• 한국가금학회지
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• 제32권1호
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• pp.29-34
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• 2005

본 연구는 한국 재래 닭의 유전적 특성을 분자표지를 이용하여 그 차이를 규명하고 초기성장에 미치는 영향을 분석하여 이를 이용한 재래닭의 개량을 목적으로 실시하였다. MHC class II B-LB 유전자 내의 염기변이체가 경제형질에 미치는 영향에 대하여 연구하였다. MHC class II B-LB유전자 내 400 bp 크기의 유전자를 증폭하여 염기서열 분석과 제한효소 처리를 이용한 다형성 분석을 실시하였다. 연구결과 두 개의 제한효소 절단지역이 발견되었으며 427 지역을 Type I 으로 651 지역은 Type II로 정하여 RFLP 분석을 실시하였다. Type I지역의 유전자형은 TT, TC, CC로 나타났으며, TypeII 지역의 유전자형은 MM, Mm, mm으로 나타났다. TC와 Mm 유전자형이 다른 유전자형과 비교하였을 때 한국재래 닭에서 높은 출현빈도를 보였다(0.8, 0.88). 유전자형이 한국 재래 닭의 150일령 체중에 미치는 영향을 분석한 결과 CC와 Mm 유전자형에서 통계적 유의성이 도출되었다 (P<0.05). 따라서 본 연구의 결과를 이용하여 한국 재래 닭의 유전적 특성을 규명할 수 있으며 초기 성장이 높은 성적을 나타내는 CC, Mm 유전자형을 개량에 이용하게 된다면 큰 효과를 얻을 수 있을 것으로 사료되어진다. 본 연구의 결과는 차후 한국 재래 닭의 과학적이고 지속적인 유전자원의 보존과 육종 전략에 있어 매우 유용하게 활용될 것으로 기대된다.

• IMMUNE NETWORK
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• 제4권4호
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• pp.205-215
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• 2004

In the early host defense system, effector function of natural killer (NK) cells results in natural killing against target cells such as microbe-infected, malignant, and certain allogenic cells without prior stimulation. NK cell cytotoxicity is selectively regulated by homeostatic prevalence between a repertoire of both activating and inhibitory receptors, and the discrimination of untransformed cells is achieved by recognition of major histocompatibility complex (MHC) class I alleles through inhibitory signals. Although it is well known that the bipotential T/NK progenitors are derived from the common precusor, functional mechanisms in terms of the development of NK cells remain to be further investigated. NK cells are mainly involved in innate immunity, but recent studies have been reported that they also play a critical role in adaptive immune responses through interaction with dendritic cells (DC). This interaction will provide effector functions and development of NK cells, and elucidation of its precise mechanism may lead to therapeutic strategies for effective treatment of several immune diseases.

• Journal of Microbiology and Biotechnology
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• 제26권12호
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• pp.2184-2191
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• 2016

The time-consuming and high-cost preparation of soluble peptide-major histocompatibility complexes (pMHC) currently limits their wide uses in monitoring antigen-specific T cells. The single-chain trimer (SCT) of peptide-${\beta}2m$-MHC class I heavy chain was developed as an alternative strategy, but its gene fusion is hindered in many cases owing to the incompatibility between the multiple restriction enzymes and the restriction endonuclease sites of plasmid vectors. In this study, overlap extension PCR and one-step cloning were adopted to overcome this restriction. The SCT gene of the $OVA_{257-264}$ peptide-$(GS_4)_3-{\beta}2m-(GS_4)_4-H-2K^b$ heavy chain was constructed and inserted into plasmid pET28a by overlap extension PCR and one-step cloning, without the requirement of restriction enzymes. The SCT protein was expressed in Escherichia coli, and then purified and refolded. The resulting $H-2K^b/OVA_{257-264}$ complex showed the correct structural conformation and capability to bind with $OVA_{257-264}$-specific T-cell receptor. The overlap extension PCR and one-step cloning ensure the construction of single-chain MHC class I molecules associated with random epitopes, and will facilitate the preparation of soluble pMHC multimers.

