• IMMUNE NETWORK
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• 제3권1호
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• pp.8-15
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• 2003

Background: CD30 is a member of TNF receptor family and expressed on lymphocytes and other hematopoietic cells following activation as well as Hodgkin and Reed-Sternberg cells in Hodgkin's lymphoma. In this study, CD30-mediated regulation of cell adhesion molecule expression on normal activated mouse T cells was investigated. Methods: Mouse T cells were activated with anti-CD3 antibody for induction of CD30, which was cross-linked by immobilized anti-CD30 antibody. Results: High level of CD30 expression on T cells was observed on day 5, but only little on day 3 even under culture condition resulting in an identical T cell proliferation, indicating that CD30 expression requires a prolonged stimulation up to 5 days. Cross-linking of CD30 alone altered neither proliferation nor apoptosis of normal activated T cells. Instead, CD30 appeared to promote cell adherence to culture substrate, and considerably upregulated ICAM-1 and, to a lesser extent, ICAM-2 expression on activated T cells, whereas CD2 and CD18 (LFA-1) expression was not affected. None of cytokines known as main regulators of ICAM-1 expression on tissue cells (IL 4, $IFN{\gamma}$ and $IFN{\alpha}$) enhanced ICAM-1 expression in the absence of CD30 signals. On the other hand, addition of $NF-{\kappa}B$ inhibitor, PDTC (0.1 mM) completely abrogated the CD30-mediated upregulation of ICAM-1 expression, but not CD2 and ICAM-2 expression. Conclusion: This results support that CD30 upregulates ICAM-1 expression of T cell and such regulation is not mediated by higher cytokine production but $NF-{\kappa}B$ activation. Therefore, CD30 may play important roles in T-T or T-B cell interaction through regulation of ICAM-1, and -2 expression.

• 대한수의학회지
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• 제42권1호
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• pp.55-64
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• 2002

HIV-1의 envelope glycoprotein은 중화항체에 의한 체액성 면역반응의 중요한 target으로 surface glycoprotein인 gp120과 transmembrane glycoprotein인 gp41로 이루어져 있다. gp120과 gp41의 ectodomain으로 이루어진 gp140 유전자를 PCR의 방법으로 증폭하고 Semliki Forest virus(SFV) 유래 expression system을 이용하여 mammalian 세포에서 발현하였다. 발현된 gp140은 natural HIV-1에서와 같이 oligomer를 형성하였다. 발현된 gp140을 정제하여 BALB/c 마우스에 접종하여 항체가 형성되었음을 확인하였다.

• Journal of Animal Science and Technology
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• 제48권6호
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• pp.933-942
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• 2006

