• Journal of Yeungnam Medical Science
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• 제30권1호
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• pp.21-24
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• 2013

Hemolytic disease in a newborn that causes early jaundice is common. It is often due to the Rh (D) and ABO incompatibility, but rarely due to unexpected antibodies. Among these unexpected antibodies, the anti-$Di^a$Dia antibody rarely occurs. The anti-$Di^a$ antibody was observed in the serum and red-cell eluate of an infant, and in the serum of his mother. The frequency of the appearance of the $Di^a$ antigen in the Korean population is estimated to be 6.4-14.5%. This paper reports a case of hemolytic disease in a newborn associated with the anti-$Di^a$ antibody. A full-term male infant was transferred to the authors' hospital due to hyperbilirubinemia the day after his birth. The laboratory data indicated a hemoglobin value of 11.6 g/dL, a reticulocyte count of 10.6%, a total bilirubin count of 14.4 mg/dL, a direct bilirubin count of 0.6 mg/dL, and a positive result in the direct Coombs' test. Due to the identification of an irregular antibody from the maternal serum, an anti-$Di^a$ antibody was detected, which was also found in the eluate made from the infant's blood. The infant had been treated with phototherapy and intravenous immunoglobulin since the second day after his birth and was discharged due to an improved condition without exchange transfusion. Therefore, in cases of iso-immune hemolytic disease in a newborn within 24 hours from birth who had a negative result in an antibody screening test, the conduct of an anti-$Di^a$ antibody identification test is recommended due to the suspicion of an anti-$Di^a$ antigen, followed by early administration of intravenous immunoglobulin.

• 약학회지
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• 제54권6호
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• pp.500-506
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• 2010

TNF-${\alpha}$ and VCAM-1 play a pivotal role in the pathogenesis of rheumatoid arthritis, and the development of drugs targeting these molecules has extended the therapeutical approaches to rheumatoid arthritis patients. Bispecific antibodies combine the antigen-binding sites of two antibodies within a single molecule and thus they are able to bind to two different epitopes simultaneously. A specific bispecific antibody format termed "Di-diabody" was made for the efficient approach to anti-inflammation. In this study, the DNA vector construct of Di-diabody was built up against two antigens, VCAM-1 and TNF-${\alpha}$. For evaluating this Di-diabody as a bispecific antibody on the efficacy of anti-inflammation, the proteins were analyzed according to each antigen binding affinity and cell based assay related separate molecules. The 7H/Humira Di-diabody produced in this study interacted with its ligands, VCAM-1 and TNF-${\alpha}$, respectively. Also, this antibody exhibited the similar functional activities as compared to 7H-IgG in respect to inhibition of hVCAM-1-induced cell adhesion and Humira-IgG in respect to inhibition of TNF-${\alpha}$ induced cytotoxicity. Further study to elucidate the pharmacological significance of the Di-diabody is warranted using experimental animals.

• 대한임상검사과학회지
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• 제54권4호
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• pp.279-284
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• 2022

환자의 긴급 상황에서 수혈 시 일부 수혈 전 검사를 수행하지 않는다. 수혈 후 항체 선별검사를 실시하여 용혈성 수혈 반응을 일으키는 비예기 항체의 실태를 조사를 비교평가하기 위해 국내·외에 보고된 공시자료를 포함하여 조사하였다. 2014~2016년 P병원에서 항체 선별 검사 68,602건과 항체 동정검사 528건을 선정하였다. 68,602건 중 1,198건(1.74%)이 양성이며 Rh계열 161건(30.49%), Lewis계열 67건(12.69%), 기타(Di (a)등) 28건(5.30%)으로 나타났다. Anti-E는 93건(17.61%), c는 13건(2.46%)이다. 국내·외 공시자료 중 국내 경우는 2007년 이전에는 Anti-E 196건(22.45%), Anti-Le(a) 82건(9.39%) Anti-E+c 60건(6.87%)으로 조사되었지만 2008년 이후에는 Anti-E 107건(17.12%), Anti-E+c 56건(8.96%), Anti-Di (a) 28건(4.48%)으로 조사되었다. 한편 다른 국내의 경우는 S병원(2012~2015년)에는 Anti-E, Anti-Le (a), Anti-E+c이며Anti-E+c이며 D병원(2016~2017년)에는 Anti-E, Anti-D Anti-E+c,Anti-E+c, Anti-C+e로 조사되었다. Saudi의 경우는 Anti-D, Anti-E, Anti-Jk (a)순이며, India 경우는 Anti-M, Anti-N, Anti-Le (a), Anti-D로 조사되었다. 권역별 외상센터 개설 전후시기에 응급수혈요청의 빈도 비교에서는 1.8배 이상이 증가한 것으로 조사되었다. 결론적으로 장기간의 항체분포로 효율성을 검증할 수는 없었지만 수혈의 안전성을 높이고 응급 수혈 상황에서 용혈성 수혈 부작용을 줄일 수 있는 대안을 위해 기초 정보를 제공하고 추가적인 검증연구가 필요할것으로 사료된다.

