• 수산해양교육연구
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• 제27권6호
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• pp.1656-1664
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• 2015

고삼 뿌리 열수 추출물을 농도별로 넙치 사료에 흡습시켜 12주 동안 어류에 급여 후 비특이적 면역 및 질병저항성에 대한 효과를 조사하였다. 고삼 추출물의 첨가는 어류 성장에는 아무런 영향을 미치지 않았다. 혈액화학적인 조사 결과에서도 고삼 첨가로 인한 넙치의 생리에는 아무런 영향을 나타내지 않았다. 라이소자임 활성은 0.05%의 고삼 추출물 농도로 흡습시킨 사료 그룹에서 라이소자임 활성이 증가됨을 알 수 있었다. 백혈구의 식세포 활성은 0.05%의 고삼 추출물로 흡습시킨 사료 그룹에서 유의적으로 증가되었다. 병리학적 결과를 통하여 고삼 추출물을 농도별로 넙치에 투여하였을때 병리조직학적 영향이 없음을 알 수 있었다. 에드와드균의 인위감염에 의한 질병저항성은 0.05%의 고삼 추출물로 흡습시킨 사료 그룹에서만 상대생존율이 높게 나타났다. 이상의 결과를 종합하여 볼 때 고삼뿌리 열수 추출물은 넙치에 대한 비특이적 면역력 증강 및 질병 저항성 증강에 효과가 있는 것으로 사료되었다.

• IMMUNE NETWORK
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• 제5권2호
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• pp.124-129
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• 2005

Background: CM1 (centrocyte/-blast marker 1) is originally defined as a germinal center B cell marker. It is known that CM1 plays a critical role on B cell development in germinal center. In addition, we have found that CM1 is expressed on lymphoma cell lines, such as Raji, Ramos and IM-9. This means that CM1 might be served as a tumor marker as well. In the present study, we examined the expression of CM1 on the surface of the other tumors and the possibility of the development of tumor screening ELISA kit by using CM1. Methods: First, we have examined the expression of CM1 on stomach cancer and hepatoma, which are predominantly (discovered) occurred in Korean, by flow cytometry analysis. After purifying of CM1 antigen from Raji and Ramos, the optimal ELISA condition was determined. And then we compared the level of CM1 between normal individuals and cancer patients by ELISA. To decrease the non-specific binding of anti-CM1 mAb with serum components except CM1 and to enhance the diagnostic accuracy, albumin depletion spin column was used. Results: CM1 was highly expressed on stomach cancer and hepatoma cell lines. In addition, we have also confirmed the increased CM1 expression on cancer patients. The difference of CM1 expression between normal individuals and cancer patients were more clearly observed, after deletion of serum albumin by using albumin depletion spin column. Conclusion: Based on the results from this study, CM1 might be a useful molecule for the early diagnosis of cancer. In addition, further studies for the increase of ELISA sensitivity and appropriate albumin depletion methods should be needed.

• 한국수정란이식학회지
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• 제33권3호
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• pp.129-137
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• 2018

Acute vascular rejection has been known as a main barrier occurring in a xenograted tissue of alpha 1,3-galactosyltransferase knock-out (GalT KO) pig into a non-human primate (NHP). Adenosine which is a final metabolite following sequential hydrolysis of nucleotide by ecto-nucleotidases such as CD39 and CD73, act as a regulator of coagulation, and inflammation. Thus xenotransplantation of CD39 and CD73 expressing pig under the GalT KO background could lead to enhanced survival of recipient NHP. We constructed a human CD39 and CD73 expression cassette designed for endothelial cell-specific expression using porcine Icam2 promoter (pIcam2-hCD39/hCD73). We performed isolation of endothelial cells (pAEC) from aorta of 4 week-old GalT KO and membrane cofactor protein expressing pig ($GalT^{-MCP/-MCP}$). We were able to verify that isolated cells were endothelial-like cells using immunofluorescence staining analysis with von Willebrand factor antibody, which is well known as an endothelial maker, and tubal formation assay. To find optimal condition for efficient transfection into pAEC, we performed transfection with GFP expression vector using four programs of nucleofection, M-003, U-023, W-023 and Y-022. We were able find that the program W-023 was optimal for pAEC with regard to viability and transfection efficiency by flow cytometry and fluorescent microscopy analyses. Finally, we were able to obtain $GalT^{-MCP/-MCP}/CD39/CD73$ pAEC expressing CD39 and CD73 at levels of 33.3% and 26.8%, respectively. We suggested that pACE isolated from $GalT^{-MCP/-MCP}$ pig might be provided as a basic resource to understand biochemical and molecular mechanisms of the rejections and as an alternative donor cells to generate $GalT^{-MCP/-MCP}/CD39/CD73$ pig expressing CD39 and CD73 at endothelial cells.

