Korean J Lab Med 2010;30: DOI /kjlm Original Article Diagnostic Immunology Allelic and Haplotypic Diversity of HLA-A, -B, -

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1 Korean J Lab Med 2010;30: DOI /kjlm Original Article Diagnostic Immunology Allelic and Haplotypic Diversity of HLA-A, -B, -C, and -DRB1 Genes in Koreans Defined by High-resolution DNA Typing Hye Yoon Chung, M.D., Jung Ah Yoon, M.S., Bok Youn Han, M.T., Eun Yung Song, M.D., and Myoung Hee Park, M.D. Department of Laboratory Medicine, Seoul National University College of Medicine, Seoul, Korea Background : In this study, we used high-resolution DNA typing to investigate the distribution of HLA alleles and haplotypes in Koreans. Methods : HLA-A, -B, -C, and -DRB1 alleles were genotyped at the allelic (4-digit) level in 474 healthy Koreans. HLA genotyping was performed in two steps. Initially, serologic typing or genericlevel DNA typing was performed using the PCR-sequence-specific oligonucleotide method, and then allelic DNA typing (exons 2 and 3 for class I, and exon 2 for DRB1) was carried out using the PCR-single-strand conformation polymorphism method or sequence-based typing. HLA allele and haplotype frequencies and linkage disequilibrium values were calculated by the maximum likelihood method using a computer program developed for the 11th International Histocompatibility Workshop. Results : A total of 21 HLA-A, 40 HLA-B, 22 HLA-C, and 29 HLA-DRB1 alleles were found in Koreans. The most frequent alleles in each locus with frequencies of 10% were, in decreasing order of frequency, as follows: A*24:02, A*02:01, A*33:03; B*51:01; C*01:02, C*03:03; and DRB1*09:01. The numbers of two- and three-locus haplotypes with frequencies of >0.5% were as follows: 44 A-C, 42 B-C, 51 A-B, 52 B-DRB1, 42 A-C-B, and 34 A-B-DRB1. Thirteen A-B-DRB1 haplotypes with frequencies of 1.0% comprised 26.0% of the total haplotypes. The six most common haplotypes were as follows: A*33:03-B*44:03-DRB1*13:02 (3.7%), A*33:03-B*44:03-DRB1*07:01 (3.0%), A*33:03-B*58: 01-DRB1*13:02 (3.0%), A*24:02-B*07:02-DRB1*01:01 (2.8%), A*30:01-B*13:02-DRB1*07:01 (2.3%), and A*11:01-B*15:01-DRB1*04:06 (2.2%). Conclusions : The information obtained in this study can be used as basic data for Koreans in the fields of organ transplantation, disease association, and anthropologic studies. (Korean J Lab Med 2010;30:685-96) Key Words : HLA, Allele, Haplotype, DNA typing, Korean 서 론 HLA 유전자는사람이갖는유전자들중에서가장심한다형 Received : June 9, 2010 Manuscript No : KJLM Revision received : October 14, 2010 Accepted : November 5, 2010 Corresponding author : Myoung Hee Park, M.D. Department of Laboratory Medicine, Seoul National University College of Medicine, 101 Daehang-no, Jongno-gu, Seoul , Korea Tel : , Fax : parkmhee@snu.ac.kr ISSN The Korean Society for Laboratory Medicine 성 (polymorphism) 을나타낸다고알려져있다. 각 HLA 유전자좌에는다수의대립유전자가존재하여 2009년 12월말까지알려진것이 HLA-A 965개, HLA-B 1,543개, HLA-C 626 개, HLA-DRB1 762개, HLA-DQB1 107개에이른다 [1, 2]. H LA 유전자의또하나의특징은 class I, II, III 유전자들이제6 번염색체의단완에아주근접해서위치하므로부모에서자식으로유전될때한염색체위에존재하는특정한조합의 HLA 대립유전자들이하나의단위로묶여서일배체형 (haplotype) 으로유전되는것이다. HLA의분포는각인종간에뚜렷한차이를보이며각인종마다특징적으로잘나타나는 HLA 대립유전 685

2 686 Korean J Lab Med 2010;30: 자와일배체형이존재한다. 각인종에서 HLA 대립유전자및일배체형의분포는조혈모세포이식등장기이식과인류유전학적연구, 질환연관성연구등에필요한기초자료이다. 현재까지 DNA 형별검사를이용한한국인의 HLA 대립유전자와일배체형의분포에대해서는 1,000명이상의비교적많은수를대상으로하여 HLA-A, -B, -DR 유전자에대한저해상도 / 혈청학적수준의다형성을분석한보고가있다 [3-5]. 대립유전자수준의다형성에대해서는 HLA class I 유전자 [6], 또는 class II 유전자 [7-9] 에대해서분석한보고가일부있으나, class I과 class II에대해동시에분석한것은 485명을대상으로한한편의보고가있을뿐이다 [10]. 따라서한국인에대한 HLA class I 과 class II에대한대립유전자수준의자료가더필요한실정이다. 이에저자들은건강한한국인 474명을대상으로 HLA -A, -B, -C, -DRB1 유전자에대해대립유전자수준의고해상도 DNA 형별검사를실시하고 HLA 대립유전자형별과일배체형의분포에대해분석한결과를보고하고자한다. 대상및방법 1. 대상건강한한국인 474명을대상으로하였는데이중에 260명은조혈모세포기증희망자등록을위해 HLA 검사가의뢰된예였고, 214명은이전에본연구진이한국인의 HLA 가계분석을위해수집한 107가족의부모 214명에해당하였다 [8, 11]. 대상은모두건강한지원자였고지원자들간에는혈연관계가없는것으로추정되었다. 