• Clinical and Experimental Pediatrics
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• 제46권5호
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• pp.467-473
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• 2003

목 적 : EBV는 전 세계적으로 다양한 질환을 일으키고, EBV는 지역과 EBV에 노출된 사람의 면역 또는 병적 상태에 따라 감염형이 달라 EBV 감염형에 관한 연구는 EBV의 역학연구와 EBV 관련 질환의 병인연구에 제일 기본적이다. EBV의 잠복감염 유전인자들에 의한 세포독성 T 림프구 면역상태는 EBV에 의한 다양한 질환의 병인적 역할이 있으며 HLA에 의하여 유발되어지는 것으로 알려져 있다. 과거에 시행한 많은 연구에서 EBV 감염과 HLA형과의 연관성은 객관적으로 입증되지 않았으나, HLA II형에 속한 항원이 EBV의 인체 내 침투에 관여한다는 사실이 최근에 보고됨으로써 이에 대한 관심이 재조명되고 있다. 이에 저자들은 EBV 감염에 의한 국내 급성 전염성 단핵구증 환아에서 주된 EBV 감염형의 분포를 확인하고, 급성 전염성 단핵구증의 발생이 HLA형과 연관성이 있는 지를 분자생물학적 분류법으로 알아보고자 이 연구를 실시하였다. 방 법 : 연구 대상은 6세에서 13세 사이의 급성 전염성 단핵구증으로 확진된 24명의 환아 이었다. 감염형의 비교 대조군은 건강한 정상 소아 중 면역혈청학적 검사로 EBV 노출이 확인된 동일한 연령의 58명이었고, HLA형 분류의 비교 대조군은 연구대상 환아와 동일한 연령의 정상 소아 200명을 대상으로 실시하였다. EBV에 의한 급성 전염성단핵구증 환아에서 중합 효소 연쇄반응법으로 EBV 감염형을 분류하여 주된 감염형을 확인하고, ARMs PCR법으로 HLA-A, B, C 항원형을, PCR-SSOP법으로 HLA-DRB 항원형을 분류하여 다음과 같은 결과를 얻었다. 결 과 : 1) EBV 감염형 결과 : 연구 대상 환아 24명에서 EBV A형이 20명(83.3%)이었고, B형은 4명(16.7%)이었다. 비교 대조군의 경우에는 58명 중 14명(24.1%)이 EBV가 검출되고 이들 중 A형은 11명(78.6%)이었고, B형이 3명(21.%)이었다. 2) HLA 형 분류 결과 : HLA I형 결과는 HLA-A24형에서 비교 대조군 200명 소아 중 69명이 양성이었고, 연구 대상 환아 24명 중 14명이 양성을 보여 RR치가 3.5724, chi-square치가 5.26, P<0.05으로 통계적 유의성을 보였다. 그러나 HLA-B, C항원형에서는 통계적으로 유의한 결과가 없었다. HLA II형 결과는 HLA-DRB1*07형에서 비교 대조군 200명 소아 중 18명이 양성이었고, 연구 대상 환아 24명 중 8명이 양성을 보여 RR치가 5.6173, chi-square치가 9.73, P<0.01로 통계적 유의성을 보였다. 결 론 : 위의 연구 결과를 통하여 국내 급성 전염성 단핵구증 환아의 EBV 감염형은 정상 비교 대조군의 소아에서 주된 감염형인 A형과 동일한 감염형임을 확인하였다. 그리고 HLA형 연구에서 HLA-A24형과 HLA-DRB1*07형이 정상 대조군과 비교하여 통계적 유의성을 보여 EBV에 의한 급성 전염성단핵구증 발생이 HLA형과 연관성이 있음을 추정하였다.