• 대한임상검사과학회지
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• 제49권4호
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• pp.390-394
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• 2017

비예기항체(unexpected antibody)는 ABO 혈액형 항체와는 달리 존재 여부를 미리 예측할 수 없는 항체로 주로 임신이나 수혈 등에 의해 다른 적혈구 항원에 노출됨으로써 발생하는 면역항체로서, 비예기항체는 해당 항원을 가진 수혈된 적혈구를 파괴하여 급성 및 지연성 용혈성부작용이나 신생아용혈성질환 등의 수혈부작용을 유발할 수 있다. 2014년 1월부터 2016년 12월까지 3년간 제주특별자치도 종합병원에 비예기항체 선별검사가 의뢰된 10,360명의 혈청 검체를 대상으로 하였으며, 의뢰된 비예기항체 선별결과 양성 결과를 보인 87 검체를 대상으로 조사하였다. 비예기항체가 선별되었던 87건 중 비예기항체 동정검사를 수탁검사기관으로 의뢰한 41 검체(0.40%) 대상으로 분석한 결과 비특이항체가 8건(19.51%), 자가항체가 3건(7.32%)이었다. 이들을 제외하고 항체가 동정된 것은 anti-E가 8건(19.51%), anti-E, anti-c 복합항체가 6건(14.63%), $anti-Le^a$, $anti-Le^b$ 복합항체가 3건(7.32%), $anti-Le^a$와 $anti-Le^b$가 각각 2건(4.88%)으로 조사되었다. 단일항체로는 anti-D, $anti-Di^a$, $anti-Fy^b$, $anti-Jk^a$, $anti-Jk^b$, anti-M, anti-P1이 각각 1건(2.44%)로 나타났으며 복합항체로는 anti-C+anti-D, anti-E+anti-c+$anti-Jk^b$가 각각 1건(2.44%)로 조사되었다. 단일항체가 동정된 검체는 19건(46.34%), 복합항체가 동정된 검체는 11건(26.83%)로 나타났다. 제주지역의 종합병원은 제주시와 서귀포시에 위치한 종합병원 7개 기관이 있으며, 모두 비예기항체 선별검사를 시행하고 있으며, 검사방법으로는 모든 검사실에서 Ortho BioVue system을 이용한 원주응집법으로 검사를 시행하고 있다. 하지만 비예기항체 동정검사는 외부수탁기관에 의뢰하고 있는 실정이다. 이번 연구는 최근 3년간 제주도내 종합병원 1개 기관에서의 동정된 비예기항체 빈도와 분포를 분석하였지만, 점차적으로 제주도내 다문화가정과 외국인 근로자가 증가 추세에 있어 향후 제주지역에 분포한 다수의 종합병원에서 동정되는 비예기항체의 빈도와 분포를 비교 분석한다면 의미가 있는 연구가 될 것이라고 사료된다.

• KSBB Journal
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• 제17권1호
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• pp.20-25
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• 2002

We performed a basic experiment for rapid, on-line, real-time measurement of HBsAg by using a surface plasmon resonance biosensor to quantify the recognition and interaction of biomolecules. We immobilized the anti-HBsAg polyclonal antibody to the dextran layer on a CM5 chip surface which was pre-activated by N-hydroxysuccinimide for amine coupling. The binding of the HBsAg to the immobilized antibody was measured by the mass increase detected by the change in the SPR signal. The binding characteristics between HBsAg and its antibody followed typical monolayer adsorption isotherm. When the entire immobilized antibody was interacted, there was no additional, non-specific binding observed, which suggested the biointeraction was very specific as expected and independent of the ligand density. No significant steric hindrance was observed at 17.6 nm/$mm^2$ immobilization density. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the chip surface was linear up to ca. $40\mu\textrm{g}$/mL, which is much wider than that of the ELISA method. It appeared the antigen-antibody binding was increased as the immobilized ligand density increased, but verification is warranted. This study showed the potential of this biosensor-based method as a rapid, simple, multi-sample, on-line assay. Once properly validated, it can serve as a more powerful method for HBsAg quantification replacing the current ELISA method.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권4호
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• pp.309-314
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• 2005

We performed a basic experiment for the rapid, on-line, real-time measurement of hepatitis B surface antigen using a surface plasmon resonance biosensor. We immobilized anti­HBsAg (hepatitis B surface antigen) polyclonal antibody, as a ligand, to the dextran layer on a CM5 chip surface that had previously been activated by N-hydroxysuccinimide. A sample solution containing HBsAg was fed through a microfluidic channel, and the reflecting angle change due to the mass increase from the binding was detected. The binding characteristics between HBsAg and its polyclonal antibody followed the typical monolayer adsorption isotherm. When the entire immobilized antibody had interacted, no additional, non-specific binding occurred, suggesting the immunoreaction was very specific. The bound antigen per unit mass of the antibody was independent of the immobilized ligand density. No significant steric hindrance was observed at an immobilization density of approximately $17.6 ng/mm^2$. The relationship between the HBsAg concentration in the sample solution and the antigen bound to the ligand was linear up to ca. $40{\mu}g$/mL. This linearity was much higher than that of the ELISA method. It appeared the anti­gen-antibody binding increased as the immobilized ligand density increased. In summary, this study showed the potential of this SPR biosensor-based method as a rapid, simple and multi­sample on-line assay. Once properly validated, it may serve as a more efficient method for HBsAg quantification for replacing the ELISA.

