• Journal of Plant Biotechnology
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• 제46권3호
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• pp.228-235
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• 2019

사과(Malus pumila)는 국내에서 가장 경제적으로 중요한 과수 중의 하나이다. 하지만 사과 바이러스 감염은 생산량을 감소시키고 수확량 손실과 과일 품질 저하와 같은 심각한 문제를 야기한다. 국내에 감염된 사과 바이러스 및 비로이드 종류는 Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV), Apple stem grooving virus (ASGV), Apple mosaic virus (ApMV)와 Apple scar skin viroid (ASSVd) 등이 알려져 있다. 사과는 바이러스나 비로이드에 감염되어 있어도 대체로 이상한 징후가 발견되지 않아 바이러스로 인해 피해가 많았다. 본 연구는 사과 왜성대목 M.9 및 M.26의 무독묘 생산을 위하여 고온처리($37^{\circ}C$, 6주), 화학처리(Ribavirin) 및 생장점 배양하여 바이러스 제거 처리를 하였다. 바이러스 검출에 일반적으로 사용되는 방법은 효소면역 측정법(ELlSA)과 중합효소연쇄반응(RT-PCR)을 이용하였는데, RT-PCR은 ELlSA방법보다 10 ~ 30% 더 민감하였다. 사과 왜성대목 바이러스 검정 결과, 바이러스 제거 효율은 생장점 배양이 가장 높았다. 생장점 배양 후 바이러스 무병묘의 획득율은 30 ~ 40%로 높게 나타났다. 생장점 배양에서 사과 왜성대목 M.9은 ACLSV, ASPV 및 ASGV의 비율이 각각 45%, 60%, 50%로 높았고, 사과 왜성대목 M.26에서는 ACLSV, ASPV 및 ASGV의 감염율은 각각 40%, 55%, 55%였다. 이상의 결과, 사과 왜성대목에서 무독묘를 생산할 수 있는 가장 효과적인 방법은 생장점 배양에 의한 것으로 판단되었다.

• Journal of Animal Science and Technology
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• 제49권3호
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• pp.309-320
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• 2007

본 연구에서는 남성 생식 기관인 efferent ductules(ED)에서 monocarboxylate transporter (MCT) isoform과 Basigin(Bsg)의 mRNA 발현을 탐구하고, MCT1의 발현이 에스트로젠 수용체 α에 의해 조절되는지를 연구하였다. 생후 여러 연령대의 백서 ED에서 MCT isoform과 Bsg의 mRNA 발현 여부는 real-time PCR에 의해 조사 되었고, 정상 생쥐와 에스트로젠 수용체 α knockout 생쥐를 사용하여 에스트로젠 수용체 α에 의한 MCT1의 발현 조절 여부는 immunohistochemistry 방법에 의해 간접적으로 알아 보았다. 본 연구 결과는 백서의 ED에서 MCT1, 2, 3, 4와 8 그리고 Bsg 유전자의 발현은 연령에 따라 다르게 나타나며, MCT1의 발현이 ED의 ciliated 세포의 basolateral 지역에서 발현되고 어떤 측면에서 ERα에 의해 조절되어 질 수 있음을 보여 주었다. 따라서 본 연구 결과는 MCT가 monocarboxylate의 세포 내, 외부로 운반을 조절함으로써 남성 생식기의 일부인 ED의 역할을 조절하는데 관여함을 시사한다.

• 대한수의학회지
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• 제47권2호
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• pp.175-185
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• 2007