본 실험은 생봉독 처리가 돼지의 체액성 면역반응에 대한 생봉독의 효과를 조사하기 위하여 실시하였다. 혈중 Immunoglobulin(Ig) G, IgA, IgM의 농도에 미치는 생봉독의 효과를 조사하기 위하여 모돈 3두에서 생산한 자돈 20두(LY ×D)를 생봉독 처리군과 대조군으로 각각 10두씩 배치하였다. 생봉독 처리군은 출생시와 3일령에 교소혈 (GV-1), 해문혈(ST-25) 및 두구혈(CV-8), 6일령에 거세, 단미 창상부위에 생봉 1마리씩 직침 시술하였고, 대조군은 생리식염수 1ml를 동일한 혈위에 주입하였다. 혈중 Ig 농도 측정을 위하여 출생시와 3일, 7일, 14일 및 21일령에 채혈하여 Immunoturbidimetric method로 IgG, IgM, IgA를 측정하였다. 지표항원으로 사용한 돈 콜레라와 위축성비염 백신에 대한 생봉독의 항체 생성효과를 조사하기 위하여 모돈 5두에서 생산된 자돈 40두(LY×D)를 생봉독 처리군과 대조군으로 각각 20두씩 공시하였다. 생봉독 처리군은 출생시에 교소혈(Jiao-Chao, GV-1), 해문혈(Hai-Men, ST-25) 및 두구혈(Du-Kou, CV-8)에, 3일령에 단미 및 거세시술 시 창상부위에 생봉독을 처리하였고, 21일령 이유시에는 교소혈(GV-1)과 백회혈(Bai- Hui, GV-20)에 생봉독을 처리하였다. 지표항원으로 위축성비염 백신은 24일령과 44일령에 접종하였고 돈콜레라 백신은 44일령과 64일령에 각각 접종하였다. 항체가 분석용 혈액은 24, 34, 44, 54 및 74일령에 채혈하여 위축성비염은 시험관응집반응, 돈콜레라는 ELISA법에 의하여 항체가를 조사하였다. IgG 농도는 처리군에서 출생시 339.52, 3일령에 366.48, 7일령에 296.52, 14일령에 242.06, 21일령에는 219.06mg/dl이었고 대조군은 각각 347.10, 333.14, 243.28, 205.18 및 191.58mg/dl 이었다. 처리전(출생시)에는 처리군과 대조군간에 유의차가 없었으나 처리군의 IgG 농도가 대조군에 비하여 3일령 10.3% (P<0.02), 7일령에는 21.9% (P<0.01), 14일령에는 18.0% (P<0.07), 21일령에는 14.3(P<0.07) 더 높게 나타났다. IgA와 IgM의 농도는 전체기간동안 처리군과 대조군간에 유의차가 인정되지 않았다. 돈콜레라 virus에 대한 항체역가는 처리군에서 대조군에 비하여 24일령 때 57.0%(P<0.03), 34일령 때 74.6% (P<0.006), 44일령에 48.6% (P<0.017), 54일령 때 45.0% (P<0.16), 74일령 때 44.4%(P<0.006)가 유의하게 높은 것으로 나타났다. 위축성비염 원인균인 Bordetella bronchiceptica에 대한 항체역가는 처리군이 대조군에 비하여 34일령 때는 39.7%(P<0.002), 44일령 때 31.9% (P<0.02), 54일령 때에 33.4%(P<0.01), 74일령 때에는 57.3%(P<0.007)가 높게 나타나 접종일인 24일령을 제외하고 전체기간에 걸쳐 높은 항체수준을 보였다. 위의 결과를 종합해 볼 때 돼지에 대한 생봉독 처리는 면역기능향상 효과가 있는 것으로 사료된다. 이와 같은 결과를 볼 때 돼지에 대한 생봉독의 처리는 양돈 생산성면에서 생봉독 처리는 돼지의 생존율을 높이고, 증체량이 높았다는 조 등(2005)의 보고를 뒷받침하고 있어 양돈 산업발전에 기여할 것으로 사료된다.

• Journal of Animal Science and Technology
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• 제64권6호
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• pp.1117-1131
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• 2022

Previous studies reported that Bifidobacterium animalis ssp. lactis HY8002 (HY8002) improved intestinal integrity and had immunomodulatory effects. Lactobacillus plantarum HY7717 (HY7717) was screened in vitro from among 21 other lactic acid bacteria (LAB) and demonstrated nitric oxide (NO) production. The aims of this study were to investigate the individual and combined ex vivo and in vivo effects of LAB strains HY8002 and HY7717 at immunostimulating mice that have been challenged with an immunosuppressant drug. The combination of HY8002 and HY7717 increased the secretion of cytokines such as interferon (IFN)-γ, interleukin (IL)-12, and tumor necrosis factor (TNF)-α in splenocytes. In a cyclophosphamide (CTX)-induced immunosuppression model, administration of the foregoing LAB combination improved the splenic and hematological indices, activated natural killer (NK) cells, and up-regulated plasma immunoglobulins and cytokines. Moreover, this combination treatment increased Toll-like receptor 2 (TLR2) expression. The ability of the combination treatment to upregulate IFN-γ and TNF-α in the splenocytes was inhibited by anti-TLR2 antibody. Hence, the immune responses stimulated by the combination of HY8002 and HY7717 are associated with TLR2 activation. The preceding findings suggest that the combination of the HY8002 and HY7717 LAB strains could prove to be a beneficial and efficacious immunostimulant probiotic supplement. The combination of the two probiotic strains will be applied on the dairy foods including yogurt and cheese.

• Parasites, Hosts and Diseases
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• 제27권1호
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• pp.23-34
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• 1989

