Chemokine CXCL4 interactions with extracellular matrix proteoglycans mediate widespread immune cell recruitment independent of chemokine receptors

Leukocyte recruitment from the vasculature into tissues is a crucial component of the immune system but is also key to inflammatory disease. Chemokines are central to this process but have yet to be therapeutically targeted during inflammation due to a lack of mechanistic understanding. Specifically, CXCL4 (Platelet Factor 4, PF4) has no established receptor that explains its function. Here, we use biophysical, in vitro, and in vivo techniques to determine the mechanism underlying CXCL4-mediated leukocyte recruitment. We demonstrate that CXCL4 binds to glycosaminoglycan (GAG) sugars on proteoglycans within the endothelial extracellular matrix, resulting in increased adhesion of leukocytes to the vasculature, increased vascular permeability, and non-specific recruitment of a range of leukocytes. Furthermore, GAG sulfation confers selectivity onto chemokine localization. These findings present mechanistic insights into chemokine biology and provide future therapeutic targets

Chemokine CXCL4 interactions with extracellular matrix proteoglycans mediate widespread immune cell recruitment independent of chemokine receptors

Leukocyte recruitment from the vasculature into tissues is a crucial component of the immune system but is also key to inflammatory disease. Chemokines are central to this process but have yet to be therapeutically targeted during inflammation due to a lack of mechanistic understanding. Specifically, CXCL4 (Platelet Factor 4, PF4) has no established receptor that explains its function. Here, we use biophysical, in vitro, and in vivo techniques to determine the mechanism underlying CXCL4-mediated leukocyte recruitment. We demonstrate that CXCL4 binds to glycosaminoglycan (GAG) sugars on proteoglycans within the endothelial extracellular matrix, resulting in increased adhesion of leukocytes to the vasculature, increased vascular permeability, and non-specific recruitment of a range of leukocytes. Furthermore, GAG sulfation confers selectivity onto chemokine localization. These findings present mechanistic insights into chemokine biology and provide future therapeutic targets

Quantification of valvular regurgitation by cardiac blood pool scintigraphy: correlation with catheterization.

The diagnosis of valvular regurgitation (R) is usually based on clinical signs. Quantification conventionally requires catheterization (C). We have quantified R with cardiac blood pool scintigraphy (CBPS) and compared the results with those obtained by C. Regurgitant fraction (RF) determined by C was calculated with the technique of Dodge. Forward output was measured by thermodilution or cardiogreen dilution. The RF at CBPS was obtained by the stroke index ratio (SIR) minus 1.2 divided by SIR, where SIR is the ratio of the stroke counts of left ventricle over those of the right ventricle. Stroke counts are calculated directly from the time-activity curves. Each time-activity curve was obtained by drawing one region of interest around each diastolic image. The correction factor (1.2) was calculated from a large normal population. 22 patients had aortic R, 7 mitral R, 12 both, 8 patients had no evidence of regurgitation. RF of the patients with R varied from 27 to 71% (x = 42%) at C and from 26 to 74% (y = 41%) at CBPS. Linear regression shows a good correlation coefficient (r = 0.82). The regression equation is y = 0.93x + 1.8. No correlation was found between RF (CBPS or C) and the severity of R assessed visually from angiography. In conclusion: CBPS, a non-invasive method, allows easy and repeatable determination of RF and correlates well with data obtained at catheterization

Conformational selection guides β-arrestin recruitment at a biased G protein–coupled receptor

G protein–coupled receptors (GPCRs) recruit β-arrestins to coordinate diverse cellular processes, but the structural dynamics driving this process are poorly understood. Atypical chemokine receptors (ACKRs) are intrinsically biased GPCRs that engage β-arrestins but not G proteins, making them a model system for investigating the structural basis of β-arrestin recruitment. Here, we performed nuclear magnetic resonance (NMR) experiments on 13CH3-ε–methionine–labeled ACKR3, revealing that β-arrestin recruitment is associated with conformational exchange at key regions of the extracellular ligand-binding pocket and intracellular β-arrestin–coupling region. NMR studies of ACKR3 mutants defective in β-arrestin recruitment identified an allosteric hub in the receptor core that coordinates transitions among heterogeneously populated and selected conformational states. Our data suggest that conformational selection guides β-arrestin recruitment by tuning receptor dynamics at intracellular and extracellular regions