• Title/Summary/Keyword: CD34 monoclonal antibody

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CD34 Monoclonal Antibody-Immobilization on Polyurethane Surface by Poly(PEGA-co-BMA) Coating (PEGA/BMA 공중합체의 코팅을 통해 CD34 단일클론항체가 고정화된 폴리우레탄 표면)

  • Joung, Yoon-Ki;Hwang, In-Kyu;Park, Ki-Dong
    • Polymer(Korea)
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    • v.33 no.6
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    • pp.602-607
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    • 2009
  • A polyurethane (PU) surface enabling in vivo endothelialization via endothelial progenitor cell (EPC) capture was prepared for cardiovascular applications. To introduce CD34 monoclonal antibody (mAb) inducing EPC adhesion onto a surface, poly (poly (ethylene glycol) acrylate-co-butyl methacrylate) and poly (PEGA-co-BMA) were synthesized and then coated on a surface of PU, followed by immobilizing CD34 mAb. $^1H$-NMR analysis demonstrated that poly(PEGA-co-BMA) copolymers with a desired composition were synthesized. Poly(PEGA-co-BMA)-coated PU was much more effective for the immobilization of CD34 mAb, comparing with PEG-grafted PU prepared in our previous study, as demonstrated by that surface density and activity of CD34 mAb increased over 32 times. Physico-chemical properties of modified PU surfaces were characterized by X-ray photoelectron spectroscopy (XPS), water contact angle, and atomic force microscopy (AFM). The results demonstrated that the poly(PEGA-co-BMA) coating was effective for CD34 mAb immobilization and feasible for applying to cardiovascular biomaterials.

Characterization of a Novel Monoclonal Antibody (27H2) Recognizing Human CD34 Class III Epitope

  • Hong, Kwon-Pyo;Kang, Sung-Hee;Lee, Kyoung-Mee;Ji, Gil-Yong;Yoon, Sang-Soon;Kim, Jong-Suk;Son, Bo-Ra;Lee, Dong-Geun;Lee, Ok-Jun;Song, Hyung-Geun
    • IMMUNE NETWORK
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    • v.10 no.6
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    • pp.239-246
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    • 2010
  • Background: Monoclonal antibodies (mAbs) recognizing Class III epitope of CD34 are essential for flow cytometric diagnosis of leukemia. Methods: 27H2 mAb was developed from a mouse alternatively immunized with human acute leukemia cell lines, KG1 and Molm-1. Using flow cytometric analysis of various leukemic cell lines and peripheral blood, immunohistochemical study of frozen tonsil, we characterized 27H2 mAb. Antigen immunoprecipitated with 27H2 mAb immunobloted with anti-CD34 mAb. A case of bone marrow sample of acute lymphoblastic leukemia (ALL) patient was obtained at CBNU Hospital. For epitope identification enzyme treatment with neuraminidase and O-sialoglycoprotein endopeptidase (OSGE) and blocking assay with known classIII mAb (HPCA-2) were done. Results: Only KG1 and Molm-1 revealed positive immunoreactivity. Immunohistochemical staining disclosed strong membranous immunoreactivity on high endothelial venules. Antigen immunoprecipitated by 27H2 mAb showed approximately 100 kDa sized band immunoblotted with anti-CD34 under non-reducing conditions. Epitope recognized by 27H2 mAb disclosed resistancy to both neuraminidase and OSGE treatment and completely blocked with known class III mAb preincubation. CD34 positive leukemic cells in BM of pre B cell ALL patient detected by FITC-conjugated 27H2 and HPCA-2 were identified with similar sensitivity. Conclusion: A novel murine mAb recognizing class III epitope of human CD34 with high affinity, which is useful for flow cytometric diagnosis of leukemia, was developed.

Enhancement of cell-mediated immunity by administration of plasma protein in pigs 1. Proportion of leukocyte subpopulations and cells expressing adhesion molecules in peripheral blood (돼지에서 plasma protein에 의한 세포성면역 증진효과에 관한 연구 1. 혈액내 백혈구 아군 및 세포별 분포율)

  • Yang, Chang-kun;Kim, Soon-jae;Moon, Jin-san;Jung, Suk-chan;Park, Yong-ho
    • Korean Journal of Veterinary Research
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    • v.34 no.2
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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.

