The Controlled Human Infection Model for Enterotoxigenic Escherichia coli

2021 ◽  
Author(s):  
Chad K. Porter ◽  
Kawsar R. Talaat ◽  
Sandra D. Isidean ◽  
Alwine Kardinaal ◽  
Subhra Chakraborty ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Human Infection ◽  
Enterotoxigenic Escherichia Coli ◽  
Infection Model

scholarly journals Hyperimmune Bovine Colostral Anti-CS17 Antibodies Protect Against Enterotoxigenic Escherichia coli Diarrhea in a Randomized, Doubled-Blind, Placebo-Controlled Human Infection Model

2019 ◽  
Vol 220 (3) ◽  
pp. 505-513 ◽  
Author(s):  
Stephen J Savarino ◽  
Robin McKenzie ◽  
David R Tribble ◽  
Chad K Porter ◽  
Aisling O’Dowd ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Clinical Trials ◽  
Human Infection ◽  
Enterotoxigenic Escherichia Coli ◽  
Watery Diarrhea ◽  
Infection Model ◽  
Significant Protection ◽  
Etec Strain ◽  
Colonization Factors ◽  
To Receive

AbstractBackgroundEnterotoxigenic Escherichia coli (ETEC) commonly cause diarrhea in children living in developing countries and in travelers to those regions. ETEC are characterized by colonization factors (CFs) that mediate intestinal adherence. We assessed if bovine colostral IgG (bIgG) antibodies against a CF, CS17, or antibodies against CsbD, the minor tip subunit of CS17, would protect subjects against diarrhea following challenge with a CS17-expressing ETEC strain.MethodsAdult subjects were randomized (1:1:1) to receive oral bIgG against CS17, CsbD, or placebo. Two days prior to challenge, subjects began dosing 3 times daily with the bIgG products (or placebo). On day 3, subjects ingested 5 × 109 cfu ETEC strain LSN03-016011/A in buffer. Subjects were assessed for diarrhea for 120 hours postchallenge.ResultsA total of 36 subjects began oral prophylaxis and 35 were challenged with ETEC. While 50.0% of the placebo recipients had watery diarrhea, none of the subjects receiving anti-CS17 had diarrhea (P = .01). In contrast, diarrhea rates between placebo and anti-CsbD recipients (41.7%) were comparable (P = 1.0).ConclusionsThis is the first study to demonstrate anti-CS17 antibodies provide significant protection against ETEC expressing CS17. More research is needed to better understand why anti-CsbD was not comparably efficacious.Clinical Trials Registration. NCT00524004

scholarly journals Improvement of the Enterotoxigenic Escherichia coli Infection Model for Post-Weaning Diarrhea by Controlling for Bacterial Adhesion, Pig Breed and MUC4 Genotype

2020 ◽  
Vol 7 (3) ◽  
pp. 106
Author(s):  
Hiroki Matsumoto ◽  
Masashi Miyagawa ◽  
Sayaka Takahashi ◽  
Ryouichi Shima ◽  
Takayuki Oosumi
Keyword(s):  
Escherichia Coli ◽  
Experimental Infection ◽  
Antibacterial Agents ◽  
Experimental Models ◽  
Enterotoxigenic Escherichia Coli ◽  
Infection Model ◽  
Swine Industry ◽  
Pig Breeds ◽  
Adhesion Ability ◽  
Escherichia Coli Infection

Enterotoxigenic Escherichia coli (ETEC) is a major cause of post-weaning diarrhea (PWD) in pigs and causes significant damage to the swine industry worldwide. In recent years, there has been increased regulation against the use of antibacterial agents in swine due to their health risks. Utilizing experimental models that consistently recapitulate PWD is important for the development of non-antibacterial agents against PWD in pigs. In this study, we established a highly reproducible PWD infection model by examining differences in adhesion of ETEC to the intestinal tissue as well as the association between MUC4 polymorphisms and sensitivity to PWD. Post-weaning diarrhea differences between pig breeds were also examined. The adhesion to enterocytes varied from 104.0 to 106.4 CFU/mL even among the F4 ETEC strains. Experimental infection revealed that PWD can be induced in all MUC4 genotypes after infection with 1010 CFU/pig of highly adherent ETEC, although there were variable sensitivities between the genotypes. Lowly adherent ETEC did not cause PWD as efficiently as did highly adherent ETEC. The incidence of PWD was confirmed for all pigs with the ETEC-susceptible MUC4 genotypes in all of the breeds. These results indicate that high-precision and reproducible experimental infection is possible regardless of pig breeds by controlling factors on the pig-end (MUC4 genotype) and the bacterial-end (adhesion ability).

