• Title/Summary/Keyword: Human Monoclonal Antibodies

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Immunohistochemical Evaluation of Angiogenesis Related Markers in Pyogenic Granuloma of Gingiva

  • Seyedmajidi, Maryam;Shafaee, Shahryar;Hashemipour, Golnarsadat;Bijani, Ali;Ehsani, Hodis
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7513-7516
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    • 2015
  • Background: Pyogenic granuloma is a common non-neoplastic connective tissue proliferation. ICAM-1 and VCAM-1 are vascular adhesion molecules and CD34 is a marker for evaluation of angiogenesis. The purpose of this study was to compare the immunohistochemical expression of ICAM-1, VCAM-1 & CD34 in oral pyogenic granuloma and normal gingiva. Materials and Methods: This study was performed on thirty five formalin-fixed, paraffin embedded samples of gingival pyogenic granuloma. Also we used thirty five paraffined blocks of normal gingiva as control group which were taken from crown lengthening surgery. We employed immunohistochemistry staining for our prepared microscopic slides using monoclonal mouse anti-human antibodies against ICAM-1 (CD54), VCAM-1 (CD106) and CD34. Slides were examined under light microscope and then the mean amount of stained vessels also known as microvascular density (MVD) in highly vascularized areas (hot spots) was measured. Paired t-test and repeated measures ANOVA were used to compare the difference between quantitative variables and Chi-square test for qualitative variables in different groups. Pearson correlation coefficient was used to compare relations between quantitative variables. P<0.05 was considered significant. Results: The mean of MVD for ICAM-1, VCAM-1 and CD34 was significantly higher in pyogenic granuloma than normal gingiva (p<0.001 & p<0.001 & p<0.001, respectively). Expression of CD34 in pyogenic granuloma was significantly higher than ICAM-1 and VCAM-1 (P<0.001). Besides, expression of ICAM-1 in normal gingiva, was significantly lower than two other markers (p<0.001). Conclusions: Regarding the results, it seems that ICAM-1, VCAM-1 and CD34 are useful biomarkers in evaluation of vascular and inflammatory lesions such as gingival pyogenic granuloma and the results indicate the role of these biomarkers in pathogenesis of oral pyogenic granuloma.

The Clinicopathological and Prognostic Impact of 14-3-3 Protein Isoforms Expression in Human Cholangiocarcinoma by Immunohistochemistry

  • Wu, Qiao;Liu, Chang-Zheng;Tao, Lian-Yuan;Yu, Lan;Liu, Wei;Chen, Song-Sen;He, Xiao-Dong
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.4
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    • pp.1253-1259
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    • 2012
  • The 14-3-3 proteins are highly conserved, ubiquitous molecules involved in a variety of biologic phenomena, such as cell cycle control, and apoptosis. However, their expression in cholangiocarcinoma has not been previously characterized. In this paper, immunohistochemistry using specific anti-14-3-3 monoclonal antibodies was performed on formalin-fixed;, paraffin embedded archival tissue from 86 patients of cholangiocarcinoma. We also examined the correlation between expression and survival rate and clinicopathologic factors such as tumor location, tumor size, pathologic differentiation, lymphatic permeation, lymph node metastasis, and tumor stage. Positive 14-3-3 proteins expression was observed for 6 isoforms (${\beta}$, ${\sigma}$, ${\gamma}$, ${\theta}$, ${\delta}$, ${\eta}$) of these proteins in 86 patients of cholangiocarcinoma. ${\beta}$ and ${\sigma}$ isoform immunoreactivity was correlated with lymph node metastasis, tumor stage and patients' survival rate. In addition, ${\delta}$ isoform immunoreactivity showed trends with tumor location, tumor size, pathologic differentiation and tumor stage, while the ${\theta}$ isoform was correlated with pathologic differentiation. These results indicated that upregulated expression of some isoforms of 14-3-3 may be a common mechanism for evading apoptosis in cholangiocarcinoma, so that targeting 14-3-3 may be a novel promising strategy for the treatment of this tumor.

