Binding of Toxic Shock Syndrome Toxin-1 to HLA-DR and HLA-DQ Molecules by L-cell Transfectants

Chang, Mee Soo,Ryu, Jiso,Lee, Mi Sook,Woo, Jun Hee 대한화학요법학회 1999 대한화학요법학회지 Vol.17 No.4

배경 : 포도구균에 의해 생산되는 TSST-1이 원인인 독성쇽증후군은 재래의 세균독소와는 달리 초항원으로 작용하는 기전에 의해 여러 장기에 병변이 합병되어 기능 부전을 초래한다. TSST-1이 초항원으로 작용하는 기전 중 TSST-1이 MHC와 결합하는 수용체를 밝히기 위하여 실험을 하였다. 방법 건강성인에서 말초혈액을 채취하여 말초혈액단핵구를 그리고 피부생검조직에서 정상 피부섬유원세포를 분리하였다. TSST-1이 결합하는 분자수용체를 동정하기 위한 방법으로 분자융합방법, 단클론항체, 결합능측정(binding assay), T cell proliferation assay을 이용하여 실험을 진행하였다. HLA-DRα, HLA-β1, HLA-DPw4β, HLA-DQ7α, HLA-DQw3β의 cDNA 클론을 expression vector에 selection amrker를 가진 plasmid pSV-neo를 Murine L-cell fibroblast에 transfection 시켜서 L-cell transfectants를 만들었고, Chloramine T를 이용한 iodination 한 TSST-1과 HLA-DR, DQ, DP의 결합능을 측정하였고 HLA-DR, DQ, DP에 대한 단클론항체로 억제한 뒤에 결합능과 비교하였다. 결과 TSST-1가 HLA-DR, DQ, DP의 결합능을 측정하고, 뒤이어 HLA-DR, DQ, DP에 대한 단클론항체로 TSST-1과의 결합을 억제시킬 수 있는지 보조세포 존재하에 증식능을 측정한 결과 HLA-DR에 대한 단클론항체는 TSST-1결합을 강력히 억제하는데 비하여 HLA-DP, HLA-DQ에 대한 단클론항체는 강하게 억제하지 못하였다. 결론 TSST-1은 HLA-DR L-cell transfectants와 결합하고, HLA-DQ L-cell transfectants에도 약간 결합하는 것으로 판단하여 TSST-1은 HLA-DR, HLA-DQ 분자의 특이부위에 결합하는 것이 관찰되었다. 초항원의 작용기전을 이해하는데 기초적인 설명을 가능하게 하는 것이다. Background : Expressible HLA class Ⅱa and cDNA were used for DNA-mediated gene transfer to produce L cell transfectants expressing single types of human class Ⅱ molecules. Cells with the highest degree of MHG class Ⅱ expression exhibited the highest number of TSST-l-binding sites. In contrast, cells with low or undetectable MHC class Ⅱ expression (resting T cells, and fibroblasts) exhibited very low or undetectable numbers of TSST-1-binding sites. Methods' The relationship between surface MHC class Ⅱ molecules and TSST-1 binding sites was assessed by examining the effect of antibodies to MHC class Ⅱ molecules on TSST-I binding. We examined TSST-I binding to mouse L-cell transfectants that expressed HLA-DR. DQ, DP gene products. Results : Immunofluorescence staining with anti-HLA-DR, -DP. and -DQ mAbs revealed comparable surface expression of the respective MHC class Ⅱ antigens by each of the three transfectants. TSST-1 bound to the HLA-DR transfectant and to the HLA-DQ transfectant but not to the HLA-DP transfectant and not to untransfected L cells. Conclusion : In summary, monoclonal antibodies to HLA-DR. but not to HLA-DP or HLA-DQ. strongly inhibited TSST-1 binding. Binding of TSST-1 was shown to HLA-DR and HLA-DQ L cell transfectants. These resu Its indicate that TSST-1 bind to distinct sites on HLA-DR and HLA-DQ molecules.

