BACKGROUND: When organ transplantation or HLA-matched platelet transfusion is considered, accu-rate identification of HLA antibody specificity in the recipient's serum is very important. In this study, we report our experience in an international quality control program. METHODS: For external quality control in a HLA antibody test, the International Serum Exchange Program distributes serum samples, generally showing polyspecific reactivity for cross-reactive epitope groups (CREGs), to participating laboratories: 4 samples per survey, 10 surveys per year. Participating in the program from May 1998 to August 2000 (24 surveys), we performed HLA antibody identification of 96 serum samples by the AHG-CDC (anti-human globulin-complement dependent cytotoxicity) method using frozen lymphocyte trays (36 lymphocyte panels). We compared the results of our laboratory with those of the total participants (all methods combined, 72 to 92 laboratories per survey) using the analyzed survey results distributed by the program organizer. RESULTS: We analyzed the survey results for the antibodies to relatively common HLA antigens in Koreans (antigen frequency >1%). For the HLA antibodies detected in >or=20% of participants, our detection rate was higher by 10-15% than that of all laboratories (HLA-A, 76% vs 65%; HLA-B, 73% vs 57%). And for the HLA antibodies detected in >or=50% of the participants, our detection rate was as high as 88% for HLA-A and 87% for HLA-B. Our detection rate for a few antibody specificities was lower than that of all laboratories, namely HLA-A1, A3, B35, and B55. Among these, A1, A3, and B55 were of lower incidence antigens in Koreans (antigen frequency 3-4%), indicating that the low detection rate was due to a limitation in the composition of lymphocyte panels. CONCLUSIONS: In general, our detection rate of HLA antibodies was superior to the average detection rate of the total participant laboratories. We would be able to improve the low detection rate for a few antibody specificities to lower incidence antigens by refining the composition of lymphocyte panels.
Antibodies ; Antibody Specificity ; HLA Antigens ; HLA-A Antigens ; HLA-A1 Antigen ; HLA-B Antigens ; Incidence ; Lymphocytes ; Organ Transplantation ; Platelet Transfusion ; Quality Control ; Transplants

BACKGROUND: Obstructive sleep apnea syndrome (OSAS) is believed to have multifactorial causes. The major risk factors for OSAS are obesity, narrowed upper airways, and abnormal cranial-facial structures. A genetic basis for OSAS has been also suggested by reports of families with many members affected. This study analyzed the HLA typing in patients with OSAS to determine the possible role of genetics in OSAS. METHODS: Twenty-five Korean patients with OSAS (1 woman and 24 men; age range 30-66 years) were enrolled in this study. A diagnosis of OSAS was made using full-night polysomnography. The control group consisted of 200 healthy Korean people. Serologic typing of the HLA-A and B alleles was performed in all patients using a standard lymphocyte microcytotoxicity test. Analysis of the polymorphic second exons of the HLA-DRB1 gene was performed using a polymerase chain reaction-sequence specific oligonucleotide probe. RESULTS: The allele frequency of HLA-A11 was significantly lower in patients with OSAS compared with the controls (p<0.05). The HLA-B allele frequencies in the patients and controls had a similar distribution. Analysis of the HLA- DRB1 gene polymorphisms showed an increased frequency of DRB1*09 in the OSA patients compared with the controls (p<0.05). When the analysis was performed after dividing the OSAS patients according to the severity of apnea, the allele frequency of HLA-DRB1*08 was significantly higher in the severe OSA patients (apnea index>45) than in the controls (p<0.05). CONCLUSION: This study revealed an association between OSAS and the HLA-A11 and DRB1*09 alleles as well as association between the disease severity and the HLA-DRB1*08 allele in Korean patients. These results suggest that genetics plays an important role in both the development and the disease severity of OSAS.
Alleles ; Apnea ; Cytotoxicity Tests, Immunologic ; Diagnosis ; Exons ; Female ; Gene Frequency ; Genetics ; Histocompatibility Testing ; HLA Antigens ; HLA-A Antigens ; HLA-A11 Antigen ; HLA-B Antigens ; HLA-DRB1 Chains ; Humans ; Lymphocytes ; Male ; Obesity ; Polysomnography ; Risk Factors ; Sleep Apnea, Obstructive*