IMMUNOHISTOCHEMICAL STUDY ON LYMPHOCYTE DISTRIBUTION IN ENDODONTICALLY TREATED AND UNTREATED PERIAPICAL LESIONS (근관치료전과 후의 치근단 병소에서 임파구의 분포에 관한 면역조직화학적 연구)

  • Oh, Tae-Seok;Lim, Sung-Sam
    • Restorative Dentistry and Endodontics
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    • v.11 no.1
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    • pp.63-75
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    • 1985
  • This study was designed to identify lymphocytes and to compare the lymphocyte distribution in endoodontically treated periapical lesions with that in endodontically untreated periapical lesions by way of immunohistochemical staining. Twenty-one human dental periapical lesions were obtained, frozened, serially sectioned to $4-5{\mu}$, and stained using the three-stage indirect immunoperoxidase technique and monoclonal antibodies for detecting the presence of B,T lymphocyte and T suppressor cell. Following results were obtained; 1. All of the examined periapical lesions had positive staining for B,T lymphocyte and T suppressor cell. 2. The concentration of T lymphocytes in 18 lesions diagnosed as periapical cyst and granuloma in both groups was greater than that of B lymphocytes and 2 periapical lesions identified as abscess in treated lesions had more positive B lymphocytes than positive T lymphocytes. 3. The average numbers of T,B lymphocytes and T suppressor cells in Endodontically treated lesions were lower than those of untreated lesions, but no statistically significant difference was noted. 4. When the distribution ratios of T lymphocytes to B lymphocytes and T suppressor cells to T lymphocytes were compared in Endodontically treated lesions by the histological aspects of the lesions and at the intervals of the duration after Endodontic treatment, a statistically significant change was not found. 5. The mean values of T lymphocytes, B lymphocytes and T suppressor cells in Endodontically treated lesions were markedly decreased in the specimens obtained at 3 month after Endodontic treatment, but no statistically significant difference was found.

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Significance of Ki67 and p27 Reactivities in Various Thyroid Disorders (갑상선 결절의 Ki67과 p27 발현도에 대한 분석)

  • Park Cheong-Soo;Chung Woung-Youn;Chang Hang-Seok;Lee Mi-Kyung
    • Korean Journal of Head & Neck Oncology
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    • v.15 no.1
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    • pp.3-8
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    • 1999
  • Objective: The expression of Ki67, a proliferation marker, and p27, a cyclin dependent kinases(CDKs) inhibitor, has been studied in various human neoplasms. This study was carried out to determine whether these markers are useful in distinguishing benign from malignant lesions of the thyroid or predicting biologic behavior of malignant lesions. Material and Methods: Using immunohistochemical techniques with monoclonal antibodies to Ki67 and p27, we analyzed the expression of Ki67 and p27 in various thyroid disorders(25 follicular adenomas, 47 follicular carcinomas, 16 papillary carcinomas, 20 adenomatous goiters and 40 normal thyroid tissues). The labeling indices(LIs) were determined by counting cells expressing these markers in 1000 cells per immunostained slide. Results: Neoplastic thyroid diseases showed higher expression of Ki67 and lower expression of p27 than non-neoplastic diseases(p<0.05). The expression of p27 was significantly different between follicular adenomas($LI=55.4{\pm}5.7$) and follicular carcinomas($LI=23.2{\pm}10.2$). There was, however, no significant correlation between the degree of Ki67 and p27labeling indices and types of carcinoma or clinical aggressiveness of diseases. Conclusion: The degree of Ki67 and p27 expression was useful in distinguishing between benign from malignant thyroid lesions, particulary between follicular adenoma and follicular carcinoma, but was not directly proportional to the tumor aggressiveness.

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Immunoscintigraphy of Colorectal and Other Gastrointestinal Cancers with Radioactive Monoclonal Antibodies to CEA and CA 19-9 (대장직장암 및 기타 소화기암에서의 단세포군항체를 이용한 방사면역신티그라피의 진단)