• IMMUNE NETWORK
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• 제2권4호
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• pp.248-255
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• 2002

Background: TAP1 and TAP2 are two ABC transporter genes located within the class II region of the human MHC. Their protein products form a heterodimer whose function is to transport peptides from the cytoplasm into the endoplasmic reticulum. This study was performed to examine the polymorphism of TAP genes and the distribution of HLA-TAP haplotypes in the Korean population through family analysis. Methods: The subjects used in this study were 50 healthy Korean families consisting of 233 individuals. TAP1 (codons 333 and 637) and TAP2 (codons 379, 565, 577, 651, 665, and 687) typings were carried out by the PCR-restriction fragment length polymorphism (RFLP) method. HLA-DRB1 and DQB1 genotyping results from a previous study were used for HLA-TAP haplotype analysis. Results: The number (gene frequency) of TAP1 and TAP2 alleles detected were 3 for TAP1 (A 81.5%, B 17.0%, and C 1.5%) and 8 for TAP2 (A1 32.0%, A2 12.5%, B 34.0%, Bky2 6.5%, C 7.0%, D 3.0%, E 4.5%, and G 0.5%). Eleven TAP1-TAP2 haplotypes were observed with $frequency{\geq}1%$, among which 4 haplotypes (A-B, B-A1, A-Bky2, and C-E) showed weak but significant positive linkage disequilibrium (P<0.05). When DRB1-DQB1 haplotypes were extended to TAP1 and TAP2 loci, much diversification of haplotypes was observed: 19 different DRB1-DQB1 haplotypes formed 58 different haplotypes extended to TAP1 and TAP2 loci. These results add more evidence to the view that recombination hotspot is present within and around TAP gene region. Conclusion: The allele frequencies of TAP1 and TAP2 genes and the distribution of TAP1-TAP2 and HLA-TAP haplotypes were studied in Koreans based on a family study.

• 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.

• BMB Reports
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• 제30권1호
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• pp.26-32
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• 1997

Of all HLA class I molecules, HLA-C gene products are most poorly understood because they express at a low level on the cell surface compared to HLA-A and -B. In order to identify serologically detectable and undetectable HLA-C antigens, we have established a DNA-based tissue typing method for the HLA-C locus by PCR-SSP (polymerase chain reaction-sequence specific primers). Genomic DNA prepared from Iymphoblastoid 21 B-cell lines and 120 Korean individuals by proteinase K digestion and pheno/chloroform extractions have been typed by PCR-SSP (23 primer mixes were used). The PCR-SSP results of control cell lines were discrepant from serology in 1 case among 21 cases: Cw6 which was negative by serology but positive by PCR-SSP (cell line: MANIKA). Twenty four HLA-Cw "blank" antigens among fifty Korean individuals were completely determined by PCR-SSP DNA typing. HLA-Cw*0101 (15.3%), Cw*1401 (12.3%) and Cw*0701 (11.7%) alleles were frequently found in 120 Korean individual samples. In conclusion. the high level of discrimination for HLA-C alleles may prove useful and informative in the study of transplant survival, and identify the importance of allelic differences, not readily detectable by serology, on host and donor compatibility.

• BMB Reports
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• 제34권2호
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• pp.176-181
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• 2001

Using a retrovirus, foreign genes can be introduced into mammalian cells. The purpose of this study is to produce a retrovirus that can make the infected cells express two genes; the human multidrug resistance gene (MDR1) and the HLA-B7 gene, which is one of the major human histocompatibility complex (MHC) class I genes. For the expression of these genes, the internal ribosome entry site (IRES) was used, which was derived from the encephalomyocarditis (EMC) virus. In order to produce retroviruses, a retroviral vector was transfected into a packaging cell line and the transfected cells were treated with vincristine, which is an anti-cancer drug and a substrate for the MDRI gene product. This study revealed that two genes were incorporated into chromosomes of selected cells and expressed in the same cells. The production of the retrovirus was confirmed by the reverse transcription (RT)-PCR of the viral RNA. The retrovirus that was produced infected mouse fibroblast cells as well as the human U937. This study showed that packaging cells produced the retroviruses, which can infect the target cells. Once the conditions for the high infectivity of retrovirus into human cells are optimized, thus virus will be used to infect hematopoietic stem cells to co-express MDRl and HLA-B7 genes, and develop the lymphocytes that can be used for the immnogene therapy.