scholarly journals Experimental Infection of Human Volunteers with the Heat-Stable Enterotoxin-Producing Enterotoxigenic Escherichia coli Strain TW11681

Pathogens ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 84 ◽  
Author(s):  
Sunniva Todnem Sakkestad ◽  
Hans Steinsland ◽  
Steinar Skrede ◽  
Elisabeth Kleppa ◽  
Kristine Lillebø ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
T Cell ◽  
Protective Efficacy ◽  
T Cell Responses ◽  
Enterotoxigenic Escherichia Coli ◽  
Cd4 T Cell ◽  
Infection Model ◽  
Serological Response ◽  
Cell Responses ◽  
Heat Stable

Infection with enterotoxigenic Escherichia coli (ETEC) producing the heat-stable enterotoxin (ST) is one of the most important causes of childhood diarrhoea in low- and middle-income countries. Here, we undertook a controlled human infection model (CHIM) study to investigate whether ST-producing ETEC strain TW11681 would be suitable for testing the protective efficacy of new ST-based vaccine candidates in vaccine challenge models. In groups of three, nine volunteers ingested 1 × 106, 1 × 107, or 1 × 108 colony-forming units (CFU) of TW11681. Flow cytometry-based assays were used to measure CD4+ T cell responses and antibody levels targeting virulence factors expressed by the strain. We found that infection with TW11681 elicited few and mild symptoms, including mild diarrhoea in two volunteers, both of whom ingested 1 × 106 CFU. Averaged across all volunteers, the CD4+ T cell responses specific for E. coli YghJ mucinase peaked 10 days after infection (3.2-fold (p = 0.016)), while the CD4+ T cell responses specific for Colonization Factor Antigen I (CFA/I) major fimbrial subunit (CfaB) peaked after 28 days (3.6-fold (p = 0.063)). The serum CfaB-specific anti-IgA and anti-IgG/IgM levels were significantly increased and peaked 3 months after infection. Both remained elevated for the duration of the 12-month follow-up. The corresponding anti-YghJ serological response was strongest after 10 days, although a significant increase was seen only for IgA levels (3.2-fold (p = 0.008)). In conclusion, due to its low diarrhoea attack risk, TW11681 is probably not suitable for testing the efficacy of new vaccines in human challenge studies at doses 1 × 106 to 1 × 108. However, the strain may still be useful in CHIMs for studying ETEC host-pathogen interactions.

scholarly journals 1028 Sleep and Enteric Disease: Sleep Now for Less Diarrhea Later

SLEEP ◽  
2020 ◽  
Vol 43 (Supplement_1) ◽  
pp. A390-A391
Author(s):  
J Mantua ◽  
R L Gutierrez ◽  
S D Isidean ◽  
A N Alaca ◽  
K J Testa ◽  
...  
Keyword(s):  
Research Program ◽  
Sleep Time ◽  
Human Infection ◽  
Enterotoxigenic Escherichia Coli ◽  
Infection Model ◽  
Enteric Disease ◽  
Us Army ◽  
Medicine Research ◽  
Severe Diarrhea ◽  
The Us

