• Title/Summary/Keyword: HLA

Search Result 372, Processing Time 0.028 seconds

Development of HLA-A, -B and -DR Typing Method Using Next-Generation Sequencing (차세대염기서열분석법을 이용한 HLA-A, -B 그리고 -DR 형별 분석법 개발)

  • Seo, Dong Hee;Lee, Jeong Min;Park, Mi Ok;Lee, Hyun Ju;Moon, Seo Yoon;Oh, Mijin;Kim, So Young;Lee, Sang-Heon;Hyeong, Ki-Eun;Hu, Hae-Jin;Cho, Dae-Yeon
    • The Korean Journal of Blood Transfusion
    • /
    • v.29 no.3
    • /
    • pp.310-319
    • /
    • 2018
  • Background: Research on next-generation sequencing (NGS)-based HLA typing is active. To resolve the phase ambiguity and long turn-around-time of conventional high resolution HLA typing, this study developed a NGS-based high resolution HLA typing method that can handle large-scale samples within an efficient testing time. Methods: For HLA NGS, the condition of nucleic acid extraction, library construction, PCR mechanism, and HLA typing with bioinformatics were developed. To confirm the accuracy of the NGS-based HLA typing method, the results of 192 samples HLA typed by SSOP and 28 samples typed by SBT compared to NGS-based HLA-A, -B and -DR typing. Results: DNA library construction through two-step PCR, NGS sequencing with MiSeq (Illumina Inc., San Diego, USA), and the data analysis platform were established. NGS-based HLA typing results were compatible with known HLA types from 220 blood samples. Conclusion: The NSG-based HLA typing method could handle large volume samples with high-throughput. Therefore, it would be useful for HLA typing of bone marrow donation volunteers.

Genotyping of HLA-B by Polymerase Chain Reaction-Sequence Specific Primer (Polymerase Chain Reaction-Sequence Specific Primer를 이용한 HLA-B 유전자의 DNA 다형성 조사)

  • Jang, Soon-Mo
    • Korean Journal of Clinical Laboratory Science
    • /
    • v.39 no.3
    • /
    • pp.147-150
    • /
    • 2007
  • Most expressed HLA (human leukocyte antigen) 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-B genotypes were determined in twenty students unrelated koreans using the PCR-SSP (polymerase chain reaction-sequence specific primer) technique. Several specific primer pairs in assigning the HLA-B gene were used ($B^{\ast}4001/4007$, $B^{\ast}4901/5001/4501$, $B^{\ast}3701$, $B^{\ast}5801$). The results of PCR-SSP, the HLA-B3701 primer was detected one (5%), the $HLA-B^{\ast}5801$ were detected four (20%), the $HLA-B^{\ast}4001/4007$ were detected nineteen (95%) and the $HLA-B^{\ast}4901/5001/4501$ were detected twenty. This study shows that the PCR-SSP technique is relatively simple, fast and a practical tool for the determination of the HLA-B genotypes. Moreover, these results genotype frequency of the HLA-B gene could be useful for database study before being applied to individual identification and transplantation immunity.

  • PDF

Knowledge based Genetic Algorithm for the Prediction of Peptides binding to HLA alleles common in Koreans (지식기반 유전자알고리즘을 이용한 한국인 빈발 HLA 대립유전자에 대한 결합 펩타이드 예측)

