• Journal of Plant Biotechnology
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• 제46권3호
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• pp.217-227
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• 2019

Production of therapeutic monoclonal antibodies (mAbs) using a plant platform has been considered an alternative to the mammalian cell-based production system. A plant-derived mAb CO17-1AK ($mAb^P$ COK) can specifically bind to various types of cancer cell lines. The target protein of $mAb^P$ COK is the epithelial cell adhesion molecule (EpCAM) highly expressed in human epithelial cancer cells, including breast and colorectal cancer cells. It has been hypothesized that its overexpression supports tumor growth and metastasis. A ganglioside is extended well beyond the surfaces of the various cell membranes and has roles in cell growth, inflammation, differentiation, and carcinogenesis. However, the regulation of EpCAM gene expression in breast cancers and the role of gangliosides in oncogenesis are unclear. Here, the purpose of this study was to determine the effects of $mAb^P$ COK on human breast cancer cell proliferation, apoptosis, and ganglioside expression patterns. Our results show that treatment with $mAb^P$ COK suppressed the growth of breast cancer cells and induced apoptotic cell death. It also upregulated the expression of metastasis-related gangliosides in breast cancer cells. Thus, treatment with $mAb^P$ COK may have chemo-preventive therapeutic effects against human breast cancer.

• 한국발생생물학회지:발생과생식
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• 제17권4호
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• pp.409-420
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• 2013

Human serum (HS) has been reported to induce aggregation of human eyelid adipose-derived stem cells (HEACs) during high-density culture in vitro. The present study focused on the role of cell adhesion molecules and gelatinases during HS-induced aggregation of HEACs. HS-induced aggregation occurred between 9-15 days of culture. Cells aggregated by HS medium (HS-agg) showed stronger expression of ${\alpha}2$, ${\alpha}2B$, ${\alpha}X$, and CEACAM1 genes compared to non-aggregated cells in HS medium (HS-ex) or in control FBS-cultured cells. HS-agg were distinctly labeled with antibodies against ${\alpha}2$, ${\alpha}2B$, and ${\alpha}X$ proteins. Western blot results demonstrated that the two integrin proteins were greatly expressed in HS-agg compared to HS-ex and control FBS-cultured cells. Treatment of HEACs with anti-integrin ${\alpha}2$ antibody during culture in HS medium delayed aggregation formation. HS-agg exhibited strong expression of MMP1 and MMP9 compared to HS-ex or FBS-cultured cells. Conditioned media from HS-culture showed remarkable increase of MMP9 gelatinolytic activity in comparison to those from FBS-culture. However, there was no change of TIMP mRNA expression in relation to the HS-induced aggregation. Based on these results, it is suggested that integrin ${\alpha}2$, ${\alpha}2B$, and ${\alpha}X$, and MMP9 might play an important role in the HS-induced aggregation of HEACs.

• Korean Journal of Acupuncture
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• 제24권4호
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• pp.111-129
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• 2007

Objectives & Methods : The purpose of this study is to observe the effects of Coicis Semen Herbal-acupuncture solution (CS-HAS) at the Joksamni (ST36) on the collagen-induced arthritis in the DBA/1J mouse. The author performed several experiments to analyze the effects of CS-HAS on arthritis; change of the weight; the spleen size and adhesion rate; serum cytokine levels; serum antibody levels; changes of immunocyte counts; the histological changes of joint. Results : In the Coicis Semen Herbal-acupuncture (CS-HA), arthritis index, the incidence of arthritis, and the degree of joint edema were decreased. In CS-HA, there was no weight loss. The size of the spleen, adhesion rate, and the edema and transformation of joint were lowered. In CS-HA, the levels of IL-1${\beta}$, IL-6, TNF-${\alpha}$, IFN-${\gamma}$, IgG, IgM, and anti-collagen II in serum and the levels of IFN-${\gamma}$, IL-4, IL-10 in spleen were significantly decreased. In CS-HA, the expression ratios of $CD45^+$ to $CD3e^+$ and $CD8^+$ to $CD4^+$ were decreased. Also, the overall $CD4^+/CD8^+$ cell ratio was lowered in spleen. Ratios of the $CD4^+/CD25^+$, $CD45^+/CD69^+$ cells were decreased in lymph nodes. In addition, ratios of the $CD3^+/CD69^+$, $CD11b^+/Gr-1^+$ cells were also decreased in synovium. In the histological study, the cartilage destruction and synovial cell proliferation, and collagen fiber destruction were decreased with CS-HA treated group. Conclusions : From the results mentioned above, it is suggested that CS-HA at the ST36 has several significant effects on the collagen-induced arthritis.

