OncohematologyOncohematologyОнкогематология1818-83462413-4023Publishing House ABV Press19610.17650/1818-8346-2016-11-2-40-50BASIC RESEARCHФУНДАМЕНТАЛЬНЫЕ ИССЛЕДОВАНИЯ В ПРАКТИЧЕСКОЙ МЕДИЦИНЕ НА СОВРЕМЕННОМ ЭТАПЕComparative analysis of minor histocompatibility antigens genotyping methodsСравнительный анализ ме тодов генотипирования минорных антигенов гистосовместимостиVdovinA. S.ВдовинА. С.
Russian Federation
PostovskayaA. M.ПостовскаяА. М.
Russian Federation
BykovaN. A.БыковаН. А.
Russian Federation
RomaniukD. S.РоманюкД. С.
Russian Federation
AlievaA. K.АлиеваА. Х.
Russian Federation
YefimovaP. R.ЕфимоваП. Р.
Russian Federation
SheetikovS. A.ШитиковС. А.
Russian Federation
JulhakyanH. L.ДжулакянУ. Л.
Russian Federation
EfimovG. A.ЕфимовГ. А.
Russian Federation
grigory@efimov.infoNational Research Center for Hematology of the Ministry of Healthcare of the Russian FederationФГБУ «Гематологический научный центр» Минздрава России4 Novy Zyikovskiy pr., Moscow, 125167, RussiaРоссия, 125167, Москва, Новый Зыковский проезд, 44 Novy Zyikovskiy pr., Moscow, 125167, Russia M.V. Lomonosov Moscow State UniversityРоссия, 125167, Москва, Новый Зыковский проезд, 4 ФГБОУ ВПО «Московский государственный университет имени М. В. Ломоносова»1 Leninskie Goryi St., Moscow, 119991, RussiaРоссия, 119991, Москва, ул. Ленинские Горы, 1The Institute of Molecular Genetics of the Russian Academy of SciencesФГБУН «Институт молекулярной генетики Российской академии наук»2, Kurchatova Sq., Moscow, 123182, RussiaРоссия, 123182, Москва, площадь академика И. В. Курчатова, 2M.V. Lomonosov Moscow State UniversityФГБОУ ВПО «Московский государственный университет имени М. В. Ломоносова»0508201611240500508201605082016https://oncohematology.abvpress.ru/ongm/article/view/196

The wide range of techniques could be employed to find mismatches in minor histocompatibility antigens between transplant recipients and their donors. In the current study we compared three genotyping methods based on polymerase chain reaction (PCR) for four minor antigens. Three of the tested methods: allele-specific PCR, restriction fragment length polymorphism and real-time PCR with TaqMan probes demonstrated 100% reliability when compared to Sanger sequencing for all of the studied polymorphisms. High resolution melting analysis was unsuitable for genotyping of one of the tested minor antigens (HA-1) as it has linked synonymous polymorphism. Obtained data could be used to select the strategy for large-scale clinical genotyping.

Для определения различий донора и реципиента по минорным антигенам гистосовместимости при трансплантации могут быть использованы разнообразные подходы. В настоящей работе нами был проведен сравнительный анализ методов генотипирования, основанных на полимеразной цепной реакции (ПЦР), для четырех минорных антигенов. Три метода: аллель-специфичная ПЦР, анализ полиморфизма длины рестрикционных фрагментов и ПЦР в реальном времени с использованием зондов TaqMan продемонстрировали 100% достоверность результатов генотипирования при сравнении с секвенированием по Сэнгеру для всех изученных полиморфизмов. Метод анализ кривых плавления оказался неприменимым для генотипирования одного минорного антигена (HA-1), имеющего сцепленный синонимичный полиморфизм. Полученная информация может быть использована при выборе метода для крупномасштабного генотипирования в клинической практике.