In order to evaluate association of particular HLA typing with certain uveitis in Korean population, HLA antigens were analyzed in 114 uneitis patients(acute anterior uveitis: 32 cases, Behcet`s disease: 25 cases, intermediate uveitis: 19 cases, Vogt-Koyanagi-Harada (V-K-H) syndrome: 10 cases, retinal vasculitis: 12 cases, Eale`s disease: 3 cases, posterior uveitis: 9 cases, pan.uveitis: 4 cases). The stronger association between acute anterior uveitis and HLA-B27 was statistically significant, and this result was similar to reports in other ethnic groups. Also, the association between V-K-H syndrome and HLA-DR4 showed same results. But the high frequency of HLA-DR7 in the patients with V-K-H syndrome was unque in patients of Korean popjlation and statistically significant. The association between HLA-A2 and posterior uveitis was high in patients of Korean population and statistically significant. Behcet`s disease was stronger association with HLA-B51 but not statistically significant and much weaker association than reports in Japanese group. Although many similarities of associations between particular uveitis and HLA typing were detected as compared with other ethnic groups, distinctive HLA associations were demonstrated in Korean population. Additional cases and long-term follow-up are required to confirm the association with HLA typing and the relationship with prognosis including clinical and laboratory variabilities.
Asian Continental Ancestry Group ; Ethnic Groups ; Follow-Up Studies ; Histocompatibility Testing ; HLA Antigens ; HLA-A2 Antigen ; HLA-B27 Antigen ; HLA-B51 Antigen ; HLA-DR4 Antigen ; HLA-DR7 Antigen ; Humans ; Prognosis ; Retinal Vasculitis ; Uveitis* ; Uveitis, Anterior ; Uveitis, Intermediate ; Uveitis, Posterior

OBJECTIVE: Behcet? disease (BD) is a chronic inflammatory disorder affecting several organs. The etiology of BD remains unclear, although genetic factors, infectious agents, and immune mechanisms have been studied. The association of BD with certain genetic factors, especially HLA-B51 antigen, is well known in some geographical areas. Nevertheless, the familial occurrence of BD is rare. In this paper, HLA phenotype was evaluated in one family member showing the clustering of BD. METHODS: The serological tissue typing of HLA class I and class II antigens was performed by standard National Institutes of Health microlymphocytotoxicity method in seven family members in which four siblings were affected by BD. The diagnosis of BD was established by the criteria of the International Study Group of BD in these four siblings. RESULTS: In this family study, all members had HLA-A2 and DQ3 antigens. Although HLA-B51 antigen was positive in six out of seven family members, BD was developed in three of the six having HLA-B51 antigen. Three siblings had the exact same HLA phenotype. However, only one person had BD among three siblings with identical HLA phenotype. In addition, all siblings who developed erythema nodosum-like lesion had HLA-B51 antigen. CONCLUSION: This family suggests that the familial clustering of BD may not be explained solely by a susceptible HLA phenotype. The environmental factor or other genetic factors besides HLA-B51 might play a role in the development of BD. Furthermore, more studies and information will be needed to clarify the role of A2 and DQ3 antigens in BD.
Diagnosis ; Erythema ; Histocompatibility Antigens Class II ; Histocompatibility Testing ; HLA Antigens* ; HLA-A2 Antigen ; HLA-B51 Antigen ; Humans ; National Institutes of Health (U.S.) ; Phenotype ; Siblings

Objective: The Human Leukocyte Antigen (HLA) Class II is the major histocompatibility complex surface glycoproteins of humans responsible for presenting exogenous antigenic peptides which help direct specificity of immune response. In immune-cell therapy, the HLA allelic variants are of particular importance as they determine the successful activation of target cells that results to a desired therapeutic response. However, HLA Class II exhibits high polymorphism and has variable distribution in population, constituting these so-called allelic variants. Specifically, the HLA Class II DRB1 is considered the predominant locus among Filipinos. This research aimed to identify the presence of HLA Class II DRB1 allelic variants in the stem cell samples of ten (10) Filipino cancer patients by reverse transcription polymerase chain reaction (RT-PCR) amplification. Method: This study employed a PCR-based HLA Class II typing to identify the HLA Class II DRB1 allelic variant in Filipino cancer patients. Design of forward and reverse primers for HLA Class II DRB1, optimization of PCR conditions for amplifying HLA Class II DRB1, and identification of HLA Class II DRB1 allelic variants from samples by sequencing and database comparison were conducted. Results: PCR optimization showed that optimum annealing temperature for HLA DRB1 was 58.8°C with 1 mM MgCl2. PCR amplification of HLA DRB1 from ten anonymized cancer patient samples and DNA sequencing revealed that Patients 1, 2, 5, 8, 9, and 10 harbor HLA DRB1 allelic variants, particularly, the HLA DRB1*04:06:01, HLA DRB1*12:01:01, HLA DRB1*0813, HLA DRB1*04:05:01, HLA DRB1*09:01:02, and HLA DRB1*16:02:01, allelic variants, respectively. Conclusion Using the designed primers and optimized RT-PCR protocol, HLA information derived from six out of ten patient samples can be used for further applications in developing personalized or generic antigenic peptides such as dendritic cell cancer vaccine.
HLA Antigens