• 대한수의학회지
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• 제34권2호
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• pp.275-286
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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. Total weight gain, daily feed intake and feed conversion rate for 10 days were significantly improved to 56%, 20% and 22% in the piglets fed plasma protein, respectively. 2. A significant increase in N (null or non T/non B) cells was also noticed. Leukocyte proportion from piglets fed plasma protein was 20.2-24.7%, otherwise that from piglets fed without plasma protein was 12.3-13.4%, respectively. 3. A significant increase in the proportion of B cells and cells expressing poCD1 was not found in piglets fed plasma protein. 4. Reaction with monoclonal antibodies specific to adhesion molecules, poCD11a, poCD11b, poCD44 and poCD45A and poCD45B, has shown that leukocyte subpopulation from piglets fed plasma protein did not significantly higher than that from piglets fed without plasma protein. 5. Total proportion of granulocytes and monocytes was about 50% in both group and the proportion after treated with Hypaque/Ficoll was 2.7% and 5.8% in each group, respectively.

• Tuberculosis and Respiratory Diseases
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• 제42권4호
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• pp.569-583
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• 1995

연구배경: 간질성 폐질환은 여러가지 다양한 원인에 의해, 또는 아직 밝혀지지 않은 원인에 의해 폐포염 및 폐간질내 염증으로 시작해 종국에는 폐섬유증으로 진행되는 질환들로 염증세포들의 이동에 필요한 접착분자들이 이들 질환의 발병기전에 중요한 역할을 할 것으로 추정되며, 현재까지는 뚜렷이 적당한 지표가 없었던 질환의 활동성을 판정하는 지표로 사용할 수 있을 가능성이 많다. 그러나 이제까지는 BAL내 AM에서 면역염색법으로 측정한 결과들로 방법 자체가 음성과 양성세포를 감별하기가 쉽지 않을 뿐 아니라 발현도 증가를 정량적으로 측정하지 못하는 단점이 있고, 따라서 발표된 결과들이 차이가 많다. 저자들은 간질성 폐질환 환자들에서 기관지 폐포세척액(BAL)내 AM과 임파구에서의 ${\beta}_2$-integrin(CD18)과 ICAM-1의 발현도를 FACS를 이용한 relative median fluorescence intenity(RMFI)를 측정하여 정상 대조군과 비교 관찰하고, 또한 혈청 및 BAL액내 가용정 ICAM-1(sICAM-1) 농도를 측정하고, BAL액내 다른 지표들과 비교하여 이들을 간질성 폐질환의 활성도를 반영할 수 있는 지표로 사용할 가능성을 알아 보았다. 방법: 대상은 조직학적으로 확인 된 미만성 간질성 폐섬유증(IPF) 환자 11명, 교원성 폐질환과 연관된 폐섬유증환자(CVD-PF) 6명, 폐유육종증 9명, 과민성 폐장염환자 2명과 대조군으후 건강한 정상인 9명이었고 BAL은 통상적인 방법을 사용하였다. ICAM-1 및 CD18은 단일항체를 사용하여 FACScan으로 median fluorescence intenity를 측정하여 RMFI를 구하고, sICAM-1농도는 ELISA법으로 측정하였다. 결과: AM에서 ICAM-1의 발현도는 $3.30{\pm}1.16$으로 대조군($0.93{\pm}0.18$)보다 증가되었고 임파구의 ICAM-1 발현도 $5.39{\pm}2.70$으로 대조군($1.06{\pm}0.21$)보다 높았다. CD18의 발현도는 임파구에서 $24.9{\pm}14.9$로 대조군($4.69{\pm}3.77$) 보다 향진되었으며, 이들 ICAM-1의 발현도는 AM 백분률과 역상관관계륜 보였고(r=-0.66, p=0.0001) 임파구의 백분율과는 정상관관계를 보였다(r=0.447, p=0.0116). 임파구가 증가되는 유육종증, 교원성질환, 과민성폐렴에서는 임파구의 ICAM-1 발현도와 BAL액내 임파구의 %(r=0.747, p=0.0006) 및 임파구농도(r=0.832, p=0.0002)와 좋은 상관관계를 보였고, 임파구의 IL-2수용체발현과도 연관관계를 나타내었다(r=0.539, p=0.0075). 또한 유육종증에서는 혈중 ACE 농도와 임파구의 ICAM-1 발현도가(r=0.905, p=0.0132) 높은 상관관계를 보여, 이들 접착분자 발현도가 병소의 활동성과 연관이 있음을 시사 하였다. 혈청내 sICAM-1농도는 환자군에서 $499.7{\pm}222.2\;ng/ml$로 대조군의 $199.0{\pm}38.9\;ng/ml$보다 높았으며(p=0.0097), BAL액내 sICAM-1농도도 환자군에서 $41.8{\pm}23.0\;ng/ml$로 대조군의 $20.1{\pm}13.6\;ng/ml$보다 증가를 보였다. 또한 혈청내 sICAM-1 농도는 AM의 ICAM-1 발현도 및 (r=0.554, p=0.0259) BAL액내 sICAM-1농도와 상관관계를 보였다. 또한 혈청 및 BAL액내의 albumin과 sICAM-1의 비율을 비교한 결과 BAL액내의 sICAM-1 농도가 훨씬 높은 것으로 미루어 대부분의 sICAM-1은 병소내에서 생성된 것임을 짐작할 수 있었다. 결론: 간질성폐질환환자들의 AM및 BAL-임파구에서 접착분자들의 발현도가 증가되었고, 혈청 및 BAL액내 sICAM-1 농도가 상승되어 있어 이들 접착분자들이 발병기전에 중요한 역할을 한다고 생각되며, 앞으로 이 활동성 판정의 지표로 사용할 기능성을 시사하였다.