This study was carried out to investigate the pathogenesis and pathogenicity of the porcine circovirus type 2 (PCV2) Korean isolate from weaned pigs. Twenty four weaned pigs, PCV2, porcine reproductive and respiratory syndrome virus (PRRSV) and porcine parvovirus (PPV) antibodies free, were allocated to 4 groups (n = 6). Six pigs were inoculated intranasally with PCV2 alone, 6 with PCV2 and PRRSV, 6 with the combined PCV2/PRRSV/PPV inoculum, and 6 were remained as a uninoculated negative control. Pigs were killed 3 and 6 weeks after inoculation and tissue samples examined for gross and microscopic lesions and for the presence of PCV2 antigens and nucleic acids. Experimentally inoculated pigs were evaluated for 3 considerations: 1. development of postweaning multisystemic wasting syndrome (PMWS), 2. distribution of viral antigens by immunohistochemistry and polymerase chain reaction (PCR), and 3. cytokine mRNA levels in lymph nodes. Pigs inoculated with PCV2/PRRSV/PPV showed typical clinical signs, gross findings, and histopathologic characteristics of PMWS. In the PCV2/PRRSV/PPV inoculated group, the PCV2 antigen was widely distributed in various parenchymal organs such as brain, spinal cord, tonsil, lymph nodes, lung, heart, liver, kidney, spleen, and peyer's patch. Lymph node mRNA expression of IL-$1{\alpha}$, IL-2R and IL-8 was determined by real-time PCR. The pigs of PCV2/PRRSV and PCV2/PRRSV/PPV inoculation group, the mRNA expression was characterized by a decrease of IL-$1{\alpha}$, IL-2R and IL-8. The decrease of cytokine mRNA represent the state of T cell immuno-suppression in pig, and nicely support the evidence for the impairment of immune system in pigs with PMWS. In conclusion, PCV2 infection and some additional infectious causes such as PRRSV and/or PPV are warranted for the presence of PMWS in weaned pigs in Korea.

• 미생물학회지
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• 제48권3호
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• pp.175-179
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• 2012

아데노바이러스는 다양한 급성호흡기감염증을 유발하며, 대부분 영유아나 어린이, 면역기능이 저하된 환자에게서 주로 나타나는 것으로 알려져 있다. 본 연구에서는 2009년부터 2011년까지 경기지역 소아청소년과 및 내과에 내원한 급성호흡기 감염증 의심환자를 대상으로 호흡기아데노바이러스의 유행양상 및 혈청형 분포양상을 분석하였다. 총 1,622명의 급성상기도 감염증이 의심되는 환자의 검체를 분석한 결과 102건(6.3%)에서 아데노바이러스를 검출하였다. 102건의 아데노바이러스 양성 검체에서 세포배양법으로 76주의 아데노바이러스를 분리하였고, 혈청형별 특이유전자인 헥손 유전자의 염기서열 분석을 통하여 혈청형을 확인하였다. 최근 3년간 경기도내에서 아데노바이러스 1형부터 6형까지 6개의 다른 혈청형이 분리되었고, 이 중 3형(n=40, 52.6%)이 가장 주류를 이루었다. 2009년에는 1형과 3형, 2010년에는 3형, 2011년에는 5형이 각각 우점하였다. 1, 2, 4, 5, 6형은 연중 산발적으로 확인되었으나, 3형은 산발적으로 발생하면서 2010년에는 큰 유행을 일으킨 것을 확인할 수 있었다. 본 연구결과를 통해 최근 3년동안 경기도내 아데노바이러스에 의한 outbreak의 주 원인 혈청형은 아데노바이러스 3형임을 알 수 있었고, 앞으로도 outbreak의 원인이 되는 특정 혈청형에 대한 지속적인 감시가 이루어져야 할 것이다.

• Journal of Veterinary Science
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• 제22권5호
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• pp.63.1-63.14
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• 2021

