• Title/Summary/Keyword: Enterotoxin

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Monoclonal Antibody Production against Piglet Diarrhea Agent (Enterotoxigenic E. coli) by Cell Fusion-Hybridoma Cell Technique (세포융합(細胞融合) 및 Hybridoma 세포작성(細胞作成)에 의한 항자돈백리(抗仔豚白痢) Monoclone항체(抗體)의 생산(生産))

  • Kim, Uh-ho;An, Soo-hwan;Yoon, Young-dhuk
    • Korean Journal of Veterinary Research
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    • v.27 no.2
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    • pp.259-267
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    • 1987
  • Enterotoxigenic E. coli (ETEC) cause an acute diarrhea (white scour) in both animals and humans. The disease process initially involves the adherence and colonization of the mucosal surface of the small intestine, followed by the elaboration of a heat-labile enterotoxin (LT) and/or heat-stable enterotoxin (ST). Intestinal adherence or colonization by ETEC is generally mediated by a specific surface-associated pilus (fimbrial) antigen that endows the bacteria with the capacity to adhere to epitherial cell surface. Fourteen monoclonal antibodies (MAbs) directed against pili antigens of ETEC were obtained by cell fusion/hybridoma technique. They were characterized by indirect immunofluorescence assay (IFA), and divided into four groups: specific to K99 antigen (group 1), cross-reactive with K99 and F41 antigens (group 2), specific to K88 antigen (group 3) and specific to 987P and K88 antigens (group 4), respectively. These MAbs demonstrated the distinct pili (K) antigens on the surface of ETEC by IFA, and could be utilized as diagnostic reagent for the identification of ETEC. When eighty-seven field isolates of E. coli from piglet with diarrhea were tested by group 3 MAb, fourty-two strains (48.3%) has K88 pilus antigen suggesting that this is one of the major pilus antigen of ETEC present in fifeld.

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Dendritic Cell-Mediated Mechanisms Triggered by LT-IIa-B5, a Mucosal Adjuvant Derived from a Type II Heat-Labile Enterotoxin of Escherichia coli

  • Lee, Chang Hoon;Hajishengallis, George;Connell, Terry D.
    • Journal of Microbiology and Biotechnology
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    • v.27 no.4
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    • pp.709-717
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    • 2017
  • Mucosal tissues are the initial site through which most pathogens invade. As such, vaccines and adjuvants that modulate mucosal immune functions have emerged as important agents for disease prevention. Herein, we investigated the immunomodulatory mechanisms of the B subunit of Escherichia coli heat-labile enterotoxin type IIa ($LT-IIa-B_5$), a potent non-toxic mucosal adjuvant. Alternations in gene expression in response to $LT-IIa-B_5$ were identified using a genome-wide transcriptional microarray that focused on dendritic cells (DC), a type of cell that broadly orchestrates adaptive and innate immune responses. We found that $LT-IIa-B_5$ enhanced the homing capacity of DC into the lymph nodes and selectively regulated transcription of pro-inflammatory cytokines, chemokines, and cytokine receptors. These data are consistent with a model in which directional activation and differentiation of immune cells by $LT-IIa-B_5$ serve as a critical mechanism whereby this potent adjuvant amplifies mucosal immunity to co-administered antigens.

Evaluation of systemic and mucosal immune responses in mice administered with novel recombinant Salmonella vaccines for avian pathogenic Esherichia coli

  • Oh, In-Gyeong;Lee, John Hwa
    • Korean Journal of Veterinary Research
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    • v.53 no.4
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    • pp.199-205
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    • 2013
  • Avian pathogenic Escherichia coli (APEC) is a causative agent for a number of extra intestinal diseases and account for significant losses to the poultry industry. Since protective immunity against APEC is largely directed to virulence antigens, we have individually expressed four different viulence antigens, papA, papG, IutA, and CS31A, using an attenuated Salmonella Typhimurium and a plasmid pBB244. Following oral immunization of mice with combination of two or four of these strains, serum IgG and mucosal IgA responses were elicited against each antigen represented in the mixture. The antigen-specific mucosal IgA responses were significantly higher in the group of mice immunized with the heat-labile Escherichia coli enterotoxin B subunit (LTB) strain than those in the group of mice immunized without the LTB strain. While, there was no significant difference between these two groups in antigen-specific serum IgG responses. The results showed that LTB could act as mucosal immune adjuvant. To assess the nature of immunity, the distribution of antigen-specific IgG isotypes was analyzed. All groups promoted Th1-type immunity as determined by the IgG2a/IgG1 ratio. Thus, our findings provided evidence that immunization with a combination of several vaccine strains is one of the strategies of developing effective vaccines against APEC.

