Journal: Journal of neuroimmune pharmacology : the official journal of the Society on NeuroImmune Pharmacology

Article Title: Extracellular microvesicles released from brain endothelial cells are detected in animal models of HIV-1 signifying unresolved inflammation

doi: 10.1007/s11481-021-10008-5

Figure Lengend Snippet: HIV viroproteins and inflammatory insults disrupt the primary human brain microvasculature endothelial barrier and corresponds to an increase in microvesicles containing the tight junction protein occludin. A. hBMVECs were grown on 96-well electrodes and electrical resistance was measured using the Z-Theta (Applied Biophysics). Cells were stimulated with 100 ng/mL of HIV-1 proteins Tat and gp120 (A) or 100 ng/mL of inflammatory insults TNF-α and LPS (B). All insults resulted in a reduction in electrical resistance equating to an increase in permeability in the endothelial monolayer. Similarly, hBMVECs grown in 100 mm dishes were treated with 100 ng/ml of LPS (B) TNF-α (C), Tat (D) and gp120 (E) for 6 h and 24 h. Media was harvested, MVs were isolated by ultracentrifugation, labeled with Occludin-FITC and analyzed by flow cytometry. Data was normalized by the basal production over the same treatment time from the same cells (paired study). (n = 7–18 all conditions, students t-test, *p < 0.5 **p < 0.01)

Article Snippet: A. hBMVECs were grown on 96-well electrodes and electrical resistance was measured using the Z-Theta (Applied Biophysics).

Techniques: Permeability, Isolation, Labeling, Flow Cytometry