Differential effects of macrophage subtypes on SARS-CoV-2 infection in a human pluripotent stem cell-derived model

Dysfunctional immune responses contribute critically to the progression of Coronavirus Disease-2019 (COVID-19), with macrophages as one of the main cell types involved. It is urgent to understand the interactions among permissive cells, macrophages, and the SARS-CoV-2 virus, thereby offering important insights into effective therapeutic strategies. Here, we establish a lung and macrophage co-culture system derived from human pluripotent stem cells (hPSCs), modeling the host-pathogen interaction in SARS-CoV-2 infection. We find that both classically polarized macrophages (M1) and alternatively polarized macrophages (M2) have inhibitory effects on SARS-CoV-2 infection. However, M1 and non-activated (M0) macrophages, but not M2 macrophages, significantly up-regulate inflammatory factors upon viral infection. Moreover, M1 macrophages suppress the growth and enhance apoptosis of lung cells. Inhibition of viral entry using an ACE2 blocking antibody substantially enhances the activity of M2 macrophages. Our studies indicate differential immune response patterns in distinct macrophage phenotypes, which could lead to a range of COVID-19 disease severity.


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All studies must disclose on these points even when the disclosure is negative. For each set of experiments, samples were prepared for all experimental arms at the same time. No data were excluded from the study design. All statistical tests are 2-sided. No adjustments were made for multiple comparisons.
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The cells or specimen were randomly separated into different groups for different treatments. stated. Used in 26 publications based on manufacturer website. 11. Anti-IL6 (Affinity, DF6087) was verified using western blot in various tissues, including rat spleen, heart, and muscle, human breast cancer. Used in 65 publications based on manufacturer website 12. Anti-IL-32 (Abcam, ab37158) was verified in human spleen. Used in 14 publications based on manufacturer website. 13. Anti-CCL2 (Abcam, ab9858) has been verified using ELISA with recombinant human MCP-1. Used in 3 publications based on manufacturer website. 14. Anti-IL-1 beta (Abcam, ab254360) has been verified in wild-type THP-1 cells and IL1B knockout THP-1 cells. Used in 3 publications based on manufacturer website. 15. Anti-ACE2 (R&D systems, AF933), Detects human ACE-2 in direct ELISAs. Detects human, mouse, and rat ACE-2 in Western blots. Detects Hamster ACE-2 in immunohistochemistry. In direct ELISAs and Western blots, less than 1% cross-reactivity with recombinant human ACE is observed. Used in 63 publications based on manufacturer website 16. Anti-CD68-PE (Biolegend, 333807) has been used in 2 publications based on manufacturer website. 17. Anti-CD11b, APC-conjugated (Biolegend, 301309) antibody inhibits heterotypic adhesion of granulocytes in response to fMLP. Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen tissue sections, immunofluorescence microscopy5, stimulation of monocytes3, blocking of heterotypic PMN aggregation8, and blocking of granulocyte activation12. This clone was tested in-house and does not work on formalin fixed paraffin embedded ( Cell lines were authenticated by DNA fingerprinting analysis.
All cell lines were confirmed to be mycoplasma free. The cells lines purchased from ATCC which certified the lots to be free of Mycoplasma contamination.
No misidentified cell lines were used in this study