Fig. 3: The VIS-associated secretome drives key immune-thrombotic components of COVID-19.
From: Virus-induced senescence is a driver and therapeutic target in COVID-19
a, The percentage of CD86+ cells in the THP-1 macrophage population exposed to supernatant from retroviral VIS compared with proliferating (Prol.) HDF IMR90 (as in Fig. 1a) and WI38 (as in Extended Data Fig. 1a) (left). Same experiment but using the supernatant of senescence-incapable IMR90 cells after mock or virus infection, as in Fig. 1e (right). b, Multiplex bead-based protein analysis of the supernatant of THP-1 exposed to supernatant as in a. SNVIS without THP-1 cells is shown on the right. c, GEP of M1-macrophage polarization-related transcripts by RNA-seq of THP-1 cells exposed to SNprol. or SNVIS and sorted for CD86 positivity (n = 2 each, left) and by oligonucleotide-based transcript hybridization of lung tissue from autopsy samples of patients with COVID-19 compared with individuals without COVID-19 (n = 3 each, right). d, SA-β-gal staining (top) and the CD86+ fraction (bottom) of THP-1 cells exposed to the supernatant of RPE1-infected VSV-ΔG* variants as indicated. e, SA-β-gal staining (top) and transcript expression by RT–qPCR (bottom) of HUVEC endothelial cells incubated with the supernatant of IMR90 cells. f, Platelet activation measured by ADP release from healthy donor platelets incubated with the supernatant of IMR90 cells as in a. n = 5 donors. g, Clotting time in whole-blood samples (n = 4 healthy donors) mixed with the indicated supernatant volumes (WI38, as in b). INTEM captures intrinsic, platelet-dependent clotting, whereas EXTEM captures extrinsic, non-platelet-dependent clotting. h, NET formation after exposing neutrophils to supernatant from HDFs as in b. n = 4 healthy donors, two measurements each. MPO, myeloperoxidase. i, Quantification of the cytolytic complement complex C5b–C9 on HUVEC cells incubated with human serum and supernatant as indicated. ina, inactivated. j, Cytotoxicity assay of HUVEC cells as in i. All quantifications (mean ± s.d. with individual values as dots) and representative photomicrographs are from n = 3 independent experiments unless otherwise noted. *P < 0.05 by unpaired two-tailed t-test (a, f, h–j), unpaired one-tailed t-test (d) or paired two-tailed t-test (g). Scale bars, 100 µm.
