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. 2020 Jan 10;10:2992. doi: 10.3389/fmicb.2019.02992

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Copyright © 2020 Pizarro, Navarrete, Mendez, Zaror, Mansilla, Tapia, Carrasco, Salazar, Murua, Padula, Otth and Rodríguez.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Human lung from lethal Hantavirus pulmonary syndrome (HPS) cases. (A) Paraffin section immunostained with anti-ANDV. Immunoreactive Hantavirus antigens are present in epithelial cells (ep) lining the alveoli, septal and luminal macrophages (M) and in cells located within blood vessels (bv) that might correspond to monocytes (Mo?). Broken and full blue arrows point to small and large immunoreactive inclusions in epithelial cells, respectively. (B) Double immunofluorescence for ANDV (green) and CF68 (red). Particulate ANDV antigens are seen in alveolar septa (arrow). There are luminal macrophages that contain virus antigens (M+) and others that do not (M–). Inset. M+ macrophage visualized only with the channel for ANDV. (C) A large blood vessel (bv) located close to an alveolar septum is shown. The alveolar epithelium (ep) contains viral granules (arrow). Cells 1 to 4 likely correspond to circulating ANDV + monocytes at different stages of migration into the alveolar lumen (al). (D) 7 μm thick paraffin section stained first with silver methenamine (SM) and then immunostained with anti-N-ANDV. The basal lamina of the epithelial and endothelial cells appears black (full black arrow). Cell located between the basal lamina and the alveolar lumen (al) contains masses (full blue arrow) and granules (broken blue arrow) of immunoreactive Hantavirus antigens (broken arrows). (E) 7 μm thick paraffin section immunostained with anti-N-ANDV and background stained with Toluidine blue (Tb). Nomarski optic. Viral antigens are seen in alveolar epithelial cells (ep) and in endothelial cells (en). Broken and full blue arrows point to small and large immunoreactive inclusions in epithelial cells, respectively. al, alveolar lumen; bv, blood vessel; ep, alveolar epithelium; en, endothelium; Mo, monocyte?; M, macrophage; Tb, toluidine blue; SM, silver methenamine. Scale bars. (A) 20 μm; (B) 14 μm; (C) 16 μm; (D) 10 μm; (E) 6 μm.