Abstract Introduction The bi-directional relationship between sleep and immune function is well-established. Sufficient sleep supports immune health and can increase vaccine efficacy. Conversely, sickness can disturb sleep quality, which can delay recovery and waking functioning. However, the bidirectional relationship between sleep and infectious diarrhea, the leading infectious disease threat to deployed military populations, has not been studied. We assessed the bi-directional relationship between sleep and enteric disease utilizing data from a recently-completed controlled human infection model (CHIM) with enterotoxigenic Escherichia coli (ETEC). Methods During a CHIM to assess the efficacy of an immunoprophylactic targeting ETEC (NCT03040687), we measured sleep via actigraphy over an 8-day inpatient period. Participants ingested prophylaxis 3 times/day during days -2 and -1 and ingested ETEC on day 0. The primary outcome was moderate-severe diarrhea following the ETEC challenge. We hypothesized better sleep pre-challenge would reduce risk of disease after the challenge (assessed using linear regression). We also hypothesized total sleep time (TST) and sleep efficiency (SE) after the challenge would be lower/poorer than baseline (assessed using paired t-test). Results Among 59 participants (aged 34.4±8.1yrs, 64% female), longer TST the night preceding ETEC challenge was associated with lower total diarrhea volume (B=-3.13,p=.001). SE was slightly but significantly poorer after the challenge (78 vs. 76%; t(55)=2.2,p=.03), but there was no significant change in TST, potentially due to low TST pre-challenge (316 vs. 329 minutes; p=0.12). Conclusion These results - in aggregation with previous work on sleep and vaccines - suggest military sleep regulations should be put in place to increase sleep prior to traveling to an area of responsibility with high risk for enteric disease. These minor behavioral changes could provide lasting benefits to readiness of military servicemembers. Support This work was supported by Joint Warfighter Medical Research Program (JWMRP) and the Military Operational Medicine Research Program (MOMRP). The opinions and assertions contained herein are the private views of the authors and are not to be construed as official or as reflecting the views of the US Army or of the US Department of Defense. This abstract has been approved for public release with unlimited distribution.

scholarly journals Technical note: novel delivery methods for an enterotoxigenic Escherichia coli infection model in MUC4-locus sequenced weaner pigs1

2019 ◽  
Vol 97 (11) ◽  
pp. 4503-4508 ◽  
Author(s):  
Samantha O Sterndale ◽  
David W Miller ◽  
Josie P Mansfield ◽  
Jae C Kim ◽  
Mark O’Dea ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Positive Control ◽  
Negative Control ◽  
Technical Note ◽  
Enterotoxigenic Escherichia Coli ◽  
Infection Model ◽  
Large White ◽  
Treatment Groups ◽  
Marker Test ◽  
Escherichia Coli Infection

Abstract An infection model with enterotoxigenic Escherichia coli (ETEC) harboring the F4 fimbriae can be used to assess the impacts that various challenges associated with weaning (e.g., dietary, psychological, environmental) have on the expression of postweaning diarrhea. The objective of this study was to develop a novel inoculation method for administering an ETEC culture that would induce a higher proportion of ETEC-F4 diarrhea, in pigs that genetically showed ETEC-F4 susceptibility or resistance. The study was designed as a factorial arrangement of treatments with the factors being 1) partially susceptible or resistant to ETEC-F4 based on genetic testing, and 2) 4 challenge treatments, being a) a conventional liquid broth method using a drenching gun [Positive control (PC)], b) a Syringe method, c) a Capsule method, and d) Negative control [pigs not challenged (NC)]. At 21 ± 3 d of age (mean ± SEM), 48 male castrate pigs (Large White × Landrace) weighing approximately 7.0 ± 1.18 kg were allocated to 4 treatment groups in 2 replicate pens (6 pigs per pen). Initial ETEC-F4 susceptibility was based on a DNA marker test and each treatment group had 9 partially susceptible and 3 resistant pigs. On days 7 and 8 after weaning, pigs were challenged with ETEC (serotype O149:K88; toxins LT1, ST1, ST2, and EAST). On each inoculation day the PC pigs were orally dosed with 9 mL 7.12 × 109 colony-forming unit (CFU), the Syringe pigs with 0.8 mL 6.72 × 109 CFU, the Capsule pigs were orally administered 2 capsules containing 0.8 mL 3.28 × 109 CFU, and the NC pigs 1 mL of phosphate-buffered saline (PBS) solution. Approximately 72 h after infection, 44, 22, 78, and 0% of partially susceptible pigs in the PC, the Syringe, the Capsule, and the NC group had developed ETEC-F4 diarrhea (P = 0.007). Partially susceptible pigs had a higher diarrhea index (DI) compared to resistant pigs (31.5 vs. 4.8, P < 0.001). The NC group had a lower DI compared to the PC and Capsule pigs (3.9, 38.1, and 40.3, respectively, P < 0.005). Following infection, genetically resistant pigs in the Capsule group had a DI of zero and the partially susceptible pigs had a DI of 55.6 (P = 0.014). This study showed that genetically screening pigs and using a Capsule to deliver ETEC-F4 can increase cases of diarrhea and the efficiency of the challenge model. Taken together, these methods have the potential to reduce the number of pigs needed in future experimental infection studies.

