Mutated C-terminal fragments of Clostridium perfringens enterotoxin have increased affinity to claudin-4 and reversibly modulate tight junctions in vitro

2011 ◽  
Vol 410 (3) ◽  
pp. 466-470 ◽  
Author(s):  
Azusa Takahashi ◽  
Masuo Kondoh ◽  
Hiroshi Uchida ◽  
Yohei Kakamu ◽  
Takao Hamakubo ◽  
...  
Keyword(s):  
Tight Junctions ◽  
Clostridium Perfringens ◽  
Clostridium Perfringens Enterotoxin

scholarly journals Innovative and Highly Sensitive Detection of Clostridium perfringens Enterotoxin Based on Receptor Interaction and Monoclonal Antibodies

Toxins ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 266
Author(s):  
Thea Neumann ◽  
Maren Krüger ◽  
Jasmin Weisemann ◽  
Stefan Mahrhold ◽  
Daniel Stern ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Clostridium Perfringens ◽  
Food Poisoning ◽  
Receptor Interaction ◽  
Clostridium Perfringens Enterotoxin ◽  
Fast Detection ◽  
Highly Sensitive ◽  
Basic And Applied Research ◽  
Highly Sensitive Detection

Clostridium perfringens enterotoxin (CPE) regularly causes food poisoning and antibiotic-associated diarrhea; therefore, reliable toxin detection is crucial. To this aim, we explored stationary and mobile strategies to detect CPE either exclusively by monoclonal antibodies (mAbs) or, alternatively, by toxin-enrichment via the cellular receptor of CPE, claudin-4, and mAb detection. Among the newly generated mAbs, we identified nine CPE-specific mAbs targeting five distinct epitopes, among them mAbs recognizing CPE bound to claudin-4 or neutralizing CPE activity in vitro. In surface plasmon resonance experiments, all mAbs and claudin-4 revealed excellent affinities towards CPE, ranging from 0.05 to 2.3 nM. Integrated into sandwich enzyme-linked immunosorbent assays (ELISAs), the most sensitive mAb/mAb and claudin-4/mAb combinations achieved similar detection limits of 0.3 pg/mL and 1.0 pg/mL, respectively, specifically detecting recombinant CPE from spiked feces and native CPE from 30 different C. perfringens culture supernatants. The implementation of mAb- and receptor-based ELISAs into a mobile detection platform enabled the fast detection of CPE, which will be helpful in clinical laboratories to diagnose diarrhea of assumed bacterial origin. In conclusion, we successfully employed an endogenous receptor and novel high affinity mAbs for highly sensitive and specific CPE-detection. These tools will be useful for both basic and applied research.

Prevalence of the Enterotoxin Gene and Clonality of Clostridium perfringens Strains Associated with Food-Poisoning Outbreaks

1998 ◽  
Vol 61 (2) ◽  
pp. 240-243 ◽  
Author(s):  
J. RIDELL ◽  
J. BJÖRKROTH ◽  
H. EISGRŰBER ◽  
B. SCHALCH ◽  
A. STOLLE ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Food Poisoning ◽  
Latex Agglutination ◽  
Pulsed Field Gel Electrophoresis ◽  
Enterotoxin Gene ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Clostridium Perfringens Enterotoxin ◽  
Polymerase Chain

The prevalence of the enterotoxin gene in a well-characterized collection of 71 Clostridium perfringens strains from 36 separate food-poisoning cases or outbreaks was analyzed with the polymerase chain reaction (PCR). The clonality of 39 strains originating from 14 outbreaks where at least two isolates were available was studied with pulsed-field gel electrophoresis (PFGE) using SmaI and ApaI restriction endonucleases. The cpe gene PCR assay was found to correlate well with Clostridium perfringens enterotoxin (CPE) production in vitro with reverse passive latex agglutination. Of the C. perfringens food and clinical food-poisoning isolates 24 (86%) and 38 (88%) were cpe-positive, respectively. Different PFGE patterns indicated that multiple cpe-positive clones are frequently present within one outbreak. The existence of cpe-positive and negative isolates with identical or nearly identical PFGE patterns in a single outbreak suggests that the cpe gene may be in a movable genetic element.