본연구는서울대학교병원의 IRB 승인을받아서진행하였다 (IRB No ). [12] 을, HLA-DR은 Dynal사 (Dynal Biotech, Wirral, U.K.) 의 PCR-SSO 키트 (Dynal RELI TM SSO HLA Test) 를이용하여실시하였다. 한국인 107가족의 214명부모에대해서는 HL A-A, -B, -C, -DR에대한첫단계검사를제11차국제조직적합성워크숍에서제공된혈청을이용하여혈청학적검사법으로시행하였고, 그결과는본연구진이이전에보고한바있다 [11]. 이중에 HLA-C 결과가혈청학적 blank를보인검체에대해서는모두 Dynal사의 PCR-SSO 키트를이용하여 DNA 검사를실시하였다. 첫단계로실시한 DNA 검사에서 DNA의증폭및역보합반응 (reverse hybridization) 은모두키트내의설명서에따라실시하였다. 두번째단계로대립유전자수준의고해상도 HLA 형별검사를실시하였는데 HLA-A, -B, -C 유전자에대해서는 exon 2 와 exon 3을, HLA-DRB1 유전자에대해서는 exon 2를분석하였다. 분석방법은기본적으로 PCR-SSCP 검사법을이용하였고, 기존에보고된방법을약간변경하여실시하였다 [13-20]. PCR-SSCP 방법으로대립유전자수준의형별판정이어려운경우 (C*08 등 ) 와일부낮은빈도의 HLA 형별 (A*68, B*08, B*47 등 ) 에대해서는기존에보고된방법과 primer를이용하여직접염기순서분석법으로형별검사를실시하였다 [19, 21, 22]. HLA 대립유전자형별은 2010년 WHO HLA 명명법 [1] 에따라부여하였다. 몇개의대립유전자형별간에구분을할수없는 ambiguity가있는경우에는한국인에가장가능성이높은대립유전자형별을부여하였다 (Table 1). 예를들면 DRB1*14: 01과 DRB1*14:54는 exon 2에대한검사만으로는구분할수없고 exon 3에차이가있는데한국인에서기존에알려진 DRB1* 14:01은 DRB1*14:54임이최근에밝혀져서 [23], DRB1*14:54 로간주하였다. 2. HLA 대립유전자의고해상도 DNA 검사 3. 통계분석 대립유전자수준의고해상도대립유전자검사결과를얻기위한 HLA 검사는두단계로실시하였다. 첫단계에서는 DNA 검사법또는혈청학적검사법을이용하여혈청학적수준의형별결과를얻었고, 이러한저해상도결과를바탕으로두번째단계의분석에서는 group 특이적인 PCR을진행한후 PCR-single-strand conformation polymorphism (SSCP) 검사법이나직접염기서열분석법을이용하여검사를진행하였다. 첫단계검사로조혈모세포기증희망자 260명의 HLA-A, -B, -C 는제13차국제조직적합성워크숍에서제공된 PCR-sequencespecific oligonucleotide (SSO) 키트인 reverse line strip (RLS) HLA-A, -B, -C, -DRB1 대립유전자형별의유전자빈도와 2-유전자좌및 3- 유전자좌일배체형빈도는 11차국제조직적합성워크숍에서사용되었던 maximum likelihood 방법에기초한전산프로그램을이용하여산출하였다 [24]. 본프로그램을이용하여각각의일배체형에대하여연쇄불평형 (linkage disequilibrium, LD) 및상대연쇄불평형 (relative linkage disequ ilibrium, RLD) 값을계산하였고, 2-유전자좌일배체형에대해서는카이제곱 (chi-square) 검정을이용하여연쇄불평형의통계적유의성을평가하였다.

3 Chung HY, et al., HLA Alleles and Haplotypes in Koreans 687 Table 1. HLA gene frequencies in Koreans (N=474) HLA-A* GF (%) HLA-B* GF (%) HLA-C* GF (%) HLA-DRB1* GF (%) A*01: B*07: C*01: DRB1*01: A*02: B*07: C*01: DRB1*03: A*02: B*08: C*02: DRB1*04: A*02: B*13: C*03: DRB1*04: A*02: B*13: C*03: DRB1*04: A*02: B*14: C*03: DRB1*04: A*02: B*15: C*04: DRB1*04: A*03: B*15: C*04: DRB1*04: A*11: B*15: C*05: DRB1*04: A*24: B*15: C*06: DRB1*07: A*24: B*15: C*07: DRB1*08: A*26: B*15: C*07: DRB1*08: A*26: B*27: C*07: DRB1*09: A*26: B*27: C*08: DRB1*10: A*29: B*35: C*08: DRB1*11: A*30: B*35: C*08: DRB1*12: A*30: B*37: C*12: DRB1*12: A*31: B*38: C*12: DRB1*13: A*32: B*39: C*14: DRB1*13: A*33: B*40: C*14: DRB1*14: A*68: B*40: C*15: DRB1*14: A blank 0.4 B*40: C*15: DRB1*14: B*40: C blank 0.0 DRB1*14: B*44: DRB1*14: B*44: DRB1*14: B*46: DRB1*14: B*47: DRB1*15: B*48: DRB1*15: B*51: DRB1*16: B*51: DRB1 blank 0.1 B*52: B*54: B*55: B*55: B*56: B*56: B*57: B*58: B*59: B*67: B blank 0.5 *HLA molecular typing was performed for the exons encoding the peptide-binding domains: exons 2 and 3 for HLA class I and exon 2 for HLA-DRB1 alleles. Listed HLA alleles are the most probable alleles in Koreans selected from particular groups of alleles designated with the suffix G in the 2010 HLA nomenclature [2]. For example, A*02:06, B*40:01, C*01:02, and DRB1*14:54 were selected from A*02:06:01G (A*02:06:01/02:126), B*40:01:01G (B*40:01:01/40:01:02/40:55), C*01:02:01G (C*01:02:01/01:02:02/01:25), and DRB1*14:01:01G (DRB1*14:01:01/14:54), respectively. Abbreviation: GF, gene frequency. 결과 1. HLA-A, -B, -C, -DRB1 대립유전자의유전자빈도 HLA-A, -B, -C, -DRB1 유전자좌에서검출된 HLA 대립유전자의유전자빈도를 Table 1에제시하였다. 한국인에서검 출된대립유전자의종류는 HLA-A 21종, HLA-B 40종, HLA-C 22종, HLA-DRB1 29종이었다. HLA-A 형별중 5% 이상으로높은빈도를보인형별을빈도순으로나열하면 A*24:02 (22.9%), *02:01, *33:03, *11:01, *02:06 등5종이었고, 전체의 73.8% 를차지하였다. 가장다양한대립유전자를가지고있는 A*02는 A*02:01, *02:03, *02:06, *02:07,