  • Cho, Yeon-Jin;Oh, Heung-Bum;Kim, Hyeon-Cheol
    • Journal of Internet Computing and Services
    • /
    • v.13 no.4
    • /
    • pp.45-52
    • /
    • 2012
  • T cells induce immune responses and thereby eliminate infected micro-organisms when peptides from the microbial proteins are bound to HLAs in the host cell surfaces, It is known that the more stable the binding of peptide to HLA is, the stronger the T cell response gets to remove more effectively the source of infection. Accordingly, if peptides (HLA binder) which can be bound stably to a certain HLA are found, those peptieds are utilized to the development of peptide vaccine to prevent infectious diseases or even to cancer. However, HLA is highly polymorphic so that HLA has a large number of alleles with some frequencies even in one population. Therefore, it is very inefficient to find the peptides stably bound to a number of HLAs by testing random possible peptides for all the various alleles frequent in the population. In order to solve this problem, computational methods have recently been developed to predict peptides which are stably bound to a certain HLA. These methods could markedly decrease the number of candidate peptides to be examined by biological experiments. Accordingly, this paper not only introduces a method of machine learning to predict peptides binding to an HLA, but also suggests a new prediction model so called 'knowledge-based genetic algorithm' that has never been tried for HLA binding peptide prediction. Although based on genetic algorithm (GA). it showed more enhanced performance than GA by incorporating expert knowledge in the process of the algorithm. Furthermore, it could extract rules predicting the binding peptide of the HLA alleles common in Koreans.

A Study of the HLA Compliance Testing System and Procedures for ROK (한국형 HLA(High Level Architecture) 적합성 인증시험 체계 및 절차 연구)

  • Jo, Hee-Jin;Lee, Chong-Ho;Kim, Jae-Man
    • Journal of the Korea Society for Simulation
    • /
    • v.27 no.1
    • /
    • pp.15-24
    • /
    • 2018
  • High Level Architecture(HLA) for modeling and simulation is an international technical standard to achieve simulation interoperability and reusability. Building federation through the interworking of federates is being used in various fields and is being used in KR(Key Resolve), UFG(Ulchi-Freedom Guardian) as a representative in domestic military. Currently, United States Department of Defense emphasizes the HLA compliance test, which confirms the compliance of Federate by using HLA. However, due to the limitations of testing tools, an HLA compliance test of DoD is not able to perform some of the functional tests on HLA/RTI services such as federation storage/recovery services and also exclude inspection of exchange data between federates. To solve these problems, this paper proposes an HLA compliance testing system of ROK suitable for domestic situations. This will enable effective HLA compliance testing and improve interoperability between federates.

A study on the correlation between Sasang Constitution and HLA type (사상체질(四象體質)과 HLA type과의 상관성(相關性)에 관(關)한 연구(硏究))

  • Koh, Byung-Hee;Song, Il-Byung;Ha, Man-Su
    • Journal of Sasang Constitutional Medicine
    • /
    • v.14 no.1
    • /
    • pp.90-99
    • /
    • 2002
  • Exact constitutional classification is especially important in Sasang Constitutional Medicine. Therefore, most studies on the Sasang Constitutional medicine have been focused on how accurately to classify Sasang Constitution. Recently, there have been reported on the studies about relationship between genetic polymorphism and Sansang Constitution. The purpose of this study is to investigate whether there is any correlation between Sasang Constitution and HLA type. Polymorphism of HLA genes have been known to be important role in transplantation, autoimmune disease, immune response against infection. This study was concentrated on a relationship of HLA-A, -C, -DR, and -DQ types with Sasang Constitution. Results are as following. 1. In the case of HLA-A, there was no significant difference. Only $HLA-A^*31$ has a tendency which Soyangin is less frequent than Soeumin and Taeumin. 2. In the case of HLA-C, there was significant difference between Sasang Constitution and HLA type. In persons having $HLA-C^*04$ allele, Taeumin is more frequent than Soyangin and Soeumin. On the other hand, Soyangin is more frequent than Taeumin and Soeumin in $HLA-C^*07$ allele. $HLA-C^*14$ has a tendency which Soeumin is more frequent than Soyangin and Taeumin. 3. In. the case of HLA-DR type and HLA-DQ type, there was no significant difference among Sansang Constitution. On the study of correlation between Sasang Constitution and HLA type, there was little statistical significance and tendency. This study was the first try about presentation of frequency on the HLA alleles corresponding with Sasang Constitution. I think that more study on the relationship between HLA gene and Sasang Constitution is necessary by method of large case research and various statistical analysis.

