• 대한임상검사과학회지
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• 제37권3호
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• pp.139-142
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• 2005

Most expressed HLA(human leukocyte antigen) loci exhibit a remarkable degree of allelic polymorphism, which is derived from sequenceing differences predominantly localized to discrete hypervariable regions of the amino-terminal domain of the molecule. In this study, the HLA-DRB1 genotypes were determined in twenty students using the PCR-SSP (polymerase chain reaction-sequence specific primer) technique. Two specific primer pairs in assigning the DRB1 gene were used. The results of PCR-SSP, the $HLA-DRB1^{\ast}0101$ primer detected nine and $HLA-DRB1^{\ast}1501$ primer detected three people. This study shows that the PCR-SSP technique is relatively simple, fast and a practical tool for the determination of the HLA-DRB1 genotypes. Moreover, these genotype frequency results of the HLA DRB1 gene could be useful for database study before being applied to individual identification and transplantation immunity.

• BMB Reports
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• 제29권1호
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• pp.45-51
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• 1996

Most expressed HLA loci exhibit a remarkable degree of allelic polymorphism, which derives from sequence differences predominantly localized to discrete hypervariable regions of the amino-terminal domain of the molecule. In this study, the HLA-DRB1 genotypes were determined in eighteen control cell lines and 112 unrelated Koreans using the PCR-SSP (Polymerase Chain Reaction-Sequence Specific Primer) technique. 29 specific primer pairs in assigning the DRB1 gene were used. The results of control cells correlated well with the data which was previously reported. The heterozygosity and homozygosity of the DRB1 gene were 0.786 and 0.214, respectively. In a total of 41 different DRB1 alleles and 83 genotypes, the most frequent allele and genotype were DRB1*04 and DRB1*0901/1501, respectively. This study shows that the PCR-SSP technique is relatively simple, fast and a practical tool for the determination of the HLA-DRBI genotypes. Moreover, these results-allele and genotype frequency and heterozygosity of the HLA DRB1 gene-could be useful for database study before being applied to individual identification and transplantation immunity.

• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.7007-7010
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• 2013

Background: In the literature, data concerning the relationship between breast cancer and HLA class II gene polymorphisms are limited, so the aim of this study was to determine if HLA-DQB1 and HLA-DRB1 MHC class-II alleles may confer susceptibility or resistance to the disease among Jordanian females. Materials and Methods: This case control study enrolled 56 Royal Hospital breast cancer patients and 60 age matched healthy controls, all of whom provided blood samples (2011-2013). A questionnaire was filled after signing a consent form and DNA was extracted, nucleic acids being amplified for assessment of HLA-DQB1 and HLA-DRB1 alleles by muliplex INNO-LiPA and allele typing carried out by reverse hybridization. Comparison of HLA-DQB1 and HLA-DRB1 allele distributions was carried out with paired t-test and chi-square statistics. Risk factors were assessed by odd ratios with 95% confidence intervals. Results: A significant negative correlation was observed between $HLADQB1^*$ 02 alleles and breast cancers (p=0.013). No significant associations were observed among $HLADQB1^*$ 03, 04, 05 and 06 or among $HLA-DRB1^*$ 01, 03, 04, 07, 08, 10, 11, 13, 14 and 15. Conclusions: $HLADQB1^*$ 02 alleles may provide positive protection against breast tumor risk among Jordanians, but not $HLADQB1^*$ 03, 04, 05 and 06 or $HLA-DRB1^*$ 01, 03, 04, 07, 08, 10, 11, 13, 14 and 15 alleles.

• Tuberculosis and Respiratory Diseases
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• 제54권4호
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• pp.367-377
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• 2003