Background: Recently, mesenchymal stem cells therapy has been performed in dogs, although the outcome is not always favorable. Objectives: To investigate the therapeutic efficacy of mesenchymal stem cells (MSCs) using dog leukocyte antigen (DLA) matching between the donor and recipient in vitro. Methods: Canine adipose-derived MSCs (cA-MSCs) isolated from the subcutaneous tissue of Dog 1 underwent characterization. For major DLA genotyping (DQA1, DQB1, and DRB1), peripheral blood mononuclear cells (PBMCs) from two dogs (Dogs 1 and 2) were analyzed by direct sequencing of polymerase chain reaction (PCR) products. The cA-MSCs were co-cultured at a 1:10 ratio with activated PBMCs (DLA matching or mismatching) for 3 days and analyzed for immunosuppressive (IDO, PTGS2, and PTGES), inflammatory (IL6 and IL10), and apoptotic genes (CASP8, BAX, TP53, and BCL2) by quantitative real-time reverse transcriptase-PCR. Results: cA-MSCs were expressed cell surface markers such as CD90⁺/44⁺/29⁺/45^- and differentiated into osteocytes, chondrocytes, and adipocytes in vitro. According to the Immuno Polymorphism Database, DLA genotyping comparisons of Dogs 1 and 2 revealed complete differences in genes DQA1, DQB1, and DRB1. In the co-culturing of cA-MSCs and PBMCs, DLA mismatch between the two cell types induced a significant increase in the expression of immunosuppressive (IDO/PTGS2) and apoptotic (CASP8/BAX) genes. Conclusions: The administration of cA-MSCs matching the recipient DLA type can alleviate the need to regulate excessive immunosuppressive responses associated with genes, such as IDO and PTGES. Furthermore, easy and reliable DLA genotyping technology is required because of the high degree of genetic polymorphisms of DQA1, DQB1, and DRB1 and the low readability of DLA 88.

• 한국해양바이오학회지
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• 제2권3호
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• pp.192-197
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• 2007

넙치의 생체방어 및 면역관련 유전자의 대량확보를 위하여, 지질다당질로 자극한 넙치의 말초백혈구 cDNA library를 suppression subtractive hybridization 방법으로 제작하여 발현유전자의 발현 양상 및 특성을 분석하였다. 총 470개의 SSH 클론 중 기존에 밝혀진 유전자와 상동성을 보인 218개의 클론을 분석한 결과, 대표적인 전염증사이토카인 IL-1의 유인수용체인 IL-1RII가 34번의 가장 높은 빈도로 발현되었으며, IL-$1{\beta}$가 14번의 높은 발현 빈도를 보였다. 그 중 펩티도글리칸 인식 단백질및 혈관활성 장 펩타이드 유전자는 어류에서는 처음으로 보고되는 것으로, 지질다당질로 자극한 넙치의 말초백혈구 SSH cDNA library는 다양한 생체방어 및 면역관련 유전자를 포함하고 있었다.

• 한국수정란이식학회지
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• 제27권4호
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• pp.245-252
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• 2012

The purpose of the present study was to investigate the effect of IFN-${\tau}$ on prostaglandin synthesis, cyclooxygenase-2 (COX-2) gene expression in vitro and concentration of progesterone (P4) in endometrial cells. Epithelial and stromal cells cultured in vitro were isolated from bovine endometrium and stimulated with increasing doses of IFN-${\tau}$ (0, 0.02, 0.2 and 2 ug/ml). Human chorionic gonadotropin (hCG, 1.5 IU/ml) was used as a positive control. Prostaglandin $E_2$ and $F_{2{\alpha}}$ levels in the culture media were analyzed by enzyme immunoassays and total RNA was extracted from the cells for RT-PCR. P4 concentrations of blood samples were assayed by chemiluminescent immuno assays system. In epithelial cells, COX-2 gene expression was increased in the presence of IFN-${\tau}$ (p<0.05), but it was not significantly different in all groups of stromal cells except for 2 ug/ml IFN-${\tau}$ group (p<0.05). Although IFN-${\tau}$ did not affect $PGE_2$ and $PGF_{2{\alpha}}$ production in epithelial cells, it decreased $PGE_2$ and $PGF_{2{\alpha}}$ production significantly in stromal cells (p<0.05). In vivo experiment, blood concentration of P4 was significantly increased after addition of IFN-${\tau}$ (1 ug/ml). The results indicate that PG production was mediated by COX-2 expression in stromal cells but it was not affected in epithelial cells and this suggest that treatment of IFN-${\tau}$ could improve the implantation environment of uterine by maintenance of high P4 concentration.

