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. 2020 Mar 20;9(1):651–663. doi: 10.1080/22221751.2020.1741326

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© 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group, on behalf of Shanghai Shangyixun Cultural Communication Co., Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — EqPV-H infects hepatocytes and results in hepatocellular necrosis with lymphocytic infiltrates. (A) An individual, necrotic hepatocyte (arrowhead) surrounded by small lymphocytes (arrow). Horse I, HE. (B) Random multifocal clusters of lymphocytes in the hepatic parenchyma (arrows). Horse J, HE. (C) Cells within the clusters of lymphocytes are CD3 positive. Horse J, CD3 IHC. (D) Horse A had the most severe biochemical and clinical hepatitis, which corresponded with the most severe liver pathology. At the beginning of hepatitis (week 7), there was marked increase in cellularity throughout the parenchyma (Di). Pathology was minimal in periportal regions (asterisk, Dii) and portal tracts contained few inflammatory cells (arrow, Dii). Increased numbers of individual necrotic hepatocytes (arrows, Diii), and clusters of lymphocytes, neutrophils, and macrophages surrounding necrotic hepatocytes (arrowhead, Diii), were found throughout zones 2 and 3 of lobules. (E) Seven days later, Horse A’s parenchymal cellularity was already reduced (Ei, Eii). Inflammatory cells and individual necrotic hepatocytes were reduced overall, although portal tracts still contained increased numbers of mixed inflammatory cells (arrow, Eii). Increased numbers of hepatocytes with mitotic figures (arrowheads, Eiii) and fewer individual necrotic cells (arrow, Eiii) were found throughout the parenchyma, compared to the week 7 biopsy (D). Inflammatory cells in portal tracts narrowly breached the limiting plate (asterisk, Eiii). (F) Three hepatocytes are shown demonstrating the range of in situ hybridization intensity with punctate nuclear (arrowhead), cytoplasmic (thin arrow), and combined nuclear and cytoplasmic (broad arrow) hybridization. Horse J, EqPV-H ISH, antisense probe. (G) An individual necrotic cell with nuclear karyorrhexis and EqPV-H hybridization within the cytoplasm and extending into the surrounding parenchyma. Horse B, EqPV-H ISH, antisense probe. (H) A focal aggregate of cells in the parenchyma with positive hybridization. Horse I, EqPV-H ISH, sense probe. (I) The biopsy with the most severe pathology had large numbers of hepatocytes with mild to strong positive nuclear and cytoplasmic hybridization (Horse A week 7, same sample as D, EqPV-H ISH, antisense probe). (J) One week later, EqPV-H hybridization was rare (Horse A week 8, same sample as E, EqPV-H ISH, antisense probe) with only mild nuclear (arrowhead) and/or cytoplasmic (arrow) hybridization.