• Asian-Australasian Journal of Animal Sciences
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• 제23권9호
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• pp.1145-1151
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• 2010

The objective of this work was to analyze the relationship between ovine major histocompatibility complex (MHC) DRB1 gene polymorphism and genetic resistance to hydatidosis in Kazakh sheep. The Ovar (ovine MHC) class II DRB1 second exon was amplified by polymerase chain reaction (PCR) from DNA samples of 702 Kazakh sheep, including 302 sheep with hydatidosis and 400 health controls. PCR products were characterized by the restriction fragment length polymorphism (RFLP) technique using five restriction enzymes, i.e., MvaI, HaeIII, SacI, SacII and Hin1I, yielding 14 alleles and 28 genotypes. Comparing the frequency of genotypes in hydatidosis sheep with the control group, it was found that the genotype frequencies of MvaIbc, Hin1Iab, SacIIab, HaeIIIde, HaeIIIdf and HaeIIIdd in control sheep were significantly (p<0.01) higher than in hydatidosis sheep, indicating that a significant correlation existed between these genotypes and resistance to hydatidosis. Genotype frequencies of MvaIbb, SacIIaa, Hin1Ibb and HaeIIIef in sheep with hydatidosis were extremely significantly (p<0.01) higher than in the control group, and the genotype frequency of HaeIIIab was significantly higher (p<0.05), indicating that a marked correlation existed between these genotypes and susceptibility to hydatidosis. By way of analyzing haplotype with these resistant genotypes, the hydatidosis resistant haplotype MvaIbc-SacIIab-Hin1Iab of Kazakh sheep was screened out, and then verified through artificial hydatid infection in sheep. The results indicated that the infection rate of sheep with the resistant haplotype of hydatidosis was significantly lower (p<0.01) than without this resistant haplotype. It showed that the genic haplotype MvaIbc-SacIIab-Hin1Iab of Ovar-DRB1 exon 2 was the resistant haplotype of hydatidosis in Kazakh sheep.

• 미생물학회지
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• 제50권2호
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• pp.169-172
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• 2014

최근 인간 간암세포주(human hepatoma cells)를 이용하여 C형 간염 바이러스(hepatitis C virus, HCV)의 복제가 가능한 세포배양모델(cell culture system)이 확립되었다. 본 연구에서는 인간 간암세포주 중 huh7.5 cell (human hepatoma 7.5 cells)과 C형 간염 바이러스인 J6/JFH1 clone (2a 유전자형)를 이용하여 감염 가능한 세포배양모델을 확립하였다. 또한, HCV 감염 간암세포주의 HCV 특이 T 림프구에 대한 항원제시(antigen presentation) 가능성을 살펴보았다. 외부에서 전달된 HCV 항원일 경우 간암세포주의 HCV 특이 T 림프구에 대한 항원제시로 T 림프구의 활성이 가능하였으나, HCV 감염 간암세포주의 경우 T 림프구의 활성을 억제하였다. 이러한 HCV 특이 T 림프구의 활성억제와 HCV 감염 간암세포주 항원제시능의 상관성을 알아보기 위해 HCV 감염 간암세포주의 주조직적합성복합체(major histocompatibility complex, MHC) 발현변화를 측정하였으나 HCV 감염은 간암세포주의 MHC 발현변화에 영향을 미치지 않았다.

• 대한수의학회지
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• 제34권2호
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• pp.287-299
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• 1994

