kjh The Korean Journal of Hematology Korean J Hematol 1738-7949 2092-9129 Korean Society of Hematology 10.5045/kjh.2007.42.1.15 kjh-42-15 Original Article Susceptibility of Leukemia according to the Genotype of Minor Histocompatibility Antigens in a Korean Population ParkMin-Ji 1 ChoiHee-Baeg 2 YoonHo-Yeol 2 ChoiEun-Jeong 2 KimSu-Yeon 2 KimHee-Je 3 EomKi-Sung 3 LeeSeok 3 KimDong-Wook 3 LeeJong-Wook 3 MinWoo-Sung 3 KimChun-Choo 3 KimTai-Gyu 1 2 Department of Microbiology, College of Medicine, Seoul, Korea Catholic Hematopoietic Stem Cell Bank, Seoul, Korea The Catholic Hematopoietic Stem Cell Transplantation Center, The Catholic University of Korea, Seoul, Korea Correspondence to：Tai-Gyu Kim Department of Microbiology and Immunology, College of Medicine, Catholic Hematopoietic Stem Cell Bank, The Catholic University of Korea 505, Banpo-dong, Seocho-gu, Seoul 137-701, Korea Tel: +82-2-590-1216, Fax: +82-2-3476-7355 E-mail: kimtg@catholic.ac.kr 032007 19032007 42115 23 10112006 14112006 16112006 Copyright © 2007 Korean Society of Hematology 2007 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Background:

In the search for susceptibility genes responsible for leukemia, genetic studies involving HLA association have been in progress extensively since the first report on its effect on the disease. Here we investigated the genetic associations of different leukemias with 4 autosomal mHags, HA-1, -2, -8 and HB-1. In particular, HB-1 is one of the leukemia-associated minor histocompatibility antigens (mHags) that is significantly expressed by Epstein-Barr virus-transformed- and tumor cells of all B lineage acute lymphoblastic leukemia (ALL).

 Methods:

A simultaneous genotyping method using PCR sequence-specific primers against HA-1, -2, -8 and HB-1 was developed, and their allelic frequencies in 139 healthy controls and 36 leukemia patients were observed. To compare genotype, phenotype, and gene frequencies of mHags with healthy controls, leukemia patients were classified into sub groups of ALL, acute myeloid leukemia (AML), and chronic myeloid leukemia (CML).

 Results:

The genotype frequencies of HA-1, -2 and -8 were not significantly different from healthy controls in every group of leukemia patients. However, the HB-1 H genotype was significantly increased in leukemia patients (P=0.03, OR=1.82, CI=1.08～3.06), particularly in AML (P=0.01, OR=2.4, CI=1.21～ 4.76) as compared with healthy controls.

 Conclusion:

Our results suggested that the genotype of HB-1 H may be associated with leukemia, particularly with AML. In further study, it is necessary to confirm the association of HB-1 with other leukemias in a larger group of patients, and to identify the underlying mechanism of HB-1 responsible for the occurrence of leukemia.

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Genotyping of HA-1, -2, -8 and HB-1 by PCR-SSP. Lane 1: H type for HA-1, lane 2: R type for HA-1, lane 3: V type for HA-2, lane 4: M type for HA-2, lane 5: R type for HA-8, lane 6: P type for HA-8, lane 7: H type for HB-1, lane 8: Y type for HB-1. Each type of mHags HA-1, -2, -8 and HB-1 of the three samples was as follows; sample 1: HH (HA-1), VM (HA-2), PP (HA-8), HH (HB-1); sample 2: RR (HA-1), VM (HA-2), RP (HA-8), YY (HB-1); sample 3: HR (HA-1), VV (HA-2), RR, (HA-8), HY (HB-1).

Allelic frequencies of HA-1, -2, -8 and HB-1 in healthy controls (n=139) and patients with leukemia (n=36) in the Korea.

Primer sets for genotyping of HA-1, HA-2, HA-8, and HB-1

Name Primer Sequence (5’-3’) Product size
HA-1 H Forward ACT TAA GGA GTG TGT GCT GCA  
R Forward ACT TAA GGA GTG TGT GTT GCG 179bp
Reverse CCT CAG AGC CTT AGC TGT CA  
HA-2 V Forward GCT CCT GGT AGG GGT TCA C  
M Forward GCT CCT GGT AGG GGT TCA T 203bp
Reverse CTT CCT TCT CCA CTC TCA GC  
HA-8 R Forward TCT AAC ACT TTG TCC AGA GTT C  
P Forward TCT AAC ACT TTG TCC AGA GTT G 257bp
Reverse ACT TGG TTG GCC TGG CTC TT  
HB-1 H Forward ATT CTT TTC TAT AGG TTC TCT GC  
Y Forward ATT CTT TTC TAT AGG TTC TCT GT 446bp
Reverse CTG TGC TTG GTA GCC ATT  
Internal∗ 　 Forward CCT TCC CAA CCA TTC CCT TA 768bp
Reverse GTC CAT GTC CTT CCT GAA GCA  

∗Homo sapiens growth hormone 2 gene (GH2, NM_022557).