• 한국염색가공학회지
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• 제30권2호
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• pp.117-129
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• 2018

Cellulose has been widely applied into various medical fields including scaffolding, tissue engineering and tissue formation. In this study, we manufactured cellulose medical fiber from Styela clava tunics(SCT-CS) and analyzed the tensile strength, elongation at break, fluid uptake and surface morphology. And then, the biocompatibility and toxicity of SCT-CS were measured in Sprague-Dawley(SD) rats after the implantation for 30, 60 and 90 days. The level of tensile strength and fluid uptake were lower in SCT-CS than chromic catgut(CCG), while elongation at break level were maintained the higher in SCT-CS. Also, the roughness with pronounced surface patterns as a result of in vivo degradation was significantly greater in CCG than this of SCT-CS although these levels gradually appeared with time in both groups. After implantation for 90 days, SCT-CS and CCG was successfully implanted around muscle of thigh without any significant immune response. Furthermore, no significant alterations were measured in serum parameters and the specific pathological features induced by most toxic compounds for liver and kidney toxicity. Therefore, these results suggest that SCT-CS showing good biocompatibility and non-toxicity can be successfully prepared from cellulose powder of SCT as well as has the potential for use as a powerful biomaterial for medical sutures.

• Clinical and Experimental Reproductive Medicine
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• 제29권3호
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• pp.179-186
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• 2002

Objective: Lectins are cell-agglutinating and sugar specific proteins or glycoproteins of non-immune origin that precipitate glycoconjugates having saccharides of appropriate complementarity. Because of these properties, plant lectins have been used to help characterize the carbohydrate moieties of glycoproteins in the zona pellucida (ZP) of several mammalian species including pigs. Treatment of oocytes with various lectins blocks sperm binding to the ZP in various mammalian species. This study was undertaken to examine the distribution of sugar residues in the ZP of pig oocytes matured in vitro and the ability of spermatozoa to bind to ZP and in vitro penetration in oocytes treated with fluorescein isothiocyanate (FITC)-labelled lectins. Materials and Methods: The lectins of Banderiaea simplicifolia (BS-II, bind to $\beta$-D-N-acetylglucosamine), Canavalin ensiformis (Con A, bind to $\alpha$-D-Mannose), Lens culinaris (LCA, bind to a-D-Mannose), Ricinus communis (RCA-I, bind to $\beta$-D-Galactose) and Ulex europaeus (UEA-I, bind to $\alpha$-L-Fucose) were examined for spermatozoa penetration, binding capacity to ZP and distribution of lectins. Results: The penetration rates were significantry (p<0.05) higher in control oocytes (63%) than those treated with all lectins, but penetration rates ($40{\sim}49%$) were simililar in group treated with lectins. The incidence of monospermy was similar in oocytes untreated and UEA-I, but it was higher in oocytes treated with BS-II, Con A, RCA-I and LCA. The porcine oocytes cultured for 48 h in TC-199 medium were freed from cumulus cells and treated for 30 min with fluorescein isothiocyanate-labelled lectins. When examined under fluorescein illumination, higher (p<0.001) proportions of oocytes showed fluorescein of zona pellucida after treatment with Con A (93%), LCA (93%) and RCA-I (100%) than BS-II (37%) and UEA-I (50%). All of the oocytes treated with RCA-I exhibited strong fluorescein in the outer region of the zona pellucida while those treated with LCA exhibited strong fluorescein throughout the zona pellucida. BS-II bounded mainly to the outer region and UEA-I bounded mainly to the inner region of the zona pellucida, with either strong or weak fluorescein. At 120 min after insemination in vitro, fewer spermatozoa were bound to the zona pellucida of the oocytes treated with BS-II, Con-A and RCA-I. Of the lectins, Con A most inhibited sperm binding. Conclusions: These results suggest that $\beta$-D-Galactose residues in the porcine zona pellucida may act as primary sperm receptors and inducers of the sperm acrosome reaction and these sugar residues may be involved in the block to polyspermy.