  • Jang, Dae-Hwan;Choi, Duck-Joo;Lee, Bum-Woo;Park, Won;Han, Chang-Soon;Kim, Hak-San;Kim, Chong-Soon
    • The Korean Journal of Nuclear Medicine
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    • v.22 no.1
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    • pp.83-92
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    • 1988
  • The cocktails of two $^{131}I$ labeled Monoclonal antibody (MCAB) (Anti CA 19-9 F$(ab')_2$ + Anti CEA $F(ab')_2$ fragment), which react specially, with human gastrointestinal cancers, were administered to 10 patients with colorectal (7), stomach(2) and pancreas(1) cancer for scintigraphic detection. All patients were known or postoperatively recurrent cases, and serum tumor markers, CA 19-9 and CEA, were measured with immunoradiometric assay, just before immunoscintigraphy (ISG). The tumor marker's level in serum is not correlated with positive tumor uptake in ISG. The sensitivity and specificity of ISG in detection of 21 tumor sites, based on surgery, CT, ultrasonography and pathology, were 90.5% and 100% One case of colon cancer showed gall bladder metastasis, which was neglected on CT study. Tumor/non tumor uptake ratio of radiolabelled antibody were progressively increased from day 3 to day 7 during study. We summerized as follows 1) The use of cocktails of CEA and CA 19-9 MCAB $F(at')_2$ increased sensitivity and specificity in ISG. 2) Delayed imaging (later than 5 days) increases sensitivitv and specificity due to exclusion of nonspecific iodine accumulation in stomach and lung. 3) Second tracer technique is essential for anatomical landmark by use of a double isotope scan, but subtraction technique, a possible source of artifacts, is no longer necessory when delayed imaging is performed. 4) It may be possible to use two MCAB cocktails of CA 19-9 and CEA in Radioimmunodetection of stomach and pancreas cancer. In conclusion, ISG using MCAB cocktails, $F(ab')_2$ fragment of anti CA 19-9 and Anti CEA, provide additional opportunity for tumor localization and detection of colorectal and other G-I cancer, such as stomach and pancreas.

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Production and Characterization of a Recombinant Antibody Neutralizing Botulinum Neurotoxin A (보툴리눔 신경독소 A를 중화하는 재조합 항체의 제조와 특성 분석)

  • Park, Hong-Gyu;Choi, Mieyoung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.1
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    • pp.295-301
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    • 2017
  • Botulinum neurotoxin (BoNT/A) is a neurotoxin that selectively attacks the peripheral cholinergic nerve endings. It is produced by Gram -positive, endospore-forming strict anaerobic bacteria, Clostridium botulinum. Since BoNT/A could be a biothreat agent, as well as a contaminator of food and water supplies, the development of sensitive assays for toxin detection and potent antitoxin for the treatment of intoxication is necessary. In this study, for the purpose of producing monoclonal antibodies (mAbs) that are capable of neutralizing Botulinum neurotoxin type A (BoNT/A), scFv (single-chain variable domain fragment) libraries from the rabbit antisera against BoNT/A was fused to a human IgG. The resulting recombinant scFvIgG antibody protein was expressed in stable cell lines and was purified using a protein A affinity chromatography. The efficacy of scFvIgG mAb was confirmed by ELISA and was evaluated for the neutralization of BoNT/A in vivo. Such an in vivo toxin neutralization assay was performed using mice. Although scFvIgG antibody proteins (10 ug) failed to fully protect the mice challenged with BoNT/A (100,000 $LD_{50}$), it significantly prolonged the survival time. These results suggest that scFvIgG mAb may be capable of neutralizing BoNT/A single-chain variable domain fragment.

An Immune-Electron Microscopic Study for Cluster Designation on the Phagocytic Synovial Cells in the Knee Joint of the Human (인체 무릎관절 윤활포식세포 cluster designation 표지에 관한 면역전자현미경적 연구)