• 대한한방내과학회지
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• 제30권2호
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• pp.375-387
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• 2009

Objective : Gyehyuldeung (GHD) has been widely used in oriental medicine for the treatments of cardiovascular and neurological disorders. Thus, its potential facilitatory activity on axonal regeneration was investigated in the rats. Methods: Sprague-Dawley rats were given crush injury at the sciatic nerve and the changes of axon growth after nerve injury on each nerve injury model were investigated with anti-NF-200 antibody, DiI, GAP-43 protein and Cdc2 protein Results : GHD-mediated enhancement of axonal regeneration after crush injury was measured in both qualitative and quantitative ways by immunofluorescence staining with anti-NF-200 antibody and retrograde tracing of fluorescence dye DiI. GAP-43 protein levels were elevated by GHD treatments in the distal injured sciatic nerve and DRG sensory neurons. The neurite outgrowth of DRG sensory neurons was facilitated by GHD treatment when co-cultured with Schwann cells and astrocytes prepared from injured sciatic nerves and injured spinal cord tissues, respectively. It was observed that Cdc2 protein was up-regulated in co-cultured Schwann cells or astrocytes and Cdc2 protein signals were co-localized to a certain extent with those of phospho-vimentin protein. Conclusions : These results suggest that GHD may play a facilitatory role in axonal regeneration by acting on the injured axons and adjacent non-neuronal cells. The current findings may be useful for the development of therapeutic targets through more specific explorations on molecular interactions between herbal components and endogenous factors.

• Molecules and Cells
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• 제26권4호
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• pp.323-328
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• 2008

Eukaryotic cells continuously integrate intrinsic and extrinsic signals to adapt to the environment. When exposed to stressful conditions, cells activate compartment-specific adaptive responses. If these are insufficient, apoptosis ensues as an organismal defense line. The mechanisms that sense stress and set the transition from adaptive to maladaptive responses, activating apoptotic programs, are the subject of intense studies, also for their potential impact in cancer and degenerative disorders. In the former case, one would aim at lowering the threshold, in the latter instead to increase it. Protein synthesis, consuming energy for anabolic processes as well as for byproducts disposal, can be a significant source of stress, particularly when difficult-to-fold proteins are produced. Recent work from our and other laboratories on the differentiation of antibody secreting cells, revealed a regulatory circuit that integrates protein synthesis, secretion and degradation (proteostasis), into cell lifespan determination. The apoptotic elimination - after an industrious, yet short lifetime - of terminal immune effectors is crucial to maintain immune homeostasis. Linking proteostasis to cell death, this paradigm might prove useful for biotechnological purposes, and the design of novel anti-cancer therapies.

• IMMUNE NETWORK
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• 제10권6호
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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.

• IMMUNE NETWORK
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• 제6권2호
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• pp.59-66
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• 2006

Background: CM1 (Centrocyte/-blast Marker I) defined by a mAb developed against concanavalin-A activated PBMC, is expressed specifically on a subpopulation of centroblasts and centrocytes of human germinal center (GC) B cells. Burkitt lymphoma (BL) is a tumor consisting of tumor cells with the characteristics of GC B cell. Previously we reported that CM1 ligation with anti-CM1 mAb induced apoptosis in Ramos $(IgM^{high})$ and Raji $(IgM^{low})$ cells. Methods & Results: In the present study, we observed that CM1 ligation with anti-CM1 mAb induced Fas ligand and Fas expression in Ramos cells, but not in Raji cells. Furthermore, anti-Fas blocking antibody, ZB4, blocked CM1-mediated apoptosis effectively in Ramos cells, but not in Raji cells. Increased mitochondrial membrane permeabilization, which was measured by $DiOC_6$, was observed only in Raji cells. In contrast to no significant change of Bax known as pro-apoptotic protein, anti-apoptotic protein Bcl-2 was significantly decreased in Raji cells. In addition, we observed that CM1 ligation increased release of mitochondrial cytochrome c and upregulated caspase-9 activity in Raji cells. Conclusion: These results suggest that apoptosis induced by CM1-ligation is mediated by Fas-Fas ligand interaction in Ramos cells, whereas apoptosis is mediated by down-regulation of Bcl-2 and subsequent decrease of mitochondrial membrane potential in Raji cells.