Genotype and phenotype frequencies of HA-1 in healthy controls and patients with leukemia in Korea

Genotype frequency n (%)
Locus   Healthy controls Leukemia patients
(n=139) Total (n=36) ALL (n=12) AML (n=20) CML (n=4)
HA-1 HH 18 (12.9) 3 (8.3) 0 (0) 3 (15) 0 (0)
HR 74 (53.3) 24 (66.7) 10 (83.3) 11 (55) 3 (75)
RR 47 (33.8) 9 (25) 2 (16.7) 6 (30) 1 (25)
H 92 27 10 14 3
R 121 33 12 17 4
Gene frequency n (%)
  H 110 (39.6) 30 (41.7) 10 (41.7) 17 (42.5) 3 (37.5)
R 168 (60.4) 42 (58.3) 14 (58.3) 23 (57.5) 5 (62.5)

Genotype and phenotype frequencies of HA-2 in healthy controls and patients with leukemia in Korea

Genotype frequency n (%)
Locus 　 Healthy controls Leukemia patients
(n=139) Total (n=36) ALL (n=12) AML (n=20) CML (n=4)
HA-2 VV 120 (86.3) 31 (86.1) 12 (100) 15 (75) 4 (100)
VM 17 (12.3) 5 (13.9) 0 (0) 5 (25) 0 (0)
MM 2 (1.4) 0 (0) 0 (0) 0 (0) 0 (0)
V 137 36 12 20 4
M 19 5 0 5 0
Gene frequency n (%)
  V 257 (92.4) 67 (93.1) 24 (100) 35 (87.5) 8 (100)
M 21 (7.6) 5 (6.9) 0 (0) 5 (12.5) 0 (0)

Genotype and phenotype frequencies of HA-8 in healthy controls and patients with leukemia in Korea

Genotype frequency n (%)
Locus 　 Healthy controls Leukemia patients
(n=139) Total (n=36) ALL (n=12) AML (n=20) CML (n=4)
HA-8 RR 21 (15.1) 7 (19.4) 3 (25) 4 (20) 0 (0)
RP 60 (43.2) 19 (52.8) 6 (50) 10 (50) 3 (75)
PP 58 (41.7) 10 (27.8) 3 (25) 6 (30) 1 (25)
R 81 26 9 14 3
P 118 29 9 16 4
Gene frequency n (%)
　 R 102 (36.7) 33 (45.8) 12 (50) 18 (45) 3 (37.5)
P 176 (63.3) 39 (54.2) 12 (50) 22 (55) 5 (62.5)

Genotype and phenotype frequencies of HB-1 in healthy controls and patients with leukemia in Korea

Genotype frequency n (%)
Locus 　 Healthy controls Leukemia patients
(n=139) Total (n=36) ALL (n=12) AML (n=20) CML (n=4)
HB-1 HH 31 (22.3) 16 (44.4)∗ 5 (41.7) 10 (50)∥ 1 (25)
YH 67 (48.2) 12 (33.3) 2 (16.6) 7 (35) 3 (75)
YY 41 (29.5) 8 (22.3) 5 (41.7) 3 (15) 0 (0)
H 98 28 7 17 4
Y 108 20† 7 10 3
Gene frequency n (%)
  H 129 (46.4) 44 (61.1)‡ 12 (50) 27 (67.5)∗∗ 5 (62.5)
Y 149 (53.6) 28 (38.9)§ 12 (50) 13 (32.5)†† 3 (37.5)

∗P-value was calculated by chi-square tests of total leukemia patients versus healthy controls (P=0.008, OR=2.8, CI=1.32～ 5.91);

P-value was calculated by chi-square tests of total leukemia patients versus healthy controls (P=0.008, OR=0.36, CI=0.17～0.76);

P-value was calculated by chi-square tests of total leukemia patients versus healthy controls (P=0.03, OR=1.82, CI=1.08～3.06);

P-value was calculated by chi-square tests of total leukemia patients versus healthy controls (P=0.03, OR=0.56, CI=0.33～0.93);

P-value was calculated by chi-square tests of AML patients versus healthy controls

P-value was calculated by chi-square test of AML patients versus healthy controls (P=0.008, OR=3.48, CI=1.39～8.77); (P=0.008, OR=0.29, CI=0.12～0.72); ∗∗P-value was calculated by chi-square test of AML patients versus healthy controls (P=0.01, OR=2.4, CI=1.21～4.76);

P-value was calculated by chi-square test of AML patients versus healthy controls (P=0.01, OR=0.42, CI=0.21～0.82).