• 대한세포병리학회지
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• 제12권2호
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• pp.81-87
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• 2001

The differentiation between reactive mesothelial and carcinoma cells in serous effusion cytology can be a diagnostic challenge based on morphology alone. The expression of some cell adhesion molecules may be helpful in the differential diagnosis. This study evaluated the usefulness of E-cadherin Immunocytochemistry for discrimination of carcinoma cells from reactive mesothelial cells. Alcohol fixed, paraffin embedded cell blocks taken from 42 reactive and 102 malignant serous effusions with histologically confirmed diagnoses were immunostained with monoclonal antibody to E-cadherin by LSAB method. E-cadherin expression was identified in only 2 benign reactive serous effusions(5%) whereas 91 malignant serous effusions(89%) expressed E-cadherin The differences in immunostaining for E-cadherin between reactive and malignant serous effusions were statistically significant(p < 0.001). The sensitivity and specificity of the E-cadherin immunostaining for carcinoma cells were 89% and 95%, respectively. In conclusion, E-cadherin is a useful diagnostic adjunct for differentiation between reactive mesothelial and carcinoma cells in serous effusions.

• 대한의생명과학회지
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• 제8권3호
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• pp.107-113
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• 2002

Enterohemorrhagic Escherichia coli (EHEC) strains of serotype O157:H7 have been shown to colonize the intestinal epithelial cell by the attaching and effacing (AE) mechanism. The AE lesion is mediated by an intimin, of which production and expression are controlled by a 3-Kb eaeA gene located EHEC chromosomal DNA. If the eaeA gene is mutated, EHEC O157:H7 strains lose capacity of adhesion to intestinal epithelial cells. In this study, a 891 bp of the 3'-end region of a gamma intimin was amplified by polymerase chain reaction (PCR). The PCR product was inserted into pSTBlue-1 cloning vector and transformed into DE3 (BL21) competent cell. After plasmid mini-preparation and restriction enzyme digestion of eaeA/891-pSTBlue-1 vector, target eaeA gene was re-inserted into pET-28a expression vector and was transformed. Then the expression of recombinant eaeA/891 (891 bp) gene was induced by isopropyl-$\beta$-D-thiogalactopyranoside (IPTG). The expression of the 40-KDa recombinant protein was identified in SDS-PAGE and confirmed by immunoblotting using the His.Tag$^{\circledR}$ and T$_{7}$.Tag$^{\circledR}$ monoclonal antibody. This recombinant protein expressed by eaeA gene could be applied in further studies on the mechanisms of E. coli O157:H7 infection and the development of recombinant vaccine.

