Allele and Haplotype Frequencies of Human Leukocyte Antigen-A, -B, and -DR Loci in Koreans: DNA Typing of 1,500 Cord Blood Units / 대한진단검사의학회지

BACKGROUND: The HLA system is known to be the most polymorphic genetic system in human, and HLA allele and haplotype distribution varies widely among different ethnic groups. This study was performed to examine the frequencies of HLA alleles and haplotypes in Koreans. METHODS: We examined HLA-A, -B, and -DR alleles at the serologic level in 1,500 cord blood units obtained from Koreans using the PCR-sequence specific oligonucleotide (SSO) method. Allele and haplotype frequencies were estimated by the maximum likelihood method using the computer program developed for the 11th International Histocompatibility Workshop. RESULTS: HLA alleles found in Koreans were 12 in A, 31 in B, and 13 in DR loci. Most frequent alleles with frequencies > or =10% in each locus in decreasing order of frequency were: A2, A24, A33, A11; B62; DR4, DR15, DR9, and DR13. Two-locus haplotypes with frequencies > or =0.1% were 104 A-B and 115 B-DR haplotypes, among which those with frequencies > or =1.0% showing significant positive linkage disequilibrium (P or =0.1% were identified. The results were similar to those of a previous study in 1,600 Koreans, although some differences were noted in the distribution of some less frequent alleles or haplotypes with frequencies < or =0.5%. CONCLUSIONS: We provided the allele and haplotype frequencies of HLA-A, -B, and -DR in cord blood units of Korean ethnicity defined by a DNA typing method, which can be used as basic data on Koreans for organ transplantation and disease association studies.

Performance Analysis of Panel Reactive Antibody Test by Lambda Antigen Tray Kit using Enzyme-Linked Immunosorbent Assay / 대한임상병리학회지

BACKGROUND: To detect human leukocyte antigen(HLA) alloantibodies, panel reactive antibody(PRA) test using complement dependent lymphocytotoxicity(CDC) has been generally used in Korea but this method lacks standardization. We analyzed the performance of Lambda Antigen Tray(LAT, One Lambda, Inc., Canoga Park, USA) kit using Enzyme-linked immunosorbent assay(ELISA). METHODS: We carried out PRA screening by ELISA with LAT10X5 kit and CDC with home-made panels for 81 sera patients. We carried out PRA identification by ELISA with LAT1240 kit for some sera with positive or discordant results of both CDC and PRA screening by ELISA. RESULTS: The agreement of CDC and PRA screening by ELISA was 85%(69/81). In 12 sera with positive results of both CDC and PRA screening by ELISA, no sera had same results of HLA antibody specificities. HLA class II antibodies were found in 13 sera(16%) and 12 sera of those had HLA class I antibodies, too. CONCLUSIONS: The agreement of CDC and PRA screening by ELISA was similar to other foreign result. Disagreement of both methods might be due to greatly difference of frequencies of HLA in used panels. New manufactured goods considered frequencies of Korean HLA should be made in order that PRA by ELISA can be applied to routine work.

CyCD3+MPO- Biphenotypic Leukemia With Unusual Presentation: A Case Report / 대한혈액학회지

No abstract available.

Peripheral Lymphocyte Subsets in Patients with Pulmonary and Extrapulmonary Tuberculosis / 대한임상병리학회지

BACKGROUND: T-cell mediated cellular immunity has been suggested as an important mechanism in mycobacterial infection. Also, it is known that there is a imbalance between helper and suppressor T cells in the pathogenesis of tuberculosis in human. This study was designed to evaluate the changes of lymphocyte subsets in patients with pulmonary and extrapulmonary tuberculosis. METHODS: Lymphocyte subset analysis was performed on 53 pulmonary tuberculosis and 21 extrapulmonary tuberculosis patients. The proportion of total T (CD3+), B (CD19+), helper T (H, CD3+CD4+) and suppressor T (S, CD3+CD8+) cells, natural killer (NK, CD16+CD56+) cells and activated T (CD3+HLA-DR+) cells were analyzed using SimultestTM (Becton-Dickinson, California, USA) by FACSortTM (Becton-Dickinson, California, USA) and each absolute cell counts and helper T/suppressor T (H/S) ratio were calculated. RESULTS: In pulmonary and extrapulmonary tuberculosis patients groups, there were no significant changes in percentage and absolute cell counts of lymphocyte subset compared to control group. But H/S ratio was significantly decreased in both groups and the H/S ratio in extapulmonary tuberculosis was lower than that in pulmonary tuberculosis (1.06+/-0.44 vs. 1.64+/-0.97). CONCLUSION: Decreased or reversed H/S ratio reflect the role of cell mediated immune response in patients with tuberculosis, expecially in the spreading of pulmonary tuberculosis. Lymphocyte subset test seems to be helpful for access the different clinical forms of tuberculosis, pulmonary and extapulmonary tuberculosis.