• 대한미생물학회지
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• 제21권2호
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• pp.233-241
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• 1986

Patients of chronic hepatic diseases(n=107) including chronic hepatitis caused by hepatitis B virus infections(n=31), liver cirrhosis(n=53), and hepatocellular carcinoma(n=23) were examined to ascertain genetic relationship between chronic hepatic diseases and histocompatibility antigen. Peripheral blood lymphocytes were separated from whole blood by the method of Ficoll/Hypaque gradient. Total 54 histocompatibility antigens(class I antigens: 41, class II antigens: 13) were analysed by performing of complement dependent microlymphocytotoxicity method using Terasaki's and Catholic Medical College tissue typing plates. HLA antigen frequencies were compared with those of 661 normal controls. The following results were obtained: 1. HLA antigen frequencies of HLA-Bw46, -Bw76, -Cw1, -Cw6, and HLA-DR8 in chronic hepatitis patients were shown to be higher than those in controls(P<0.01). 2. HLA antigen frequencies of HLA-Bw46, -Cw7(P<0.01), and HLA-B37, -Bw58, -Cw1, -MT1(P < 0.05) in liver cirrhosis patients were shown relatively higher frequencies than those in controls. 3. In patients with hepatocellular carcinoma, antigens of HLA-A1, -A26, -Cw3, -Cw7 and HLA-DR6 were dominantly detected. 4. There were negative associations with HLA-Cw4, and -DR4 in patients of chronic hepatic diseases(P < 0.05).

• Asian Pacific Journal of Cancer Prevention
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• 제14권7호
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• pp.4167-4175
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• 2013

A najor current challenge and constraint in cervical cancer research is the development of vaccines against human papilloma virus (HPV) epitopes. Although many studies are done on epitope identification on HPVs, no computational work has been carried out for high risk forms which are considered to cause cervical cancer. Of all the high risk HPVs, HPV 16, HPV 18 and HPV 45 are responsible for 94% of cervical cancers in women worldwide. In this work, we computationally predicted the promiscuous epitopes among the E6 proteins of high risk HPVs. We identified the conserved residues, HLA class I, HLA class II and B-cell epitopes along with their corresponding secondary structure conformations. We used extremely precise bioinformatics tools like ClustalW2, MAPPP, NetMHC, Epi,Jen, EpiTop 1.0, ABCpred, BCpred and PSIPred for achieving this task. Our study identified specific regions 'FAFR(K)DL' followed by 'KLPD(Q)LCTEL' fragments which proved to be promiscuous epitopes present in both human leukocyte antigen (HLA) class I, class II molecules and B cells as well. These fragments also follow every suitable character to be considered as promiscuous epitopes with supporting evidences of previously reported experimental results. Thus, we conclude that these regions should be considered as the important for design of specific therapeutic vaccines for cervical cancer.

• Journal of Periodontal and Implant Science
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• 제30권4호
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• pp.895-913
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• 2000