4 688 Korean J Lab Med 2010;30: *02:10, *02:41 등 6종류의대립유전자를보였다. 이외에도 A*24, A*26, A*30 형별이 2개이상의대립유전자를보였다. HLA-B 형별중 5% 이상으로높은빈도를보인형별은 B* 51:01 (10.2%), *15:01, *44:03, *35:01, *58:01, *54:01, *40: 02, *46:01 등 8종이었고, 전체의 54.8% 를차지하였다. 가장 많은수의대립유전자를가지고있는 B*15는 B*15:01, *15:02, *15:07, *15:11, *15:18, *15:27 등 6종류의대립유전자를보였고, 그외에도 B*07, B*13, B*27, B*35, B*40, B*44, B*51, B*55, B*56 형별이 2개이상의대립유전자를보였다. HLA-C 형별중 5% 이상으로높은빈도를보인형별은 C* Table 2. Two-locus HLA haplotypes (A-C and B-C) in Koreans (N=474) A-C haplotype* HF (%) LD RLD c 2 B-C haplotype* HF (%) LD RLD c 2 A*01:01 C*06: B*07:02 C*07: A*02:01 C*01: B*13:01 C*03: A*02:01 C*03: B*13:02 C*06: A*02:01 C*03: B*14:01 C*08: A*02:01 C*07: B*15:01 C*01: A*02:01 C*08: B*15:01 C*03: A*02:01 C*14: B*15:01 C*03: A*02:01 C*15: B*15:01 C*04: A*02:03 C*07: B*15:07 C*03: A*02:06 C*01: B*15:11 C*03: A*02:06 C*03: B*15:18 C*07: A*02:06 C*03: B*15:18 C*08: A*02:06 C*08: B*27:05 C*01: A*02:06 C*14: B*27:05 C*02: A*02:07 C*01: B*35:01 C*03: A*03:01 C*05: B*35:01 C*04: A*11:01 C*01: B*35:01 C*08: A*11:01 C*03: B*37:01 C*06: A*11:01 C*04: B*38:02 C*07: A*11:01 C*07: B*39:01 C*07: A*11:01 C*08: B*40:01 C*03: A*24:02 C*01: B*40:02 C*03: A*24:02 C*03: B*40:02 C*03: A*24:02 C*03: B*40:03 C*03: A*24:02 C*04: B*40:06 C*08: A*24:02 C*07: B*44:02 C*05: A*24:02 C*08: B*44:03 C*07: A*24:02 C*12: B*44:03 C*14: A*24:02 C*14: B*46:01 C*01: A*26:01 C*01: B*46:01 C*01: A*26:01 C*03: B*48:01 C*08: A*26:01 C*03: B*48:01 C*08: A*26:02 C*03: B*51:01 C*03: A*26:03 C*03: B*51:01 C*14: A*30:01 C*06: B*51:01 C*15: A*30:04 C*08: B*51:02 C*15: A*31:01 C*03: B*52:01 C*12: A*31:01 C*03: B*54:01 C*01: A*31:01 C*14: B*55:02 C*01: A*33:03 C*01: B*57:01 C*06: A*33:03 C*03: B*58:01 C*03: A*33:03 C*07: B*59:01 C*01: A*33:03 C*07: A*33:03 C*14: *Haplotypes with frequencies of >0.5% are listed. Haplotypes showing strong positive linkage disequilibrium (chi-square>10.8; P<0.001) are underlined and those with RLD values of 0.7 are in bold; RLD values of 0.7 are in bold. Abbreviations: HF, haplotype frequency; LD, linkage disequilibrium; RLD, relative linkage disequilibrium.

5 Chung HY, et al., HLA Alleles and Haplotypes in Koreans 689 Table 3. Two-locus HLA haplotypes (A-B and B-DR) in Koreans (N=474) A-B haplotype* HF (%) LD RLD c 2 B-DR haplotype* HF (%) LD RLD c 2 A*01:01 B*37: B*07:02 DRB1*01: A*02:01 B*13: B*07:05 DRB1*08: A*02:01 B*15: B*13:01 DRB1*12: A*02:01 B*15: B*13:02 DRB1*07: A*02:01 B*27: B*14:01 DRB1*08: A*02:01 B*35: B*15:01 DRB1*04: A*02:01 B*40: B*15:01 DRB1*09: A*02:01 B*40: B*15:01 DRB1*14: A*02:01 B*40: B*15:01 DRB1*15: A*02:01 B*48: B*15:07 DRB1*04: A*02:01 B*51: B*27:05 DRB1*01: A*02:01 B*54: B*35:01 DRB1*04: A*02:03 B*38: B*35:01 DRB1*09: A*02:06 B*27: B*35:01 DRB1*11: A*02:06 B*35: B*35:01 DRB1*12: A*02:06 B*40: B*35:01 DRB1*15: A*02:06 B*40: B*37:01 DRB1*10: A*02:06 B*48: B*38:02 DRB1*15: A*02:06 B*51: B*39:01 DRB1*08: A*02:06 B*54: B*40:01 DRB1*04: A*02:07 B*46: B*40:01 DRB1*08: A*03:01 B*44: B*40:01 DRB1*09: A*11:01 B*15: B*40:02 DRB1*04: A*11:01 B*27: B*40:02 DRB1*09: A*11:01 B*40: B*40:02 DRB1*14: A*11:01 B*51: B*40:02 DRB1*14: A*11:01 B*54: B*40:02 DRB1*15: A*24:02 B*07: B*40:06 DRB1*09: A*24:02 B*15: B*40:06 DRB1*12: A*24:02 B*35: B*44:02 DRB1*04: A*24:02 B*39: B*44:03 DRB1*07: A*24:02 B*40: B*44:03 DRB1*13: A*24:02 B*40: B*46:01 DRB1*08: A*24:02 B*40: B*46:01 DRB1*09: A*24:02 B*46: B*48:01 DRB1*12: A*24:02 B*51: B*48:01 DRB1*15: A*24:02 B*52: B*51:01 DRB1*04: A*24:02 B*54: B*51:01 DRB1*04: A*24:02 B*55: B*51:01 DRB1*08: A*24:02 B*59: B*51:01 DRB1*09: A*26:01 B*15: B*51:01 DRB1*12: A*26:01 B*27: B*51:01 DRB1*13: A*26:01 B*35: B*51:01 DRB1*14: A*26:01 B*40: B*51:01 DRB1*14: A*29:01 B*07: B*51:01 DRB1*15: A*30:01 B*13: B*52:01 DRB1*15: A*30:04 B*14: B*54:01 DRB1*04: A*31:01 B*48: B*54:01 DRB1*15: A*31:01 B*51: B*57:01 DRB1*07: A*33:03 B*44: B*58:01 DRB1*03: A*33:03 B*58: B*58:01 DRB1*13: B*59:01 DRB1*04: *Haplotypes with frequencies of >0.5% are listed. Haplotypes showing strong positive linkage disequilibrium (chi-square>10.8; P<0.001) are underlined and those with RLD values of 0.7 are in bold; RLD values of 0.7 are in bold. Abbreviations: HF, haplotype frequency; LD, linkage disequilibrium; RLD, relative linkage disequilibrium.