• 대한의생명과학회지
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• 제5권1호
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• pp.1-10
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• 1999

본 연구에서는 혈장이나 양수에 있는 $\alpha$-fetoprotein (AFP)을 인식 할 수 있는 모노클로날 항체를 제조하고, 모노클로날 항체를 이용한 효소면역분석법을 개발하고자 하였다. 양수로부터 얻은 AFP를 쥐에 주사한 후 비장을 분리하여 종양세포 (Sp2/O-Ag-14)와 융합하였고, 하이브리도마 기술을 이용하여 모노클로날 항체를 제조하였다. 모노클로날 항체를 클로닝하였으며, 생성된 항체를 MabF22로 명명하였고, IgG1 중사슬과 k 경사슬의 isotype을 나타냈다. 또한 immunoblotting 방법과 ELISA로 특이도를 조사한 결과 모노클로날 항체는 AFP와만 반응하였고, 결합 친화상수는 0.8$\times$$10^{-10}$M이었다. 두 종류의 효소면역분석법 -경쟁적 또는 비경쟁적 분석 -을 이용하여 항체의 효용성을 조사하였으며, 두 방법 모두 AFP와 농도에 비례하여 반응하였다. 따라서 본 연구에서 생산된 모노클로날 항체는 연구목적으로 뿐만 아니라 AFP 농도를 측정하기 위한 면역진단시약의 개발에도 유용할 것으로 생각된다.

• Parasites, Hosts and Diseases
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• 제30권1호
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• pp.43-48
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• 1992

폐흡충(Paragonimus westermani) 피낭유충을 마우스에 감염시킨 후 in vitro상에서 비장 림프구에 폐흡충 항원, phytohemagglutinin(PHA) 및 혈청 첨가시 림프구 아세포화 반응을 관찰하였다. PHA로 자극시킨 감염 마우스의 비장 림프구는 비감염군에 비해 감염 1 주 후에 평균 53.6%의 유의한 증식 억제를 관찰하였고, 폐흡충 성충항원으로 자극시킨 경우 감염 마우스의 비장 림프구의 증식은 감염 1 주 후부터 6 주까지 비감염군에 비해 중가되었으며 특히 감염 1 주와 4 주 후에는 각각 평균 200.9% 및 280.2%의 유의한 수준으로 증가되었다. 또한 피낭유충 항원으로 감염 마우스의 비장 림프구를 자극시에도 비감염군의 림프구의 증식에 비해 전반적으로 증가되었으며 특히 감염 4 주 째에는 평균 180.5%의 유의한 수준으로 증가하였다. 폐홉충에 감염된 마우스의 감염 4 주 째 혈청을 PHA로 자극시킨 비감염 마우스 비장 림프구에 첨가하였을 때 유의한 수준의 증식의 억제를 관찰하였다. 이러한 결과들로 보아 폐흡충 감염 마우스에서 림프구가 세포 매개성 면역에 관여하며 감염 혈청이 PHA에 대한 T 림프구의 증식을 억제함을 알 수 있었다.

• 수산해양교육연구
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• 제28권6호
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• pp.1715-1723
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• 2016

복합 생약재 열수추출물(6종 혼합)을 농도별 기간별로 사료에 흡습시켜 12주 동안 넙치에 급여 후 비특이적 면역 및 질병저항성에 대한 효과를 조사하였다. 복합 생약재 열수추출물의 첨가는 어류 성장에는 아무런 영향을 미치지 않았으며 혈액생화학적인, 결과도 넙치의 생리에는 아무런 영향을 미치지 않았다. 라이소자임 활성은 0.01%의 복합 생약재 열수추출물 농도로 흡습시킨 사료 그룹에서 라이소자임 활성이 증가됨을 알 수 있었다. 복합 생약재 열수추출물의 투여에 따른 넙치 장기의 병리조직에 영향이 없음을 알 수 있었다. 에드와드균의 인위감염에 의한 질병저항성은 0.01% 복합 생약재 열수추출물 농도에서 상대생존율이 높게 나타났다. 이상의 결과를 종합하여 볼 때 넙치에 복합 생약재 열수추출물을 투여시에는 어류의 비특이적 면역력 증강 및 질병 저항성 증강을 나타내는 것으로 사료되었다.