Prediction of HLA-DQ in Deceased Donors and its Clinical Significance in Kidney Transplantation

Kim Soo-Kyung,Yang John Jeongseok,Hwang Sang-Hyun,Sung Heungsup,Shin Sung,Ko Sun-Young,Oh Heung-Bum 대한진단검사의학회 2021 Annals of Laboratory Medicine Vol.41 No.2

Background: HLA-DQ typing in deceased donors is not mandatory in Korea. Therefore, when patients develop DQ antibodies after kidney transplantation (KT) from deceased donor, it is impossible to determine whether they are donor-specific antibodies (DSA). We developed DQ prediction programs for the HLA gene and evaluated their clinical utility. Methods: Two HLA-DQ prediction programs were developed: one based on Lewontin’s linkage disequilibrium (LD) and haplotype frequency and the other on an artificial neural network (ANN). Low-resolution HLA-A, -B, -DR, and -DQ typing data of 5,603 Korean patients were analyzed in terms of haplotype frequency and used to develop an ANN DQ prediction program. Predicted DQ (pDQ) genotype accuracy was analyzed using the typed DQ data of 403 patients. pDQ DSA agreement, sensitivity, specificity, and false-negative rate was evaluated using 1,970 single-antigen bead assays performed on 885 KT recipients. The clinical significance of DQ and pDQ DSA was evaluated in 411 KT recipients. Results: pDQ genotype accuracies were 75.4% (LD algorithm) and 75.7% (ANN). When the second most likely pDQ (LD algorithm) was also considered, the genotype accuracy increased to 92.6%. pDQ DSA (LD algorithm) agreement, sensitivity, specificity, and false-negative rate were 97.5%, 97.3%, 98.6%, and 2.4%, respectively. The antibody-mediated rejection treatment frequency was significantly higher in DQ or pDQ DSA-positive patients than in DQ or pDQ DSA-negative patients (P<0.001). Conclusions: Our DQ prediction programs showed good accuracy and could aid DQ DSA detection in patients who had undergone deceased donor KT without donor HLA-DQ typing.

Analysis of HLA DQαusing Polymerase Chain Reaction and Allele Specific Oligonucleotide Method in Korean Insulin Dependent Diabetes Mellitus Patients

Yoon, Kun Ho,Yoo, Soon Jib,Kang, Moo-Il,Son, Hyun Shik,Hong, Kwan Soo CATHOLIC MEDICAL CENTER 1992 Bulletin of the Clinical Research Institute Vol.20 No.2

Genetic predispostition to the autoimmune disease insulin dependant diabetes mellitus is determined, in part, by a gene within the major histocompatibility complex (MHC) on human chromosome 6. After amplification of exon 2 HLA-DQα chain gene with polymerase chain reaction (PCR) procedure, we used non-radioactive HLA-DQα ASO probes which labelled with horseradish peroxidase (HRP) coupled to the 5’ end of the oligonucleotide for the HLA-DQα typing. The results were as follows: 1. The first domain elf the DQα chain gene was amplified from minimum 1 ng of genomic DNA with use of the PCR. And from 1 ㎍ DNA samples, all the samples were successfully amplified which have a product of approximately 242 bp. 2. On oligonulcleotide dot blot analysis of study subjects and standard control cell lines, TAB (HLA-DQAI homozygosity) cell line was hybridized to RH 83 probe only and DKB (HLA-DQA3 homozygosity) cell line to GH 67 probe and LUY (HLA-DQA4 homozygosity) cell line to GH 66. According to these results, we accurately determined HLA-DQα subtypes in study subjects. 3. Frequency of DQA3 allele in Korean was high and DQA2 allele was low compared to other populations. In IDDM patients, DQA3 allele w as significantly increased (relative risk 6.6, P=0.03) compared with controls. Conversely the frequencies of DQA1 allele was decreased among patients (relative risk 0.25, P=0.02). These results suggest that HLA-DQA3 allele is positively associated with IDDM as in other populations and HLA-DQA1 allele is negatively associated with IDDM in Korean population.