It is becoming increasingly clear that human gingival fibroblasts(HGF) may play a role in regulating immune responsiveness in inflammatory periodontal lesions. Stimulation of HGF with locally-secreted T cell cytokine $IFN_{\gamma}$ induces human leukocyte antigen class II(HLA II) expression on HGF, which is one of the characteristic feature of professional antigen presenting cells(pAPC). However, $IFN_{\gamma}$-treated HGF and other nonprofessional antigen presenting cells(npAPC) are known to be ineffective or less effective antigen presenter to resting T cells. This study, therefore, was undertaken in an effort to elucidate the differences in expression of cell surface molecules between npAPC in periodontal tissues, such as HGF and periodontal ligament fibroblasts(PDLF), and pAPC such as monocytes/macrophages. Using flow cytometry, the levels of cell surface expression of HLA-D, ICAM-1, LFA-3, and B7-1, which are involved in antigen presentation, were determined in HGF, PDLF and human myelomonocytic cell line THP-1. $IFN_{\gamma}$ clearly induced HLA-D expression on both of fibroblasts and monocytes dose dependently. However, expression level on monocytes were 4 to 5 times higher than that on fibroblasts, and induction rate was faster in monocytes than in fibroblasts. The levels of ICAM-1 expression on fibroblasts and monocytes were enhanced by $IFN_{\gamma}$ in a dose dependent manner. On the other hand, the expression of LFA-3 molecule, which could be detected in fibroblasts and monocytes without cytokine stimulation, was no more enhanced by addition of $IFN_{\gamma}$. B7-1, important costimulatory molecule in T cell activation and proliferation, was not detected on both of fibroblasts and monocytes even when stimulated with $IFN_{\gamma}$, except on monocytes fully differentiated by pretreatment of PMA and treated by $IFN_{\gamma}$. These results suggest that delayed expression of HLA-D and absence of B7-1 on $IFN_{\gamma}$ - treated fibroblasts may at least in part be involved in the ineffectiveness of fibroblasts as primary APC. And it is postulated that although periodontal fibroblasts may not serve as primary APC in normal periodontium, sustained expression of HLA II on ubiquitous fibroblasts in inflammatory lesions may perpetuate immune responses and produce chronic inflammation and tissue injury.

• IMMUNE NETWORK
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• 제3권2호
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• pp.96-102
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• 2003

Background: Granulocyte-macrophage colony-stimulating factor (GM-CSF) gene-transduced tumor cell vaccines induce very potent systemic anti-tumor immunity in preclinical and clinical models. Our previous phase I clinical trial in patients with metastatic renal cell carcinoma (RCC) has demonstrated both immune cell infiltration at vaccine sites and T cell-mediated delayed-type hypersensitivity (DTH) response to whole tumor cell vaccines. Methods: To investigate the immune responses to autologous genetically- modified tumor cell vaccines, tumor-specific $CD8^+$ T cell lines were generated from peripheral blood lymphocytes (PBL) of a RCC patient 1.24 by repeated in vitro stimulation with either B7.1-transduced autologous RCC tumor cells or B7.1-transduced autologous tumor cells treated with interferon gamma ($IFN{\gamma}$), and cloned by limiting dilution. Results: Among several RCC-specific cytotoxic T lymphocytes (CTLs), a $CD4^+/CD8^+$ double positive T cell clone (17/A2) appeared to recognize $IFN{\gamma}$-treated autologous RCC restricted by HLA-B39. The 17/A2 also recognized other HLA-B39 positive RCC tumor cells after $IFN{\gamma}$ treatment. Conclusion: These results demonstrate that autologous RCC vaccination successfully generates the tumor-specific CTL 17/A2, and suggest that the presentation and recognition of the tumor antigen by the 17/A2 might be upregulated by $IFN{\gamma}$.

• 대한치매학회지
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• 제17권4호
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• pp.131-136
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• 2018

Alzheimer's disease (AD) related genes have been elucidated by advanced genetic techniques. Familial autosomal dominant AD genes founded by linkage analyses are APP, PSEN1, PSEN2, ABCA7, and SORL1. Genome-wide association studies have found risk genes such as ABCA7, BIN1, CASS4, CD33, CD2AP, CELF1, CLU, CR1, DSG2, EPHA1, FERMT2, HLA-DRB5-HLA-DRB1, INPP5D, MEF2C, MS4A6A/MS4A4E, NME8, PICALM, PTK2B, SLC24A4, SORL1, and ZCWPW1. ABCA7, SORL1, TREM2, and APOE are proved to have high odds ratio (>2) in risk of AD using next generation sequencing studies. Thanks to the promising genetic techniques such as CRISPR-CAS9 and single-cell RNA sequencing opened a new era in genetics. CRISPR-CAS9 can directly link genetic knowledge to future treatment. Single-cell RNA sequencing are providing useful information on cell biology and pathogenesis of diverse diseases.