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Effect of factor XIII on endothelial barrier function.

Noll T, Wozniak G, McCarson K, Hajimohammad A, Metzner HJ, Inserte J, Kummer W, Hehrlein FW, Piper HM - J. Exp. Med. (1999)

Bottom Line: The thrombin-activated plasma factor XIII (1 U/ml) reduced albumin permeability of endothelial monolayers within 20 min by 30 +/- 7% (basal value of 5.9 +/- 0.4 x 10(-6) cm/s), whereas the nonactivated plasma factor XIII had no effect.This study shows that activated factor XIII reduces endothelial permeability.Its effect seems related to a modification of the paracellular passageways in endothelial monolayers.

View Article: PubMed Central - PubMed

Affiliation: Physiologisches Institut, Justus-Liebig-Universität, D-35392 Giessen, Germany. thomas.noll@physiologie.med.uni-giessen.de

ABSTRACT
The effect of factor XIII on endothelial barrier function was studied in a model of cultured monolayers of porcine aortic endothelial cells and saline-perfused rat hearts. The thrombin-activated plasma factor XIII (1 U/ml) reduced albumin permeability of endothelial monolayers within 20 min by 30 +/- 7% (basal value of 5.9 +/- 0.4 x 10(-6) cm/s), whereas the nonactivated plasma factor XIII had no effect. Reduction of permeability to the same extent, i.e., by 34 +/- 9% could be obtained with the thrombin-activated A subunit of factor XIII (1 U/ml), whereas the iodoacetamide-inactivated A subunit as well as the B subunit had no effect on permeability. Endothelial monolayers exposed to the activated factor XIII A exhibited immunoreactive deposition of itself at interfaces of adjacent cells; however, these were not found on exposure to nonactivated factor XIII A or factor XIII B. Hyperpermeability induced by metabolic inhibition (1 mM potassium cyanide plus 1 mM 2-deoxy-D-glucose) was prevented in the presence of the activated factor XIII A. Likewise, the increase in myocardial water content in ischemic-reperfused rat hearts was prevented in its presence. This study shows that activated factor XIII reduces endothelial permeability. It can prevent the loss of endothelial barrier function under conditions of energy depletion. Its effect seems related to a modification of the paracellular passageways in endothelial monolayers.

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Immunostaining of factor XIII  A in endothelial monolayers. (A) Endothelial cells were incubated for 20 min in the  presence of activated factor XIII A (1 U/ml).  Factor XIII A–positive staining is seen along  the interfaces of adjacent endothelial cells.  (B) Endothelial cells were exposed to nonactivated factor XIII A (10 μg/ml). No positive staining for factor XIII A is observed.  (C) Endothelial cells not preincubated with  factor XIII A were exposed to anti-factor  XIII A and FITC-coupled anti–rabbit IgG  antibody (first and second antibody control).  Only background fluorescence is seen. (D)  Endothelial cells not preincubated with factor XIII A were exposed only to the FITC-coupled anti–rabbit IgG antibody (second  antibody control). Again, only background  fluorescence is apparent. Bar, 10 μm.
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Figure 6: Immunostaining of factor XIII A in endothelial monolayers. (A) Endothelial cells were incubated for 20 min in the presence of activated factor XIII A (1 U/ml). Factor XIII A–positive staining is seen along the interfaces of adjacent endothelial cells. (B) Endothelial cells were exposed to nonactivated factor XIII A (10 μg/ml). No positive staining for factor XIII A is observed. (C) Endothelial cells not preincubated with factor XIII A were exposed to anti-factor XIII A and FITC-coupled anti–rabbit IgG antibody (first and second antibody control). Only background fluorescence is seen. (D) Endothelial cells not preincubated with factor XIII A were exposed only to the FITC-coupled anti–rabbit IgG antibody (second antibody control). Again, only background fluorescence is apparent. Bar, 10 μm.

Mentions: For immunostaining, a polyclonal rabbit anti-factor XIII A antibody was used which recognizes the activated as well as the nonactivated factor XIII A (32). Immunostaining of endothelial monolayers incubated for 20 min in the presence of thrombin-activated factor XIII A (1 U/ml) revealed factor XIII A–positive staining along the interface of adjacent endothelial cells (Fig. 6 A). In monolayers that were exposed to nonactivated factor XIII A at equivalent protein concentration (10 μg protein/ml), immunostaining for factor XIII A remained absent (Fig. 6 B). As control, endothelial monolayers that had not been incubated in the presence of factor XIII A were exposed to either the first anti-factor XIII A and second antibody (FITC-coupled anti–rabbit IgG; Fig. 6 C) or the second antibody alone (Fig. 6 D). No specific staining was observed with these protocols.


Effect of factor XIII on endothelial barrier function.