6 690 Korean J Lab Med 2010;30: :02 (17.4%), *03:03, *03:04, *08:01, *14:02, *07:02, *03: 02, *04:01, *06:02, *14:03 등 10종이었고, 전체의 84.8% 를차지하였다. C*02, C*05, C*06 형별에는각각 1개의대립유전자만존재하였으나그외의다른 C 형별은모두 2개이상의대립유전자를보였다. HLA-DRB1 형별중 5% 이상으로높은빈도를보인형별은 DRB1*09:01 (10.4%), *04:05, *15:01, *13:02, *01:01, *07: 01, *08:03, *04:06, *12:01 등 9종이었고, 전체의 67.0% 를차지하였다. DRB1*04와 DRB1*14는각각 7개의대립유전자를가지고있어서대립유전자수준에서가장높은다양성을보였다. 그외에 DRB1*08, DRB1*12, DRB1*13, DRB1*15 형별이각각 2개씩의대립유전자를가지고있었다. Table 4. Three-locus HLA haplotypes (A-C-B and A-B-DR) in Koreans (N=474) A-C-B haplotype* HF (%) LD A-B-DR haplotype* HF (%) LD A*33:03 C*03:02 B*58: A*33:03 B*44:03 DRB1*13: A*33:03 C*14:03 B*44: A*33:03 B*44:03 DRB1*07: A*24:02 C*14:02 B*51: A*33:03 B*58:01 DRB1*13: A*30:01 C*06:02 B*13: A*24:02 B*07:02 DRB1*01: A*33:03 C*07:01 B*44: A*30:01 B*13:02 DRB1*07: A*11:01 C*04:01 B*15: A*11:01 B*15:01 DRB1*04: A*24:02 C*07:02 B*07: A*33:03 B*58:01 DRB1*03: A*02:07 C*01:02 B*46: A*24:02 B*52:01 DRB1*15: A*24:02 C*12:02 B*52: A*02:07 B*46:01 DRB1*08: A*02:01 C*01:02 B*27: A*02:01 B*27:05 DRB1*01: A*24:02 C*01:02 B*54: A*02:06 B*27:05 DRB1*01: A*02:01 C*01:02 B*54: A*02:01 B*54:01 DRB1*04: A*24:02 C*03:04 B*40: A*24:02 B*54:01 DRB1*04: A*24:02 C*03:03 B*35: A*01:01 B*37:01 DRB1*10: A*31:01 C*14:02 B*51: A*02:01 B*51:01 DRB1*12: A*30:04 C*08:02 B*14: A*24:02 B*15:01 DRB1*04: A*02:01 C*14:02 B*51: A*02:06 B*51:01 DRB1*09: A*02:01 C*03:04 B*40: A*02:06 B*54:01 DRB1*15: A*02:06 C*14:02 B*51: A*02:01 B*13:01 DRB1*12: A*02:06 C*03:04 B*40: A*24:02 B*59:01 DRB1*04: A*02:06 C*03:03 B*35: A*24:02 B*46:01 DRB1*08: A*01:01 C*06:02 B*37: A*02:01 B*54:01 DRB1*15: A*02:01 C*08:01 B*35: A*24:02 B*40:01 DRB1*04: A*02:06 C*08:01 B*48: A*24:02 B*40:06 DRB1*09: A*11:01 C*01:02 B*54: A*02:07 B*46:01 DRB1*09: A*02:01 C*03:03 B*15: A*24:02 B*15:01 DRB1*15: A*24:02 C*01:02 B*59: A*24:02 B*13:01 DRB1*12: A*02:01 C*03:04 B*13: A*02:03 B*38:02 DRB1*15: A*24:02 C*08:01 B*40: A*24:02 B*51:01 DRB1*11: A*02:06 C*08:01 B*40: A*29:01 B*07:05 DRB1*08: A*02:01 C*01:02 B*15: A*30:04 B*14:01 DRB1*08: A*02:01 C*03:03 B*15: A*31:01 B*51:01 DRB1*08: A*02:06 C*01:02 B*54: A*33:03 B*51:01 DRB1*13: A*03:01 C*05:01 B*44: A*02:06 B*35:01 DRB1*11: A*11:01 C*14:02 B*51: A*24:02 C*01:02 B*46: A*24:02 C*04:01 B*15: A*02:06 C*01:02 B*27: A*02:03 C*07:02 B*38: A*24:02 C*03:04 B*13: A*33:03 C*03:02 B*51: A*24:02 C*03:03 B*15: *Haplotypes with frequencies of >0.5% are listed. Abbreviations: HF, haplotype frequency; LD, linkage disequilibrium.

7 Chung HY, et al., HLA Alleles and Haplotypes in Koreans HLA 일배체형의빈도통계프로그램을통해산출된 2-유전자좌일배체형의빈도를 Table 2, 3에제시하였고, 3-유전자좌일배체형의빈도를 Table 4에제시하였다. 1) 2-유전자좌일배체형한국인에서 maximum likelihood 방법으로산출된빈도 0.5% 이상인 A-C 일배체형은총 44 종이었다 (Table 2). 이중에유의한양성연쇄불평형 (chi-square >10.8, P<0.001) 을나타내는일배체형은 18종 (40.9%) 이었고, RLD 값이 0.7 이상의 Table 5. HLA gene frequencies (%) in Koreans (N=474) and Japanese (N=371) HLA-A* Korean Japanese HLA-B* Korean Japanese HLA-C* Korean Japanese HLA-DRB1* Korean Japanese A*01: B*07: C*01: DRB1*01: A*02: B*07: C*01: DRB1*03: A*02: B*08: C*02: DRB1*04: A*02: B*13: C*03: DRB1*04: A*02: B*13: C*03: DRB1*04: A*02: B*14: C*03: DRB1*04: A*02: B*15: C*04: DRB1*04: A*02: B*15: C*04: DRB1*04: A*03: B*15: C*05: DRB1*04: A*03: B*15: C*06: DRB1*07: A*11: B*15: C*07: DRB1*08: A*11: B*15: C*07: DRB1*08: A*24: B*27: C*07: DRB1*09: A*24: B*27: C*08: DRB1*10: A*24: B*35: C*08: DRB1*11: A*26: B*35: C*08: DRB1*12: A*26: B*37: C*12: DRB1*12: A*26: B*38: C*12: DRB1*13: A*26: B*39: C*14: DRB1*13: A*26: B*39: C*14: DRB1*14: A*29: B*39: C*15: DRB1*14: A*30: B*40: C*15: DRB1*14: A*30: B*40: DRB1*14: A*31: B*40: DRB1*14: A*32: B*40: DRB1*14: A*33: B*44: DRB1*14: A*68: B*44: DRB1*15: B*46: DRB1*15: B*47: DRB1*16: B*48: B*51: B*51: B*51: B*52: B*54: B*55: B*55: B*56: B*56: B*56: B*57: B*58: B*59: B*67: *HLA alleles with significantly different gene frequencies between Koreans and Japanese are underlined (P<0.001) or in bold (P< ). Koreans: present study. Japanese: from reference [26]. DRB1*1401 (reported) is converted to DRB1*14:54 in this table.