• 대한핵의학회지
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• 제24권1호
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• pp.29-36
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• 1990

Most of the diagnostic methods currently used for the detection of neoplastic masses provide indirect evidence. To obtain greater specificity in the interpretation of neoplasias by in vivo methods, the immunological approach appears to be most promising. Two problems that interfered with progress in this field were the lack of tumor specific antigen and the lack of well-defined and reproducible antibodies. To improve the sensitivity and specificity of radioimmunoscintigraphy as a technique for tumor localization, the use of monoclonal antibodies, fragments of antibodies and single photon emission computerized tomography (SPECT) are reasonable. The obvious advantages of monoclonal antibodies are their homogeneity, their specificity for the immunizing antigen and the reaction with a single determinant-thus no large immunecomplexes with antigen are formed. Monoclonal antibody technique has recently provided an opportunity to reevaluate the role of nuclear medicine for the diagnosis of malignant diseases by using the immunological approach. Out first results by means of radioimmunoscintigraphy of CEA and CA 19-9 producing tumors using a cocktail of fragments F $(ab')_2$, of mocolonal antibodies to CA 19-9 and CEA labeled with $^{131}I$ (IMACIS-1) are reported. The aims of this investigation was to evaluate the role of immunoscintigraphy in patients with colorectal and other cancers for diagnosis of local recurrences and metastasis. This report contains results of the first 8 colorectal and pancreas cancer patients with the elevation of the level of serum CEA and/or CA 19-9. IMACIS-1 was injected intravenously during 30 minutes in 100 ml saline solution after skin test. Planar scintigrams were recorded 3, 5 and 7 days after the injection of the IMACIS-1. Anterior, lateral and posterior views of the liver as well as anterior and posterior views of the pelvis were obtained in each patients as an $^{131}I-antibody$ image. We were able to localize exactly the malignant process with the double-nuclide double-compound $^{99m}Tc\;^{131}I$ (Tc+l) scintigrams. In Tc & I double-nuclide scintigraphy, computer subtraction display provided more clear localization of the tumor. We compared the results of radioimmunoscintigraphy with CT, ultrasonograms, conventional scintigrams. The results were as follows: 1) The sensitivity and specificity of radioimmunoscintigraphy using the fragments $F(ab')_2$ of the cocktails of CEA and CA 19-9 monoclonal antibodies were 80% and 100% respectively. 2) Tumor detection rate was not proportionated to the level of serum tumor markets. 3) Second tracer technique was essential for tumor localization as an anatomic landmark using double-nuclide scintigraphy. 4) A slow infusion of the antibodies was necessary to prevent the formation of large immune complexes. 5) Tumor/non-tumor radioactivity was most elevated at 7 days delayed imaging. 6) Using planar scintigraphic technique of $^{131}I$ labeled monoclonal antibodies are possible for imaging most of the tumors.

• 생명과학회지
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• 제31권8호
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• pp.771-777
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• 2021

인간 내생 레트로바이러스(Human Endogenous Restrovirus, HERV)는 수백만년전 인간의 유전체에 삽입되었으며 이후 오랜 세월을 거치며 재조합, 결실 및 돌연변이 등 여러 원인에 의해 더 이상 활성화된 바이러스로 역할을 하지 못하고 감염되지 않는다. 하지만 HERV는 최근 연구들은 HERV 유래 인자들이 실제 생리현상 및 암을 비롯한 특정 질환에 관여 하고 있다는 것을 보여 주었다. HERV와 관련된 여러가지 생리 현상 중 염증반응에 초점을 맞추어 고찰해 볼 필요가 있다. HERV는 류마티스, 다발성 경화증, 근위축성 측삭경화증, 쇼그렌 증후군 같은 자가면역질환을 비롯한 여러 염증질환에 직접적으로 관여하는 것으로 보고 되고 있다. HERV의 염증 조절 기작으로는 HERV 유래 인자들이 비특이적 선천성 면역과정을 유발할 가능성과 HERV 유래의 RNA와 단백질이 특정 수용체를 통해 선택적 신호전달기작을 유발할 가능성을 고려 할 수 있다. 하지만 어떠한 방식으로 잠재되어 있던 HERV가 염증반응에서 활성화 되는지 또한 HERV와 관여된 인자들과 신호기작들이 어떠한 것들이 있는지 등 HERV의 인자들이 염증반응을 조절하는 기작에는 아직 많은 것들이 밝혀지지 않아 질병 발병에 대한 연구에 어려움이 있는 실정이다. 본 리뷰에서는 HERV 관련 자가 면역질환을 소개하고 염증반응 조절 기작에 관한 HERV의 분자수준에서의 작용 메커니즘을 제안 하고자 한다.