  • Lim, Hyoung-Soo;Cho, Kook-Hyeung;Kim, Yong-Wook;Park, Kyeong-Han;Hwang, Young-Il;Chang, Ka-Young;Hwang, Douk-Ho
    • Applied Microscopy
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    • v.30 no.2
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    • pp.173-183
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    • 2000
  • This study was designed to observe the ultrastructural localization of synoviocytes, which are concerned with the function of phagocytic synovial cells (type A synoviocytes, macrophage-like synoviocytes), in the knee joint of the human for CD14 and CD105 by cryo-immune-electron microscopic technique. The synovium were dissected and fixed for two hours (in 4% paraformaldehyde and 0.1% glutaraldehyde mixture), and were immerged in 2.3 M sucrose and 20% PVP solution. Finally, they were cut with the cryoultramicrotome and labelled with primary antibodies (monoclonal mouse anti-human CD14, monoclonal mouse anti-human CD105 (endoglin) and secondary (donkey anti-mouse IgG) tagged with 6 nm colloidal gold particles. The tissues were observed under transmission electron microscope. This study was resulted as follows. 1. In the synovium of the human knee joint, CD14+ cells were identified. These cells showed phagocytic synovial cell's features. In the phagocytic synoviocyte, the distributions of CD14 were marked in the cytoplasm, around vacuoles, and in cytoplasmic process, but not detected inside of vacuoles. 2. In the synovium of the human knee joint, CD105+ cells were identified. These cells were recognized endothelial cells and phagocytic synovial cells. In the phagocytic synovial cells, the distributions of CD105 (endoglin) were marked in cytoplasic process, around vacuoles, and in cell membrane, but not detected inside of vacuoles. On the basis of above findings, it is obvious that phagocytic synovial cells were marked at CD 14 and CD 105, and might be play the role of activated macrophages or phagocytes in the synovial membrane.

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Human Papillomavirus Infections and p53 Expression in Prostatic Carcinoma (전립선 암조직에서의 사람파필로마바이러스의 감염과 p53단백질의 발현에 대한 연구)

  • Kwon, Dur-Han;Jin, Seung-Won;Kang, Byung-Tae;Yoon, Hee-Sik;Yoo, Wang-Don;Kim, Hyeun-Soo;Lee, Sang-Sook;Lee, Ho-Sa;Park, Sue-Nie
    • The Journal of Korean Society of Virology
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    • v.26 no.2
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    • pp.227-234
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    • 1996
  • Prostatic carcinoma is the leading second cause of cancer in men. Previous epidermiological studies implicated human papillomavirus as an infectious agent. Since there are only limited studies on the association of HPV to prosate cancer, we examined the prevalence of HPV infections in korean prostate cancer patients. We observed that out of 26 cases, 4 cases and 5 cases were infected by HPV 16(27%) and HPV 18 (31%), respectively and 3 cases by both (46%) and at least 18 were positive for HPV (69%). For these samples, immunohistochemical detection of the p53 and proliferative cell nuclear antigen (PCNA) were also studied, using monoclonal antibodies. Sixteen of 26 (61%) showed immunostaining for p53 protein. While 8 samples with no HPV infection (100%) showed all positive for p53 protein staining, less than half of the 18 patients with any HPV infection (44%) showed p53 protein staining. These findings indicate that altered expression of p53 protein occurs in the more than half of prostate cancers, however, p53 expression is less frequent in HPV infected tissues. This implies that there might be an inverse correlation in general between HPV infection and p53 amplification. However, while 50% (4 of 8) of HPV negative prostate cancer was positive for PCNA staining, 13 out of 18 HPV infected patients (72%) were positive. Therefore HPV infection is more strongly associated with increase proliferation. In addition HPV infected cancer patients are generally in more advanced status implying that HPV infection plays a role in the development of highly malignant prostatic carcinomas, eventhough the statistical significance of this interpretation might be waited for the analysis of more cases.

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Detection of HHV6 and EBV in histiocytic necrotizing lymphadenitis (Histiocytic necrotizing lymphadenitis에서 HHV6과 EBV의 검출)