HLA-A2 is present at high frequency in most populations, as identified by serological and biochemical means. The values of these methods are limited by their failures to discriminate the products of the known allelic HLA-A02 variants. The great majority of genetic polymorphism which defines the allelic variants is found in exons 2 and 3 of the HLA-A02 gene. These exons encode the a-1 and a-2 domains of the HLA class I molecules, and the variation within the genes may influence the peptide binding specificity of the gene products of each allele. To determine the 17 known alleles of HLA-A02 an ARMS-PCR was developed. We applied this ARMS-PCR to 10 standard cell lines and we confirmed the specificity and sensitivity of this method. We defined the HLA-A 02 subtypes in 146 healthy Koreans who were serologically identified as HLA-A2. Five subtypes out of the 17 known A02 alleles were detected (A'0201, 0203, 0206, 0207, '0210) and A'0201 was most frequent (53.4%) and A'0206, '0207, '0203, 0210 (37.0%, 18.5%, 2.7%, 2.1%), were followed respectively. By linkage disequilibrium analysis with HLA-B alleles, A*02 subtypes were defined to be associated with many B alleles (B27, 35, 38, 39, 46, 52, 60, and 61). It is suggested that these findings may be helpful for the selection of patients for the specific immunotherapy with HLA-A02 restricted peptide vaccines and for the unrelated bone marrow transplantation in Korean.
Alleles ; Bone Marrow Transplantation ; Cell Line ; Exons ; HLA-A Antigens ; HLA-A2 Antigen ; HLA-B Antigens ; Humans ; Immunotherapy ; Linkage Disequilibrium ; Polymorphism, Genetic ; Sensitivity and Specificity ; Vaccines, Subunit

PURPOSE: The Epstein-Barr virus(EBV), gamma herpesvirus, is an important pathogen that is widespread around the world. The EBV causes various diseases depending on the geographic location, and on the immunity or the premorbid condition of the person exposed to EBV. To evaluate EBV typing may be the most important step to figure out the pathogenesis of EBV associated diseases, and we need to re-evaluate the pathologic role of human leukocyte antigen(HLA) in developing Epstein-Barr virus associated acute infectious mononucleosis by using newly developed methods. METHODS: This study included 24 children(age range : 6 to 13 years), serologically confirmed with acute infectious mononucleosis. The control group for the HLA type consisted of 200 age-matched healthy children. To classify HLA I, modified ARMs-PCR was used, while modified PCR-SSOP was utilized in typing of HLA II. Also, we performed EBV typing in study patients by using a one-step PCR. RESULTS: The results of HLA types : In HLA class I, HLA-A24 was positive in 69 of 200 healthy children and positive in 14 of 24 patients in the study group(relative risk : 3.5724, chi-square; 5.26, P<0.05). In HLA class II, HLA-DRB1*07 was detected in 18 of 200 healthy children, and eight of 24 patients in the study group(relative risk; 506173, chi-square; 9.73, P<0.01). The results of EBV types : In the research group, 20(83.8%) of 24 patients were shedding type A virus, while 4(16.7%) were type B. CONCLUSION: We conclude that development of infectious mononucleosis may be associated with HLA types, and these results suggest that acute infectious mononucleosis could have hereditary traits. And we confirm that type A EBV is highly prevalent in patients with acute infectious mononucleosis in Korea. Also, our results suggest that further large scale studies, including adult groups, regarding the association between pathogenesis of EBV with HLA-DP or HLA-DQ will be warranted.
Adult ; Child* ; Herpesvirus 4, Human* ; HLA-A24 Antigen ; HLA-DP Antigens ; HLA-DQ Antigens ; Humans* ; Infectious Mononucleosis* ; Korea ; Leukocytes* ; Polymerase Chain Reaction