scholarly journals Antibacterial and Antibiofilm Activities of Novel Antimicrobial Peptides against Multidrug-Resistant Enterotoxigenic Escherichia Coli

2021 ◽  
Vol 22 (8) ◽  
pp. 3926
Author(s):  
Kang-Chi Wu ◽  
Kuo-Feng Hua ◽  
Yu-Hsiang Yu ◽  
Yeong-Hsiang Cheng ◽  
Ting-Ting Cheng ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Peptides ◽  
Cellular Localization ◽  
Membrane Integrity ◽  
Multidrug Resistant ◽  
Enterotoxigenic Escherichia Coli ◽  
Infection Model ◽  
Common Disease ◽  
Selective Toxicity ◽  
Minimal Inhibition Concentration

Post-weaning diarrhea due to enterotoxigenic Escherichia coli (ETEC) is a common disease of piglets and causes great economic loss for the swine industry. Over the past few decades, decreasing effectiveness of conventional antibiotics has caused serious problems because of the growing emergence of multidrug-resistant (MDR) pathogens. Various studies have indicated that antimicrobial peptides (AMPs) have potential to serve as an alternative to antibiotics owing to rapid killing action and highly selective toxicity. Our previous studies have shown that AMP GW-Q4 and its derivatives possess effective antibacterial activities against the Gram-negative bacteria. Hence, in the current study, we evaluated the antibacterial efficacy of GW-Q4 and its derivatives against MDR ETEC and their minimal inhibition concentration (MIC) values were determined to be around 2~32 μg/mL. Among them, AMP Q4-15a-1 with the second lowest MIC (4 μg/mL) and the highest minimal hemolysis concentration (MHC, 256 μg/mL), thus showing the greatest selectivity (MHC/MIC = 64) was selected for further investigations. Moreover, Q4-15a-1 showed dose-dependent bactericidal activity against MDR ETEC in time–kill curve assays. According to the cellular localization and membrane integrity analyses using confocal microscopy, Q4-15a-1 can rapidly interact with the bacterial surface, disrupt the membrane and enter cytosol in less than 30 min. Minimum biofilm eradication concentration (MBEC) of Q4-15a-1 is 4× MIC (16 μg/mL), indicating that Q4-15a-1 is effective against MDR ETEC biofilm. Besides, we established an MDR ETEC infection model with intestinal porcine epithelial cell-1 (IPEC-1). In this infection model, 32 μg/mL Q4-15a-1 can completely inhibit ETEC adhesion onto IPEC-1. Overall, these results suggested that Q4-15a-1 may be a promising antibacterial candidate for treatment of weaned piglets infected by MDR ETEC.

scholarly journals Interleukin-8 Response in an Intestinal HCT-8 Cell Line Infected with Enteroaggregative and Enterotoxigenic Escherichia coli

2004 ◽  
Vol 11 (3) ◽  
pp. 548-551 ◽  
Author(s):  
David B. Huang ◽  
Herbert L. DuPont ◽  
Zhi-Dong Jiang ◽  
Lily Carlin ◽  
Pablo C. Okhuysen
Keyword(s):  
Escherichia Coli ◽  
Cell Line ◽  
Virulence Factors ◽  
Interleukin 8 ◽  
Enterotoxigenic Escherichia Coli ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infection Model ◽  
E Coli ◽  
Heat Stable ◽  
Line Infection

