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Autocrine regulation of endothelial barrier integrity by CAP37
Author(s) -
Fox Elizabeth,
Pereira Anne,
Reichner Jonathan
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.646.13
Subject(s) - endothelial stem cell , barrier function , autocrine signalling , umbilical vein , human umbilical vein endothelial cell , microbiology and biotechnology , endothelium , monolayer , chemistry , biology , biophysics , endocrinology , biochemistry , receptor , in vitro
BACKGROUND CAP37 is a peptide released by neutrophils upon stimulation of β2 integrins. Its effects include increased ICAM‐1 expression and endothelial cell cytoskeleton reorganization. Previous work has shown that endothelial cells may produce CAP37. We sought to determine the effect of neutralization of CAP37 on endothelial monolayer resistance. METHODS Human umbilical vein endothelial cells (HUVECs) were grown to confluence and activated with TNFα for six hours. Two mouse‐derived monoclonal anti‐CAP37 antibodies (B1B5 and D5F10) were applied to the monolayer. The effect on endothelial barrier function was determined using Electric Cell‐substrate Impedance Sensing (ECIS). Barrier function of the monolayer was expressed as a fraction of pre‐treatment resistance values. RESULTS Both antibodies caused a biphasic change in monolayer resistance. Within 30 minutes, resistance increased to a peak of 1.16+/−0.04 (B1B5 [p<0.0001 vs. IgG control]) and 1.12+/−0.11 (D5F10 [p=0.03 vs. IgG control]). After this initial peak, resistance progressively decreased, with endothelial barrier integrity falling below 50% of the pre‐treatment value within 5 hours. CONCLUSIONS Neutralization of CAP37 causes a change in endothelial barrier function. This occurs in the absence of leukocytes, demonstrating the role of CAP37 in the autocrine regulation of endothelial monolayer integrity, and suggesting a potential therapeutic target. Future work will include further characterization of endothelial cell CAP37. (NIH GM066194 )