OBJECTIVE: This study was conducted to elucidate the associations of HLA with systemic sclerosis (SSc) in Koreans. METHODS: HLA associations with SSc according to SSc-specific autoantibody status and clinical subsets (diffuse and limited) were investigated. HLA-A, B, and C antigens were typed by the serological method using microlymphocytotoxicity test, and HLA-DR by DNA typing method using PCR-reverse hybridization and PCR-SSCP in 56 Korean patients with SSc and 226 healthy controls. For SSc patients, anti-Scl-70 and anicentromere antibodies were tested by double immunodiffusion and indirect immunofluorescence, respectively. RESULTS: The results of HLA class I antigen typing showed that the frequencies of HLA-A24, B52 and B62 were increased, whereas those of A33, B44 and B58 were decreased in SSc patients compared to healthy controls. The frequency of HLA-DR2 was significantly increased, whereas that of HLA-DR13 was decreased in patients with SSc compared to controls. Among HLA-DR2 alleles, both HLA-DRB1*1501 and *1502 were increased in SSc patients compared to controls. According to clinical status, HLA-DRB1*1501 was increased in limited SSc patients and that of DRB1*1502 was increased both in diffuse and limited SSc patients compared to controls. According to autoantibody status, HLA- DRB1 1502 was significantly increased in anti-Scl-70-positive SSc patients and that of DRB1 1501 was increased in anti-Scl-70-negative SSc patients compared to controls. The association of HLA-DR2 alleles with SSc according to clinical subsets and anti-Scl-70 antibody status revealed that the frequency of HLA- DRB1 *1501 was significantly increased in anti-Scl-70-negative limited SSc patients compared to controls. CONCLUSIONS: These results suggest that different HLA-DR2 alleles are associated with different types of SSc in Koreans. HLA-DRB1 1502 shows strong association with anti-Scl-70-positive SSc, and DRB1 1501 with anti-Scl-70-negative limited SSc. It is concluded that the pathogenesis of SSc in Koreans is in part, based on the same genetic background.
Alleles ; Antibodies ; Asian Continental Ancestry Group ; DNA Fingerprinting ; Fluorescent Antibody Technique, Indirect ; HLA-A Antigens ; HLA-A24 Antigen ; HLA-DR Antigens ; HLA-DR2 Antigen ; HLA-DRB1 Chains ; Humans ; Immunodiffusion ; Scleroderma, Systemic*

The gene complex of Human Leukocyte Antigen (HLA) is located of chromosome 6p21, which is the most complicated dominant polymorphic genetic system. The HLA system has 108 genotypes. It is the best human genetic marker. It has been applied to forensic paternity test and individual identification. This article discusses the research development of HLA polymorphism and its application in forensic medicine.
Forensic Medicine ; Gene Frequency ; HLA Antigens/genetics* ; HLA-A3 Antigen/genetics* ; Humans ; Polymorphism, Genetic

To study the gene polymorphism of HLA-A, B, DRB1 alleles in patients with chronic myelogenous leukemia and to explore the correlation of HLA with chronic myelogenous leukemia, the polymerase chain reaction-reverse sequence specific oligonucleotide (PCR-RSSO) was used to analyze the polymorphism of HLA-A, B, DRB1 alleles of 293 CML Patients and 406 randomized and synchronous blood donors (healthy and unrelated with patients) from Guangdong Han population. The results indicate that the gene frequency of HLA-A*24 in CML group was 15.53% lower than that of control group (22.09%, RR = 0.63, p = 0.005); the gene frequency of HLA-B*13 in CML group was 10.41% higher than that of control group (6.74%, RR = 1.68, p = 0.016). The gene frequency of HLA- DRB1*14 in CML group was 7.51% lower than that of control group (11.89%, RR = 0.58, p = 0.008). The differences were all statistically significant. It is concluded that the gene frequency of HLA-A*24, HLA- DRB1*14 in CML patients is significantly lower than normal people in Guangdong. The gene frequency of HLA-B*13 in CML patients is significantly higher than normal people in Guangdong. Further study is needed to make sure whether HLA-A*24 and HLA- DRB1*14 are protective gene markers for CML acquisition on Guangdong Chinese Han population and whether HLA-B*13 is a gene marker for CML susceptibility on this population.
Adolescent ; Adult ; Aged ; Blood Donors ; Child ; Child, Preschool ; China ; Female ; HLA-A Antigens ; genetics ; metabolism ; HLA-A24 Antigen ; HLA-B Antigens ; genetics ; metabolism ; HLA-B13 Antigen ; HLA-DR Antigens ; genetics ; metabolism ; HLA-DRB1 Chains ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; immunology ; Male ; Middle Aged ; Young Adult