• Journal of Chest Surgery
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• 제34권9호
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• pp.729-732
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• 2001

49세 남자 환자가 객혈을 주소로 입원하였다. 단순 흉부방사선 소견과 흉부단층촬영 소견에서 우측 폐첨부에 연부조직 덩어리가 있는 큰 공동이 관찰되었으며 또한 혈청진균검사상 국균증에 대해 양성이었다. 폐국균종 의심하에 폐우상엽절제술을 계획하였으나 폐첨부와 종격동 흉막의 심한 유착으로 페엽절제술이 불가능하여 공동절개술과 전거근을 이용한 근육충진술을 동시에 시행하였다. 환자는 수술직후 객혈이 멈추었고 특별한 합병증없이 술후 20일만에 퇴원하였으며 8개월간 추적관찰 중에 객혈의 소견은 없었다.

• 농업생명과학연구
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• 제46권2호
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• pp.107-114
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• 2012

The present study evaluated the potential use of immunoglobulin prepared from egg yolk of chickens immunized with Escherichia coli K88 (IgY-Ec) in the control of E. coli K88 infection in RAW 264.7 murine macrophage. The binding activity of IgY-Ec against E. coli K88 surface protein was more specific and increased than control IgY. In infection assay of E. coli in macrophage, the specific IgY-Ec to E. coli K88 remarkably inhibited the phagocytic activity comparing to nonspecific IgY (p<0.001). In adherence assay, bacterial adhesion on macrophage cells was definitely reduced by preincubation of IgY-Ec compared with nonspecific IgY (p<0.05). These findings suggested that IgY-Ec have the protective effects against pathogens and IgY-based diets may have potential benefits for preventing or treating various infections in domestic animals.

• Journal of Korean Neurosurgical Society
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• 제29권4호
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• pp.477-484
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• 2000

Objective : Despite advances in the understanding of tumor biology and the tumor immunology, there has been no effective treatment. The Intercellular adhesion molecule-1(ICAM-1) has been shown to be important in interaction involving cells of the immune system and to be upregulated in a number of cell culture systems by cytokines, including immune interferon($IFN-{\gamma}$) and tumor necrosis $factor-{\alpha}$($TNF-{\alpha}$). ICAM-1 has been identified as one of the ligands for lymphocyte function-associated antigen-1(LFA-1). The effectiveness of various cytokines to ICAM-1 induction on cultured human glioblastoma cell lines and potential efficacy of immunotherapy were studied. Method : Human glioblastoma cell lines, U-251 MG, U-373 MG were trypsinized and suspended at $1{\times}10^5cells/ml$ and grown on 8 well chamber slide, the cells were incubated in 0.3ml medium alone or medium containing $IFN-{\gamma}$(1000U/ml) or $TNF-{\alpha}$(250U/ml) or $IFN-{\gamma}$ plus $TNF-{\alpha}$ for 6, 12, 24, 48 and 72 hours. The coverslip were then removed and stained with a 1/30 dilution of anti-ICAM-1 antibody. Result : Surface antigen expression of ICAM-1 was increased by incubating glioblastoma cell lines with $IFN-{\gamma}$ and $TNF-{\alpha}$. Combined effect of $IFN-{\gamma}$ and $TNF-{\alpha}$ has induced more ICAM-1 expression on glioblastoma cell lines. Upregulation of ICAM-1 expression in an established glioblastoma cell line was of greater magnitude and more rapid following incubation with $IFN-{\gamma}$ plus $TNF-{\alpha}$. Surface antigen expression of ICAM-1 was increased for up to 48 hours after cytokine treatment on both cell lines(p<0.05). There was no difference on both cell lines(p>0.05). Conclusion : The results of the present study indicate that ICAM-1 expression in glioblastoma cell lines, U-251 MG and U-373 MG, are induced and enhanced after treatment with $IFN-{\gamma}$ and $TNF-{\alpha}$. Combined effect of $IFN-{\alpha}$ and $TNF-{\gamma}$ is stronger and more rapid than $IFN-{\gamma}$ or $TNF-{\alpha}$ alone.