Polarized rotavirus entry and release from differentiated small intestinal cells

Area of Science:

• Virology
• Cell Biology
• Gastroenterology

Background:

• Rotaviruses are a leading cause of severe diarrheal disease in infants.
• Most rotavirus cellular entry studies use non-intestinal cell lines, limiting understanding of in vivo mechanisms.
• Porcine rotavirus (PRV) and its interactions with intestinal cells require further investigation.

Purpose of the Study:

• To investigate the cellular entry mechanism of the porcine rotavirus YM strain into the IPEC-J2 small intestinal cell line.
• To compare rotavirus entry and egress pathways from the apical versus basolateral cell surfaces.
• To elucidate the role of cholesterol and specific cellular processes in rotavirus infection.

Main Methods:

• Utilized the IPEC-J2 small intestinal cell line for infection studies.
• Compared infection efficiency from apical and basolateral surfaces.
• Assessed the impact of cholesterol depletion and enzymatic treatments (neuraminidase, bafilomycin) on viral entry.
• Investigated virus egress pathways.

Main Results:

• Porcine rotavirus YM, similar to human rotavirus Wa, preferentially infects IPEC-J2 cells from the basolateral surface.
• Cellular infection from both surfaces is cholesterol-dependent.
• Neuraminidase, sucrose, and bafilomycin treatments indicate distinct receptor usage and entry mechanisms for apical and basolateral infections.
• While entry is more efficient from the basolateral side, rotavirus egress is predominantly apical.

Conclusions:

• Rotavirus exhibits distinct entry and egress strategies depending on the cellular surface of intestinal cells.
• Basolateral entry is more efficient but apical egress is favored, suggesting complex viral trafficking.
• Cholesterol plays a critical role in rotavirus binding and/or entry into intestinal epithelial cells.