![]() ![]() ( E) Distribution of S protein concentration according to the global COVID-19 severity. ( D) Distribution of S protein concentration according to patients’ gender. ( C) Distribution of S protein concentration according to patients’ age. ( B) Distribution of S protein concentration in patients according to the time from the onset of disease symptoms. ![]() ( A) Serological S protein concentration in pre-pandemic controls ( n=14) vs COVID-19 patients ( n=64). Abbreviations: LV, left ventricle RA, right atrium RV, right ventricle. ( D) Table showing the row data of the PC infection and summarising the patients’ characteristics and cell source. ( C) Analysis of ACE2 and CD147 protein levels in cardiac PCs using Western blotting. The bar graph reports individual values and means ± SEM. ( B) Quantification of the percentage of PCs positive for dsRNA in the three experimental groups. Because of the high magnification used (20×), the images shown for patient #2 are not representative of the real % of infection, but we aimed to provide examples of infected PCs in each experimental group. For PCs, we show example images from two patients. ( A) Immunofluorescence images show SARS-CoV-2 nucleocapsid protein (N, magenta) and double-stranded RNA (dsRNA, orange) indicative of virus replication. Primary cardiac PCs ( n=6 patients) and the Caco-2-ACE2 cell line were either mock-infected or inoculated with SARS-CoV-2 isolated early in the pandemic (REMRQ001) or the α (B.1.1.7) or δ (B.1.617.2) variants, all at an MOI = 10, and incubated for 24 h before immunostaining for viral and PC markers. si BSG = PC in which BSG (CD147) was silenced, used as a negative control for CD147. ( F) Expression of CD147 and ACE2 determined using Western blotting. ( E) Expression of CD147 (green) in cardiac PCs and control human CAECs assessed by immunostaining. ( D) Expression of ACE2 and TMPRSS2 assessed using Western blotting. VeroE6/ACE2/TMPRSS2: African green monkey kidney cell line engineered to overexpress the human ACE2 and TMPRSS2. ( C) Expression of ACE2 (green) and TMPRSS2 (red) in cardiac PCs and control cells assessed by immunostaining. ( B) Immunofluorescence images of human cardiac fibroblasts and CAECs employed as negative or positive controls for markers in (A). PDGFRα: platelet derived growth factor receptor α. In blue nuclei (DAPI), NG2: Neural/glial antigen 2. In green fluorescence, the antigens as indicated. ( A) Contrast-phase and immunofluorescence images showing the characteristic shape and antigenic phenotype of cultured cardiac PCs. This mechanism may have clinical and therapeutic implications.ĬD147 COVID-19 Microvascular disease Spike protein angiotensin converting enzyme 2 pericyte. In conclusion, our findings suggest that the S protein may prompt PC dysfunction, potentially contributing to microvascular injury. Immunoreactive S protein was detected in the peripheral blood of infected patients. The neutralisation of CD147, either using a blocking antibody or mRNA silencing, reduced ERK1/2 activation, and rescued PC function in the presence of the S protein. Next, adopting a blocking strategy against the S protein receptors angiotensin-converting enzyme 2 (ACE2) and CD147, we discovered that the S protein stimulates the phosphorylation/activation of the extracellular signal-regulated kinase 1/2 (ERK1/2) through the CD147 receptor, but not ACE2, in PCs. Exposure to the recombinant S protein alone elicited signalling and functional alterations, including: (1) increased migration, (2) reduced ability to support endothelial cell (EC) network formation on Matrigel, (3) secretion of pro-inflammatory molecules typically involved in the cytokine storm, and (4) production of pro-apoptotic factors causing EC death. Here we newly show that the in vitro exposure of primary human cardiac PCs to the SARS-CoV-2 wildtype strain or the α and δ variants caused rare infection events. We tested this hypothesis in pericytes (PCs), which are reportedly reduced in the heart of patients with severe coronavirus disease-2019 (COVID-19). Additionally, the SARS-CoV-2 Spike (S) protein may act as a ligand to induce non-infective cellular stress. The capacity of SARS-CoV-2 to infect vascular cells is still debated. The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes a broad range of clinical responses including prominent microvascular damage. ![]()
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