3B, red line) Accordingly, an HBVpreS1-receptor is present on th

3B, red line). Accordingly, an HBVpreS1-receptor is present on the hepatocytes of mice and rats. We extended this analysis and tested HBVpreS-peptide binding to primary hepatocytes from rabbits, dogs, cynomolgus monkeys, rhesus monkeys, and pigs. As depicted in Fig. 3C, we found specific binding of HBVpreS/2-48myr-K-FITC to rat, rabbit,

and dog hepatocytes but surprisingly not to hepatocytes from cynomolgus and rhesus monkey, despite their closer evolutionary relation to humans. Binding was also not observed on pig hepatocytes. Thus, differentiated hepatocytes from some HBV nonsusceptible species do express the HBVpreS-specific receptor (mouse, rat, dog, and rabbit), while others do not (pig, cynomolgus, and rhesus monkey), indicating that a step downstream specific preS-binding must restrict HBV/HDV-buy RG7204 infection in these species. The susceptibility of HepaRG cells to HBV infection depends on a differentiated state of the cells.7, 30 Moreover, PHH lose their

susceptibility to HBV when DMSO is withdrawn from the medium.31-34 In order to test if this correlates with the presence of the HBVpreS-receptor we analyzed undifferentiated (5 days after seeding) and differentiated HepaRG cells (28 days after seeding, including a 2-week DMSO treatment) for their ability to bind HBVpreS/2-48myr-K-FITC. As seen in Fig. 4A (upper panel, left), HBVpreS/2-48myr-K-FITC did not stain the PM of

undifferentiated cells, whereas binding was seen after differentiation (lower panel, left). This implies a cell state-dependent induction of HBVpreS/2-48myr-receptor expression during HepaRG-differentiation. Accordingly, we investigated whether dedifferentiation of binding competent primary hepatocytes result in an opposite effect. We cultivated PMH in the absence of DMSO and followed their ability to bind HBVpreS/2-48myr-K-FITC Sulfite dehydrogenase over time (Fig. 4A, right panels). While freshly isolated PMH specifically accumulated the peptide at the PM, hepatocytes from the same preparation lost their ability to bind HBVpreS/2-48myr-K-FITC within a few days of cultivation. Loss of binding could be prevented by addition of DMSO (data not shown). This correlates with the fact that PHH lose their susceptibility for HBV during several days of cultivation in the absence of DMSO.28 Thus, expression of an HBVpreS-receptor is linked to pathways controlling the differentiation state of hepatocytes. To investigate whether HuH7 and HepG2 express detectable amounts of the HBVpreS-receptor following DMSO-induced differentiation, we performed binding experiments with these cells under differentiation conditions. As shown in Fig. 4B (upper panel) dividing cultures of HuH7 and HepG2 cells showed no significant binding of HBVpreS/2-48myr-K-FITC. Cultivation in the presence of 0.

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