Journal: Clinical Immunology (Orlando, Fla.)

Article Title: SARS-CoV-2 proteins regulate inflammatory, thrombotic and diabetic responses in human arterial fibroblasts

doi: 10.1016/j.clim.2021.108733

Figure Lengend Snippet: Adventitial fibroblast viability (A), density (B) and metabolic activity (C) after the exposure to SARS-CoV-2 Spike (S) Protein, Nucleocapsid (N) Protein and Membrane-Envelope (M) Protein for 24 h (A and B) or 1 h (C). All data are reported as the mean + standard error of the mean for 9 independent experiments (for viability/density) and 3 independent experiments (for metabolic activity). *Significantly different than negative control (ANOVA, Duncan method).

Article Snippet: Human aortic adventitial fibroblasts (AFs) were purchased from ScienCell Research Laboratories (Carlsbad, CA) and maintained in fibroblast medium-2 supplemented with 5% fetal bovine serum, antibiotics (penicillin/streptomycin) and growth supplement (as suggested by the manufacturer) at 37 °C and 5% CO 2 .

Techniques: Activity Assay, Membrane, Negative Control

Journal: Clinical Immunology (Orlando, Fla.)

Article Title: SARS-CoV-2 proteins regulate inflammatory, thrombotic and diabetic responses in human arterial fibroblasts

doi: 10.1016/j.clim.2021.108733

Figure Lengend Snippet: gC1qR expression on the cell membrane of adventitial fibroblasts after exposure to SARS-CoV-2 Spike (S) Protein, Nucleocapsid (N) Protein and Membrane-Envelope (M) Protein for 1 h. We observed different protein locations that are associated with different structural and functional locations of gC1qR: including 60.11 (A, the C1q globular head binding domain), 74.5.2 (B, activation of kinin and coagulation systems) and 83.13 (C, the HK binding domain). All data are reported as the mean + standard error of the mean for a minimum of 4 independent experiments (range 4–14). *Significantly different than negative control (ANOVA, Duncan method).

Article Snippet: Human aortic adventitial fibroblasts (AFs) were purchased from ScienCell Research Laboratories (Carlsbad, CA) and maintained in fibroblast medium-2 supplemented with 5% fetal bovine serum, antibiotics (penicillin/streptomycin) and growth supplement (as suggested by the manufacturer) at 37 °C and 5% CO 2 .

Techniques: Expressing, Membrane, Functional Assay, Binding Assay, Activation Assay, Coagulation, Negative Control

Journal: Clinical Immunology (Orlando, Fla.)

Article Title: SARS-CoV-2 proteins regulate inflammatory, thrombotic and diabetic responses in human arterial fibroblasts

doi: 10.1016/j.clim.2021.108733

Figure Lengend Snippet: ICAM-1 (A), Tissue Factor (B), RAGE (C) and GLUT-4 (D) expression on the cell membrane of adventitial fibroblasts after exposure to SARS-CoV-2 Spike (S) Protein, Nucleocapsid (N) Protein and Membrane-Envelope (M) Protein for 1 h. All data are reported as the mean + standard error of the mean for a minimum of 4 independent experiments (range 4–5 for ICAM-1, 7 for tissue factor, 6–7 for RAGE and 6–7 for GLUT-4). *Significantly different than negative control (ANOVA, Duncan method).

Article Snippet: Human aortic adventitial fibroblasts (AFs) were purchased from ScienCell Research Laboratories (Carlsbad, CA) and maintained in fibroblast medium-2 supplemented with 5% fetal bovine serum, antibiotics (penicillin/streptomycin) and growth supplement (as suggested by the manufacturer) at 37 °C and 5% CO 2 .

Techniques: Expressing, Membrane, Negative Control

Journal: Clinical Immunology (Orlando, Fla.)

Article Title: SARS-CoV-2 proteins regulate inflammatory, thrombotic and diabetic responses in human arterial fibroblasts

doi: 10.1016/j.clim.2021.108733

Figure Lengend Snippet: Proposed model of SARS-CoV-2 structural protein induced adventitial fibroblast inflammation, thrombosis, and diabetes progression. SARS-CoV-2 that has entered the vasculature can localize to the endothelium via gC1qR or transmigrate into the sub-endothelial space where viral structural proteins (Spike, Nucleocapsid, Membrane and/or Envelope Proteins) can interact with adventitial fibroblast gC1qR. The interaction of SARS-CoV-2 structural proteins with fibroblast gC1qR may activate internal signal transduction pathways (such as the MAPK pathway), which can lead to the enhanced expression or activation of GLUT-4, RAGE (receptor for advanced glycation end products), tissue factor, ICAM-1, and gC1qR's high molecular weight kininogen (HK) binding site. As each of these receptors interact with their specific ligands, vascular responses such as hypoglycemia, chronic inflammation, edema, coagulation, and thrombosis may be observed or promoted leading to vascular dysfunction. Further, each of these receptors and their heightened responses have been implicated in vascular disease progression. Importantly, these vascular disease responses have been shown to be heightened during COVID-19 progression, thus this figure proposes a possible mechanism to link SARS-CoV-2 infection and vascular disease progression.

Article Snippet: Human aortic adventitial fibroblasts (AFs) were purchased from ScienCell Research Laboratories (Carlsbad, CA) and maintained in fibroblast medium-2 supplemented with 5% fetal bovine serum, antibiotics (penicillin/streptomycin) and growth supplement (as suggested by the manufacturer) at 37 °C and 5% CO 2 .

Techniques: Membrane, Transduction, Expressing, Activation Assay, High Molecular Weight, Binding Assay, Coagulation, Biomarker Discovery, Infection

Journal: Gels

Article Title: Developing Biomimetic Hydrogels of the Arterial Wall as a Prothrombotic Substrate for In Vitro Human Thrombosis Models

doi: 10.3390/gels9060477

Figure Lengend Snippet: Ascorbic acid supplementation of the TEML hydrogels enhances its pro-aggregatory properties under physiological flow. ( A , B ) Washed human platelets were exposed to NASC-treated and ASC-treated TEMLs, and their respective acellular controls, under magnetic stirring for 7 min at 37 °C. Samples were then transferred to a light transmission aggregometer ( A ) A representative aggregometry trace. ( B ) A bar chart summarising the extent of aggregation elicited by the TEML hydrogels and collagen hydrogel following the 7-min incubation period. ( C ) 3D printed chamber utilised to perfuse washed DiOC 6 -labelled human platelets at physiological arterial shear stresses (14 dynes/cm 2 ) over the ASC-treated and NASC-treated TEMLs, as well as an acellular collagen hydrogel control. Representative images are shown from samples perfused with blood from three different donors.—Scale bar = 5 mm. ( D ) A summary bar chart showing the mean fluorescent intensity of the TEML surface after platelet perfusion. * indicates p < 0.05 compared to acellular control, † indicates p < 0.05 compared to NASC.

Article Snippet: This can be achieved by creating a biomimetic TEAL hydrogel through culturing human aortic adventitial fibroblasts (HAoAFs) in a 3D collagen hydrogel.

Techniques: Transmission Assay, Incubation, Shear, Control