8 692 Korean J Lab Med 2010;30: 강한연관을보이는일배체형이 7종 (15.9%) 이었다. 특히 A*02:03-C*07:02, A*30:04-C*08:02 등 2종의일배체형은 RLD 값 1.0으로매우강한양성연쇄불평형을보였다. 반면에 A*33:03-C*01:02는유의한음성연쇄불평형 (RLD -0.78, P<0.001) 을보였다. 빈도 0.5% 이상인 B-C 일배체형은총 42 종이었다 (Table 2). 이중에유의한양성연쇄불평형 (P<0.001) 을나타내는일배체형은 37종으로 88.1%, 그리고 RLD 값이 0.7 이상의강한연관을보이는일배체형이 29종으로 69.0% 에달했다. RLD 값이 1.0으로매우강한연관을보이는일배체형도 B*13:02- C*06:02, B*14:01-C*08:02 등을비롯해 12종이나되었다. 빈도 0.5% 이상인 A-B 일배체형은총 51 종이었다 (Table 3). 이중에유의한양성연쇄불평형 (P<0.001) 을나타내는일배체형은 18종 (35.3%) 이었고, RLD 값이 0.7 이상의강한연관을보이는일배체형이 9종 (17.6%) 이었다. 특히 A*02:03-B*38:02, A*29:01-B*07:05, A*30:04-B*14:01 등 3종의일배체형은 RLD 값 1.0으로매우강한양성연쇄불평형을보였다. 빈도 0.5% 이상인 B-DRB1 일배체형은총 52 종이었다 (Table 3). 이중에유의한양성연쇄불평형 (P<0.001) 을나타내는일배체형은 29종 (55.8%) 이었고, RLD 값이 0.7 이상의강한연관을보이는일배체형이 10종 (19.2%) 이었다. 특히 B*07:05-DR- B1*08:03, B*13:01-DRB1*12:02, B*57:01-DRB1*07:01 등 3종의일배체형은 RLD 값 1.0으로매우강한양성연쇄불평형을보였다. 2) 3-유전자좌일배체형빈도 0.5% 이상인 A-C-B 일배체형은총 42 종이었고빈도순으로 Table 4에제시하였다. 이중 1.0% 이상의빈도를보이는일배체형은 20종으로전체일배체형의 44.3% 를차지하였다. 빈도 2.0% 이상으로가장흔한일배체형은 A*33:03- C*03:02-B*58:01 (5.6%), A*33:03-C*14:03-B*44:03 (4.1%) 등을비롯해 9종이었다. 빈도 0.5% 이상인 A-B-DRB1 일배체형은총 34 종이었고, 이중 1.0% 이상의빈도를보이는일배체형은 13종으로전체일배체형의 26.0% 를차지하였다 (Table 5). 빈도 2.0% 이상으로가장흔한일배체형 6종은 A*33:03-B*44:03-DRB1*13:02 (3.7%), A*33:03-B*44:03-DRB1*07:01 (3.0%), A*33:03- B*58:01-DRB1*13:02 (3.0%), A*24:02-B*07:02-DRB1* 01:01 (2.8%), A*30:01-B*13:02-DRB1*07:01 (2.3%), A*11: 01-B*15:01-DRB1*04:06 (2.2%) 이었다. 고찰본연구에서는 474명의건강한한국인을대상으로 HLA class I 과 class II의 4자리수고해상도대립유전자형별을분석한결과, HLA-A, -B, -C, -DRB1 유전자좌에각각 21, 40, 22, 29종의대립유전자가검출되었다. 한국인에서검출된대립유전자형별의종류는 2010년 WHO 명명법에포함된 HLA-A - B, -C, -DRB1 대립유전자수의각각 2.2%, 2.6%, 3.5%, 3.8% 에불과하여한국인에서는비교적제한된 HLA 다형성을나타냄을알수있었다. 이러한결과는 Lee 등 [10] 이한국인 485명을대상으로 PCR-SSO 방법으로분석하여보고한결과 (HLA- A, -B, -C, -DRB1에각각 20, 43, 21, 31종 ) 와유사하다. 같은인종이라도더많은수를대상으로분석하면드물게존재하는여러종류의대립유전자형별이추가로발견될가능성이높다. Yi 등 [25] 이한국인제대혈 4,108단위를대상으로직접염기서열분석법으로분석하여보고한결과 ( 초록 ) 를보면 HLA-A, -B, -DRB1 유전자좌에각각 46, 62, 46종의 HLA 대립유전자형별이검출되어위의두연구에비해훨씬많은종류가존재함을알수있다. 이대규모제대혈연구에서추가로검출된 HLA 대립유전자의대부분은 0.1% 또는그이하의매우낮은빈도를보이는드문형별이었다. 본연구에서만검출된 A*02:41은골수기증희망자중 1명에서직접염기순서분석법 (A*02:06:01, *02:41) 으로확인된예로서위의대규모제대혈분석에서도검출되지않은매우드문형별로생각된다. 이렇게분석대상규모가커지면검출되는대립유전자형별의수가상당히증가되지만, 0.5% 이상의비교적드물지않은형별의수는거의차이가없다. 즉 0.5% 이상빈도를보이는 HLA-A, -B, -DRB1 대립유전자형별의수는본연구 ( 각각 17, 30, 23종 ) 와 Lee 등 [10] 의연구 ( 각각 18, 30, 26종 ) 결과가대규모제대혈연구 ( 각각 17, 31, 25종 ) 와매우유사한결과를보였다. 한가지특기할점은본연구에서 A*26 아형인 A*26:01, *26:02, *26:03의빈도가각각 3.8%, 2.2%, 1.1% 로 Lee 등 [10] 의연구 ( 각각 6.0%, 0.6%, 1.0%) 와차이를보였으나, Yi 등 [25] 의대규모제대혈연구 ( 각각 3.8%, 2.0%, 0.9%) 와는매우유사한결과를보였다. 각인종에서발견되는 HLA 대립유전자형별의다양성은인종간에큰차이를보이는데한국인의결과를다른동부아시아인종과비교해보았다. 각인종에서 4자리수의대립유전자수준의 HLA 형별자료는많지않은데, 현재까지보고된결과중에서비교적대표성이있는것으로생각되는일본인 (N=371)[26], 북부한족 (N=618)[27], 남부한족 (N=264)[28], 타이완족 (N=364) [29] 에대한자료와비교해보았다. 동부아시아인종에대한연

