4-15.
10) Vela Ojeda J. Trasplante alogénico de células hematopoyéticas no mieloablativo. Gac Méd Méx 2002; 138: 142-4.
11) Van Deerlin VMD, Leonard DGB. Bone marrow engraftment analysis after allogeneic bone marrow transplantations. Clin Lab Med 2000; 20: 197-225.
12) Wood K, Sachs DH. Chimerism and transplantation tolerance: cause and effect. Immunol Today 1996; 17: 584-9.
13) Zinkernagel RM, Doherty PC. Restriccion of in vitro T-cell-mediated cytotoxicity in lymphocytic choriomeningitis within a syngeneic or semiallogeneic system. Nature 1974; 248: 701-2.
14) Newton-Nash D. The molecular basis of allorecognition. Hum Immunol 1994; 41: 105-11.
15) The MHC sequence consortium. Complete sequence and gene map of a human major histocompatibility complex. Nature 1999; 40: 921-3.
16) Jordan F, McWhinnie A, Turner S, Gavira N, Calvert A, Cleaver S, et al. Comparison of HLA DRB1 typing by DNA-RFLP, PCR-SSO and PCR-SSP methods and their application in providing matched unrelated donors for bone marrow transplantation. Tissue Antigens 1995; 45: 103-10.
17) Little A, Marsh S, Madrigal A. Current methodologies of human leukocyte antigen typing utilized for bone marrow donor selection. Curr Opin Hematol 1998; 5: 419-28.
18) Marsh SG. The HLA Sequence Database. HLA Informatics group. http://www.anthonynolan.com (visto el 14/10/2002).
19) Mori M, Beatty P, Graves M, Boucher K, Mildford E. HLA gene and haplotype frequencies in the North American population. Transplantation 1987; 64: 1017-27.
20) Shipper RF, D'Amaro J, Bakker JT, Bakker J, Van Rood JJ, Oudshoorn M. HLA gene and haplotype frequencies in bone marrow donors worldwide registries. Hum Immunol 1997; 52: 54-71.
21) Zachary AA, Steinberg AG, Bias WB, Leffell MS. The frequencies of HLA alleles and haplotypes and their distribution among donors and renal patients in the UNOS registry. Transplantation 1996; 62: 272-83.
22) Bengochea M, Álvarez I, Toledo R, Cabrera A, Abilleira D, Silva E, et al. Frecuencias HLA A y B estudiadas con anticuerpos monoclonales. Libro de Resúmenes. Congreso Latinoamericano de Antropología Biológica, 4, Piriápolis: ALAB, 2000: 48.
23) Petersdorf E, Longton G, Anasetti C, Mickelson E, McKinney S, Smith A. Association of HLA-C disparity with graft failure after marrow transplantation from unrelated Donors. Blood 1997; 89: 1818-23.
24) Petersdorf E, Longton G, Anasetti C, Mickelson E, Smith A, Martin P, et al. Definition of HLA-DQ as a transplantation antigen. Proc Natl Acad Sci USA 1996; 93: 15358-63.
25) Madrigal A, Scott I, Arguello R, Szydlo R, Little A, Goldman J, et al. Factors influencing the outcome of bone marrow transplants using unrelated donors. Immunol Rev 1997; 157: 153-66.
26) Petersdorf E, Mickelson E, Anasetti C, Martin P, Woolfrey A, Hansen J. Effect of HLA mismatches on the outcome of hematopoietic transplants. Curr Opin Immunol 1999; 11: 521-6.
27) Scott I, O'Shea J, Bunce M, Tiercy J, Arguello J, Firman H, et al. Molecular typing shows a high level of HLA class I incompatibility in serologically well matched donor/patient pairs: Implications for unrelated bone marrow donor selection. Blood 1998; 92: 4864-71.
28) Petersdorf E, Gooley T, Anasetti C, Martin P, Smith A, Mickelson E, et al. Optimizing outcome after unrelated marrow transplantation by comprehensive matching of HLA class I and II alleles in the donor and Recipient. Blood 1998; 92: 3515-20.
29) Henslee-Downe PJ, Abhyankar S, Parrish R, Pati A, Godder K, Neglia W, et al. Use of partially mismatched related donors extends access to allogeneic marrow transplant. Blood 1997; 89: 3864-72.
30) Barker JN, Krepski T, DeFor T, Davies S, Wagner J, Weisdorf D, et al. Searching for unrelated donor haemopoietic stem cell: Avaibility and speed of umbilical cord blood versus Bone marrow. Biol Blood Marrow Transplant 2002; 8: 257-60.
31) Bone Marrow Donor Worldwide. Annual Reports 2001. Leiden: Drukkerij Groen. http://www.bmdw.org (visto el 28/8/2002).
32) O'Shea J, Cleaver S, Little A, Madrigal A. Searching for an unrelated haemopoietic stem cell donor- A United Kingdom Perspective. In: Cecka JN, Terasaki PI, ed. Clinical Transplants 1999. Los Angeles: UCLA Immunogenetics Center, 1999: 129-37.
33) Terasaki PI, Pernoco F, Park MS, Ozturk G, Iwaki Y. Microdroplet testing for HLA A,B,C and D antigens. Am J Clin Pathol 1978; 69: 103-20.
34) Gustincich S, Manfiolett G, Del Sal G, Schneider C, Carminci P. A fast method for high-quality genomic DNA extraction from whole human blood. BioTechniques 1991; 11: 298.
35) Bugawan T, Erlich H. A method for typing polymorphism at HLA A locus using PCR amplification and immobilized oligonucleotide probes.Tissue Antigens 1994; 44: 137-47.
36) Marsh SGE. Nomenclature for factors of the HLA system. http:// www.anthonynolan.com/HIG/nomenc.html (visto el 30/12/2002).
37) Gjertson DW, Geer L, Lee S-H, Kawata J, Sutrino R. Populations studies. In: Terasaki PI, Gjertson DW, ed. HLA 1997. Los Angeles: UCLA Tissue Typing Laboratory. 1997: 171-3.
38) Weir BS. Genetic Data Analysis II. 2da. ed. Sunderland: Sinauer, 1996: 105-33.
39) Schneider S, Roessli D, Excoffier L. Arlequin ver. 2.000. A software for population genetic data analysis. Genetics and Biometry Laboratory. Geneva: University of Geneva, 2000.
40) Lewontin RC. On measures of gametic disequilibrium. Genetics 1988; 120: 849-52.
41) Thomson G, Baur MP. Third order linkage disiquilibrium. Tissue Antigens 1984; 24: 250-5.
42) Guo SW, Thompson EA. Performing the exact test of Hardy-Weinberg proportion for multiple alleles. Biometrics 1992; 48: 361-72.
43) Álvarez I, Sans M, Toledo R, Sosa M, Bengochea M, Salzano FM. Hla gene and haplotype frequencies in Uruguay. Int J Anthrop 1993; 8: 163-8.
44) Lonjou C, Clayton J, Cambon-Thomsen A, Raffoux C. HLA -A, -B, -DR haplotype frequencies in France-Implications for recruitment of potential bone marrow donors. Transplantation 1995; 60: 375-83.
45) Cao K, Hollenbach J, Shi X Shi W, Chopek M, Fernández-Viña MA. Analysis of the frequencies of HLA-A, B, and C alleles and haplotypes in the five major ethnic groups of the United States reveals high levels of diversity in these loci and constrasting distribution pattern in these populations. Hum Immunol 2001; 62: 1009-30.