연구배경 : 결핵균에 감염된 사람 중 약 10%만이 임상적으로 발병하는 것으로 보아 결핵의 발병 감수성에 숙주의 유전적 인자가 작용할 것으로 생각되고 있다. 저자들은 한국인 폐결핵 환자를 대상으로 고해상도 HLA 형별검사를 이용한 대립유전자 수준의 HLA-DR 및 DQ 유전자의 연관성에 대해 분석해 보았다. 방 법 : 이전 결핵 치료력이 없는 결핵 환자 67명(약제감수성군 38명, 다제내성군 29명)과 200명의 정상대조군을 대상으로 하였으며 HLA-DRB1 형별검사는 reverse SSO (sequence specific oligonucleotide)와 PCR-SSCP (single strand conformational polymorphism) 방법으로, DQB1 형별검사는 PCR-RFLP (restriction fragment length polymorphism), PCR-SSCP 및 PCR-SSP (sequence specific primer) 방법으로 시행하였다. 결 과 : 결핵환자군 중 다제내성군에서 대조군에 비해 DRB1*0701과 *08032의 빈도가 약 2배 정도 증가되는 경향을 보였고 이들 대립유전자와 연관된 DQB1*0202와 *0601(15.5% vs 34.5%, p=0.01)이 증가되었다. DQB1*0609는 결핵환자군에서 대조군에 비해 유의하게 증가되었고(4.0% vs 14.9%, p=0.004), 약제감수성군과 다제내성군에서 유사한 정도의 증가를 보였다. 폐병변의 중증도와 HLA의 연관성을 보면 덜 진행된 군(약제감수성군의 경증, 다제내성군의 중등증)에 비해 더 진행된 군(약제감수성군의 중등증+중증, 다제내성군의 중증)에서 DRB1*08032(4.2% vs 32.6%, p=0.007)와 DQB1*0601(12.5% vs 34.9%, p=0.047)이 유의하게 증가되었다. DQB1*0609는 다제내성군에서는 중증군에서, 약제감수성군에서는 경증군에서 대조군에 비해 유의하게 증가되었고 DRB1*0701과 DQB1*0202는 다제내성군의 중등증군에서만 유의하게 증가되어 이들 대립유전자는 폐병변의 중증도와 일정한 연관을 보이지는 않았다. 결 론 : HLA-DRB1*08032 및 DQB1*0601 대립유전자는 한국인에서 다제내성 결핵의 유전적 감수성인자로 생각되며 질환의 중증도와도 연관을 보이는 것으로 나타났다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권5호
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• pp.2491-2497
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• 2016

Background: Human leukocyte antigen (HLA) genes have been implicated in cervical cancer in several populations. Objectives: To study the predispositions of HLA alleles/haplotypes with cervical cancer. Materials and Methods: Clinically diagnosed and PAP smear confirmed cervical cancer patients (n 48) and age matched controls (n 47) were genotyped for HLA-A,-B,-DRB1* and DQB1* alleles by PCR-SSP methods. Results: The frequencies of alleles DRB1*04 (OR=2.57), DRB1*15 (OR=2.04), DQB1*0301 (OR=4.91), DQB1*0601 (OR=2.21), B*15 (OR=13.03) and B*07 (OR=6.23) were higher in cervical cancer patients than in the controls. The frequencies of alleles DRB1*10 (OR=0.22) and B*35 (OR=0.19) were decreased. Strong disease associations were observed for haplotypes DRB1*15-DQB1*0601 (OR=6.56; p< $3.5{\times}10^{-4}$), DRB1*14-DQB1*0501 (OR=6.51; p<0.039) and A*11-B*07 (OR=3.95; p<0.005). The reduced frequencies of haplotypes DRB1*10-DQB1*0501 (OR=0.45), A*03-B*35 (OR=0.25) and A*11-B*35 (OR= 0.06) among patients suggested a protective association. HLA-C* typing of 8 patients who possessed a unique three locus haplotype 'A*11-B*07-DRB1*04' (8/48; 16.66%; OR=6.51; p<0.039) revealed the presence of a four locus haplotype 'A*11-B*07-C*01-DRB1*04' in patients (4/8; 50%). Amino acid variation analysis of susceptible allele DQB1*0601 suggested 'tyrosine' at positions ${\beta}9$ and ${\beta}37$ and tyrosine-non-tyrosine genotype combination increased the risk of cervical cancer. Conclusions: Strong susceptible associations were documented for HLA alleles B*15, B*07, DRB1*04, DRB1*15, DQB1*0301, DQB1*0601 and haplotypes DRB1*15-DQB1*0601 and DRB1*14-DQB1*0501. Further, protective associations were evidenced for alleles B*35 and DRB1*10 and haplotypes A*11-B*35 and DRB1*10-DQB1*0501 with cervical cancer in South India.