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Flow cytometric immunophenotyping of canine adipose-derived mesenchymal stem cells (ADMSCs) and feline ADMSCs using anti-human antibodies

  • Ko, Minho;Lee, Kwon Young;Kim, Sae Hoon;Kim, Manho;Choi, Jung Hoon;Im, Wooseok;Chung, Jin Young
    • Korean Journal of Veterinary Research
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    • v.58 no.1
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    • pp.33-37
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    • 2018
  • Various trials have been conducted to develop therapies for serious untreatable diseases. Among these, those using stem cells have shown great promise, and adipose-derived mesenchymal stem cells (ADMSCs) are easier to obtain than other types of stem cells. Prior to clinical trials, characterization of ADMSCs with monoclonal antibodies should be performed. However, it is difficult to use species-specific antibodies for veterinarians. This study was conducted to confirm the panel of human antibodies applicable for use in immunophenotypic characterization of canine adipose-derived stem cells and feline ADMSCs extracted from subcutaneous adipose tissue collected during ovariohysterectomy. For flow cytometric immunophenotyping, the third passages of canine ADMSC and feline ADMSC and human CD31, CD34, CD42, CD44, CD62 and CD133 antibodies were used. Of these, CD133 reacted with canine cells (3.74%) and feline cells (1.34%). CD133 is known as a marker related with more primitive stem cell phenotype than other CD series. Because this human CD133 was not a species-specific antibody, accurate percentages of immunoreactivity were not confirmed. Nevertheless, the results of this study confirmed human CD133 as a meaningful marker in canine and feline ADMSCs.

Enzymatic study on lymphocyte CD38 (임파구 CD38의 효소학적 연구)

  • Park, Hyang Ran;Kim, Jong Ju;An, Nyeon Hyoung
    • Korean Journal of Clinical Pharmacy
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    • v.8 no.1
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    • pp.29-34
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    • 1998
  • Murine CD38 is a 42 kDa type II glycoprotein expressed on cell surface of both B and T lymphocytes. CD38 is a multifunctional enzyme that catalyzes the formation and hydrolysis of cyclic adenosine diphosphoribose (cADPR): ADP-ribosyl cyclase activity of CD38 catalyzes the formation of cADPR from NAD and cADPR hydrolase activity of CD38 catalyzes the hydrolysis of cADPR to ADP-ribose (ADPR). And also, CD38 has the catalytic activity of NAD glycohydrolase (NADase) which catalyzes the hydrolysis of catalyzes the formation and hydrolysis of cyclic adenosine diphosphoribose (cADPR): ADP-ribosyl cyclase activity of CD38 catalyzes the formation of cADPR from NAD to ADPR. In this study, we attempted to purify CD38 from mouse lymphocytes by using the immobilized anti-CD38 monoclonal antibody. The single step immuno-affinity column chromatography resulted in homogeneous purification, showing a single protein of 42 kDa on a SDS polyacrylamide gel. We have investigated the effects of various inhibitors on the enzyme activities of the purified CD38. Cibacron blue (0.5 mM) inhibited all three enzyme activities of CD38, NADase, ADP-ribosyl cyclase and cADPR hydrolase activities. ADPR (2 mM) showed inhibitory effect on both cADPR hydrolase activity and NADase, but not on ADP-ribosyl cyclase activity. However, ATP (2 mM) inhibited only cADPR hydrolase activity. $Zn^{2+}$ (1 mM) showed similar inhibitory effect as that of ADPR, but activated cyclase activity These results suggest that CD38 has three different catalytic activity domains which might be differentially regulated by their specific inhibitors.

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Changes in Immunogenicity of Preserved Aortic Allograft (보존된 동종동맥편 조직의 면역성 변화에 관한 연구)

  • 전예지;박영훈;강영선;최희숙;임창영
    • Journal of Chest Surgery
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    • v.29 no.11
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    • pp.1173-1181
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    • 1996
  • The causes of degenerative changes in allograft cardiac valves are not well known to this day. Today's preserved allografts possess highly viable endothelial cells and degeneration of allografts can be facilitated by immune reaction which may be mediated by these viable cells. To test the antigenicity of endothelial cells, pieces from aortic wall were obtained from fresh and cryo-preserved rat allograft. Timings of sampling were prior to sterilization, after sterilization, after 1, 2, 7, 14 days of fresh preservation and cryopreservation. Endothelial cells were tested by immunohistochemical methods using monoclonal antibodies to MHC class I(MRC OX-18), class II(MRC OX-6) and ICAM-1 antigens. After transplantation of each group of aortic allograft at the subcutaneous layers of rats, population of CD4$^{+}$ T cell and CD8$^{+}$ T cell were analyzed with monoclonal antibodies after 1, 2, 3, 4, 6 and 8 weeks. MHC class I expression was 23.95% before preservation and increased to 35.53~48.08% after preservation(p=0.0183). MHC Class II expression was 9.72% before preservation and 10.13~13.39% after preservation(P=0.1599). ICAM-1 expression was 15.02% before preservation and increased to 19.85~35.33% after preservation(P=0.001). The proportion of CD4$^{+}$ T-cell was 42.13% before transplantation. And this was 49.23~36.8% after transplantation in No treat group (p=0.955), decreased to 29.56~32.80% in other group(p=0.0001~0.008). In all the groups, the proportion of CD8$^{+}$ T-cell increased from 25.57% before transplantation to 42.32~58.92% after transplantation(p=0.000l~0.0002). The CD4$^{+}$/CD8$^{+}$ ratio decreased from 1.22~2.28 at first week to 0.47~0.95 at eighth week(p=0.0001). The results revealed that the expression of MHC class I and ICAM-1 in aortic allograft endothelium were increased but that of MHC class II were not changed, despite the different method of preservation. During 8 weeks after transplantation of aortic allograft, the subpopulations of CD4$^{+}$ T cell were not changed or only slightly decreased but those of CD8$^{+}$ T cell were progressively increased.ely increased.

