Fig. 3: ISH of AAV2 in liver tissue.
From: Adeno-associated virus 2 infection in children with non-A–E hepatitis
a–g, RNA ISH for the detection of AAV2 RNA in sections of formalin-fixed and paraffin-embedded liver tissues from children (one section per patient) with non A–E hepatitis. a, AAV2 RNA (red signal, indicated by an arrow) was detected in the endothelial cells of arteries in an explant liver section from patient CVR35. The vascular lumen is highlighted with by an asterisk. b, A positive AAV2 signal was detected in the nuclei of hepatocytes with vacuolated morphology from patient CVR4 (indicated by arrows) and in a negative cell (indicated by the circle). c,d, A liver section from patient CVR1 showed AAV2 RNA both in the nucleus and in the cytoplasm (c), whereas for patient CVR9 (d), AAV2 RNA was found only in the nucleus (indicated by arrows). e, A high percentage of hepatocytes with a positive signal for AAV2 was present predominantly in the nucleus of hepatocytes in the samples from patient CVR1. f, AAV2 was not detectable in liver sections from samples from healthy individuals in either the endothelial cells or hepatocytes. g, Samples from patient CVR35 showed inclusion bodies in hepatocytes. Left, small, dark basophilic intranuclear inclusions next to the nucleolus (indicated by arrows). Right, a large, pale basophilic, diffuse intranuclear inclusion body (suggestive of adenovirus infection; indicated by an arrow) next to a multinucleated giant cell in the liver (indicated by the asterisk). h, AAV2-positive cells were quantified using QuPath in biopsy samples from five patients with non-A–E hepatitis (cases) and from controls. Patient CVR35 (who received a liver transplant) is highlighted in red. Using the entire section, cells were segmented to identify the nuclei and cytoplasm, and the algorithm was tuned to detect red signals. All samples were analysed using the same algorithm. Scale bars, 25 μm (insets of c,d), 50 μm (a–d,f,g) or 200 μm (e).
