Evidence for Clostridium perfringens Enterotoxin (CPE) Inducing a Mitogenic and Cytokine Response In Vitro and a Cytokine Response In Vivo

1999 ◽  
Vol 38 (2) ◽  
pp. 96-100 ◽  
Author(s):  
F. Morgan Wallace ◽  
Annette S. Mach ◽  
Andreas M. Keller ◽  
James A. Lindsay
Keyword(s):  
Clostridium Perfringens ◽  
Cytokine Response ◽  
Clostridium Perfringens Enterotoxin

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.

scholarly journals Effective Oncoleaking Treatment of Pancreatic Cancer by Claudin-Targeted Suicide Gene Therapy with Clostridium perfringens Enterotoxin (CPE)

Cancers ◽  
2021 ◽  
Vol 13 (17) ◽  
pp. 4393
Author(s):  
Jessica Pahle ◽  
Dennis Kobelt ◽  
Jutta Aumann ◽  
Diana Behrens ◽  
Ole Daberkow ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Gene Therapy ◽  
Clostridium Perfringens ◽  
Suicide Gene ◽  
Suicide Gene Therapy ◽  
Selective Toxicity ◽  
Pancreas Carcinoma ◽  
Clostridium Perfringens Enterotoxin

Pancreatic cancer (PC) is one of the most lethal cancers worldwide, associated with poor prognosis and restricted therapeutic options. Clostridium perfringens enterotoxin (CPE), is a pore-forming (oncoleaking) toxin, which binds to claudin-3 and -4 (Cldn3/4) causing selective cytotoxicity. Cldn3/4 are highly upregulated in PC and represent an effective target for oncoleaking therapy. We utilized a translation-optimized CPE vector (optCPE) for new suicide approach of PC in vitro and in cell lines (CDX) and patient-derived pancreatic cancer xenografts (PDX) in vivo. The study demonstrates selective toxicity in Cldn3/4 overexpressing PC cells by optCPE gene transfer, mediated by pore formation, activation of apoptotic/necrotic signaling in vitro, induction of necrosis and of bystander tumor cell killing in vivo. The optCPE non-viral intratumoral in vivo jet-injection gene therapy shows targeted antitumoral efficacy in different CDX and PDX PC models, leading to reduced tumor viability and induction of tumor necrosis, which is further enhanced if combined with chemotherapy. This selective oncoleaking suicide gene therapy improves therapeutic efficacy in pancreas carcinoma and will be of value for better local control, particularly of unresectable or therapy refractory PC.

The role of enterotoxin in Clostridium perfringens type A enteritis

1971 ◽  
Vol 17 (7) ◽  
pp. 987-991 ◽  
Author(s):  
A. H. W. Hauschild ◽  
L. Niilo ◽  
W. J. Dorward
Keyword(s):  
Clostridium Perfringens ◽  
Causative Agent ◽  
Type A ◽  
Rabbit Serum ◽  
Vegetative Cells ◽  
Clostridium Perfringens Enterotoxin ◽  
Immune Rabbit

Vegetative cells of three strains of Clostridium perfringens type A, free of erythemal activity, were suspended in fresh medium and injected into ligated intestinal loops of lambs. Examination of the loop contents after 6.5 h showed significant accumulation of fluid, multiplication and sporulation of C. perfringens, and erythemal activity in both the supernatant fluids and the sediments.The erythemal factor produced in vivo was identical with the erythemal factor of sporulated cells of C. perfringens grown in vitro, and again caused accumulation of fluid when transferred into ligated intestinal loops of recipient lambs. Immune rabbit serum prepared against extracts from sporulated cells of C. perfringens, and absorbed with extracts from vegetative cells of the same strain, completely neutralized the enterotoxic and erythemal activities of the in vivo-produced factor.It is concluded that the erythemal factor is the causative agent in C. perfringens type A enteritis. The term "Clostridium perfringens enterotoxin" is proposed to characterize the erythemal factor.

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.

scholarly journals Using More Than 1 (Path)Way to Kill a Host Cell: Lessons From Clostridium perfringens Enterotoxin

2020 ◽  
Vol 13 ◽  
pp. 117863612093151
Author(s):  
Bruce McClane ◽  
Archana Shrestha
Keyword(s):  
Recent Work ◽  
Clostridium Perfringens ◽  
Caspase 3 ◽  
Kinase Domain ◽  
Intermediate Step ◽  
Clostridium Perfringens Enterotoxin ◽  
Mixed Lineage Kinase ◽  
Intestinal Infections ◽  
Apoptosis And Necrosis

Clostridium perfringens enterotoxin (CPE) is responsible for the symptoms of common intestinal infections due to C. perfringens type F isolates. CPE is a pore-forming toxin that uses certain claudins as a receptor. Previous studies showed that, in enterocyte-like Caco-2 cells, low CPE concentrations cause caspase 3-mediated apoptosis but high CPE concentrations cause necrosis. The recent work published in mBio by Shrestha, Mehdizadeh Gohari, and McClane determined that RIP1 and RIP3 are involved in both CPE-mediated apoptosis and necrosis in Caco-2 cells. Furthermore, mixed lineage kinase-domain (MLKL) oligomerization was shown to be important for necrosis caused by CPE, identifying this necrosis as programmed necroptosis. In addition, calpain activation due to Ca2+ influx through the CPE pore was identified as a critical intermediate step for MLKL oligomerization and, thus, CPE-induced necroptosis. These findings may have applicability to understand the action of some other pore-forming toxins that induce necroptosis and may also be important for understanding CPE action in vivo.

scholarly journals A Novel Panel of Rabbit Monoclonal Antibodies and Their Diverse Applications Including Inhibition of Clostridium perfringens Epsilon Toxin Oligomerization

Antibodies ◽  
2018 ◽  
Vol 7 (4) ◽  
pp. 37 ◽  
Author(s):  
Jennifer Linden ◽  
Kiel Telesford ◽  
Samantha Shetty ◽  
Paige Winokour ◽  
Sylvia Haigh ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Death ◽  
Monoclonal Antibodies ◽  
Western Blot ◽  
Clostridium Perfringens ◽  
Specificity And Sensitivity ◽  
Epsilon Toxin ◽  
Blocking Antibodies

The pore-forming epsilon toxin (ETX) produced by Clostridium perfringens is among the most lethal bacterial toxins known. Sensitive antibody-based reagents are needed to detect toxin, distinguish mechanisms of cell death, and prevent ETX toxicity. Using B-cell immuno-panning and cloning techniques, seven ETX-specific monoclonal antibodies were generated from immunized rabbits. ETX specificity and sensitivity were evaluated via western blot, ELISA, immunocytochemistry (ICC), and flow cytometry. ETX-neutralizing function was evaluated both in vitro and in vivo. All antibodies recognized both purified ETX and epsilon protoxin via western blot with two capable of detecting the ETX-oligomer complex. Four antibodies detected ETX via ELISA and three detected ETX bound to cells via ICC or flow cytometry. Several antibodies prevented ETX-induced cell death by either preventing ETX binding or by blocking ETX oligomerization. Antibodies that blocked ETX oligomerization inhibited ETX endocytosis and cellular vacuolation. Importantly, one of the oligomerization-blocking antibodies was able to protect against ETX-induced death post-ETX exposure in vitro and in vivo. Here we describe the production of a panel of rabbit monoclonal anti-ETX antibodies and their use in various biological assays. Antibodies possessing differential specificity to ETX in particular conformations will aid in the mechanistic studies of ETX cytotoxicity, while those with ETX-neutralizing function may be useful in preventing ETX-mediated mortality.

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.

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