요 약 : 마우스에 있어서 왜소조충(Hymenolepis nana) 감염이 숙주의 비특이 면역반응에 미치는 영향을 조사하기 위하여 ICR계 마우스(20 g 정도)를 인공감염 군과 자연감염 군으로 나누어 인공감염 군의 일부는 최초 감염 후 15일째에, 자연감염 군은 실험 시작일에 프라지관텔(25mg/kg/day)을 3일 간 연속 투여하고 경시적으로 50일까지 면양 적혈구(sRBC) 감작에 대한 지연형 과민반응, 적혈구 응집소가 및 용혈소가, 말초혈액 내 호산구 출현률, 비장세포의 sRBC 로제트(로제트) 형성능, 항체처리 로제트 형성능 및 소장 점막 비만세포 출현빈도에 대하여 관찰하였다. 인공감염군에 있어서 지연형 과민반응은 감염 10일에 대조보다 일시적으로 현저히 저하하였다가 그 후 점차 상승하여 거의 정상으로 복귀하였다. 호산구 출현률은 왜소조충이 감염됨에 따라 대조보다 다소 상승하는 경향이었다. 적혈구 응집소가 및 용혈소가는 감염 초기에는 대조보다 다소 높은 값을 나타내었으나 감염 15일에 일시적으로 낮아졌다가 다시 거의 정상으로 복귀하였다. 로제트 및 항체 처리 로제트 형성능은 감염 15일에 대조보다 일시적으로 낮아졌다가 그 이후에는 점차 높아졌다. 비만세포 출현빈도는 감염후 시일 경과에 따라 점차 높아져 10일에 피이크에 이른 다음 그보다 약간 낮은 수준으로 계속 유지하였다. 인공감염군의 치료례에 있어서 지연형 과민반응과 호산구 출현률은 인공감염군에 비해 전반적으로 약간 떨어지는 경향이었으나, 로제트 형성능과 비만세포 출현빈도는 현저하게 저하하였다. 적혈구 응집소가, 용혈소가 및 항체처리 로제트 형성능은 인공감염 군에 비해 다소 높은 경향이었다. 한편, 자연감염 군에 있어서는 왜소조충 감염에 의해 높았던 지연형 과민반응, 비만세포 출현빈도 및 호산구 출현률 등이 치료 후에 점차 저하되어 대조와 거의 일치하였다. 또, 감염에 의해 낮았던 적혈구 응집소가 및 용혈소가는 치료 후 점차 상승하여 대조와 거의 일치하였는데 로제트 및 항체처리 로제트 형성능은 낮았던 값이 치료 후에도 계속 낮게 유지되었다. 이상의 실험결과로 미루어 보아 ICR계 마우스에 있어서 sRBC에 대한 면역능은 왜소조충 감염에 의해 일반적으로 활성화되는데 감염 후 10∼15일에는 일시적으로 저하된다는 것을 알 수 있었다.

• IMMUNE NETWORK
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• 제8권2호
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• pp.53-58
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• 2008

Background: Molecular mechanisms of natural killer (NK) cell development from hematopoietic stem cells (HSCs) have not been clearly elucidated, although the roles of some genes in NK cell development have been reported previously. Thus, searching for molecules and genes related NK cell developmental stage is important to understand the molecular events of NK cell development. Methods: From our previous SAGE data-base, Gpnmb (Glycoprotein non-metastatic melanoma protein B) was selected for further analysis. We confirmed the level of mRNA and protein of Gpnmb through RT-PCR, quantitative PCR, and FACS analysis. Then we performed cell-based ELISA and FACS analysis, to know whether there are some molecules which can bind to Gpnmb. Using neutralizing antibody, we blocked the interaction between NK cells and OP9 cells, and checked IFN-${\gamma}$ production by ELISA kit. Results: Gpnmb expression was elevated during in vitro developmental stage and bound to OP9 cells, but not to NK precursor cells. In addition, we confirmed that the levels of Gpnmb were increased at NK precursor stage in vivo. We confirmed syndecan4 as a candidate of Gpnmb's binding molecule. When the interaction between NK cells and OP9 cells were inhibited in vitro, IFN-${\gamma}$ production from NK cells were reduced. Conclusion: Based on these observations, it is concluded that Gpnmb has a potential role in NK cell development from HSCs.

• Journal of Microbiology and Biotechnology
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• 제16권2호
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• pp.318-324
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• 2006

In order to develop an oral vaccine to prevent H. pylori infection, we have expressed the hpaA gene of H. pylori K51 isolated from Korean patients, encoding 29-kDa HpaA that is known to be localized on the cell surface and flagella sheath, in a live delivery vector system, Lactococcus lactis. The hpaA gene, amplified by PCR using the genomic DNA of H. pylori K51, was cloned in the pGEX-2T vector, and the DNA sequence analysis revealed that the hpaA gene of H. pylori K51 had 99.7% and 94.8% identity with individual hpaA genes of the H. pylori 26695 strain (U.K) and the J99 strain (U.S.A). A polyclonal anti-HpaA antibody was raised in rats using GST-HpaA fusion protein as the antigen. The hpaA gene was inserted in an E. coli-L. lactis-shuttle vector (pMG36e) to express in L. lactis. Western blot analysis showed that the expression level of HpaA in the L. lactis transformant remained constant from the exponential phase to the stationary phase, without extracelluar secretion. These results indicate that the HpaA of H. pylori K51 was successfully expressed in L. lactis, and suggest that the recombinant L. lactis expressing HpaA may be applicable as an oral vaccine to induce a protective immune response against H. pylori.