• Tuberculosis and Respiratory Diseases
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• 제59권3호
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• pp.298-305
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• 2005

연구배경 : 폐쇄성 수면무호흡증후군은 다양한 원인에 의해 발생하게 된다. 주된 위험요소로는 비만과 좁은 상기도, 비정상적인 머리-얼굴 구조 등이 알려져 있으며 유전적 요인 또한 가족내 집단적 발생하였다는 보고들에 의해 뒷받침되고 있다. 본 연구에서 저자들은 HLA검사를 통하여 폐쇄성 수면무호흡증후군에서 유전학적인 배경을 규명하고자 하였다. 방 법 : 철야 수면다원검사로 진단한 25명의 폐쇄성 수면무호흡증후군 환자 (여자 1명과 남자 24명, 연령 30-66세)를 대상으로 하였으며 대조군은 200명의 건강한 한국인으로 하였다. HLA-A와 -B 대립유전자의 검사는 미세세포독성검사로 시행하였고 HLA-DRB1 유전자의 두번째 엑손의 다형성에 대한 분석은 PCRSSOP방법을 이용하여 시행하였다. 결 과 : HLA-A11 대립유전자의 빈도는 폐쇄성 수면무호흡증후군 환자군에서 대조군에 비해 유의하게 감소되어 있었다 (p<0.05). HLA-B 대립유전자의 빈도는 양군간에 유의한 차이가 없었다. HLA-DRB1 유전자의 다형태 분석에서는 DRB1*09의 빈도가 폐쇄성 수면무호흡환자군에서 대조군에 비해 유의하게 증가되어 있었다 (p <0.05). 환자군을 무호흡지수 45를 기준으로 경-중등증군과 중증군으로 나누어 대조군과 비교하였을 때 중증군에서 HLA-DRB1*08의 빈도가 유의하게 증가되어 있었다 (p <0.05). 결 론 : 한국의 폐쇄성 수면무호흡증후군 환자에서 HLAA11과 DRB1*09가 폐쇄성 수면무호흡증후군과 관련되어 있고, HLA-DRB1*08이 이 질환의 중증도와 연관되어 있음을 알 수 있었다. 이상의 결과는 폐쇄성 수면 무호흡 증후군의 발생뿐 아니라 경중도 여부에도 유전적인 요소가 중요한 역할을 한다는 것을 시사한다.

• IMMUNE NETWORK
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• 제2권4호
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• pp.227-232
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• 2002

Background: Systemic lupus erythematosus (SLE) is a complex autoimmune disease characterized by diverse clinical manifestations and autoantibody production, which is known to be strongly influenced by genetic factors. Previous studies have revealed the associations of SLE with HLA class II alleles and antiphospholipid antibody system (anticardiolipin antibody (aCL) and anti-${\beta}_2$ glycoprotein I antibody (anti-${\beta}_2$ GPI)). Therefore, we studied the associations of HLA class II alleles with SLE and antiphospholipid antibody system. Methods: The genotyping for HLA-DRB1 and DQB1 alleles were performed in 61 SLE patients and 100 controls by the polymerase chain reaction (PCR)-sequence specific oligonucleotide probe method. ELISA tests for aCL and anti-${\beta}_2$ GPI were performed in 39 of the 61 SLE patients. The results were evaluated statistically by Chi-square test. Results: The frequencies of the HLA-$DRB1^*15$ and $DQB1^*06$ in SLE patients were significantly higher than those in controls. HLA-$DRB1^*12$ was significantly lower in SLE patients than controls. Nine of 39 patients were positive for aCL (IgG) and three were positive for aCL (IgM). One of 39 patients were positive for anti-${\beta}_2$ GPI (IgG) and none of them positive for anti-${\beta}_2$ GPI (IgM). Association of aCL with HLA class II alleles was not observed in our study. Conclusion: According to our results, it was found that HLA-$DRB1^*15$ and $DQB1^*06$ were associated with genetic susceptiblility and $DRB1^*12$ was associated with resistance to SLE in Korean population. No Association of aCL with HLA class II alleles was observed and the positive rate for anti-${\beta}_2$ GPI was very low.