minor histocompatibility antigensingle nucleotide polymorphismallogeneic hematopoietic stem cells transplantationgenotypingallelic discriminationминорные антигены гистосовместимостиоднонуклеотидные полиморфизмыаллогенная трансплантация гемопоэтических стволовых клетокгенотипированиеаллельная дискриминация1. Talmadge J. E. Hematopoietic stem cell graft manipulation as a mechanism of immunotherapy. Int Immunopharmacol. 2003;3(8):1121–43. DOI: 10.1016/S1567– 5769(03) 00014–6; PMID: 12860168.Talmadge J. E. Hematopoietic stem cell graft manipulation as a mechanism of immunotherapy. Int Immunopharmacol. 2003;3(8):1121–43. DOI: 10.1016/S1567– 5769(03) 00014–6; PMID: 12860168.2. Marsh S. G. and WHO Nomenclature Committee for Factors of HLA System. System, nomenclature for factors of the HLA system, update December 2015. HLA, 2016;87(2):126– 30 DOI: 10.1111/tan. 12745; PMID: 26889910.Marsh S. G. and WHO Nomenclature Committee for Factors of HLA System. System, nomenclature for factors of the HLA system, update December 2015. HLA, 2016;87(2):126– 30 DOI: 10.1111/tan. 12745; PMID: 26889910.3. Felix N. J., Allen P. M. Specificity of T-cell alloreactivity. Nat Rev Immunol. 2007;7(12):942– 53. DOI: 10.1038/nri2200; PMID: 18007679.Felix N. J., Allen P. M. Specificity of T-cell alloreactivity. Nat Rev Immunol. 2007;7(12):942– 53. DOI: 10.1038/nri2200; PMID: 18007679.4. Goulmy E., Termijtelen A., Bradley B. A., van Rood J. J. Alloimmunity to human H-Y. Lancet 1976;2(7996):1206. PMID: 63036.Goulmy E., Termijtelen A., Bradley B. A., van Rood J. J. Alloimmunity to human H-Y. Lancet 1976;2(7996):1206. PMID: 63036.5. den Haan J. M., Sherman N. E., Blokland E. et al. Identification of a graft versus host disease-associated human minor histocompatibility antigen. Science 1995;268(5216):1476– 80. PMID: 7539551.den Haan J. M., Sherman N. E., Blokland E. et al. Identification of a graft versus host disease-associated human minor histocompatibility antigen. Science 1995;268(5216):1476– 80. PMID: 7539551.6. Ефимов Г. А., Вдовин А. С., Григорьев А. А. и др. Иммунобиология острой реакции «трансплантат против хозяина. Медицинская иммунология 2015;17(6):499–516. [EfimovG. A., Vdovin A.S., Grigoryev A.A. et al. Immunobiology of acute graft-versus-host disease. Meditsinskaya immunologiya = Medical immunology 2015;17(6):499–516 (In Russ.)]. DOI:10.157 89/1563‑0625‑2015‑6‑499‑516.Ефимов Г. А., Вдовин А. С., Григорьев А. А. и др. Иммунобиология острой реакции «трансплантат против хозяина. Медицинская иммунология 2015;17(6):499–516. [EfimovG. A., Vdovin A.S., Grigoryev A.A. et al. Immunobiology of acute graft-versus-host disease. Meditsinskaya immunologiya = Medical immunology 2015;17(6):499–516 (In Russ.)]. DOI:10.157 89/1563‑0625‑2015‑6‑499‑516.7. Murata M., Warren E. H., Riddell S. R. A human minor histocompatibility antigen resulting from differential expression due to a gene deletion. J Exp Med. 2003;197(10): 1279–89. DOI: 10.1084/jem. 20030044; PMID: 12743171.Murata M., Warren E. H., Riddell S. R. A human minor histocompatibility antigen resulting from differential expression due to a gene deletion. J Exp Med. 2003;197(10): 1279–89. DOI: 10.1084/jem. 20030044; PMID: 12743171.8. Griffioen M., van Bergen C. A., Falkenburg J. H. Autosomal minor histocompatibility antigens: how genetic variants create diversity in immune targets. Front Immunol. 2016;7:100. DOI: 10.3389/ fimmu. 2016.00100; PMID: 27014279.Griffioen M., van Bergen C. A., Falkenburg J. H. Autosomal minor histocompatibility antigens: how genetic variants create diversity in immune targets. Front Immunol. 2016;7:100. DOI: 10.3389/ fimmu. 2016.00100; PMID: 27014279.9. Быкова Н. А., Малько Д. Б., Вдовин А.