ABSTRACT This study examined the interleukin-8 (IL-8) response of the intestinal adenocarcinoma HCT-8 cell line to infection with enteroaggregative and enterotoxigenic Escherichia coli pathotypes isolated from patients with travelers' diarrhea. Individual diarrheagenic E. coli strains (enteroaggregative E. coli [EAEC]; n = 30), heat-stable enterotoxin (ST)-producing enterotoxigenic E. coli (ETEC ST; n = 11), heat-labile enterotoxin (LT)-producing enterotoxigenic E. coli (ETEC LT; n = 10), and ST- and LT-producing enterotoxigenic E. coli (ETEC ST:LT; n = 8) were coincubated with HCT-8 cells for 3 h. Tissue culture supernatants were assayed for IL-8 content by enzyme-linked immunosorbent assay. Fifty percent of EAEC (72% of those EAEC carrying the virulence factors aggR, aggA, and aspU and 40% of those EAEC not carrying virulence factors) and 64% of ETEC ST elicited IL-8 production. In contrast, 10% of ETEC LT elicited the production of IL-8 above baseline. These results suggest that (i) the HCT-8 cell line infection model can be used as a tool to differentiate proinflammatory E. coli from noninflammatory isolates; (ii) EAEC has a heterogeneous ability to induce the production of IL-8, and this may be associated with the presence of virulence factors; and (iii) ETEC ST can elicit an inflammatory response and helps explain our earlier findings of increased fecal IL-8 in patients with ETEC diarrhea.

scholarly journals An infection-induced RhoB-Beclin 1-Hsp90 complex enhances clearance of uropathogenic Escherichia coli

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Chunhui Miao ◽  
Mingyu Yu ◽  
Geng Pei ◽  
Zhenyi Ma ◽  
Lisong Zhang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Treatment Strategies ◽  
Uropathogenic Escherichia Coli ◽  
Beclin 1 ◽  
Host Cells ◽  
Infection Model ◽  
Intracellular Bacteria ◽  
Bladder Epithelium ◽  
Binding Residue

AbstractHost cells use several anti-bacterial pathways to defend against pathogens. Here, using a uropathogenic Escherichia coli (UPEC) infection model, we demonstrate that bacterial infection upregulates RhoB, which subsequently promotes intracellular bacteria clearance by inducing LC3 lipidation and autophagosome formation. RhoB binds with Beclin 1 through its residues at 118 to 140 and the Beclin 1 CCD domain, with RhoB Arg133 being the key binding residue. Binding of RhoB to Beclin 1 enhances the Hsp90-Beclin 1 interaction, preventing Beclin 1 degradation. RhoB also directly interacts with Hsp90, maintaining RhoB levels. UPEC infections increase RhoB, Beclin 1 and LC3 levels in bladder epithelium in vivo, whereas Beclin 1 and LC3 levels as well as UPEC clearance are substantially reduced in RhoB+/− and RhoB−/− mice upon infection. We conclude that when stimulated by UPEC infections, host cells promote UPEC clearance through the RhoB-Beclin 1-HSP90 complex, indicating RhoB may be a useful target when developing UPEC treatment strategies.

scholarly journals Structure of the master regulator Rns reveals an inhibitor of enterotoxigenic Escherichia coli virulence regulons

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Charles R. Midgett ◽  
Kacey Marie Talbot ◽  
Jessica L. Day ◽  
George P. Munson ◽  
F. Jon Kull
Keyword(s):  
Escherichia Coli ◽  
Small Molecules ◽  
Family Member ◽  
Shigella Flexneri ◽  
Decanoic Acid ◽  
Enterotoxigenic Escherichia Coli ◽  
Gram Negative Bacteria ◽  
Enteric Infections ◽  
First Time ◽  
Arac Family

AbstractEnteric infections caused by the gram-negative bacteria enterotoxigenic Escherichia coli (ETEC), Vibrio cholerae, Shigella flexneri, and Salmonella enterica are among the most common and affect billions of people each year. These bacteria control expression of virulence factors using a network of transcriptional regulators, some of which are modulated by small molecules as has been shown for ToxT, an AraC family member from V. cholerae. In ETEC the expression of many types of adhesive pili is dependent upon the AraC family member Rns. We present here the 3 Å crystal structure of Rns and show it closely resembles ToxT. Rns crystallized as a dimer via an interface similar to that observed in other dimeric AraC’s. Furthermore, the structure of Rns revealed the presence of a ligand, decanoic acid, that inhibits its activity in a manner similar to the fatty acid mediated inhibition observed for ToxT and the S. enterica homologue HilD. Together, these results support our hypothesis that fatty acids regulate virulence controlling AraC family members in a common manner across a number of enteric pathogens. Furthermore, for the first time this work identifies a small molecule capable of inhibiting the ETEC Rns regulon, providing a basis for development of therapeutics against this deadly human pathogen.

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