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Published on: February 17, 2023
Feline Tritrichomonas foetus adhere to intestinal epithelium by receptor-ligand-dependent mechanisms
M K Tolbert1, S H Stauffer, J L Gookin
1North Carolina State University, Center for Comparative Medicine and Translational Research, College of Veterinary Medicine, Department of Clinical Sciences, Raleigh, NC, USA.
Abstract:
Tritrichomonas foetus (TF) is a protozoan that infects the feline ileum and colon resulting in chronic diarrhea. Up to 30% of young purebred cats are infected with TF and the infection is recognized as pandemic. Only a single drug, characterized by a narrow margin of safety and emerging development of resistance, is effective for treatment. While the venereal pathogenicity of bovine TF is attributed to adherence to uterovaginal epithelium, the pathogenesis of diarrhea in feline TF infection is unknown. The aim of this study was to establish an in vitro model of feline TF adhesion to intestinal epithelium. Confluent monolayers of porcine intestinal epithelial cells (IPEC-J2) were infected with axenic cultures of feline TF that were labeled with [(3)H] thymidine or CFSE and harvested at log-phase. The effect of multiplicity and duration of infection, viability of TF, binding competition, formalin fixation and cytoskeletal inhibitors on adherence of feline TF to IPEC-J2 monolayers was quantified by liquid scintillation counting and immunofluorescence. [(3)H] thymidine and CFSE-labeled TF reproducibly adhered to IPEC-J2 monolayers. Clinical isolates of feline TF adhered to the intestinal epithelium in significantly greater numbers than Pentatrichomonas hominis, the latter of which is a presumably nonpathogenic trichomonad. Adhesion of TF required viable trophozoites but was independent of cytoskeletal activity. Based on saturation and competition binding experiments, adherence of feline TF to the epithelium occurred via specific receptor-ligand interactions. The developed model provides a valuable resource for assessing pathogenic mechanisms of feline TF and developing novel pharmacologic therapies for blocking the adhesion of feline TF to the intestinal epithelium.
Insights
Tritrichomonas foetus (TF) causes pandemic feline diarrhea. This study developed an in vitro model showing TF adheres to intestinal cells via specific interactions, paving the way for new treatments.
Area of Science:
- Veterinary Parasitology
- Feline Infectious Diseases
- Cell Biology
Background:
- Tritrichomonas foetus (TF) is a protozoan causing chronic diarrhea in cats, with up to 30% of young cats infected.
- Current treatments for feline TF are limited by narrow safety margins and emerging resistance.
- The pathogenesis of diarrhea in feline TF infection remains largely unknown.
Purpose of the Study:
- To establish an in vitro model to study feline TF adhesion to intestinal epithelium.
- To investigate the mechanisms underlying feline TF adherence to intestinal cells.
- To provide a platform for developing novel therapeutic strategies against feline TF.
Main Methods:
- Developed an in vitro model using porcine intestinal epithelial cells (IPEC-J2) infected with axenic feline TF cultures.
- Labeled TF with [(3)H] thymidine or CFSE for quantification via liquid scintillation counting and immunofluorescence.
- Assessed the effects of infection parameters, TF viability, competition, and cytoskeletal inhibitors on TF adherence.
Main Results:
- Reproducible adhesion of labeled feline TF to IPEC-J2 monolayers was demonstrated.
- Clinical TF isolates showed significantly greater adherence than non-pathogenic Pentatrichomonas hominis.
- TF adhesion requires viable trophozoites and specific receptor-ligand interactions, independent of host cell cytoskeletal activity.
Conclusions:
- The developed in vitro model effectively simulates feline TF adhesion to intestinal epithelium.
- Adherence involves specific molecular interactions, offering potential targets for therapeutic intervention.
- This model is crucial for understanding feline TF pathogenesis and developing new anti-adhesion therapies.
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