• IMMUNE NETWORK
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• 제14권4호
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• pp.219-225
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• 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.

• IMMUNE NETWORK
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• 제23권2호
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• pp.17.1-17.16
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• 2023

Latent membrane protein 2A (LMP2A), a latent Ag commonly expressed in Epstein-Barr virus (EBV)-infected host cells, is a target for adoptive T cell therapy in EBV-associated malignancies. To define whether individual human leukocyte antigen (HLA) allotypes are used preferentially in EBV-specific T lymphocyte responses, LMP2A-specific CD8⁺ and CD4⁺ T cell responses in 50 healthy donors were analyzed by ELISPOT assay using artificial Ag-presenting cells expressing a single allotype. CD8⁺ T cell responses were significantly higher than CD4⁺ T cell responses. CD8⁺ T cell responses were ranked from highest to lowest in the order HLA-A, HLA-B, and HLA-C loci, and CD4⁺ T cell responses were ranked in the order HLA-DR, HLA-DP, and HLA-DQ loci. Among the 32 HLA class I and 56 HLA class II allotypes, 6 HLA-A, 7 HLA-B, 5 HLA-C, 10 HLA-DR, 2 HLA-DQ, and 2 HLA-DP allotypes showed T cell responses higher than 50 spot-forming cells (SFCs)/5×105 CD8⁺ or CD4⁺ T cells. Twenty-nine donors (58%) showed a high T cell response to at least one allotype of HLA class I or class II, and 4 donors (8%) had a high response to both HLA class I and class II allotypes. Interestingly, we observed an inverse correlation between the proportion of LMP2A-specific T cell responses and the frequency of HLA class I and II allotypes. These data demonstrate the allele dominance of LMP2A-specific T cell responses among HLA allotypes and their intra-individual dominance in response to only a few allotypes in an individual, which may provide useful information for genetic, pathogenic, and immunotherapeutic approaches to EBV-associated diseases.

• 대한의생명과학회지
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• 제4권1호
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• pp.11-25
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• 1998

HLA 유전자군은 인간의 유전자 중에서 가장 높은 유전적 다형성을 보이며, 분석방법에 따라 판정할 수 있는 대립유전자의 수에 많은 차이가 있다. 현재까지 혈청학적 방법을 이용하여 HLA항원형 구분을 하였으나, 최근 골수이식 등 여러 HLA활용분야에서 HLA유전자형 분석이 요구되고 있어, 많은 수의 HLA유전자형을 쉽고 정확하게 구분할 수 있는 HLA DNA typing 방법이 필요한 실정이다. 본 연구에서는 HLA-A, B, C, DRBI 유전자형 구분은 PCR-SSP 방법을, HLA-DQAl, DQBl, DPBl 유전자형 구분은 PCR-RFLP 방법을 사용한 HLA DNA typing 방법으로 한국인에서 HLA 유전자형을 구분하고자 하였다. 본 방법을 이용하여 HLA형이 규명된 B-임파아구 표준세포에서 DNA typing을 실시하였을 때, 11차 국제 조직적 합성학회에서 발표된 결과와 모두 일치하였다. 한국인에서 HLA-A, -B, -C 대 립 유전자는 17종, 23종, 16종이 확인되었으며, HLA-DQAl, -DQBl, -DPBl, -DRBl 대립유전자는 8종, 16종, 13종, 37종이 확인되었다. 한국인에서 빈도가 높은 HLA-class I 유전자는 HLA-A 유전자에서 $A^*$02가 27.0%였으며 HLA-B 유전자에서 는 $B^*$40이 17.6%를 나타내 었고 HLA-C 유전자에서는 Cw$^*$0101이 19.2%로 가장 높은 빈도를 나타내었다. 한국인에서 가장 빈도가 높은 HLA-class II 유전자는 DQAl 유전자에서 DQAl$^*$0301이 32.1%였고, DQBl 유전자에서는 DQBl$^*$0303이 12.9%를 나타내었으며, DPBl 유전자에서는 DPBl$^*$0501이 31.3%였고 DRBl 유전자에서는 DRBl$^*$1501이 9.2%를 나타내었다. 본 연구에서 실시한 HLA DNA typing 방법은 비교적 빠른 시간 내에 많은 종류의 HLA 대립 유전자형을 정확하게 구분할 수 있으므로 앞으로 tissue typing 실험실에서 유용하게 활용될 수 있을 것으로 생각된다. 또한 DNA typing방법을 이용하여 분석한 한국인의 HLA-class I, II 유전자군의 유전자형 빈도는 골수이식을 비롯한 각종 이식검사, 특수 질환 관련검사나 인류유전학연구 등 HLA 유전자의 임상적 활용을 위한 자료로 사용될 수 있을 것으로 기대된다.