Celiac Disease in a Predisposed Subject (HLA-DQ2.5) with Coexisting Graves’ Disease

황인경,김선혜,이언주,진상욱,이상열,오승준,우정택,김성운,김영설,전숙 대한내분비학회 2015 Endocrinology and metabolism Vol.30 No.1

Celiac disease is an intestinal autoimmune disorder, triggered by ingestion of a gluten-containing diet in genetically susceptible individuals. The genetic predisposition is related to human leukocyte antigen (HLA) class II genes, especially HLA-DQ2-positive patients. The prevalence of celiac disease has been estimated to be ~1% in Europe and the USA, but it is rarer and/or underdiagnosed in Asia. We report a case of celiac disease in a predisposed patient, with a HLA-DQ2 heterodimer, and Graves’ disease that was treated successfully with a gluten-free diet. A 47-year-old woman complained of persistent chronic diarrhea and weight loss over a 9 month period. Results of all serological tests and stool exams were negative. However, the patient was found to carry the HLA DQ2 heterodimer. Symptoms improved after a gluten-free diet was initiated. The patient has been followed and has suffered no recurrence of symptoms while on the gluten-free diet. An overall diagnosis of celiac disease was made in a genetically predisposed patient (HLA-DQ2 heterodimer) with Graves’ disease.

한국인 전신성홍반성루푸스 환자에서 HLA-DRB1, DQB1 대립유전자의 연관성 및 항인지질 항체와 항β₂ Glycoprotein I 항체에 관한 연구

이상곤,차훈석,양윤선,Lee, Sang Gon,Cha, Hoon Suk,Yang, Yoon Sun 대한면역학회 2002 Immune Network Vol.2 No.4

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.

한국인 집단의 HLA-DQχ 대립유전자와 유전자형 빈도

선문숙,한면수,박기원,이양한,최상규,강순자 이화여자대학교 생명과학연구소 1993 생명과학연구논문집 Vol.4 No.-

Allele and genotype frequencies at the HLA-DQa locus were determined by the use of PCR amplification and non-radioactive allele specific oligonucleotides(ASOs). The probes defined six alleles and 21 genotypes in a dot-blot format. A total of 206 individuals were typed using this method. The HLA-DQa genotype frequencies did not deviate significantly from Hardy-Weinberg equilibrium. The allele frequencies ranged from 5.3% to 33.7% and the frequency of DQ A3 allele was highest(33.7%). The observed frequency of DQa 3-4 genotype was highest(13.6%). The allelic diversity value was 0.79. The heterozygosity at the HLA-DQa locus was 83.5% These population data will allow the use of the HLA-DQa marker in paternity determination and the analysis of individual identity in forensic samples.

Association of HLA-DR and -DQ Genes with Familial Moyamoya Disease in Koreans

Hong, Seok-Ho,Wang, Kyu-Chang,Kim, Seung-Ki,Cho, Byung-Kyu,Park, Myoung-Hee The Korean Neurosurgical Society 2009 Journal of Korean neurosurgical society Vol.46 No.6

Objective : Moyamoya disease (MMD) is an uncommon cerebrovascular disorder, characterized by progressive occlusion at the terminal portion of the internal carotid artery. Incidence of the disease is high in East Asia and familial MMD accounts for about 15% of the disease. Although the pathogenesis is unknown, association of HLA class I or II alleles with MMD has been reported with conflicting results. We investigated whether there is a difference in HLA class II association between familial and non-familial forms of the disease. Methods : A total of 70 Korean children with MMD, including 16 familial cases (10 probands), and 207 healthy controls were studied. Among familial cases, only 10 probands were used for the HLA frequency analysis. High resolution HLA-DRB1 and DQB1 genotyping was performed using polymerase chain reaction (PCR)-sequence specific oligonucleotide hybridization and PCR-single strand conformation polymorphism methods. Results : The phenotype frequencies of HLA-DRB1*1302 (70.0%) and DQB1*0609 (40.0%) were significantly increased in familial MMD compared to both controls [vs. 15.5%, corrected p ($p_c$) = 0.008, odds ratio (OR) = 12.76; vs. 4.3%, $p_c\;=\;0.02$, OR = 14.67] and non-familial MMD patients (vs. 14.8%, $p_c\;=\;0.02$, OR = 13.42; vs. 1.9%, $p_c\;=\;0.02$, OR = 35.33). The frequencies of DRB1 and DQB1 alleles in non-familial MMD patients were not significantly different from those in controls. Conclusion : Our findings suggest that the genetic polymorphism of HLA class II genes or other closely linked disease relevant gene(s) could be a genetic predisposing factor for familial MMD.

