Regulation of cytokine and chemokine release in PBMC elicited by SARS-CoV-2 VOCs.

Unpublished data, available soon for sharing

For further details regarding the image, please refer to the figure legend in the paragraph below.

a) A schematic representation of isolation of peripheral blood mononuclear cells (PBMC) of healthy volunteers exposed to 24 hour (h) treatment with different SARS-CoV-2 viral strains (D614G, Alpha, Beta, Gamma, Delta, Omicron BA.1, Omicron XBB.1) at a MOI of 0.04 is shown. Cells were then analyzed by spectral flow cytometry and supernatants analyzed by next-generation ELISA. The scheme was created with the licenced version of BioRender.  

b, c) Cytokine (b) and chemokine (c) analysis in culture supernatants of SARS-CoV-2 exposed PBMCs. 

SARS-CoV-2 stimulation induces a TLR7/8-dependent cytokine and chemokine production in PBMC.

Full article: https://doi.org/10.1371/journal.ppat.1009878

Peripheral blood mononuclear cells (PBMC) were left untreated (not stimulated, ns) or Mock-treated as negative controls, or stimulated for 24 hours with the TLR7/8 agonist R848 (5μM) as positive control, with SARS-CoV-2 (CoV2) at different multiplicity of infection (MOI; 0.02, 0.04 and 0.1) in presence or absence of a 30 minute pre-treatment with a specific TLR7/8 inhibitor (I-TLR7/8, 1μM).

Production of cytokines (A)

and chemokines (B)

was tested by multiparametric cytokine bead arrays in collected cell culture supernatants.

SARS-CoV-2 stimulation regulates cytokine release in plasmacytoid Dendritic Cells in a TLR7- and Neuropilin-1-dependent manner.

Full article: https://doi.org/10.1371/journal.ppat.1009878

For further details regarding the image, please refer to the figure legend in the paragraph below.

A. Purified plasmacytoid dendritic cells (pDC) were left untreated (not stimulated, ns) or Mock-treated as negative controls. Cells were also stimulated for 24 hours with the TLR7/8 agonist R848 (5μM) or with SARS-CoV-2 (CoV2; MOI = 0.1) with increasing doses of anti-Neuropilin 1 monoclonal antibody (anti-NRP1, 1 or 2 μg/ml) or anti-human IgG control (IgG ctrl, 2 μg/ml). IFN-α production was tested by ELISA in culture supernatants.

B. CoV2-stimulated pDC were also pre-treated with a specific TLR7/8 inhibitor (I-TLR7/8, 1μM) and the inflammatory cytokines TNF-α and IL-6 measured by cytokine bead array.

COVID-19 asymptomatic and hospitalized patients display a specular anti-viral and pro-inflammatory profile.

Full article: https://doi.org/10.1371/journal.ppat.1009878

Peripheral blood mononuclear cells (PBMC) and sera were collected from asymptomatic (CP-AS, n = 8) and hospitalized COVID-19 patients (CP, n = 6) as well as matched healthy donors (HD, n = 5).

(A) Relative expression of Mx1 gene was measured by quantitative real time PCR analysis and normalized to TBP level by using the equation 2-ΔCt in total RNA isolated from ex vivo PBMC.

(B) Production of IFN-αs was tested in serum samples by a specific ELISA kit. Production of cytokines (IL-6, TNF-α, IL-1β, IL-10)

(C) and chemokines (CXCL-10, CCL-2, CXCL-8)

(D) was tested by multiparametric cytokine bead arrays in collected serum samples.

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