• 대한수혈학회지
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• 제24권3호
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• pp.233-240
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• 2013

배경: 수혈의 기왕력이 항HLA항체의 형성과 연관이 있다는 사실은 잘 알려져 있다. 하지만, 특발성 혈소판 감소성 자반증의 주 표적은 혈소판의 당단백이며, 당단백 항체와 항 HLA class I 항체의 형성기전은 완전히 다른 것이다. 이번 연구의 목적은 특발성 혈소판 감소성 자반증 환아의 항HLA class I 항체의 임상적 의미를 찾아 보고자 함에 있다. 방법: 1990년부터 2010년까지 경상대학교병원을 방문한 48명의 대조군과 48명의 특발성 혈소판감소성자반증 환아들을 대상으로 하였다. 각각의 환자들의 혈청을 Modified Antigen Capture Enzyme-linked immunosorbent assay (MACE) kit를 이용하여 항 당단백 항체와 항 HLA class I 항체를 검사하였다. 결과: 두 그룹간의 항 HLA class I항체의 양성률은 의미 있는 차이를 보였으며[36/39 (92.3%) vs 29/46 (63.0%)][혈소판감소성자반증 환자군 vs 정상대조군] (P=0.002), 항HLA class I 평균항체 S/C ratio 또한 의미 있는 차이를 보였다(3.55 vs 1.51) [혈소판감소성자반증 환자군 vs 정상대조군] (P=0.0000). 하지만 혈소판 감소성 자반증 환자군에서 수혈을 받은 환자들과 수혈을 받지 않은 환자들 간의 항HLA항체 양성률은 의미 있는 차이를 보이지 않았으며[12/12 (100%) vs 24/27 (88%)][수혈을 받은 환자군 vs 비수혈 환자군](P=0.22), 평균 항체 S/C ratio 또한 통계적으로 의미 있는 차이를 보이지 않았다(4.30 vs 3.25)[수혈을 받은 환자군 vs 비수혈 환자군](P=0.22). 수혈 이후에 추적검사를 위해 채혈을 했던 환아들의 샘플을 이용한 항체 양성률 및 항체가 검사에서도 의미 있는 차이는 없는 것으로 나타났다. 결론: 항 HLA class I 항체는 혈소판 감소성 자반증과 연관이 있을 수 있으나, 수혈은 혈소판 감소성 자반증 환아의 임상적 경과에 영향을 미치지 못한다.

• 대한의생명과학회지
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• 제20권4호
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• pp.250-255
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• 2014

Mycobacterium tuberculosis (MTB) resides and replicates inside macrophages. In our previous report, we reported that CD8+ T cell-mediated immune responses specific for the peptide derived from MTB RNA polymerase beta-subunit ($RpoB_{127-135}$) could be induced in TB patients expressing HLA-$A^*0201$ subtype. In order to examine whether $RpoB_{127-135}$ specific CD8+ T cells can recognize MTB infected macrophages in vitro, CD8+ T cell lines specific for $RpoB_{127-135}$ peptide were generated from peripheral blood mononuclear cells (PBMCs) of healthy HLA-$A^*0201$ subjects by in vitro immunization technique. In this study, we observed $RpoB_{127-135}$ specific CD8+ T cells could recognize and destroy macrophages infected with MTB for 2 to 4 days. $RpoB_{127-135}$ specific CD8+ T cell immune response was inducible from PBMC of healthy subjects expressing HLA-$A^*0206$ subtype, one of HLA-A2 supertype members. Next, we investigated the HLA-I processing mechanism of $RpoB_{127-135}$ peptide in MTB infected macrophages. As a result, the presentation of the MTB derived epitope peptide, $RpoB_{127-135}$, to CD8+ T cells was not inhibited by the treatment with brefeldin-A (ER-Golgi transport inhibitor) or lactacystin (proteasome inhibitor), which blocks the classical HLA-I processing pathway. However, $RpoB_{127-135}$ specific CD8+ T cell activity was blocked either by the blocking agent for the endocytosis (cytochalasin D) or by the blocking antibody (W6/32) for HLA-I molecules. Therefore, the $RpoB_{127-135}$ peptide may be processed by accessing the alternate HLA-I processing pathway. Understanding the processing and presentation mechanisms of the MTB derived proteins will help to improve the efficacy of vaccines and the efficiency of therapeutic agents for TB.