9 Chung HY, et al., HLA Alleles and Haplotypes in Koreans 693 구에서발견된 HLA-A, -B, -DRB1 대립유전자형별의종류는한국인 ( 각각 21, 40, 29 종 ), 일본인 (20, 38, 27종 ), 남부한족 (20, 50, 28종 ), 타이완족 (19, 44, 32종 ) 등네인종사이에는큰차이가없었으나, 북부한족 ( 각각 84, 148, 122종 ) 에서는거의 4배가까운대립유전자종류가검출되어 HLA 분포에있어서훨씬높은다양성을보인다는사실을확인하였다. 한국인은 HLA 분포에있어서일본인과가장유사한것으로알려져있다. 본연구에서얻은한국인의 HLA 대립유전자형별의빈도를일본인 [26] 과비교하여 Table 5에제시하였다. HLA 대립유전자의분포가두인종간에상당히유사하나일부형별에서는유의한차이가관찰되었다 (P<0.001, Fisher s exact test). 한국인에서일본인에비해유의하게높은빈도를보이는것은 A*30:01, *33:03; B*13:02, *27:05, *58:01; C*03:02, *06:02, *0701; DRB1*03:01, *07:01, *14:05 등이었고, 반대로일본인에서한국인에비해유의하게높은빈도를보이는것은 A*24:02, *26:01; B*39:01, 52:01; C*07:02, *12:02; DRB1*1406, *15:02 등이었다. 통계적유의성은없었으나, 한국인에 0.5% 이상존재하는대립유전자중에일본인에는검출되지않은것이 A*02:03, *29:01, *30:04, 32:01; B*07:05, *14:01, *57:01; C*02:02, *07:01, *08:02; DRB1*03:01 등이었다. 더큰규모의일본인 (N=1,028) 을대상으로 PCR-SSO Luminex 방법으로분석한 Itoh 등 [30] 의연구에서도이들대립유전자는매우드문빈도로존재하거나 (A*02:03 0.1%, DRB1*03: %) 검출되지않는것으로나타났다. 따라서이들대부분의대립유전자형별이일본인에는없거나극히드문것으로생각된다. HLA 대립유전자의분포도인종간에큰차이를보이지만 HLA 일배체형의분포와 HLA 대립유전자간의연쇄불평형은인종간에더욱큰차이를보이는것으로알려져있다. 특징적인 HLA 일배체형은지역적으로제한된분포를보이고, 인구집단의이동경로추적에유용한정보를제공하여인류유전학적연구에도움을준다 [31]. 한국인에서 maximum likelihood 방법으로산출한 2-유전자좌 HLA 일배체형중에빈도 0.5% 이상인것은 A-C, B-C, A-B, B-DR 일배체형에서각각 44, 42, 51, 52종이있었다. HLA-A, -B, -C, -DRB1 유전자좌간에연쇄불평형의강도를알아보기위해이들일배체형의 RLD 값의총합과카이제곱검정값의총합으로비교해보았다. HLA 일배체형중에 RLD 값의총합은 B-C가가장큰값 (29.65) 을, A -C가가장작은값 (9.83) 을보였으며 A-B와 B-DR은그사이 ( 각각 13.47, 17.76) 에속했다. 카이제곱검정값의총합에서도이와유사한결과를보여 (B-C 10212, A-B 4927, B-DR 4460, A-C 2936) 연쇄불평형의정도는 B-C 유전자간에가장강하 고, A-C 유전자간에가장약한것을알수있었다. 이러한경향은 RLD 값이 0.7 이상의강한연관을보이는일배체형의비율에서도알수있었는데, 이러한비율이 B-C 일배체형에서가장높고 (69.0%, 29/42종 ), A-C 일배체형에서가장낮은것으로 (15.9%, 7/44종 ) 나타났다. Lee 등 [10] 의한국인에대한연구 ( 빈도 0.9% 이상의일배체형에대한분석 ) 에서도 2-유전자좌일배체형의 RLD 값의분포가대체로본연구와유사한결과를보였으나, 소수의일배체형에서는차이를보였다. 즉, 본연구에서 2% 이상의빈도와비교적강한연관을보이는것으로관찰된 B*40:01-C*03:04 ( 빈도 2.5%, RLD 0.54), B*40:02- C*03:04 ( 빈도 3.8%, RLD 0.72) 는이들의연구에서는관찰되지않았고, 대신에 B*40:01-C*03:02 ( 빈도 1.6%, RLD 0.32), B*40:02-C*03:02 ( 빈도 1.4%, RLD 0.30) 가 1% 이상의빈도를보였다. 이는이들의연구에서본연구에비해 C*03:04의빈도가낮고 (3.9% vs. 9.9%), 대신에 C*03:02의빈도가높은 (10.8% vs. 6.3%) 것과관련이있는것으로생각된다. 한편이들두종류의일배체형은일본인 ( 각각빈도 2.5%, RLD 0.55; 빈도 7.4%, RLD 0.84) 에서도빈도 2% 이상이고 RLD 0.5 이상의강한연관을보여본연구결과와유사하였다. HLA 일배체형중에 A-B-DR 일배체형의분포는장기이식, 특히비혈연조혈모세포이식에서각인종에필요한공여자풀의규모를산정하는데매우중요하다. 본연구결과한국인에서 0.5% 이상의빈도를보이는 A-B-DR 일배체형은 34종이었고, 이중에 13종은 1% 이상의빈도를가지는흔한일배체형에속했다 (Table 4). 본연구에서관찰된 A-B-DR 일배체형의분포는 Lee 등 [10] 의연구결과 ( 빈도 1.0% 이상, 14 종 ) 와대체로유사하였다. 한국인에서관찰된 A-B-DR 일배체형의분포를동부아시아인종들과비교해보았다. 빈도 1% 이상의흔한 A- B-DR 일배체형이한국인, 일본인 [32], 타이완족 [29] 에서는각각 10 종이상씩있었고이들일배체형이총일배체형의 25% 이상을차지했으나, 북부한족 [27] 에서는 4종뿐이었고이들이총일배체형의 8% 정도에불과해북부한족은 HLA 일배체형의분포에있어서도훨씬높은다양성을나타냄을확인할수있었다. A-B-DR 일배체형의분포에있어서도한국인은일본인과가장가까운것으로나타났는데, 일본인 (N=516)[32] 에서는빈도 0.5% 이상이 31종, 이중에 1% 이상이 11종으로본연구에서한국인의결과와유사하였다. 또한일본인에 1% 이상빈도로존재하는일배체형 11종중에 6종이한국인에도 1% 이상빈도로관찰되어일본인에흔한일배체형종류의반이상이한국인에도흔하게존재함을알수있었다. 조혈모세포이식에서환자와공여자간에 HLA가일치하는정

