• Title/Summary/Keyword: antigen free

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Current Trends in Cancer Vaccines - a Bioinformatics Perspective

  • Sankar, Shanju;Nayanar, Sangeetha K.;Balasubramanian, Satheesan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4041-4047
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    • 2013
  • Cancer vaccine development is in the process of becoming reality in future, due to successful phase II/III clinical trials. However, there are still problems due to the specificity of tumor antigens and weakness of tumor associated antigens in eliciting an effective immune response. Computational models to assess the vaccine efficacy have helped to improve and understand what is necessary for personalized treatment. Further research is needed to elucidate the mechanisms of activation of antigen specific cytotoxic T lymphocytes, decreased TREG number functionality and antigen cascade, so that overall improvement in vaccine efficacy and disease free survival can be attained. T cell epitomic based in sillico approaches might be very effective for the design and development of novel cancer vaccines.

Ginsenosides Promote Proliferation of Cultured Ovarian Germ Cells Involving Protein Kinase C-mediated System in Embryonic Chickens

  • Liu, Hongyun;Zhang, Caiqiao
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.7
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    • pp.958-963
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    • 2006
  • The effect of ginsenosides (GS) on germ cell proliferation was evaluated with a chicken ovarian germ-somatic cell coculture model and the mechanism involving protein kinase C (PKC) pathway was investigated. Ovarian cells were cultured in serum-free McCoy's 5A medium and challenged with GS alone or in combinations with PKC activator (phorbol 12-myristate 13-acetate, PMA) or inhibitor ($H_7$) for 48 h. The number of germ cells was counted and the proliferating cells were identified by immunocytochemistry of proliferating cell nuclear antigen (PCNA). Results showed that GS significantly increased germ cell proliferation and this stimulating effect was further increased by PMA, but inhibited by H7, in a dose-dependent manner. Moreover, GS-elevated PCNA expression and the PCNA -labeling index of germ cells displayed similar changes with the increased numbers of germ cells. These results indicated that GS stimulated proliferation of ovarian germ cells with involvement of the PKC-mediated system.

The Principles and Practices of Immunocytochemical method in Light Microscopic Level (광학 현미경적 수준에서의 면역조직화학적 방법의 원리 및 실제)

  • Kim Jin-Sang
    • The Journal of Korean Physical Therapy
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    • v.3 no.1
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    • pp.229-250
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    • 1991
  • The study was carried out to investigate and review the principles and practices of immunocytochemical method in light microscopic level. The results were as follows. 1. Immunocytochemistry is the method to search out the intracellular position of the specific meterials using antigen -antibody reaction. 2. The chief items in immunocytochemistry are antigen, antibody and chromogen. 3. The identifical fixation is cardiac perfusion fixation. 4. The tissue slides must be prepared by vibratomy. 5. All stainings are carried out with free floating staining method. 6. There are polyclonal and monoclonal antibodies used in immunocytochemistry.

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Enhancement of Hybridoma Cell Growth and Anti-Hepatitis B Surface Antigen Monoclonal Antibody Production in Enriched Media with Low Serum (저혈청농축배지에서 세포성장 및 간염표면항원에 대한 단일클론항체 생산의 증가)

  • 전복환;조의철김동일백승복
    • KSBB Journal
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    • v.5 no.1
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    • pp.87-94
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    • 1990
  • Enhancement of hybridoma cell growth and monoclonal antibody(MAb) production by the addition of a small amount of serum into both serum-free medium and enriched medium was studied. The enriched medium was constructed by mixing a basal serum-free medium and a nutrient-fortified RPMI 1640 medium. It was supplemented with human serum albumin, insulin, transferrin, and monoethanolamine. It was found that addition of low concentration of serum with other serum-free supplements was favorable for growth of a mouse hybridoma 2c3.1 cells. The concentration of serum was determined to 0.5%. The maximum cell concentration obtained in this enriched medium supplemented with 0.5% fetal bovine serum (FBS) was $3.06{\times}10^6$ cells/ml and the concentration of secreted anti-Hepatitis surface antigen (antiHBsAg) MAb was $159.7{\mu\textrm{g}}\;/\;ml$ compared to $43{\mu\textrm{g}}\;/\;ml$ in RPMI 1640 medium with 10% FBS and $50{\mu\textrm{g}}\;/\;ml$ in previously-developed serum-free medium. The 2c3.1 cell growth and MAb production could be enhanced considerably by using the enriched medium supplemented with 0.5% FBS and serum-free supplements instead of RPMI 1640 medium or serum-free medium. The enhancement in MAb production in the enriched medium was more noticeable.

