Parechovirus A Infection of the Intestinal Epithelium: Differences Between Genotypes A1 and A3

Figures

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  Figure 1. Schematic representation of the model used in this study. Dissociated enteroids are seeded on a porous Transwell ® insert where they form a polarized monolayer containing the different cell types of the intestinal epithelium.

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  Figure 2. Replication kinetics of lab-adapted strain PeV-A1 (Harris) and two lab-adapted PeV-A3 strains (PeV-A3 152037 and A308-99) in intestinal monolayers. (A–C) Represents the relative increase in RNA copies in samples collected from the apical compartment after both apical and basolateral inoculation. (D) Represents the total amount of viral copies at 0 dpi and 4 dpi in the basolateral compartment, after either apical or basolateral inoculation. In all cases, data represent the mean ± SEM of two technical replicates in three biological replicates. * p-value < 0.05. dpi = days post infection.

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  Figure 3. Virus titer at 0 and 4 dpi of apically collected samples. Titers were determined by TCID50. In all cases, data represent the mean ± SEM of two technical replicates in three biological replicates. * p-value < 0.05. dpi = days post infection.

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  Figure 4. Replication kinetics of clinical isolates of PeV-A1 and PeV-A3 in intestinal monolayers. (A–D) Represent the relative increase in RNA copies in samples collected from the apical compartment after both apical and basolateral inoculation; (A, B) are PeV-A1 clinical isolates from stool samples; (C, D) are PeV-A3 clinical isolates from stool samples. (E) Represents the total amount of viral copies at 0 dpi and 4 dpi in the basolateral compartment, after either apical or basolateral inoculation. In all cases, data represent the mean ± SEM of two technical replicates in three biological replicates. * p-value < 0.05. dpi = days post infection.

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  Figure 5. Virus titer at 0 and 4 days post infection of apically collected samples for (A) PeV-A1 clinical isolates and (B) PeV-A3 clinical isolates. Titers were determined by TCID50. In all cases, data represent the mean ± SEM of two technical replicates in three biological replicates. * p-value < 0.05. dpi = days post infection.

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  Figure 6. Confocal images of infected cultures with clinical isolates of PeV-A1 and PeV-A3. In panel (A), cultures were stained with Paneth cell marker (LYZ) in yellow and PeV-A antibody in red; in panel (B), they were stained with enterocyte marker (villin) in yellow and PeV-A antibody in red, and in panel (C), cultures were stained with goblet cell marker (MUC2) in yellow and PeV-A antibody in red. In all cases, nuclei were stained with DAPI in cyan. Scale bars in white represent 20 µm, and the boxes indicate positively stained cells for the cell marker in the MOCK cases and for viral and cell marker staining in the infected images.