  • PDF

A Study of Genetic Polymonhisms of HLA-class I and II Genes Using Polymerase Chain Reaction (중합효소연쇄반응을 이용한 HLA-class I, II 유전자군의 유전적 다형성에 관한 연구)

  • Kyung-Ok Lee
    • Biomedical Science Letters
    • /
    • v.4 no.1
    • /
    • pp.11-25
    • /
    • 1998
  • The HLA genes located in the short arm of chromosome 6 specify heterodimeric glycoproteins involved in the regulation of the immune response. Recently, in the elucidation of HLA polymorphism, serological and cellular typing methods have been replaced by DNA typing using polymerase chain reaction (PCR). The purpose of this study was to establish the HLA DNA typing methods and determine gene frequencies of HLA molecules in Koreans. PCR-SSP (sequence specific primers) and PCR-RFLP (restriction fragment length polymorphism) techniques were used for the analysis of HLA-A, -B, -C, DRBl genes and HLA-DQAl, DQBl, DPBl genes, respectively. The results of B-lymphoblastoid cells used for control experiment were consistent with the previous data identified in the 11th International Histocompatibility Workshop. Seventeen, 23, 16, 8, 16, 13 and 37 types of HLA-A, B, C, DQAl, DQBl, DPBl and DRBl alleles were found, respectively, in a total of unrelated 120 Korean individuals. The most frequent HLA alleles were $A^*$02 (27.0%), B$^*$40 (17.6%), Cw$^*$01 (19.2%), DQAl$^*$0301 (32.1%), DQBl$^*$0303 (12.9%), DPBl$^*$0501 (31.3%) and DRBl$^*$1501 (9.2%) among Koreans. This study shows that DNA typing method using PCR technique is a relatively simple, fast and practical tool for the determination of the HLA-class I and II genes. Moreover, the data of HLA gene frequencies could be useful for the Korean database before clinical applications, including organ and unrelated bone marrow transplantation, anthropological study, disease association and individual identification.

  • PDF

Thyrocyte HLA-DR Expression in Variety of Thyroid Diseases (갑상선 질환들에서 여포세포의 HLA-DR발현)

  • Chang Eun-Sook;Sohn Soo-Sang;Cho Seung-Che;Kim In-Ho;Kim Sang-Pyo;Kang Joong-Shin
    • Korean Journal of Head & Neck Oncology
    • /
    • v.13 no.1
    • /
    • pp.16-23
    • /
    • 1997
  • Although normal thyroid epithelial cells do not constitutively express HLA-DR antigen, their expression in wide spread within thyroid glands obtained from the human with autoimmune thyroid disease and with many neoplastic thyroids. We have, therefore, studied immunohistochemically with regard to the expression of HLA-DR antigen of thyroidectomy specimens from 50 patients of various thyroid diseases with use of paraffin-embedded tissue. One or two sections from each case were stained with commercially available mouse monoclonal antibody for class II HLA-DR antigen(HLA-DR/Alpha, DAKO) and examined by semiquantitative counting system for thyrocytes, neoplastic thyrocytes and other cells expressing HLA-DR antigen. All patients with lymphocytic thyroiditis(2/2) and diffuse hyperplasia(Graves' disease)(5/5), most patients with Hashimoto's disease(9/ll) expressed HLA-DR antigens in thyrocyte with abundant HLA-DR expressing lymphocytic infiltrates with lymph follicle formation in its vicinity or adjacent to the lesion. Most patients with papillary carcinoma(9/1l) had HLA-DR antigen detected in malignant thyrocytes ; while follicular carcinoma(0/3) and follicular adenoma(0/5) did not have detactable HLA-DR immunoreactivity. Adenomatous goiter(3/7) had HLA-DR antigen detected focally in lesser than half cases. Conversely, in four papillary carcinomas and three adenomatous goiters, HLA-DR expression of thyrocytes was found in the absence of HLA-DR expressing lymphoid infiltrates. In such cases therefore other factors more than thyroid autoimmunity must be causative for HLA-DR immunoreactivity. The results of this study indicate as follows. 1) The expression of HLA-DR on thyrocytes involved in autoimmune reactions appeared to be secondary to cytokine release from associated lymphocytic infiltrates. 2) Thyrocytes in thyroid lesions with equal degrees of lymphocytic infiltration without HLA­DR expression exhibited no HLA-DR immunoreactivity. 3) In neoplastic thyrocytes, most papillary carcinoma(9/11) exhibited detactable HLA-DR expression, while follicular carcinoma/adenoma(0/3/0/5) exhibited no detactable HLA-DR immunoreactivity which suggest the existence of divergent mechanisms inducing and modulating HLA-DR expression of different types of neoplastic thyrocytes.