Isolation and characterization of Clostridium Perfringens on bovine and porcine carcass (소와 돼지의 도체에서 Clostridium perfringens의 분리와 특성)

  • Chae Hee-Sun;Kim Youn-Ha;Kim Ju-Young;Kim Jong-Hwa;Kim Gyu-Hyeon;Choi Tae-Seok;Shin Bang-Woo;Lee Duck-Joo;Lee Jung-Hark
    • Korean Journal of Veterinary Service
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    • v.29 no.2
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    • pp.97-102
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    • 2006
  • A total of 1,848 samples was taken from bovine and porcine from January 2003 to November 2005. They were examined for the presence of Clostridiuml perfringens. The properties of the isolates were characterized for Gram staining, biochemical features and enterotoxin production. Forty-one strains (2.2%) of C perfringens were isolated from the 1,848 bovine and porcine carcass using selective media. 30 (3.2%) C perfringens were isolated from the 925 of bovine cacasses, and 11(1.2%) were isolated from the 923 of porcine cacasses. In TSC agar, all isolates showed lecithinase activity. The isolation rate was higher in spring and summer than in autumn and winter. Among 41 isolates, only 1 isolate detected the cpe gene in PCR. The PCR amplified band was observed 233 bp.

Inhibitory Effect of Carbamylated Staphylococcal Enterotoxins B on Inflammatory Response in HL-60 Cells

  • Chang, Jeong Hyun
    • Biomedical Science Letters
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    • v.20 no.2
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    • pp.96-102
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    • 2014
  • Staphylococcal enterotoxin B (SEB) is bacterial toxin that induces the activation of immune cells. Because the inhibition of pro-inflammatory effect of SEB can resolve the inflammation, I determined the influence of functional or structural change of SEB on immune cells. The post translational modification of protein occurs through carbamylation. Carbamylation can change the structure of proteins and can modify the biological activity of protein. In the present study, I investigated the effect of carbamylated SEB (CSEB) on the inflammatory response mediated by LPS in HL-60 cells. To determine the anti-inflammatory effect of CSEB, I produced carbamylated SEB using potassium cyanate (KCN) and then examined whether CSEB involved in cytokine releases and apoptosis of LPS-stimulated HL-60 cells. Although CSEB had not any effect on the LPS-stimulated HL-60 cells, the protein levels of IL-8, TNF-${\alpha}$ and IL-$1{\beta}$ were significantly decreased by CSEB without cytotoxicity. CSEB also blocked Akt and NF-${\kappa}B$ activation. These results indicate that the suppressive effect of CSEB in LPS-stimulated cytokine releases is occurred by inhibition of Akt and NF-${\kappa}B$ activity. Through further studies, CSEB may be used as anti-inflammatory molecule that makes the immune system more efficient.

Construction of Novel Bifunctional Chimeric Proteins Possessing Antitumor and Thrombolytic Activities

  • Hui, Jing;Dai, Youjin;Bian, Yuanyuan;Li, Hui;Cui, Xiaojin;Yu, Xiaojie;You, Song;Hu, Fengqing
    • Journal of Microbiology and Biotechnology
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    • v.22 no.7
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    • pp.894-901
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    • 2012
  • Based on their respective antitumor and thrombolytic activities, the superantigen staphylococcal enterotoxin C2 (SEC2) and staphylokinase (Sak) were chosen for the construction of the novel chimeric proteins Sak-linker-SEC2 and SEC2-linker-Sak using a linker composed of nine Ala residues. Both chimeric proteins possessed nearly the same PBMC proliferation stimulating activity and antitumor activity as SEC2 and thrombolytic activity as Sak. Neither the SEC2 or Sak component of each chimeric protein affected the activity of the other component. The results presented in this study provide a possible strategy to prevent and cure tumor thrombus.