Plasma protein which has been known as one of nonspecific immunostimulators was added to feedstuff to examine its effect on the enhancement of cellular immune response in porcine immune system. A total of 40 piglets, 20 male and 20 female each, were fed for 30 days with or without plasma protein. The peripheral blood were collected and analyzed for the investigation of leukocyte subpopulations and their activities by using a panel of monoclonal antibodies specific to porcine leukocyte differentiation antigens and flow cytometry. The results obtained as follows. 1. Subpopulations expressing major histocompatibility complex(MHC) class I antigen were $96.2{\pm}3.1%$ and $86.6{\pm}3.8%$ in piglets fed with plasma protein and in piglets fed without plasma protein, respectively. 2. Proportion of leukocyte subpopulation expressing MHC class II antigens were significantly higher in the piglets fed with plasma protein than ones without plasma protein. The proportion was $27.6{\pm}3.6%$ and $16.6{\pm}2.2%$ in MHC class II DQ antigen, and $28.1{\pm}2.0%$ and $20.0{\pm}0.3%$ in MHC class II DR antigen, respectively. 3. A significant increase in the proportion of cells expressing poCD2 was not found in piglets fed plasma protein. 4. Proportion of subpopulation expressed porcine(Po) CD4 antigens which specific to helper T lymphocytes were not increased (18.3-19.1% vs. 25.6-28.8%), rather slightly decreased, in plasma protein-treated group. 5. The most important increase of proportion in plasma protein-treated group was the leukocyte subpopulation specific to $poCD8^+$ T cytotoxic/suppressor lymphocytes. The expression level was significantly higher up to 45.9-47.1% in plasma protein-treated group in comparing with 29.7-33.0% in non-plasma protein-treated group. 6. Lymphoblastogenetic responses using different concentrations of Con A mitogen and plasma protein has found that the responses of lymphocyte from piglets fed plasma protein was significantly activated (p<0.01). The activities measured by 3[H]-thymidine incorporation showed 3-6 times stronger in plasma protein-treated group than those in non-plasma protein-treated group. The study has concluded that plasma protein, which has known as a nonspecific immunostimulator, may have an immunoenhancing activities in porcine lymphoid system by increase the activated cell proportions and their blastogenetic properties which is critical to host immune responses.

• Asian-Australasian Journal of Animal Sciences
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• 제22권1호
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• pp.20-25
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• 2009

Xenotransplantation of pig organs into humans is a potential solution for the shortage of donor organs for transplantation. However, multiple immune barriers preclude its clinical application. In particular, the initial type of rejection in xenotransplantation is an acute cellular rejection by host $CD8^+$ cytotoxic T lymphocyte (CTL) cells that react to donor major histocompatibility complex (MHC) class I. The human cytomegalovirus (HCMV) glycoprotein Unique Short (US) 2 specifically targets MHC class I heavy chains to relocate them from the endoplasmic reticulum (ER) membrane to the cytosol, where they are degraded by the proteasome. In this study we transfected the US2 gene into minipig fetal fibroblasts and established four US2 clonal cell lines. The integration of US2 into transgenic fetal cells was confirmed using PCR and Southern blot assay. The reduction of Swine Leukocyte Antigen (SLA)-I by US2 was also detected using Flow cytometry assay (FACS). The FACS analysis of the US2 clonal cell lines demonstrated a substantial reduction in SLA-I surface expression. The level (44% to 76%) of SLA-I expression in US2 clonal cell lines was decreased relative to the control. In cytotoxicity assay the rate of $CD8^+$ T cell-mediated cytotoxicity was significantly reduced to 23.8${\pm}$15.1% compared to the control (59.8${\pm}$8.4%, p<0.05). In conclusion, US2 can directly protect against $CD8^+$-mediated cell lysis. These results indicate that the expression of US2 in pig cells may provide a new approach to overcome the CTL-mediated immune rejection in xenotransplantation.

• Clinical and Experimental Pediatrics
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• 제52권5호
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• pp.567-575
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• 2009

목적 : 간질은 전세계인구의 0.5%에서 발병하며 유전적 성향이 많고, 이는 중추신경계의 과 흥분성에 기인한다고 알려져 있다. 최근 프로테오믹스기법의 발달로 질병관련 단백질 동정이 활발히 연구되어지고 있다. 더불어, 간질의 진단은 영상기법 및 뇌파 분석 등이 이용되고 있으나, 가장 손쉽고 경제적인 혈청단백질을 이용한 진단법은 확립되어 있지 못하다. 그러므로 본 연구에서는 측두엽 간질환자의 혈장 단백질을 분석하여 간질의 진단 표지단백질 및 질병관련단백질을 발굴하고자 하였다. 방 법 : 저자들은 8명의 측두엽 간질환자와 8명의 정상인 혈청을 비교하였다. 결 과 : 간질환자의 혈청에서 정상 혈청단백질과 유의하고 일관성 있는 차이를 보이는 12개의 단백질을 발견하였다. 그 중, 6개의 단백질을 동정하였고, 6개의 단백질은 동정하지 못하였다. 더불어, haptoglobin Hp2, PRO2675, immunoglobulin heavy chain constant region gamma 2와 1개의 명명되지 않은 단백질 및 3개의 미지의 단백질을 포함한 7개의 단백질은 간질환자의 혈액에서 증가하였다. 반면, MHC class I antigen, plasma retinol-binding protein precursor 및 3개의 미지의 단백질을 포함한 5개의 단백질은 감소하였다. 결 론 : MHC class I antigen, immunoglobulin heavy chain constant region gamma 2 및 수술 전에 증가하였던 3개의 미지의 단백질 중에서 1개, 감소하였던 3개의 미지의 단백질 중에서 2개를 포함한 모두 5개의 단백질은 간질을 일으키는 뇌 부위 절제 후 정상으로 회복되었다. 이는 이런 단백질들을 측두엽 간질의 진단 및 경과관찰인자로서, 활용할 수 있음을 시사한다. 나아가, 이러한 단백질들은 간질의 병태 생리 연구 및 새로운 치료약물개발의 표적 단백질로 활용될 수 있을 것이다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권22호
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• pp.9667-9672
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• 2014