Clinical Spectrum and Prognostic Factors of Acute Necrotizing Encephalopathy in Children

Seo, Hye-Eun,Hwang, Su-Kyeong,Choe, Byung Ho,Cho, Min-Hyun,Park, Sung-Pa,Kwon, Soonhak The Korean Academy of Medical Sciences 2010 JOURNAL OF KOREAN MEDICAL SCIENCE Vol.25 No.3

<P>This study was conducted to investigate the etiology, the clinical characteristics and prognosis of acute necrotizing encephalopathy (ANE) in Korean children. Six children (1 yr to 7 yr) patients with ANE were enrolled. They were diagnosed by clinical and radiological characteristics and their clinical data were retrospectively analyzed. In a search of clinically plausible causes, brain MRI in all patients, mitochondrial DNA studies for mitochondrial encephalomyopathy, lactic acidosis, and strokelike episodes (MELAS) and myoclonus epilepsy and ragged red fibers (MERRF) in four patients, and genomic typing on HLA DRB/HLA DQB genes in three patients were performed. All had precedent illnesses and the main initial symptoms included mental change (83%), seizures (50%), and focal deficits (50%). MRI revealed increased T2 signal density in the bilateral thalami and/or the brainstem in all patients. Mitochodrial DNA studies for MELAS and MERRF were negative in those children and HLA-DRB1*1401, HLA-DRB3*0202, and HLA-DQB1*0502 seemed to be significant. A high dose steroid was given to all patients, which seemed to be partly effective except for 2 patients. In conclusion, ANE is relatively rare, but can result in serious neurological complication in children. Early detection and appropriate treatment may lead to a better neurological outcome.</P>

한국소아 그레이브스 병 및 하시모투 갑상선염의 발생기전에서 HLA-DQ유전자의 역할

심계식,최규철,양세원 凡石學術奬學財團 2000 凡石學術論文集 Vol.4 No.-

성인에서 발생하는 인슐린의존성 당뇨병 유병율 추정

박용수(Y . S . Park),이경진(K . J . Lee),김태화(T . W . Kim),김목현(M . H . Kim),이홍규(H . K . Lee),김성연(S . Y . Kim),고창순(C . S . Hoh),민헌기(H . K . Min) 대한내과학회 1997 대한내과학회지 Vol.52 No.5

Objective: IDDM is an autoimmune disease, which occurs among genetically susceptible individuals. In the Asian populations, it is not uncommon for adult patients with NIDDM to eventually lose beta cell function and develop IDDM. These individuals may be characterized by autoantibodies to GAD and high risk HLA-DQ alleles, which are unlikely to be prevalent among true NIDDM cases or in the general population. The objective of the present study was to evaluate and compare the prevalence of these immunogenetic markers in NIDDM patients and healthy non-diabetic individuals from Korea. Methods: The prevalence of anti-GAD antibodies and HLA-DQA1 and DQB1 alleles among 121 newly diagnosed NIDDM cases identified from a population-based study in Yonchon, Korea and 100 matohed healthy control subjects were evaluated and compared. Results: The overall prevalence of anti-GAD antibodies was 1.7% (2 of 121) in patients with previously undiagnosed NIDDM, whereas 1 of 100 controls had positive antibodies. Among those who were positive, their titer of antibodies to GAD were not high. No statistically significant differences in the distributions of either mean levels of anti-GAD or DQA1 and DQB1 alleles were found comparing NIDDM patients to controls. Conclusion: The low prevalence of anti-GAD antibodies and HLA-DQA1 and DQB1 susceptibility alleles among recent-onset NIDDM patients, which was similar to observations in controls, suggests that diabetes in Korean adults is unlikely to have an autoimmune component to its pathogenesis.