10 694 Korean J Lab Med 2010;30: 도가이식성적에큰영향을미친다는사실은잘알려져있는데, 최근에는비혈연이식에서환자와공여자간에 HLA-A, -B, -C, -DRB1을대립유전자 ( 고해상도 ) 수준까지맞추어서이식을하는경우이식편대숙주반응 (GVHD) 의발생이나생존율에있어서더좋은성적을보인다는사실이보고되었다 [33-36]. 그런데, 대립유전자수준에서불일치를나타내는 HLA 형별은인종에따라상당히차이가있을수있다. 본연구진은한국인에서 A2가포함된 A-B-DR 일배체형인경우 A2가대립유전자수준에서흔히불일치를보인다는사실을보고한바있다 [13]. 본연구에서도 0.5% 이상의빈도를보이는 A-B-DR 일배체형중에 A2 아형이다른것이두종류가있었다. A*02-B*27:05-DRB1*01:01과 A*02-B*54:01-DRB1*15:01 일배체형의경우 A*02:01, A*02: 06과연관된것이비슷한비율로존재하여 (Table 4) 이런일배체형을갖는개체간에는 B와 DR이대립유전자수준에서일치하여도 A*02아형에서불일치할가능성이높은것으로나타났다. 이외에도동일한일배체형이서로다른 A*02 아형과연관된것이여러종류있었는데예를들면, A*02-B*46:01-DRB1*09:01 의경우 A*02:01, *02:06, *02:07과연관된것이각각 0.3%, 0.2%, 0.6% 로나타났다. 최근에본연구진이비혈연조혈모세포이식이필요한환자와혈청학적수준에서일치하는 HLA-A, -B, -DR 형별을갖고있는기증자간에대립유전자수준에서불일치를많이나타내는형별을조사해본결과 A*02 외에도 A*26, B*15 ( 혈청학적 B62), B*40 ( 혈청학적 B61), DRB1*04, DRB1* 14 등이가장흔하게불일치를보이는것으로나타났다. 본연구의제한점으로는대립유전자형별판정을위해 HLA class I에대해서는 exon 2와 exon 3만을, HLA-DRB1에대해서는 exon 2만을분석하였고, 구분할수없는 allelic ambiguity 에대해서는한국인에서가장가능성이높은대립유전자형별로판정하였다는점이다 (Table 1). 최근에는직접염기서열분석의경우 HLA class I에서 exon 4도추가하여분석을하는경향이지만 class I 유전자에서 exon 4의변이는비교적제한적이기때문에 exon 4를분석하여도 ambiguity의상당부분은그대로남아있게되고앞으로새로운대립유전자가추가됨에따라이런경향은더욱증가될것이다. 그러나특정한인종에나타나는대립유전자다형성은비교적제한되어있는것으로알려졌다 [37]. 본연구에서검출된 HLA 대립유전자형별의종류와그빈도가한국인제대혈을대상으로직접염기서열분석법으로분석한결과 [25] 와유사한것으로보아형별판정에별문제는없었을것으로생각된다. 또하나의문제점은각 HLA 유전자좌에서검출된 blank 빈도가 HLA-A, -B, -C, -DRB1 에서각각 0.4%, 0.5%, 0.0%, 0.1% 로, HLA-A와 -B에서약 간높은결과를보였다는점이다. 이는분석에포함된대상자중에가족검체에서 HLA-A, -B에대한일차검사를혈청학적방법으로실시하였으므로실제로있는대립유전자를일부검출하지못했을가능성이있고, HLA-C는혈청학적검사에서 blank가나온모든검체를 DNA 검사로다시확인하였기때문에 blank가 0% 로낮게나온것으로생각된다. 실제로통계분석에서 blank 가포함된일배체형중에 0.2% 의빈도를보인것이 A blank 2개 (A blank-b*40:03-drb1*14:05, A blank-b*67:01-drb1* 12:01), B blank 1개 (A*24:02-B blank-drb1*08:03) 가관찰되었고 0.1% 이하의일배체형도일부관찰되었다. 본연구에서는건강한한국인 474명을대상으로고해상도 DNA 검사법을이용하여 HLA-A, -B, -C, -DRB1 형별을분석하고통계적방법을이용하여대립유전자 (4자리수) 형별빈도와일배체형분포에대한자료를산출하여제시하였다. 본연구의결과는앞으로한국인에서장기이식, 특히조혈모세포이식, 질환연관성연구, 인류유전학적연구등에유용한자료로이용될수있을것으로생각된다. 요약배경 : HLA 형별은혈청학적수준 (generic level) 에서도다형성이심하지만대립유전자수준에서는더욱심한다형성을보이고인종간에큰차이를나타내는것으로알려졌다. 본연구에서는고해상도 DNA 검사법을이용하여한국인에서 HLA 대립유전자형별과일배체형의종류및빈도를알아보고자하였다. 방법 : 건강한한국인 474명을대상으로 HLA-A, -B, -C, -DRB1 유전자에대해두단계의검사로대립유전자 (4자리수) 형별분석을실시하였다. 1단계로혈청학적수준의형별검사를혈청학적검사법이나 sequence-specific oligonucleotide (PCR-SSO) 방법으로시행하였고, 그다음단계로대립유전자형별검사를 class I은 exon 2와 exon3, DRB1은 exon 2에대해 single-strand conformation polymorphism (PCR-SSCP) 또는직접염기서열분석법을이용하여실시하였다. HLA 대립유전자의유전자빈도, 일배체형빈도, 연쇄불평형값은 maximum likelihood 원리에근거한제11차국제조직적합성워크숍컴퓨터프로그램을이용하여산출하였다. 결과 : 한국인에서검출된 HLA-A, -B, -C, DRB1 대립유전자형별은각각 21, 40, 22, 29종이었다. 이중에유전자빈도 10% 이상을보인대립유전자형별 ( 빈도순나열 ) 은 A*02:01, A*24:02, A*33:03; B*51:01; C*01:02, C*03:03; DRB1*09:01

11 Chung HY, et al., HLA Alleles and Haplotypes in Koreans 695 등이었다. HLA 일배체형의분석결과 0.5% 이상의빈도를나타내는 2-유전자좌일배체형은 A-C 44종, B-C 42종, A-B 51종, B-DRB1 52종이었고, 3-유전자좌일배체형은 A-C-B 42종, A-B-DRB1 34종이었다. 한국인에서빈도 1% 이상의 A-B-DR 일배체형은 13종으로, 전체일배체형의 26.0% 를차지하였고, 2% 이상으로가장흔한 A-B-DR 일배체형은 A*33: 03-B*44:03-DRB1*13:02 (3.7%), A*33:03-B*44:03-DRB1* 07:01 (3.0%), A*33:03-B*58:01-DRB1*13:02 (3.0%), A*24: 02-B*07:02-DRB1*01:01 (2.8%), A*30:01-B*13:02-DRB1* 07:01 (2.3%), A*11:01-B*15:01-DRB1*04:06 (2.2%) 등 6종이었다. 결론 : 본연구를통해한국인의대립유전자수준의 HLA 형별과 HLA 일배체형빈도에대한자료를제시하였으며, 본연구의결과는한국인에서장기이식, 질환연관성연구, 인류유전학적연구등에서중요한기초자료로이용될수있을것으로기대된다. 