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Stimulation of Macrophage Differentiation by Bu-Zhong-Yi-Qi- Tang Extract (보중익기탕 추출물의 대식세포 분화 유도 효과)

  • Kang, Hye-In;Shin, Sung-Hae;Cho, Young-Sook;Cho, Sung-Ki;Byun, Mung-Woo;Yee, Sung-Tae
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.3
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    • pp.330-335
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    • 2005
  • We have investigated the effects of Bu-Zhong-Yi-Qi- ang on the differentiation of murine bone marrow cells in methylcellulose culture. GM-CSF and IL-3 supported primarily the formation of granulocyte/mac-rophage colony formation. However, the addition of Bu-Zhong-Yi-Qi-Tang extract yielded a significant increase in the numbers of colonies and differentiated cells in the presence of GM-CSF and IL-3. We have analyzed CD11b (Mac-1) expression of differentiated cells from bone marrow by staining with monoclonal anti-CD11b antibody. The majority of colony-forming cells were in CD11b/sup +/ population. Also Bu-Zhong-Yi-Qi-Tang extract promoted the production of IL-6 and nitric oxide by macrophages. These results demonstrate that extract of Bu-Zhong-Yi-Qi-Tang, a prescription of traditional oriental medicine, is effective in supporting macrophage potential of the primary colonies.

Infliximab: The Benefit for Refractory Crohn Disease and Top-down Induction Therapy in Severe Crohn Disease (Infliximab: 불응성 크론병 치료법으로서의 유용성과 Top-down 관해 유도 요법으로서의 가능성)

  • Lee, Jee-Hyun;Lee, Hae-Jeong;Park, Sung-Eun;Choe, Yon-Ho
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.11 no.1
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    • pp.28-35
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    • 2008
  • Purpose: The aim of this study is to report the efficacy of infliximab, a monoclonal antibody directed against tumor necrosis factor alpha which is used for both treatment of refractory pediatric Crohn disease (CD) and induction of remission. Methods: Among pediatric patients who were diagnosed with CD at Samsung Medical Center between March 2001 and August 2007, a total of 16 patients were given infliximab to treat conventional therapyresistant refractory CD and severe active CD for induction of remission. Patients needing maintenance therapy were treated with an infliximab infusion every 8 weeks, and fistulizing CD patients occasionally received the infusion upon the condition that a fistula developed. The efficacy of treatment was assessed by comparing the Pediatric Crohn Disease Activity Index (PCDAI), Hct, ESR, CRP, and serum albumin levels using paired t-test. Results: The male/female ratio was 13:3, and the median age was 13 years (range, 21 months~15 years). The patients included 7 cases of therapy-resistant refractory CD, 7 cases of severe active CD, and 2 cases of fistulizing CD. Mean PCDAI before infliximab therapy was 34.19${\pm}$14.96, and mean follow-up PCDAI within 2 to 4 weeks after the last infusion was significantly lower, at 6.88${\pm}$10.31 (p=0.000). Hematological markers such as ESR (p=0.000), serum albumin (p=0.016), and CRP (p=0.009) also improved significantly after infusion. Remission was achieved in 2 of 4 patients refractory to conventional therapy. Among 3 steroid-dependent patients, 2 were able to discontinue steroid therapy, and dose reduction was possible in 1 patient. Remission after top-down therapy without prior use of other immunomodulators was achieved in 6 weeks in all 7 of the patients who had severe CD. Nine of ten refractory fistulizing CD patients also showed improvement after infliximab therapy. Conclusion: Infliximab was effective in pediatric refractory CD for induction of remission and maintenance therapy, as well as in severe CD for top-down induction therapy. Furthermore, infliximab has contributed to steroid cessation and dose reduction. Long-term follow-up evaluation is needed to determine safety and efficacy of infliximab in the future.