Noll T, Wozniak G, McCarson K, Hajimohammad A, Metzner HJ, Inserte J, Kummer W, Hehrlein FW, Piper HM - J. Exp. Med. (1999)

Immunostaining of factor XIII  A in endothelial monolayers. (A) Endothelial cells were incubated for 20 min in the  presence of activated factor XIII A (1 U/ml).  Factor XIII A–positive staining is seen along  the interfaces of adjacent endothelial cells.  (B) Endothelial cells were exposed to nonactivated factor XIII A (10 μg/ml). No positive staining for factor XIII A is observed.  (C) Endothelial cells not preincubated with  factor XIII A were exposed to anti-factor  XIII A and FITC-coupled anti–rabbit IgG  antibody (first and second antibody control).  Only background fluorescence is seen. (D)  Endothelial cells not preincubated with factor XIII A were exposed only to the FITC-coupled anti–rabbit IgG antibody (second  antibody control). Again, only background  fluorescence is apparent. Bar, 10 μm.
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Related In: Results  -  Collection

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Figure 6: Immunostaining of factor XIII A in endothelial monolayers. (A) Endothelial cells were incubated for 20 min in the presence of activated factor XIII A (1 U/ml). Factor XIII A–positive staining is seen along the interfaces of adjacent endothelial cells. (B) Endothelial cells were exposed to nonactivated factor XIII A (10 μg/ml). No positive staining for factor XIII A is observed. (C) Endothelial cells not preincubated with factor XIII A were exposed to anti-factor XIII A and FITC-coupled anti–rabbit IgG antibody (first and second antibody control). Only background fluorescence is seen. (D) Endothelial cells not preincubated with factor XIII A were exposed only to the FITC-coupled anti–rabbit IgG antibody (second antibody control). Again, only background fluorescence is apparent. Bar, 10 μm.
Mentions: For immunostaining, a polyclonal rabbit anti-factor XIII A antibody was used which recognizes the activated as well as the nonactivated factor XIII A (32). Immunostaining of endothelial monolayers incubated for 20 min in the presence of thrombin-activated factor XIII A (1 U/ml) revealed factor XIII A–positive staining along the interface of adjacent endothelial cells (Fig. 6 A). In monolayers that were exposed to nonactivated factor XIII A at equivalent protein concentration (10 μg protein/ml), immunostaining for factor XIII A remained absent (Fig. 6 B). As control, endothelial monolayers that had not been incubated in the presence of factor XIII A were exposed to either the first anti-factor XIII A and second antibody (FITC-coupled anti–rabbit IgG; Fig. 6 C) or the second antibody alone (Fig. 6 D). No specific staining was observed with these protocols.

Bottom Line: The thrombin-activated plasma factor XIII (1 U/ml) reduced albumin permeability of endothelial monolayers within 20 min by 30 +/- 7% (basal value of 5.9 +/- 0.4 x 10(-6) cm/s), whereas the nonactivated plasma factor XIII had no effect.This study shows that activated factor XIII reduces endothelial permeability.Its effect seems related to a modification of the paracellular passageways in endothelial monolayers.

View Article: PubMed Central - PubMed

Affiliation: Physiologisches Institut, Justus-Liebig-Universität, D-35392 Giessen, Germany. thomas.noll@physiologie.med.uni-giessen.de

ABSTRACT
The effect of factor XIII on endothelial barrier function was studied in a model of cultured monolayers of porcine aortic endothelial cells and saline-perfused rat hearts. The thrombin-activated plasma factor XIII (1 U/ml) reduced albumin permeability of endothelial monolayers within 20 min by 30 +/- 7% (basal value of 5.9 +/- 0.4 x 10(-6) cm/s), whereas the nonactivated plasma factor XIII had no effect. Reduction of permeability to the same extent, i.e., by 34 +/- 9% could be obtained with the thrombin-activated A subunit of factor XIII (1 U/ml), whereas the iodoacetamide-inactivated A subunit as well as the B subunit had no effect on permeability. Endothelial monolayers exposed to the activated factor XIII A exhibited immunoreactive deposition of itself at interfaces of adjacent cells; however, these were not found on exposure to nonactivated factor XIII A or factor XIII B. Hyperpermeability induced by metabolic inhibition (1 mM potassium cyanide plus 1 mM 2-deoxy-D-glucose) was prevented in the presence of the activated factor XIII A. Likewise, the increase in myocardial water content in ischemic-reperfused rat hearts was prevented in its presence. This study shows that activated factor XIII reduces endothelial permeability. It can prevent the loss of endothelial barrier function under conditions of energy depletion. Its effect seems related to a modification of the paracellular passageways in endothelial monolayers.

Show MeSH
Related in: MedlinePlus