Enterotoxigenic Escherichia coli in Korean Children with and without Diarrhea (소아 설사증에서 분리한 대장균 장독소의 병원적 역할)

  • Ahn, Byung-Soo;Kim, Kyung-Hee;Han, Wang-Soo;Suh, Inn-Soo
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.2
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    • pp.131-137
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    • 1987
  • The incidnce of enterotoxigenic Esherichia coli(ETEC) was investigated in E. coli strains isolated from Korean infants less than two years old. Over a period of 12 months, ETEC strains have been isolated from 45(45.0%) of 100 children with acute diarrhea and from 9(20.5%) of 44 children without diarrhea. In the group with diarrhea, 41(41.0%) strains produced heat-stable toxin, 3(3.1%) produced heat-labile toxin, and 1(1.0%) produced both heat-stable and heat-labile toxins. In the control group, 7(15.9%) released heat-stable toxin, 2(4.5%) released heat-labile toxin and none released both. A statistical association of strains releasing heat-stable toxin was significant(P<0.025).

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Expression of Functional Pentameric Heat-Labile Enterotoxin B Subunit of Escherichia coli in Saccharomyces cerevisiae

  • Lim, Jung-Gu;Kim, Jung-Ae;Chung, Hea-Jong;Kim, Tae-Geum;Kim, Jung-Mi;Lee, Kyung-Ryul;Park, Seung-Moon;Yang, Moon-Sik;Kim, Dae-Hyuk
    • Journal of Microbiology and Biotechnology
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    • v.19 no.5
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    • pp.502-510
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    • 2009
  • Although the Escherichia coli heat-labile enterotoxin B subunit (LTB) has already been expressed in several different systems, including prokaryotic and eukaryotic organisms, studies regarding the synthesis of LTB into oligomeric structures of pentameric size in the budding yeast Saccharomyces cerevisiae have been limited. Therefore, this study used a functional signal peptide of the amylase 1A protein from rice to direct the yeast-expressed LTB towards the endoplasmci reticulum to oligomerize with the expected pentameric size. The expression and assembly of the recombinant LTB were confirmed in both the cell-free extract and culture media of the recombinant strain using a Western blot analysis. The binding of the LTB pentamers to intestinal epithelial cell membrane glycolipid receptors was further verified using a GM1-ganglioside enzyme-linked inmmunosorbent assay (GM1-ELISA). On the basis of the GM1-ELISA results, pentameric LTB proteins comprised approximately 0.5-2.0% of the total soluble proteins, and the maximum quantity of secreted LTB was estimated to be 3 mg/l after a 3-day cultivation period. Consequently, the synthesis of LTB monomers and their assembly into biologically active aligomers in a recombinant S. cerevisiae strain demonstrated the feasibility of using a GRAS microorganism-based adjuvant, as well as the development of carriers against mucosal disease.

Application of a Peptide Nucleic Acid-Based Asymmetric Real-Time PCR Method for Rapid Detection of Vibrio cholerae (비브리오 콜레라 신속 검출을 위한 펩티드 핵산 기반 비대칭 real-time PCR 방법의 적용)

  • Kang, Mingyeong;Lee, Taek-Kyun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.12
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    • pp.117-124
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    • 2019
  • Vibrio cholerae is a very important pathogenic bacterium that has to be monitored in seafood and ships' ballast water. Various methods have been developed to identify this bacterium, yet these methods are time-consuming and have limitations for their sensitivity to detect contamination. The purpose of the present study was to develop a robust and reliable method for identifying V. cholerae. Peptide nucleic acid (PNA) probes were developed to use for PNA-based asymmetrical real-time PCR techniques. The toxigenic Cholera enterotoxin subunit B (ctxB) gene was selected as a target for detecting V. cholerae and the gene was synthesized as a positive template for conventional and real-time PCR. Real-time PCR primers and PNA probes were designed and standard curves were produced for the quantitative analysis. The selected PNA probes reacted specifically to V. cholerae without any ambiguity, even among closely related species, and the detection limit was 0.1 cfu/100 mL. Taken together, the PNA probes and asymmetrical qPCR methods developed in this present study could contribute to the rapid, accurate monitoring of V. cholerae in marine environments, and as well as in seafood and ships' ballast waters.