참고문헌 1. Marsh SG, Albert ED, Bodmer WF, Bontrop RE, Dupont B, Erlich HA, et al. Nomenclature for factors of the HLA system, Tissue Antigens 2010;75: Lee KW and Park MH. New HLA nomenclature (2010) and its clinical application in Koreans. Korean J Lab Med 2010;30: Roh EY, Kim HS, Kim SM, Lim YM, Han BY, Park MH. HLA-A, -B, -DR allele frequencies and haplotypic associations in Koreans defined by generic-level DNA typing. Korean J Lab Med 2003;23: ( 노은연, 김현수, 김선미, 임영미, 한복연, 박명희. DNA 형별검사에의한한국인의 HLA-A, -B, -DR 형별및일배체형의분포. 대한진단검사의학회지 2003;23: ) 4. Whang DH, Yang YS, Hong HK. Allele and haplotype frequencies of human leukocyte antigen-a, -B, and -DR loci in Koreans: DNA typing of 1,500 cord blood units. Korean J Lab Med 2008;28: ( 황동희, 양윤선, 홍혜경. 한국인에서 human leukocyte antigen-a, -B, -DR 대립유전자와일배체형빈도 : 제대혈 1,500단위의 DNA 형별검사. Korean J Lab Med 2008;28: ) 5. Yoon JH, Shin S, Park MH, Song EY, Roh EY. HLA-A, -B, -DRB1 allele frequencies and haplotypic association from DNA typing data of 7,096 Korean cord blood units. Tissue Antigens 2010;75: Hwang SH, Oh HB, Yang JH, Kwon OJ, Shin ES. Distribution of HLA -A, B, C allele and haplotype frequencies in Koreans. Korean J Lab Med 2004;24: ( 황상현, 오흥범, 양진혁, 권오중, 신은순. 한국인의 HLA-A, -B, -C 대립유전자와일배체형분포. 대한진단검사의학회지 20 04;24: ) 7. Park MH, Kim HS, Kang SJ. HLA-A, -B, -DRB1 allele and haplotype frequencies in 510 Koreans. Tissue Antigens 1999;53: Song EY, Park MH, Kang SJ, Park HJ, Kim BC, Tokunaga K, et al. HLA class II allele and haplotype frequencies in Koreans based on 107 families. Tissue Antigens 2002;59: Song EY, Park H, Roh EY, Park MH. HLA-DRB1 and -DRB3 allele frequencies and haplotypic associations in Koreans. Hum Immunol 2004;65: Lee KW, Oh DH, Lee C, Yang SY. Allelic and haplotypic diversity of HLA-A, -B, -C, -DRB1, and -DQB1 genes in the Korean population. Tissue Antigens 2005;65: Park MH, Hwang YS, Park KS, Tokunaga K, Akaza T, Juji T, et al. HLA haplotypes in Koreans based on 107 families. Tissue Antigens 1998;51: Mack SJ, Jani AJ, Geyer LN, Erlich HA. Using the reverse line strip system for studies of human diversity. 13th IHWS Technology Joint Report. In: Hansen JA, ed. Immunobiology of the human MHC: Proceedings of the 13th international histocompatibility workshop and conference. Vol. 1. Seattle: IHWG Press, 2006: Park MH, Whang DH, Kang SJ, Han KS. HLA-A*02 allele frequencies and haplotypic associations in Koreans. Tissue Antigens 2000; 55: Whang DH and Park MH. Diversity of HLA-A26 alleles and haplotypes in Koreans. Korean J Lab Med 2003;23:52-6. ( 황동희및박명희. 한국인에서 HLA-A26 대립유전자및관련일배체형의분포양상. 대한진단검사의학회지 2003;23:52-6.) 15. Bannai M, Tokunaga K, Lin L, Ogawa A, Fujisawa K, Juji T. HLA- B40, B18, B27, and B37 allele discrimination using group-specific amplification and SSCP method. Hum Immunol 1996;46: Bannai M, Tokunaga K, Tanaka H, Lin L, Kashiwase K, Tokunaga K, et al. Five HLA-B22 group alleles in Japanese. Tissue Antigens 1997;49: Song EY, Whang DH, Hur M, Kang SJ, Han KS, Park MH. HLA- B*44 allele frequencies and haplotypic associations in Koreans. Hum Immunol 2001;62: Wang H, Tokunaga K, Ishikawa Y, Tanaka H, Kashiwase K, Shibata Y, et al. A high-resolution genotyping method for HLA-C alleles and possible shared HLA-C-B haplotypes between Japanese and

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