Globoside and the mucosal pH mediate parvovirus B19 entry through the epithelial barrier

Parvovirus B19 (B19V) is transmitted primarily via the respiratory route, however, the mechanism involved remains unknown. B19V targets a restricted receptor expressed in erythroid progenitor cells in the bone marrow. However, B19V shifts the receptor under acidic conditions and targets the widely expressed globoside. The pH-dependent interaction with globoside may allow virus entry through the naturally acidic nasal mucosa. To test this hypothesis, MDCK II cells and well-differentiated human airway epithelial cell (hAEC) cultures were grown on porous membranes and used as models to study the interaction of B19V with the epithelial barrier. Globoside expression was detected in polarized MDCK II cells and the ciliated cell population of well-differentiated hAEC cultures. Under the acidic conditions of the nasal mucosa, virus attachment and transcytosis occurred without productive infection. Neither virus attachment nor transcytosis was observed under neutral pH conditions or in globoside knockout cells, demonstrating the concerted role of globoside and acidic pH in the transcellular transport of B19V. Globoside-dependent virus uptake involved VP2 and occurred by a clathrin-independent pathway that is cholesterol and dynamin-dependent. This study provides mechanistic insight into the transmission of B19V through the respiratory route and reveals novel vulnerability factors of the epithelial barrier to viruses. Author summaryThe mechanism of entry of parvovirus B19 through the respiratory epithelium remains elusive. By using polarized MDCK II and well-differentiated primary human airway epithelial cell cultures, we revealed that the virus exploits the natural acidic environment of the nasal mucosa and the expression of globoside in the ciliated cell population of the airway to breach the respiratory epithelium by transcytosis. Mechanistically, virus entry and intracellular trafficking in the epithelial cells differ substantially from that in the permissive erythroid progenitor cells.