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Characteristics of Protein G-modified BioFET

  • Sohn, Young-Soo
    • Journal of Sensor Science and Technology
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    • v.20 no.4
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    • pp.226-229
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    • 2011
  • Label-free detection of biomolecular interactions was performed using BioFET(Biologically sensitive Field-Effect Transistor) and SPR(Surface Plasmon Resonance). Qualitative information on the immobilization of an anti-IgG and antibody-antigen interaction was gained using the SPR analysis system. The BioFET was used to explore the pI value of the protein and to monitor biomolecular interactions which caused an effective charge change at the gate surface resulting in a drain current change. The results show that the BioFET can be a useful monitoring tool for biomolecular interactions and is complimentary to the SPR system.

Overview on Molecular Toxicological Aspects of Helicobacter pylori Virulence Factor, Cytotoxin-associated Antigen A (CagA) (헬리코박터 파이로리의 병원성 단백질, CagA에 대한 분자 독성학적 측면에서의 고찰)

  • Kim Byung J.;Jung Hwa Jin;Hwang Jee Na;Kang Seok Ha;Oh Se-Jin;Seo Young Rok
    • Toxicological Research
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    • v.20 no.3
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    • pp.179-185
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    • 2004
  • Helicobacter pylori (H. pylori) infects more than half of the people in the world as a major microbe to cause most of gastric diseases. Recently, cytotoxin associated-antigen A (CagA) is believed as one of the most important virulence factors of H. pylori. Molecular toxicological pathway of CagA is necessary to investigate for understanding the pathological and toxicological aspects of H. pylori, since this virulence protein harasses intercellular processes of host cells to get profit for the survival of H. pylori. CagA is coded from cag pathogenicity island (cag PAI) and translocated into host cells by Type 4 secretion system (TFSS). Tyrosine phosphorylation of CagA targets Src homology 2-containing phosphotyrosine phosphatase (SHP-2) to form a CagA-SHP-2 complex. This complex depends on the similarity of sequence between EPIYA motif and Src homology 2 domain (SH2 domain) of CagA. The generation of growth factors is an essential role of CagA in protecting and healing gastric mucosa for the survival of H. pylori. On the other hand, the activation of IL-8 by CagA induces neutrophils generating inflammation and free radicals. Indeed, free radicals are well known carcinogen to induce DNA damage. In addition, the transduction of mitogen-activation signal by CagA is one of the interesting features to understand how to cause cancer. The relationship between cancer and inflammation with CagA was mainly discussed in this review.

Human Kallikrein-2, Prostate Specific Antigen and Free-Prostate Specific Antigen in Combination to Discriminate Prostate Cancer from Benign Diseases in Syrian Patients

  • Bachour, Dala-Maria;Chahin, Emil;Al-Fahoum, Sahar
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7085-7088
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    • 2015
  • Background: The high incidence of prostate cancer as the most common malignancy in males in many countries raises the question of developing reliable detection tests. The prostate specific antigen (PSA) test is the most widely used for screening for prostate cancer; however, its low specificity elevates the number of unnecessarily biopsies. Serum human kallikrein-2 (hK2) is considered as a promising marker, and especially its ratio to fPSA, for predicting the presence of malignancy to select the best choice referring to biopsy or surveillance. In this study, we investigated the role of hK2 and its combinations with other markers to discriminate prostate cancer from benign diseases in Syrian patients. Materials and Methods: In this prospective oriented cross-sectional cohort study, serum samples were collected from patients referred to many Hospitals in Damascus, Syria, between May 2011 and March 2012, and diagnosed with biopsy proven benign prostate hyperplasia (BPH) or prostate cancer (PCa). Serum was analyzed for hK2, PSA and fPSA, and the ratios of fPSA/PSA and hK2/fPSA were calculated. Results: We found that mean hK2/fPSA ratios were significantly higher (P=0.01) in prostate cancer patients than in the BPH or control groups. Also the ratio hk2/fPSA gave the largest area under the curve (AUC:0.96) which was significantly larger than for fPSA/PSA (AUC:0.41) indicative of higher specificity. Conclusions: Our results demonstrate that the ratio of hK2/fPSA might be superior to the use of fPSA/PSA alone. The hK2 could be shown to enhance the early detection of prostate cancer; especially the ratio hK2/fPSA improves specificity and hence may reduce the number of negative biopsies.