• Asian Pacific Journal of Cancer Prevention
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• 제15권10호
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• pp.4125-4128
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• 2014

The median ages at death from cancers between 1985 and 2005 were calculated to demonstrate that inherent anticancer mechanisms may be a common pathway for different cancers. Seventy-eight patients with gastric, liver and lung cancers, were recruited in the solid cancer group. The leukemia group consisted of 31 patients with three main types of leukemia. The controls were 100 healthy individuals. The samples were typed using an HLA-DR/DQ PCR-SSP typing kit. The results showed that the median ages at death from all causes were 64.7 years in 1985 and 70.1 years in 2005. The range of the median ages at death from all cancers was similar to the corresponding value for deaths attributed to all causes. The frequency of $DRB1^*03$ was 9.6% in the solid cancer group and 3.0% in the control group (p<0.05). The frequency of $DRB1^*04$ in the leukemia group were significantly lower than that of the control (p<0.05). $DRB1^*13$ and $DRB1^*06$ frequencies in the leukemia group were significantly higher than those of the controls (p<0.05). It is suggested that inherent anti-cancer mechanisms may be a common pathway for different cancers and are associated with the immune system and HLA.

• IMMUNE NETWORK
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• 제2권4호
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• pp.242-247
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• 2002

Background: Tumor necrosis factor-alpha (TNF-$\alpha$) is known to play an important role in various conditions such as inflammation, autoimmunity, apoptosis, insulin resistance and sleep induction. Five single nucleotide polymorphisms (SNPs) have been known to affect the transcriptional activities of TNF-$\alpha$: -1,031T/C, -863C/A, -857C/T, -308G/A and -238G/A. Methods: We have investigated 5 SNPs of the promoter region of TNF-$\alpha$ gene, the distribution of 5-locus TNF-$\alpha$ haplotypes, and their haplotypic associations with previously typed HLA-A, -B and -DRB1 loci in 107 healthy unrelated Koreans. TNF-$\alpha$ SNPs were typed using PCR-single-strand conformation polymorphism (SSCP) and PCR-restriction fragment length polymorphism (RFLP) methods. Results: The allele frequencies of -1,031C, -863A, -857T, -308A, and-238A, which are known as the high-producer-type, were 19.3%, 15.9%, 14.0%, 5.9%, and 2.9%, respectively. The frequency of -308A allele, known to be associated with autoimmune diseases, was 5.9% in Koreans which was lower than Caucasians (14~17%) and somewhat higher than Japanese (1.7%). Five most common TNF-$\alpha$ haplotypes (-1,031/-863/-857/-308/-238) comprised over 95% of total haplotypes: TCCGG (58.4%), CACGG (14.8%), TCTGG (13.7%), TCCAG (5.3%), and CCCGA (3.1%). Strong positive associations (P<0.001) were observed between TCCGG and B62; between CACGG and B51, $DRB1^*0901$; between TCTGG and B35, B54, B59, $DRB1^*1201$; and between TCCAG and A33, B58, $DRB1^*0301$, $DRB1^*1302$. Five most common extended haplotypes (>3%) comprised around 16% of total haplotypes: A33-B58-TCCAG-$DRB1^*1302$, A24-B52-TCCGG-$DRB1^*1502$, A33-B44-TCCGG-$DRB1^*1302$, A24-B7-TCCGG-$DRB1^*0101$, and A11-B62-TCCGG-$DRB1^*0406$. The distribution of extended HLA and TNF-$\alpha$ haplotypes showed that most of HLA haplotypes were almost exclusively associated with particular TNF-$\alpha$ haplotypes. Conclusion: The results obtained in this study would be useful as basic data for anthropologic studies and disease association studies in Koreans.

• 사상체질의학회지
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• 제13권1호
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• pp.97-103
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• 2001

Purpose This study was designed co determine the possibility of HLA typing in the objeccification of constitution. Methods We selected 100 patients who showed Taeyang characteristics, and divided them into Soum-inclined group and Soyang-inclined group. HLA-A, B, DRBI typing was perfomed by ARMS-PCR and PCR-SSOP method. Results Taeyang characteristic group as a whole showed significant difference in A1, A11, B37, B70/71, DRBI*15,DRB1*14 alleles in comparison with normal control group. Soum-inclined group showed significant difference in All, B70/71, Soyang-inclined group in DRBl*15, hard drinker group in DRBl*15, DRBl*13, drink-rejecting group in All, B37, DRBl*7, DRB1*14 in comparison with normal control group. Conclusion : There were significant relations between constitutional information and HLA types.