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A Video Protocol of Retroviral Infection in Primary Intestinal Organoid Culture
Published on: August 11, 2014
Modeling rotavirus infection and antiviral therapy using primary intestinal organoids
Yuebang Yin1, Marcel Bijvelds1, Wen Dang1
1Department of Gastroenterology and Hepatology, Erasmus MC-University Medical Center, Rotterdam, The Netherlands.
Abstract:
Despite the introduction of oral vaccines, rotavirus still kills over 450,000 children under five years of age annually. The absence of specific treatment prompts research aiming at further understanding of pathogenesis and the development of effective antiviral therapy, which in turn requires advanced experimental models. Given the intrinsic limitations of the classical rotavirus models using immortalized cell lines infected with laboratory-adapted strains in two dimensional cultures, our study aimed to model infection and antiviral therapy of both experimental and patient-derived rotavirus strains using three dimensional cultures of primary intestinal organoids. Intestinal epithelial organoids were successfully cultured from mouse or human gut tissues. These organoids recapitulate essential features of the in vivo tissue architecture, and are susceptible to rotavirus. Human organoids are more permissive to rotavirus infection, displaying an over 10,000-fold increase in genomic RNA following 24h of viral replication. Furthermore, infected organoids are capable of producing infectious rotavirus particles. Treatment of interferon-alpha or ribavirin inhibited viral replication in organoids of both species. Importantly, human organoids efficiently support the infection of patient-derived rotavirus strains and can be potentially harnessed for personalized evaluation of the efficacy of antiviral medications. Therefore, organoids provide a robust model system for studying rotavirus-host interactions and assessing antiviral medications.
Insights
Three-dimensional organoid cultures effectively model rotavirus infection and antiviral treatment. This advanced model aids in understanding rotavirus pathogenesis and developing new therapies for this deadly childhood virus.
Area of Science:
- Virology
- Gastroenterology
- Biotechnology
Background:
- Rotavirus remains a significant cause of mortality in children under five, despite oral vaccine availability.
- Current rotavirus research models have limitations, hindering the development of effective antiviral therapies.
- Understanding rotavirus pathogenesis requires advanced experimental models beyond traditional cell lines.
Purpose of the Study:
- To develop and validate a novel three-dimensional (3D) organoid model for rotavirus infection and antiviral therapy.
- To compare rotavirus infection dynamics in primary intestinal organoids versus traditional cell culture models.
- To assess the efficacy of antiviral agents in a human organoid model using patient-derived rotavirus strains.
Main Methods:
- Primary intestinal organoids were successfully cultured from mouse and human gut tissues.
- Organoids were infected with rotavirus strains to model infection and assess viral replication.
- The effect of antiviral treatments, including interferon-alpha and ribavirin, was evaluated in infected organoids.
Main Results:
- Intestinal organoids recapitulated in vivo tissue architecture and supported rotavirus infection.
- Human organoids demonstrated high permissiveness to rotavirus, with over 10,000-fold genomic RNA increase.
- Antiviral treatments significantly inhibited rotavirus replication in both mouse and human organoids.
- Human organoids supported patient-derived rotavirus strains, showing potential for personalized antiviral evaluation.
Conclusions:
- Three-dimensional intestinal organoids provide a robust and relevant model for studying rotavirus-host interactions.
- This organoid system facilitates the assessment of antiviral medications against diverse rotavirus strains.
- The model holds promise for advancing rotavirus pathogenesis research and personalized medicine approaches.