• 대한수의학회지
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• 제45권1호
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• pp.45-53
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• 2005

GroE that is a heat shock protein composed of GroEL and GroES is known as an immunodominant target of both the humoral and cellular immune responses in bovine brucellosis. This study was carried out to characterize groE gene encoding heat shock proteins of B. abortus isolated in Korea and to evaluate the immunogenicity of the GroE protein expressed in E. coli system. In PCR the specific signals with the size of 2,077 bp were detected in five strains isolated from the mammary lymphnodes of the dairy cattle that were serologically positive and the reference strains. In comparison of the sequences of nucleotides and amino acids among the strains, GroES showed 100% identity in both sequences. GroEL was evaluated 99.0~99.9% in nucleotides and 98.0~100% homology in amino acids. The groE gene including groES and groEL was inserted into pET29a vector and constructed pET29a-GroE recombinant plasmids. The inserted groE was confirmed by digestion with Nco1 and EcoR1 endonucleases and nucleotide sequencing. E. coli BL (DE3) was transformed with pET29a-GroE, named as E. coli BL (DE3)/pET29a-GroE. In SDS-PAGE, it was evident that the recombinant plasmid effectively expressed the polypeptides for GroES (10 kDa) and GroEL (60 kDa) in 0.5, 1 and 2 hours after IPTG induction. The immuno-reactivity of the expressed proteins were proved in mouse inoculation and Western blot analysis.

• 대한본초학회지
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• 제32권1호
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• pp.1-13
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• 2017

Objective : Soybeans of Canavalia gladiata(CG) and root of Arctium lappa(AL) have been reported to have anti-inflammatory, antioxidant effect. However, the immunoregulatory mechanisms of its combinational prescription remain a matter of considerable debate. In the current study, we investigated whether CG and AL and its combinational prescription(CG+AL) regulate immune system using chronic immobilization-stress mouse model. Methods : C57BL/6J mice fixed for 2 hours into immobilization tube after CG, AL, CG+AL oral administration after 2 hours daily for 21 days. After every experiment has ended the C57BL/6J mice were sacrificed on 22 days. The production of Serotonin and Cortisol, lgA were observed by ELISA method, The proportion of immune cells such as T/B cell and macrophage, NK cell were measured by FACS. Then, Real-time PCR was performed to measure the mRNA expression of Inflammatory cytokines(IL-1beta, IL-6, TNF-a) and T cell activation cytokines(IL-2, IL-10, IFN-gamma, IL-12p35 / p40). Result : When chronic immobilization-stress mouse model were treated with CG+AL(1:4), the expression of mRNA were significantly decreased at the Inflammatory cytokines(IL-1beta, IL-6, TNF-a). While, the levels of mRNA were significantly increased at immune T cell activation cytokines. Additionally, CG+AL(1:4) combinational prescription group enhanced immune cells such as T/B cell and macrophage, NK cell. Furthermore, the Immuno-fluorescence result of brain tissue can confirm that CG+AL(1:4) group significantly increased the BDNF expression. Conclusion : These result suggest that CG+AL(1:4) combinational prescription has Immune System enhancement via stress-mediated immunocyte.

• 한국수정란이식학회지
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• 제32권3호
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• pp.73-79
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• 2017

Transplantation is considered to be a very useful approach to improve human welfare and to prolong life-span. Heterologous organ transplantation using pig organs which are similar to human beings and easy to make mass-production has known as one of the alternatives. To ensure potential usage of the pig organ for transplantation application, it is essentially required to generate transgenic pig modifying immuno-related genes. Previously, we reported production of heterozygous ${\alpha}1,3$-galactosyltransferase (GalT) knock-out and human membrane cofactor protein (MCP) expressing pig ($GalT^{-MCP/+}$), which is enforced for suppression of hyperacute and acute immunological rejection. In this study, we reported generation of homozygous pig ($GalT^{-MCP/-MCP}$) by crossbreeding $GalT^{-MCP/+}$ pigs. Two female founders gave birth to six of $GalT^{-MCP/-MCP}$, and seven $GalT^{-MCP/+}$ pigs. We performed quantitative real-time PCR, western blot, and flow cytometry analyses to confirm GalT and MCP expression. We showed that fibroblasts of the $GalT^{-MCP/-MCP}$ pig do not express GalT and its product Gal antigen, while efficiently express MCP. We also showed no expression of GalT, otherwise expression of MCP at heart, kidney, liver and pancreas of transgenic pig. Taken together, we suggest that the $GalT^{-MCP/-MCP}$ pig is a useful candidate to apply xenotransplantation study.