  • Park, Kyung Hee;Park, Sung Shik;Kim, Ji Yeon;Park, Su Eun
    • Clinical and Experimental Pediatrics
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    • v.51 no.9
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    • pp.987-991
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    • 2008
  • Purpose : Kikuchi-Fujimoto disease (KFD), also known as histiocytic necrotizing lymphadenitis (HNL), is a self-limited disease characterized by cervical lymphadenopathy and fever. The etiology of KFD remains unknown; however, the self-limiting nature of HNL suggests the cause of this disease could be viral infection. For this reason, several viruses have been evaluated as possible etiologies of HNL, including Epstein-Barr virus (EBV), human herpesvirus 6 (HHV6), human herpesvirus 8 (HHV8), and cytomegalovirus (CMV). The aim of this study was to examine the relationship of EBV and HHV6 to HNL. Methods : Data pertaining to 51 cases with biopsy-confirmed HNL were collected between January 1999 and December 2005, from the Department of Pathology, College of Medicine, Pusan National University, Busan, Korea. The clinical records-including data regarding age, gender, duration of fever, and lymph node involvementwere reviewed retrospectively. The in situ hybridization (ISH) assay was performed by EBER PNA probe (Dako, Capinteria, CA, USA), and immunohistochemistry testing was performed with anti-HHV type 6 monoclonal antibodies (Chemicon, Temecula, CA, USA). Results : The HNL patients in this study were 24 males and 27 females, ranging in age from seven to 61 years (median: 25.9). ISH for EBV was positive in 8/51 (15.7%) biopsies, and immunohistochemistry for HHV6 was positive in 15/51 (29.4%) biopsies. Serologic analysis of EBV IgM was performed in 23 cases; only one patient was positive for EBV IgM and EBV ISH. Conclusion : Our study could not provide supportive evidence of a viral pathogenesis for HNL; therefore, cases of HNL may not have a dominant viral cause. However, some rare exceptional cases may have been caused by viral infection.

Comparative Expression of Bcl-2 and NOS2 in Oral White Lesions and Squamous Cell Carcinoma (구강내 백색병소와 편평상피세포암종에서 bcl-2와 NOS2 비교발현에 관한 연구)

  • Shin, Min;Kim, Eun-Cheol
    • Journal of Oral Medicine and Pain
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    • v.24 no.2
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    • pp.145-161
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    • 1999
  • The proto-oncogene bcl-2 confers a survival advantage to cells by blocking programmed cell death (apoptosis). Overexpression of bcl-2 probably plays a role in tumorigenesis, and the expression of the bcl-2 protein has been investigated in many kinds of tumors. An increased expression of nitric oxide synthetase(NOS) has been observed in human colon cancer cell lines as well as in human gynecological, breast, and CNS tumors. However there have been only a few reports on the expression of bcl-2 and $NOS_2$ in oral white lesions and cancer. The aim of this study was to investigate the relationship between the expression of Bcl-2 and $NOS_2$ and several pathological parameters such as histological types and layers. We reported desregulation of bcl-2 and $NOS_2$ expression during progression from oral white lesion, lichen planus and leukoplakia to squamous cell carcinoma. The obtained results were as follows: 1. Immunohistochemical analysis with monoclonal antibodies to bcl-2 oncoprotein and $NOS_2$ in formalin-fixed paraffin-embedded tissue sections revealed that bcl-2 expression is restricted to the basal cell layer and $NOS_2$ was mild expressed only in subepithelial inflammatory cells in normal human mucosa. There wasn't specific finding of those in lichen planus and leukoplakia. 2. Bcl-2 immunoreactivity in severe epithelial dysplasia or CIS occurs throughout the epithelium, $NOS_2$ reactivity in most superficial layer were noted. 3. In well-differentiated squamous cell carcinomas, mostly bcl-2 was overexpressed. In moderated and poor squamous cell carcinomas, the expression of $NOS_2$ was increased and that of bcl-2 was decreased. 4. The immunoreactivity of bcl-2 was 12.5% of normal mucosa, 30% of leukoplakia, 44% of lichen planus and 67% of carcinoma in situ. In carcinoma, those were 43%, 50% and 67% according to differentiation, respectively. 5. The immunoreactivity of $NOS_2$ was 25% of normal mucosa, 70% of leukoplakia, 78% of lichen planus and 100% of carcinoma in situ and epithelial dysplasia. In carcinoma, those were higher in moderated(100%) and poor(83%) squamous cell carcinomas than in well differentiated type(71%). 6. The expression of bcl-2 and $NOS_2$ by Western blot was increased highly in lichen planus and leukoplakia. Therefore, the expression of bcl-2 was increased in the white and precancerous lesions and that was decreased by differentiation of carcinoma. However, $NOS_2$ immunoreactivity in carcinoma in situ was lower than those in moderated and poor squamous cell. These findings suggest that the interaction of bcl-2 and $NOS_2$ may be roled importantly in growth and development of carcinoma.

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