scholarly journals Use of Modified Clostridium perfringens Enterotoxin Fragments for Claudin Targeting in Liver and Skin Cells

2019 ◽  
Vol 20 (19) ◽  
pp. 4774 ◽  
Author(s):  
Laura-Sophie Beier ◽  
Jan Rossa ◽  
Stephen Woodhouse ◽  
Sophia Bergmann ◽  
Holger Kramer ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Clostridium Perfringens ◽  
Hepatoma Cell ◽  
Human Epidermis ◽  
Hepatoma Cell Line ◽  
Dependent Manner ◽  
Clostridium Perfringens Enterotoxin ◽  
Epidermal Barrier ◽  
Reconstructed Human Epidermis

Claudins regulate paracellular permeability in different tissues. The claudin-binding domain of Clostridium perfringens enterotoxin (cCPE) is a known modulator of a claudin subset. However, it does not efficiently bind to claudin-1 (Cldn1). Cldn1 is a pharmacological target since it is (i) an essential co-receptor for hepatitis C virus (HCV) infections and (ii) a key element of the epidermal barrier limiting drug delivery. In this study, we investigated the potential of a Cldn1-binding cCPE mutant (i) to inhibit HCV entry into hepatocytes and (ii) to open the epidermal barrier. Inhibition of HCV infection by blocking of Cldn1 with cCPE variants was analyzed in the Huh7.5 hepatoma cell line. A model of reconstructed human epidermis was used to investigate modulation of the epidermal barrier by cCPE variants. In contrast to cCPEwt, the Cldn1-binding cCPE-S305P/S307R/S313H inhibited infection of Huh7.5 cells with HCV in a dose-dependent manner. In addition, TJ modulation by cCPE variant-mediated targeting of Cldn1 and Cldn4 opened the epidermal barrier in reconstructed human epidermis. cCPE variants are potent claudin modulators. They can be applied for mechanistic in vitro studies and might also be used as biologics for therapeutic claudin targeting including HCV treatment (host-targeting antivirals) and improvement of drug delivery.

Radioimmunoassay for Clostridium perfringens Enterotoxin and Its Use in Screening Isolates Implicated in Food-Poisoning Outbreaks

1983 ◽  
Vol 46 (12) ◽  
pp. 1069-1073 ◽  
Author(s):  
GERARD N. STELMA ◽  
JOHN C. WIMSATT ◽  
PETER E. KAUFFMAN ◽  
DHIRENDRA B. SHAH
Keyword(s):  
Clostridium Perfringens ◽  
Food Poisoning ◽  
Soluble Starch ◽  
Optimum Conditions ◽  
Ileal Loop ◽  
Clostridium Perfringens Enterotoxin ◽  
Highly Active ◽  
Enterotoxin Production

Fourteen isolates of Clostridium perfringens obtained from food-poisoning outbreaks were screened for enterotoxigenicity using a radioimmunoassay (RIA) that detects 1.0 ng of enterotoxin/ml. Only four of the isolates produced enterotoxin in concentrations too low to be detected by counterimmunoelectrophoresis when grown in Duncan-Strong sporulation (D-S) medium. Substitution of raffinose for soluble starch or addition of theobromine to the medium stimulated enterotoxin production by three of the four enterotoxin-positive isolates. Raffinose and theobromine did not stimulate enterotoxin production by isolates that were enterotoxin-negative in D-S medium. Enterotoxin production by the RIA-positive strains correlated with the numbers of heat-resistant spores they produced. The RIA-negative isolates produced approximately the same numbers of spores/ml as the high enterotoxin producers, and more spores/ml than strain H8 produced under optimum conditions. Therefore, inability to sporulate is not the cause for failure of these isolates to produce enterotoxin. Rabbit ileal loop assays showed that the two isolates that were lowest enterotoxin producers in vitro were highly active in vivo.

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