• IMMUNE NETWORK
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• 제2권2호
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• pp.102-108
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• 2002

Background: This study was aimed to differentiate two forms of CTLA-4 (CD152) in activated peripheral blood lymphocyte and clarify the mechanism how cytoplasmic form of this molecule is targeted to cell surface. Methods: For this purpose we generated 2 different anti-human CD152 peptide antibodies and 5 different N'-terminal deletion mutant CTLA4Ig fusion proteins and carried out a series of Western blot and ELISA analyses. Antipeptide antibodies made in this study were anti-CTLA4pB and anti-CTLA4pN. The former recognized a region on extracellular single V-like domain and the latter recognized N'-terminal sequence of leader domain of human CD152. Results: In Western blot, the former antibody recognized recombinant human CTLA4Ig fusion protein as an antigen. And this recognition was completely blocked by preincubating antipeptide antibody with the peptide used for the antibody generation at the peptide concentration of 200 ug/ml. These antibodies were recognized human CD152 as a cytoplasmic sequestered- and a membrane bound- forms in phytohemagglutinin (PHA)-stimulated peripheral blood lymphocyte (PBL). These two forms of CD152 were further differentiated by using anti-CTLA4pN and anti-CTLA4pB antibodies such that former recognized cytosolic form only while latter recognized both cytoplasmic- and membraneforms of this molecule. Furthermore, in a transfection expression study of 5 different N'-terminal deletion mutant CTLA4Ig, mutated proteins were secreted out from transfected cell surface only when more than 6 amino acids from N'-terminal were deleted. Conclusion: Our results implies that cytosolic form of CTLA-4 has leader sequence while membrane form of this molecule does not. And also suggested is that at least N'-terminal 6 amino acid residues of human CTLA-4 are required for regulation of targeting this molecule from cytosolic- to membrane- area of activated human peripheral blood T lymphocyte.

• Asian Pacific Journal of Cancer Prevention
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• 제16권9호
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• pp.3901-3905
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• 2015

Objective: Our aim was to investigation the roles of MHC class I chain-related gene A(MICA) and natural killer cell group 2D(NKG2D) in human renal cancer cells. Materials and Methods: The expression of membrane MICA (mMICA) on renal cells and NKG2D on NK cells were detected by flow cytometry (FCM); the content of sMICA were detected by enzyme linked immunosorbent assay (ELISA) and the distribution of mMICA on renal tumor tissues by immunohistochemistry; the interaction between MICA and NKG2D was observed by antibody closed method. Results: Our results showed that the expression of mMICA in renal cancer tissues was significantly higher than in controls, where the soluble MICA was not expressed. Cytotoxic activity of NK cells was significantly reduced after exposure to NKG2D and MICA antibodies (P<0.05), and serum containing sMICA can obviously lower the function of NKG2D (P<0.05). Conclusions: The interaction of mMICA and NKG2D play important roles in mediation of cytotoxicity of NK cells in RCC. On the other hand, sMICA may mediate tumor immune escape through down- regulated NKG2D expression.

• Molecules and Cells
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• 제38권5호
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• pp.441-451
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• 2015

Recognition of cytosolic DNA initiates a series of innate immune responses by inducing IFN-I production and subsequent triggering JAK1-STAT1 signaling which plays critical roles in the pathogenesis of infection, inflammation and autoimmune diseases through promoting B cell activation and antibody responses. The stimulator of interferon genes protein (STING) has been demonstrated to be a critical hub of type I IFN induction in cytosolic DNA-sensing pathways. However, it still remains unknown whether cytosolic DNA can directly activate the JAK1-STAT1 signaling or not. And the role of STING is also unclear in this response. In the present study, we found that dsDNA directly triggered the JAK1-STAT1 signaling by inducing phosphorylation of the Lyn kinase. Moreover, this response is not dependent on type I IFN receptors. Interestingly, STING could inhibit dsDNA-triggered activation of JAK1-STAT1 signaling by inducing SHP-1 and SHP-2 phosphorylation. In addition, compared with normal B cells, the expression of STING was significantly lower and the phosphorylation level of JAK1 was significantly higher in B cells from MRL/lpr lupus-prone mice, highlighting the close association between STING low-expression and JAK1-STAT1 signaling activation in B cells in autoimmune diseases. Our data provide a molecular insight into the novel role of STING in dsDNA-mediated inflammatory disorders.