Development of ELISA for detection of canine brucellosis (Canine brucellosis 검출을 위한 ELISA 진단법 확립)

  • Hur, Jin;Baek, Byeong-Kirl
    • Korean Journal of Veterinary Service
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    • v.34 no.2
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    • pp.159-166
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    • 2011
  • This study was carried out to develope enzyme-linked immunosorbent assay (ELISA) for detection of canine brucellosis in dogs experimentally inoculated with Brucella abortus 1119-3 and B. canis RM666. Groups A, B and C of dogs (each group consisting of three dogs) were orally inoculated with approximately $5{\times}10^9$ colony-forming units of B. abortus and B. canis, and with sterile pyrogen-free PBS, respectively. The animals were monitored at regular intervals upto the 12th week post inoculation (PI) by standard tube agglutination test (STAT), plate agglutination test (PAT), Rose Bengal test (RBT), 2-mercaptoethanol rapid slide agglutination test (2ME-RSAT) and ELISA. The induced antibody titers in group A dogs were detected from the first week PI to the eighth week PI in STAT, PAT and RBT using the inactivated whole cells of B. abortus 1119-3 as antigens, while no sera in groups B and C dogs reacted with the antigens. In 2ME-RSAT using whole cells of B. canis M-strain as antigens, the induced antibody titers in group B dogs were observed at the second week PI and persisted for the 12th week PI, while sera of groups A and C dogs did not react with the whole cells. In ELISA using cytoplasmic fractions antigen of B. abortus 1119-3, the mean optical density of antibodies in groups A and B was detected from the first and second weeks PI, respectively, and persisted for 12th week PI, while sera of group C did not cross-react with the fractions antigen. However, in ELISA using the hot saline extracts of B. canis M- as an antigen, the induced antibody titers in only group B dogs were detected from second week PI and persisted for until the end of this study. These results indicate that the ELISA using B. abortus 1119-3 cytoplasmic fractions as antigens can be a good candidate for detection of brucellosis by B. abortus as well as B. canis in dogs.

Multilevel Precision-Based Rational Design of Chemical Inhibitors Targeting the Hydrophobic Cleft of Toxoplasma gondii Apical Membrane Antigen 1 (AMA1)

  • Vetrivel, Umashankar;Muralikumar, Shalini;Mahalakshmi, B;K, Lily Therese;HN, Madhavan;Alameen, Mohamed;Thirumudi, Indhuja
    • Genomics & Informatics
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    • v.14 no.2
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    • pp.53-61
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    • 2016
  • Toxoplasma gondii is an intracellular Apicomplexan parasite and a causative agent of toxoplasmosis in human. It causes encephalitis, uveitis, chorioretinitis, and congenital infection. T. gondii invades the host cell by forming a moving junction (MJ) complex. This complex formation is initiated by intermolecular interactions between the two secretory parasitic proteins-namely, apical membrane antigen 1 (AMA1) and rhoptry neck protein 2 (RON2) and is critically essential for the host invasion process. By this study, we propose two potential leads, NSC95522 and NSC179676 that can efficiently target the AMA1 hydrophobic cleft, which is a hotspot for targeting MJ complex formation. The proposed leads are the result of an exhaustive conformational search-based virtual screen with multilevel precision scoring of the docking affinities. These two compounds surpassed all the precision levels of docking and also the stringent post docking and cumulative molecular dynamics evaluations. Moreover, the backbone flexibility of hotspot residues in the hydrophobic cleft, which has been previously reported to be essential for accommodative binding of RON2 to AMA1, was also highly perturbed by these compounds. Furthermore, binding free energy calculations of these two compounds also revealed a significant affinity to AMA1. Machine learning approaches also predicted these two compounds to possess more relevant activities. Hence, these two leads, NSC95522 and NSC179676, may prove to be potential inhibitors targeting AMA1-RON2 complex formation towards combating toxoplasmosis.

Development of serodiagnostic surface plasmon resonance imaging assay for the detection of antibodies to porcine circovirus type 2

  • Park, Chul;Kim, Bum-Seok;Kim, Yong-Hwan;Cho, Ho-Seong
    • Korean Journal of Veterinary Service
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    • v.34 no.1
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    • pp.1-4
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    • 2011
  • A surface plasmon resonance imaging (SPRI) assay was developed for measuring porcine circovirus type 2 (PCV2) antibody using a recombinant capsid protein as an antigen. The diagnostic potential of SPRI for detecting antibodies to the PCV2 capsid protein was compared with that of a conventional enzyme-linked immunosorbent assay (ELISA) using 70 pig serum samples taken from 6 pig farms. There was a strong positive correlation between the SPRI and ELISA (n = 70, r = 0.911, P<0.01). Therefore, this recombinant capsid protein can be used as an antigen for serological studies, and the SPRI, a label-free and high-throughput method, is expected to be a valuable tool in the serodiagnosis of PCV2 infection.