Background: Pancreatic adenocarcinoma is a malignant gastrointestinal cancer with significant morbidity and mortality. Despite severe side effects of chemotherapy, the use of immunotherapy combined with chemotherapy has emerged as a common clinical treatment. In this study, we investigated the efficacy of the combined doxorubicin and interferon-${\alpha}$ (IFN-${\alpha}$) therapy on murine pancreatic cancer Panc02 cells in vitro and in vivo and underlying mechanisms. Materials and Methods: A Panc02-bearing mouse model was established to determine whether doxorubicin and interferon-${\alpha}$ (IFN-${\alpha}$) could effectively inhibit tumor growth in vivo. Cytotoxicity of natural killer (NK) cells and cytotoxic T lymphocytes (CTLs) was evaluated using a standard LDH release assay. To evaluate the relevance of NK cells and CD8 T cells to the combination therapy-mediated anti-tumor effects, they were depleted in tumor-bearing mice by injecting anti-asialo-GM-1 antibodies or anti-CD8 antibodies, respectively. Finally, the influence of doxorubicin+interferon-${\alpha}$ (IFN-${\alpha}$) on the ligands of NK and T cells was assessed by flow cytometry. Results: The combination therapy group demonstrated a significant inhibition of growth of Panc02 in vivo, resulting from activated cytotoxicity of NK cells and CTLs. Depleting CD8 T cells or NK cells reduced the anticancer effects mediated by immunochemotherapy. Furthermore, the doxorubicin+IFN-a treatment increased the expression of major histocompatibility complex class I (MHC I) and NKG2D ligands on Panc02 cells, suggesting that the combined therapy may be a potential strategy for enhancing immunogenicity of tumors. All these data indicate that the combination therapy using doxorubicin and interferon-${\alpha}$ (IFN-${\alpha}$) may be a potential strategy for treating pancreatic adenocarcinoma.

• Asian-Australasian Journal of Animal Sciences
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• 제30권6호
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• pp.773-780
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• 2017

Objective: "Hatchability" is an important economic trait in domestic poultry. Studies on poultry hatchability focus mainly on the genetic background, egg quality, and incubation conditions, whereas the molecular mechanisms behind the phenomenon that some ducklings failed to break their eggshells are poorly understood. Methods: In this study, the transcriptional differences between the livers of normally hatched and assisted ducklings were systematically analyzed. Results: The results showed that the clean reads were de novo assembled into 161,804 and 159,083 unigenes (${\geq}200-bp$ long) by using Trinity, with an average length of 1,206 bp and 882 bp, respectively. The defined criteria of the absolute value of log2 fold-change ${\geq}1$ and false discovery rate${\leq}0.05$ were differentially expressed and were significant. As a result, 1,629 unigenes were identified, the assisted ducklings showed 510 significantly upregulated and 1,119 significantly down-regulated unigenes. In general, the metabolic rate in the livers of the assisted ducklings was lower than that in the normal ducklings; however, compared to normal ducklings, glucose-6-phosphatase and ATP synthase subunit alpha 1 associated with energy metabolism were significantly upregulated in the assisted group. The genes involved in immune defense such as major histocompatibility complex (MHC) class I antigen alpha chain and MHC class II beta chain 1 were downregulated in the assisted ducklings. Conclusion: These data provide abundant sequence resources for studying the functional genome of the livers in ducks and other poultry. In addition, our study provided insight into the molecular mechanism by which the phenomenon of weak embryos is regulated.