  • PDF

Genotyping of HLA-DRB1 by Polymerase Chain Reaction-Sequence Specific Primer (Polymerase Chain Reaction-Sequence Specific Primer를 이용한 HLA-DRB1 유전자의 DNA 다형성)

  • Jang, Soon-Mo
    • Korean Journal of Clinical Laboratory Science
    • /
    • v.37 no.3
    • /
    • pp.139-142
    • /
    • 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.

  • PDF

HLA-B27 DNA Typing using Group Specific Polymerase Chain Reaction (중합효소연쇄반응을 이용한 HLA-B27 유전자분석)

  • Kyung Ok Lee;Sung Hoi Hong;Moom Ju Oh;Kyung In Kim;Min Jung Kim
    • Biomedical Science Letters
    • /
    • v.2 no.2
    • /
    • pp.223-229
    • /
    • 1996
  • HLA-B27 gene, one of the HLA-class I molecule, is strongly associated with ankylosing spondylitis. It has been most frequently used as a disease-correlated HLA gene by clinicians. In most laboratories, conventional HLA-B27 typing is still performed by cell cytotoxicity tests or fluorescence serology with specific antibodies. In this study, DNA typing method for HLA-B27 was developed by using group specific Polymerase Chain Reaction (PCR). Four HLA-B27 cell lines (HOM-2, JESTHOM, WT24 and BTB) and fifty six B27 Korean individuals defined by serology were used. The results of control cell and B-27 positive individual samples were correlated well with the data which was performed by serological method. All of B27 positive PCR products gave positive signals on Southern blot hybridization with B27 specific probe. This study shows that the HLA-B27 DNA typing is a relatively simple, fast and practical tool for the determination of the HLA-B27 gene in routine clinical laboratory work.

  • PDF

Relationship between Poor Immunogenicity of HLA-A2-Restricted Peptide Epitopes and Paucity of Naïve $CD8^+$ T-Cell Precursors in HLA-A2-Transgenic Mice

  • Choi, Yoon Seok;Lee, Dong Ho;Shin, Eui-Cheol
    • IMMUNE NETWORK
    • /
    • v.14 no.4
    • /
    • pp.219-225
    • /
    • 2014
  • We examined the immunogenicity of H-2 class I-restricted and HLA-A2-restricted epitopes through peptide immunization of HLA-A2-transgenic mice that also express mouse H-2 class I molecules. All four of the tested epitopes restricted by H-2 class I robustly elicited T-cell responses, but four of seven epitopes restricted by HLA-A2 did not induce T-cell responses, showing that HLA-A2-restricted peptide epitopes tend to be poorly immunogenic in HLA-A2-transgenic mice. This finding was confirmed in HLA-A2-transgenic mice infected with a recombinant vaccinia virus expressing hepatitis C virus proteins. We examined the precursor frequency of epitope-specific naïve $CD8^+$ T cells in HLA-A2-transgenic and conventional C57BL/6 mice and found that the poor immunogenicity of HLA-A2-restricted peptide epitopes is related to the paucity of naïve $CD8^+$ T-cell precursors in HLA-A2-transgenic mice. These results provide direction for the improvement of mouse models to study epitope repertoires and the immunodominance of human T-cell responses.