The epithelial lining of large airways (bronchi) has a basal layer containing progenitor cells characterized by cytoplasmic expression of cytokeratin 5 (green) and nuclear expression of p63 (red). cells. Aggregates of related inflammatory cells are present subpleurally (arrow). C,D: Aerosol AGM, remaining anterior lobe. C) The pleura is definitely segmentally thickened (arrows). D) The pleura is definitely lined by hypertrophic mesothelial cells (arrow) and there is infiltration of the subpleural parenchyma by histiocytes. E,F: IT/IN RM, remaining lower lobe. E) The pleura is definitely segmentally thickened (pleuritis, arrows). F) The pleura is definitely thickened by fibrosis and infiltrated by mononuclear cells, predominately lymphocytes. G,H: IT/IN AGM, right lower lobe. G) There is slight congestion and rare perivascular swelling (arrow). H) Perivascular swelling is characterized by infiltration of the tunica adventitia by mononuclear cells.(TIF) ppat.1010618.s004.tif (12M) GUID:?C910139E-3F4A-4A1A-8964-750618CDCEB1 S5 Fig: BAL flow cytometry gating strategy. Representative gating strategy to classify alveolar (CD163+CD206+), interstitial Eplivanserin mixture (CD163+CD206-), monocyte-derived (CD163+CD206+CD16+CCR2+), and resident alveolar (CD163+CD206+CD16-) macrophages in BAL.(TIF) ppat.1010618.s005.tif (8.0M) GUID:?A77F429C-0771-4CB3-A962-85C2EF95A34D S6 Fig: Hematology-Based Guidelines of SARS-CoV-2 Challenge. Complete blood counts were performed at indicated occasions and were compared for counts of RBCs, platelets, neutrophils, Eplivanserin mixture lymphocytes, WBCs and monocytes (A, B, C, D, E, and F, respectively), as well as neutrophil/lymphocytes percentage (G). Comparisons were made via two-way ANOVA with Tukeys multiple comparisons test. Asterisks represent significant comparisons (****, p 0.0001).(TIF) ppat.1010618.s006.tif Eplivanserin mixture (2.0M) GUID:?2AFDFDD1-8E93-414F-8535-0659946B077B S7 Fig: Clinical Chemistry-Based Parameters of SARS-CoV-2 Challenge. Clinical chemistries were performed at the indicated times post challenge. Comparisons between each group were made for log2 fold change from baseline of creatinine (A), ALT (B), BUN (D) and concentrations of CRP (C) and AST (E). Comparisons were made via two-way ANOVA with Tukeys multiple comparisons test. Asterisks represent significant comparisons (**, p 0.01).(TIF) ppat.1010618.s007.tif (1.7M) GUID:?4C8BB283-1280-4023-A802-8C6FF24F5641 S1 Table: Study Animal Characteristics. (TIF) ppat.1010618.s008.tif (3.0M) GUID:?8FA55F3E-C0E7-4AE7-89B3-B1AEE4438657 S2 Table: RT-qPCR Primers and Probes. (TIF) ppat.1010618.s009.tif (1.3M) GUID:?CFB0ACF2-EDB8-44BD-8FC3-DA66C330F10B S3 Table: Antibodies used for fluorescent immunochemistry. (TIF) ppat.1010618.s010.tif (3.3M) GUID:?EA0124C2-6430-4B80-A4CD-7C08AB92029B S4 Table: Antibodies used for flow cytometry analysis. (TIF) ppat.1010618.s011.tif (1.5M) GUID:?A77AEF4D-9BE0-4AE1-84D2-9F2B3E893158 Data Availability StatementAll relevant data are within the manuscript and its Supporting Information files. Abstract The novel coronavirus SARS-CoV-2 emerged in late 2019, rapidly reached pandemic status, and has maintained global ubiquity through the emergence of variants of Eplivanserin mixture concern. Efforts to develop animal models have mostly fallen short of recapitulating severe disease, diminishing their utility for research focusing on severe disease pathogenesis and life-saving medical countermeasures. We tested whether route of experimental contamination substantially changes COVID-19 disease characteristics in two species of nonhuman primates ((African green monkeys, AGMs) or (Rhesus macaques, RMs). Other examples of species studied include (Southern pigtail macaque) [7], (Northern pigtail macaque) [8], (Cynomolgus macaque) [9], (Common marmoset) and (Baboon) [10]. Most of these models involve installation of the virus directly to mucosal surfaces [11,12], though some have included the aerosol modality of exposure [13]. Overwhelmingly, the NHP model of SARS-CoV-2 contamination results in a moderate to moderate disease, with only one study reporting euthanasia criteria being met post challenge [14]. The RM model of disease, utilized for vaccination [15,16], re-challenge [17], and therapeutic [18] Rabbit Polyclonal to MAPK1/3 studies, results in disease resolving within three weeks post challenge [9], though some evidence of longer term viral replication has also been reported [19]. AGMs have been utilized for many comparable respiratory-based viral diseases including SARS-CoV-1[20], parainfluenza virus [21] and Nipah virus [22]. Their use as SARS-CoV-2 contamination models has resulted in observed moderate respiratory disease like RMs, but with prolonged shedding of viral RNA [11,13]. Severe disease COVID-19 disease in humans can result in ARDS, characterized by the destruction of the alveolar epithelial lining and significant impairment of gas exchange. Upon survival, the pulmonary space requires substantial repair to regain function, commonly resulting in fibroproliferative disease due to dysregulation of these repair mechanisms. Up to 61% of human autopsies after fatal ARDS show signs of pulmonary fibrosis and 25% of survivors show evidence of restrictive lung disease with long lasting morbidity [23C25]. Within the COVID patient cohort, 42% who develop severe pneumonia will progress to ARDS, with fatal cases generally presenting with pulmonary fibrosis [26,27] or pleuritis [28C31] at autopsy. COVID is usually characterized by a myeloid cell migration into the lung with a striking accumulation of CD16 expressing monocytes and macrophages within the lungs of humans [32]. Comparable myeloid cell infiltration into the lungs has previously been reported in NHP models, and was associated with both a more.