HLA-B60 and HLA-B61 Discrimination by PCR using Sequence-specific Primers (PCR-SSP) Method / 대한임상병리학회지

BACKGROUND: HLA-B40 is the most frequently identified HLA-B type in Koreans. Also HLA-B60 and B61 are the serologic split antigens of HLA-B40. But because of the lack of mono-specific alloantisera, cross reactivity of sera used as typing reagents, and poor antigenicity of some specific cells such as cord blood lymphocytes, discrimination between HLA-B60 and B61 has been often problematic in laboratories. In this study, authors evaluated whether the PCR-SSP method can be useful for accurate assignments of HLA-B60 and B61 or not. METHODS: Twenty-nine lymphocytes samples which were suspected as heterozygotes or homozygotes of HLA-B60 or B61 and three samples typed as HLA-B40 are selected from stored cord blood and organ transplantation donors. HLA types of these samples were defined by serologic method using a commercial typing kit. PCR that amplified exons 2 and 3 of the HLA-B gene using sequence specific primer pairs exactly matched to HLA-B60 or B61 allele making up a serological specificity was done. RESULTS: A clear discrimination between B60 and B61 was possible in all samples including 9 serologically ambiguous samples. Discrepancy between serologic typing and molecular typing was seen in three cases identified serologically as B40 positive but inable to define a split. Among three samples, two were identified as HLA-B61 and one was identified as HLA-B60. CONCLUSIONS: Molecular typing was useful in discriminating between HLA-B60 and B61. The PCR-SSP method for HLA-B60 and B61 including other cross-reactive HLA types will be helpful as a supplemental method of the serologic typing.

Comparison of Complement-Dependent Cytotoxicity, Enzyme-Linked Immunosorbent Assay and Flow Cytometric Assay for the Detection of HLA Class I Alloantibodies / 대한임상병리학회지

BACKGROUND: To detect anti-HLA class I alloantibodies in potential solid organ recipients, panel reactive antibody (PRA) test using complement-dependent lymphocytotoxicity (CDC) has been widely utilized. Recently, enzyme-linked immunosorbent assay (ELISA) and flow cytometry based methods (Flow PRA) were developed. We have compared these three methods for the detection of anti-HLA class I alloantibodies. METHODS: A total of 30 sera of various PRA reactivity determined using home-made CDC PRA panel were compared with ELISA PRA by QuikScreen Solid Phase HLA Antibody Screening Kit (GTI, Brookfield, USA). Among these samples, 19 sera were also compared with Flow PRA by FlowPRA I Screening Test (One Lambda, Inc., Canoga Park, USA). Qualitative as well as semiquantitative results using CDC %PRA, ELISA OD ratio (ratio of the test OD value divided by the x2 the value obtained for the mean of the negative controls) and Flow %PRA were compared. RESULTS: Sixteen out of 19 sera (84%) showed concordant results among all three methods, and 11 sera tested by both CDC PRA and ELISA PRA but not by Flow PRA showed perfect concordance. CDC %PRA, ELISA OD ratio, and Flow %PRA showed excellent correlation between each other (Pearson's correlation coefficient >0.67, P<0.005). CONCLUSIONS: Results obtained in this study indicate that ELISA PRA and Flow PRA tests might be good alternatives of conventional CDC PRA for the screening of anti-HLA class I alloantibodies in potential solid organ recipients.