С., Ефимов Г. А. In silico анализ иммуногеннного потенциала однонуклеотидных полиморфизимов при полностью HLA- совместимой трансплантации. Российский иммунологический журнал 2016; 10(19)(1):39–49. [Bykova N. A., Malko D. B., Vdovin A. S., Efimov G. A. In silico analysis of single nucleotide polymorphism immunogenic potential in fully HLAmatched transplantation. Rossiiskii immunologicheskii zhurnal = Russian Immunology Journal 2016; 10(19)(1): 39–49 (In Russ.)].Быкова Н. А., Малько Д. Б., Вдовин А.С., Ефимов Г. А. In silico анализ иммуногеннного потенциала однонуклеотидных полиморфизимов при полностью HLA- совместимой трансплантации. Российский иммунологический журнал 2016; 10(19)(1):39–49. [Bykova N. A., Malko D. B., Vdovin A. S., Efimov G. A. In silico analysis of single nucleotide polymorphism immunogenic potential in fully HLAmatched transplantation. Rossiiskii immunologicheskii zhurnal = Russian Immunology Journal 2016; 10(19)(1): 39–49 (In Russ.)].10. Granados D. P., Sriranganadane D., Daouda T. et al. Impact of genomic polymorphisms on the repertoire of human MHC class I-associated peptides. Nat Commun. 2014;5:3600. DOI: 10.1038/ncomms4600; PMID: 24714562.Granados D. P., Sriranganadane D., Daouda T. et al. Impact of genomic polymorphisms on the repertoire of human MHC class I-associated peptides. Nat Commun. 2014;5:3600. DOI: 10.1038/ncomms4600; PMID: 24714562.11. Li J. M., Giver C. R., Lu Y. et al. Separating graft-versus-leukemia from graftversus- host disease in allogeneic hematopoietic stem cell transplantation. Immunotherapy 2009;1(4): 599–621. DOI: 10.2217/imt. 09.32; PMID: 20191089.Li J. M., Giver C. R., Lu Y. et al. Separating graft-versus-leukemia from graftversus- host disease in allogeneic hematopoietic stem cell transplantation. Immunotherapy 2009;1(4): 599–621. DOI: 10.2217/imt. 09.32; PMID: 20191089.12. Wilke M., Poo J., den Haan J. M., Goulmy E. Genomic identification of the minor histocompatibility antigen HA-1 locus by allele-specific PCR. Tissue Antigens 1998;52(4):312–7. PMID: 9820596.Wilke M., Poo J., den Haan J. M., Goulmy E. Genomic identification of the minor histocompatibility antigen HA-1 locus by allele-specific PCR. Tissue Antigens 1998;52(4):312–7. PMID: 9820596.13. Wilke M., Pool J., Goulmy E. Allele specific PCR for the minor Histocompatibility antigen HA-2. Tissue Antigens 2002;59(4):304–7. PMID: 12135429.Wilke M., Pool J., Goulmy E. Allele specific PCR for the minor Histocompatibility antigen HA-2. Tissue Antigens 2002;59(4):304–7. PMID: 12135429.14. Spierings E., Brickner A. G., Caldwell J. A. et al. The minor histocompatibility antigen HA- 3 arises from differential proteasomemediated cleavage of the lymphoid blast crisis(Lbc) oncoprotein. Blood 2003;102(2):621–9. DOI: 10.1182/ blood-2003‑01‑0260; PMID: 12663445.Spierings E., Brickner A. G., Caldwell J. A. et al. The minor histocompatibility antigen HA- 3 arises from differential proteasomemediated cleavage of the lymphoid blast crisis(Lbc) oncoprotein. Blood 2003;102(2):621–9. DOI: 10.1182/ blood-2003‑01‑0260; PMID: 12663445.15. Simsek M., Adnan H. Effect of single mismatches at 3’-end of primers on polymerase chain reaction. J Sci Res Med Sci 2000;2(1):11–4. PMID: 24019700.Simsek M., Adnan H. Effect of single mismatches at 3’-end of primers on polymerase chain reaction. J Sci Res Med Sci 2000;2(1):11–4. PMID: 24019700.16. Spierings E., Drabbels J., Hendriks M. et al. A uniform genomic minor histocompatibility antigen typing methodology and database designed to facilitate clinical applications. PLoS One 2006;1: e42. DOI: 10.1371/journal. pone. 0000042; PMID: 17183671.Spierings E., Drabbels J., Hendriks M. et al. A uniform genomic minor histocompatibility antigen typing methodology and database designed to facilitate clinical applications. PLoS One 2006;1: e42. DOI: 10.1371/journal. pone. 0000042; PMID: 17183671.17. Saiki R. K., Scharf S., Faloona F. et al. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 1985;230(4732):1350–4. PMID: 2999980.Saiki R. K., Scharf S., Faloona F. et al. Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia. Science 1985;230(4732):1350–4. PMID: 2999980.18. Krypuy M., Newnham G. M., Thomas D. M. et al. High resolution melting analysis for the rapid and sensitive detection of mutations in clinical samples: KRAS codon 12 and 13 mutations in non-small cell lung cancer. BMC Cancer 2006;6:295. DOI: 10.1186/1471‑2407‑6‑295; PMID: 17184525.Krypuy M., Newnham G. M., Thomas D. M. et al. High resolution melting analysis for the rapid and sensitive detection of mutations in clinical samples: KRAS codon 12 and 13 mutations in non-small cell lung cancer. BMC Cancer 2006;6:295. DOI: 10.1186/1471‑2407‑6‑295; PMID: 17184525.19. Gundry C. N., Vandersteen J. G., Reed G. H. et al. Amplicon melting analysis with labeled primers: a closed-tube method for differentiating homozygotes and heterozygotes. Clin Chem. 2003;49(3):396– 406. PMID: 12600951.Gundry C. N., Vandersteen J. G., Reed G. H. et al. Amplicon melting analysis with labeled primers: a closed-tube method for differentiating homozygotes and heterozygotes. Clin Chem. 2003;49(3):396– 406. PMID: 12600951.20. Turpeinen H., Volin L., Partanen J. Domestic and foreign donor candidates result in differential probability of matching minor histocompatibility antigens–relevance of selection for hematopoietic stem celltransplantation. Tissue Antigens 2009;73(3):236–41. DOI: 10.1111/j. 1399– 0039.2008.01210. x; PMID: 19254253.Turpeinen H., Volin L., Partanen J. Domestic and foreign donor candidates result in differential probability of matching minor histocompatibility antigens–relevance of selection for hematopoietic stem celltransplantation. Tissue Antigens 2009;73(3):236–41. DOI: 10.1111/j. 1399– 0039.2008.01210. x; PMID: 19254253.21. Di Terlizzi S., Zino E., Mazzi B. et al. Therapeutic and diagnostic applications of minor histocompatibility antigen HA-1 and HA-2 disparities in allogeneic hematopoietic stem cell transplantation: a survey of different populations. Biol Blood Marrow Transplant 2006;12(1):95–101. DOI: 10.1016/j. bbmt. 2005.09.017; PMID: 16399573.Di Terlizzi S., Zino E., Mazzi B. et al. Therapeutic and diagnostic applications of minor histocompatibility antigen HA-1 and HA-2 disparities in allogeneic hematopoietic stem cell transplantation: a survey of different populations. Biol Blood Marrow Transplant 2006;12(1):95–101. DOI: 10.1016/j. bbmt. 2005.09.017; PMID: 16399573.22. Ryazantsev D. Y., Rogozhin E. A., Dimitrieva T. V. et al. A novel hairpin-like antimicrobial peptide from barnyard grass (Echinochlo acrusgalli L.) seeds: structurefunctional and molecular-genetics characterization. Biochimie 2014;99:63–70. DOI: 10.1016/j. biochi. 2013.11.005; PMID: 24275143.Ryazantsev D. Y., Rogozhin E. A., Dimitrieva T. V. et al. A novel hairpin-like antimicrobial peptide from barnyard grass (Echinochlo acrusgalli L.) seeds: structurefunctional and molecular-genetics characterization. Biochimie 2014;99:63–70. DOI: 10.1016/j. biochi. 2013.11.005; PMID: 24275143.23. Untergasser A., Cutcutache I., Koressaar T. et al. Primer3 – new capabilities and interfaces. Nucleic Acids Res 2012;40(15): e115. DOI: 10.1093/nar/gks596; PMID: 22730293.Untergasser A., Cutcutache I., Koressaar T. et al. Primer3 – new capabilities and interfaces. Nucleic Acids Res 2012;40(15): e115. DOI: 10.1093/nar/gks596; PMID: 22730293.