概要: Growing evidence suggests that conventional dendritic cells (cDCs) undergo aberrant maturation in COVID-19, which negatively affects T-cell activation. The presence of effector T cells in patients with mild disease and dysfunctional T cells in severely ill patients suggests that adequate T-cell responses limit disease severity. Understanding how cDCs cope with SARS-CoV-2 can help elucidate how protective immune responses are generated. Here, we report that cDC2 subtypes exhibit similar infection-induced gene signatures, with the upregulation of IFN-stimulated genes and IL-6 signaling pathways. Furthermore, comparison of cDCs between patients with severe and mild disease showed severely ill patients to exhibit profound downregulation of genes encoding molecules involved in antigen presentation, such as MHCII, TAP, and costimulatory proteins, whereas we observed the opposite for proinflammatory molecules, such as complement and coagulation factors. Thus, as disease severity increases, cDC2s exhibit enhanced inflammatory properties and lose antigen presentation capacity. Moreover, DC3s showed upregulation of anti-apoptotic genes and accumulated during infection. Direct exposure of cDC2s to the virus in vitro recapitulated the activation profile observed in vivo. Our findings suggest that SARS-CoV-2 interacts directly with cDC2s and implements an efficient immune escape mechanism that correlates with disease severity by downregulating crucial molecules required for T-cell activation.
项目整体设计: Single-cell RNA sequencing of CD11c+ MHCII+ cells isolated from PBMCs of 3 COVID-19 patients and 2 healthy donors.
Myeloid cells were sorted (CD11c+ MHCII+) from 3 COVID-19 patients and 2 healthy donors using a MACSQuant Tyto (Miltenyi). 10,000 cells per sample were loaded on a Chromium Next GEM Chip G (10x Genomics). A Chromium controller (10x Genomics, Pleasanton, CA, USA) was used to generate single-cell GEMs, according to Chromium Next GEM Single Cell 5’ Library & Gel Bead Kit v1.1 protocol (PN-1000165; 10x Genomics).
建库方案:
Full-length cDNA amplification and 5’ gene expression library construction were performed according to manufacturers’ instructions in a Veriti 96-well Thermal Cycler (Thermo Fisher Scientific).
测序信息
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polyA(+) RNA
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PAIRED
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-
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ILLUMINA
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Illumina NovaSeq 6000
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-
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文章
Maturation signatures of conventional dendritic cell subtypes in COVID-19 suggest direct viral sensing.
European journal of immunology . 2021-10-01 [PMID:
34333764]