• BMB Reports
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• 제29권4호
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• pp.286-293
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• 1996

Association of antigenic peptide with class I MHC is believed to be crucial for maintaining stable conformation of class I molecules. T2 cells that are defective in TAP gene function mainly express class I molecules with an unstable conformation due to little or no association with antigenic peptides, whereas T1 cells that are normal in TAP gene function mainly express the stable form of class I molecules. In this work, attempts were made to determine the molecular stability of stable and unstable class I molecules. Dissociation of HLA-A2 molecules on T1 and T2 cells was monitored by flow cytometry using anti-HLA-A2 antibody after the cells were treated with brefeldin A to shut down the transport of newly-assembled HLA-A2. Estimated dissociation rate constants for the stable and unstable forms of HLA-A2 were 0.076 $h^{-1}$ and 0.66 $h^{-1}$, respectively. It appeared that both T1 and T2 cells express stable and unstable class I complex, but with different ratios of the two forms. Furthermore, $interferon-{\gamma}$ treatment of T1 cells appeared to induce the expression of both the stable and unstable class I molecules. These results demonstrate that class I MHC molecules can be divided into two groups in terms of structural stability and that they exist on the cell surface in both forms in a certain ratio.

• 대한의생명과학회지
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• 제2권2호
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• pp.223-229
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• 1996

HLA-class I 항원의 HLA-B 유전자좌에 존재하는 HLA-B27 유전자는 임상적으로 강직성 척수염과 강한 관련성이 있음이 보고되고 있으며, 현재 HLA 유전자중 질병과의 관련성을 보기 위한 검사로 임상에서 가장 널리 사용되고 있다. 대부분의 검사실에서는 현재까지 혈청학적 검사방법을 이용하여 HLA-B27 검사를 실시하고 있는데, 이 방법은 시약이 고가이고, 검체의 안정성과 보관이 어려우며, 분석시간이 오래 걸리는 등 불편한 점이 있고, 또한 현재에도 계속 새로운 HLA-B27 대립유전자가 발견되고 있으므로 위음성의 가능성도 배제할 수 없어, 보다 정확한 검사방법이 요구되고 있다. 최근 HLA-B27 대림유전자의 염기배열이 대부분 밝혀져 혈청학적 방법 대신 DNA를 이용한 typing방법이 보고되고 있다. 저자들은 HLA-B2l 대립 유전자에 공통으로 존재하는 염기배열 부분을 선택하여 group specific PCR(Polymerase Chain Reaction)을 실시하고 그 유용성을 검토하였다. 혈청학적 방법으로 HLA B-27 형이 확인된 검체 56 개와 4 개의 표준세포주 (HOM-2, JESTHOM, WT24, BTB)를 이용하여 혈청학적 방법과 DNA typing을 비교한 결과, 두 방법사이에 완벽한 일치를 나타내었다. 따라서 group specific PCR을 이용한 HLA-B27 DNA typing은 검체 및 시약의 안정성이 높고, 경제적이며 신속한 검사가 가능하므로 임상에서 활용성이 매우 클 것으로 사료된다.

• 대한임상검사과학회지
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• 제40권2호
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• pp.94-97
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• 2008

The human leukocyte antigen (HLA) is the name of the major histocompatibility complex (MCH) in humans. The superlocus contains a large number of genes related to immune system function in humans. This group of genes resides on chromosome 6. and encode cell surface antigen-presenting proteins and many other genes. HLA class I antigen (A, B & C) present peptides from inside the cell. These peptides are produced from digested proteins that are broken down in the lysozymes. 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 sutdy, the HLA-A 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-A gene were used (A*0201, A*33, A*2401). The results of PCR-SSP, the HLA-A*0201 primer was detected eleven (55%), the HLA-A*33 were detected seven (35%) and the HLA-A*2401 were detected seven (35%). This study shows that the PCR-SSP technique is relatively simple, fast and a practical tool for the determination of the HLA-A genotypes.

• 생물정신의학
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• 제1권1호
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• pp.79-87
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• 1994

For the purpose of evaluating the human leukocyte antigen(HLA) disease-association with bipolar disorder, HLA class I and class II allelic frequencies were assessed in 37 bipolar patients and were compared to the data from normal population. HLA class 1 typing was performed with microlymphocytotoxicity method while class II(DRB1) genotyping with reverse dot blot hybridization and sandwich method. Statistical analysis consisted of relative risk, Haldane's modified relative risk, Fisher's exact test and Bonferoni's corrected P. The results were as follows : 1) Bipolar patients showed increased allelic frequency of HLA A3 which has statistical significance. 2) Allelic frequencies of HLA B7, B14 and B54 were higher, while those of B51 and B55 were lower in bipolar patients, but they were not statistically significant. 3) Both of increased frequencies of DR2 in bipolar patients and DR15 in normal controls had statistical significance. The results of the present study suggested that some of HLA allelic types might be associated with bipolar disorder. To clarify the genetic influence of HLA to bipolar disorder, we should do consecutive study of bipolar disorder with new information about HLA system including alleles.

• 사상체질의학회지
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• 제14권1호
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• pp.90-99
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• 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.