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Effect of Humidified High Flow CO2 Gas Insufflation on the Coronary Endothelium (가습화한 고유량의 이산화탄소가스 통기가 관상동맥 내피세포층에 미치는 영향)

  • 최재성;김준성;서정욱;김기봉
    • Journal of Chest Surgery
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    • v.37 no.2
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    • pp.131-138
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    • 2004
  • High-flow gas insufflation to get a bloodless field during off-pump coronary artery bypass may have adverse effects on the coronary endothelium. This study was designed (1) to elucidate the effect of carbon dioxide gas insufflations on the coronary endothelium at different flow rates and (2) to assess the protective effect of humidifcation against the coronary endothelial damage. Material and Method: In nine pigs, the left anterior descending coronary artery (LAD) was exposed after a median sternotomy. The LAD was divided into 4 segments and a coronary arteriotomy was made in each LAD segment in the beating heart. The far distal arteriotomy was exposed to room air for 10 minutes and was harvested as a control. Non-humidified carbon dioxide gas at a continuous flow rate of 5 L/min (Group I), humidified carbon dioxide gas at a continuous flow rate of 5 L/min (Group II), and humidified carbon dioxide gas at a continuous flow rate of 10 L/min (Group III) were insufflated for 10 minutes on each coronary arteriotomy site, respectively. After harvesting the coronary segments, hematoxylin-eosin staining, elastic fiber staining, and immunostaining with a CD34 monoclonal antibody were performed to evaluate the depth of endothelial damage and to count the residual endothelial cells, Result: In all three groups (Group I, II, and III), internal elastic laminae were preserved, however, the endothelial layers were significantly damaged by carbon dioxide gas insufflation. The mean percentages of remaining endothelial cells were 20,9$\pm$16.7%, 39.3$\pm$19.6%, and 6.8$\pm$5.3%, in groups I, II, and III, respectively. The percentages of remaining cells were significantly higher in group II than in groups I and III (p=0.008). The percentages of remaining cells were significantly higher in group I than in group III (p=0.008). Conclusions: The harmful effect of carbon dioxide gas insufflation on the coronary endothelium was dependent on the flow rate. The addition of humidification did not protect the coronary endothelium from denudation injury caused by high flow carbon dioxide gas insufflations.

Image-Based Assessment and Clinical Significance of Absorbed Radiation Dose to Tumor in Repeated High-Dose $^{131}I$ Anti-CD20 Monoclonal Antibody (Rituximab) Radioimmunotherapy for Non-Hodgkin's Lymphoma (반복적인 $^{131}I$ rituximab 방사면역치료를 시행 받은 비호지킨 림프종 환자 군에서 종양 부위의 영상기반 방사선 흡수선량 평가와 임상적 의의)

  • Byun, Byung-Hyun;Kim, Kyeong-Min;Woo, Sang-Keun;Choi, Tae-Hyun;Kang, Hye-Jin;Oh, Dong-Hyun;Kim, Byeong-Il;Cheon, Gi-Jeong;Choi, Chang-Woon;Lim, Sang-Moo
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.1
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    • pp.60-71
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    • 2009
  • Purpose: We assessed the absorbed dose to the tumor ($Dose_{tumor}$) by using pretreatment FDG-PET and whole-body (WB) planar images in repeated radioimmunotherapy (RIT) with $^{131}I$ rituximab for NHL. Materials and Methods: Patients with NHL (n=4) were administered a therapeutic dose of $^{131}I$ rituximab. Serial WB planar images alter RIT were acquired and overlaid to the coronal maximum intensity projection (MIP) PET image before RIT. On registered MIP PET and WB planar images, 2D-ROls were drawn on the region of tumor (n=7) and left medial thigh as background, and $Dose_{tumor}$ was calculated. The correlation between $Dose_{tumor}$ and the CT-based tumor volume change alter RIT was analyzed. The differences of $Dose_{tumor}$ and the tumor volume change according to the number of RIT were also assessed. Results: The values of absorbed dose were $397.7{\pm}646.2cGy$ ($53.0{\sim}2853.0cGy$). The values of CT-based tumor volume were $11.3{\pm}9.1\;cc$ ($2.9{\sim}34.2cc$), and the % changes of tumor volume before and alter RIT were $-29.8{\pm}44.3%$ ($-100.0%{\sim}+42.5%$), respectively. $Dose_{tumor}$ and the tumor volume change did not show the linear relationship (p>0.05). $Dose_{tumor}$ and the tumor volume change did not correlate with the number of repeated administration (p>0.05). Conclusion: We could determine the position and contour of viable tumor by MIP PET image. And, registration of PET and gamma camera images was possible to estimate the quantitative values of absorbed dose to tumor.