Effect of recombinant equine interleukin-1β on function of equine endothelial colony-forming cells in vitro

Claudia L. Reyner Department of Clinical Sciences, College of Veterinary Medicine, Auburn University, Auburn, AL 36849.

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Randolph L. Winter Department of Clinical Sciences, College of Veterinary Medicine, Auburn University, Auburn, AL 36849.

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Kara L. Maneval Department of Clinical Sciences, College of Veterinary Medicine, Auburn University, Auburn, AL 36849.

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Lindsey H. Boone Department of Clinical Sciences, College of Veterinary Medicine, Auburn University, Auburn, AL 36849.

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Anne A. Wooldridge Department of Clinical Sciences, College of Veterinary Medicine, Auburn University, Auburn, AL 36849.

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Abstract

OBJECTIVE

To investigate the effects of recombinant equine IL-1β on function of equine endothelial colony-forming cells (ECFCs) in vitro.

SAMPLE

ECFCs derived from peripheral blood samples of 3 healthy adult geldings.

PROCEDURES

Function testing was performed to assess in vitro wound healing, tubule formation, cell adhesion, and uptake of 1,1′-dioctadecyl-3,3,3′,3′ tetramethylindocarbocyanine perchlorate–labeled acetylated low-density lipoprotein (DiI-Ac-LDL) by cultured ECFCs. Cell proliferation was determined by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide assay. Effects on function test results of different concentrations and exposure times of recombinant equine IL-1β were assessed.

RESULTS

Challenge of cultured ECFCs with IL-1β for 48 hours inhibited tubule formation. Continuous challenge (54 hours) with IL-1β in the wound healing assay reduced gap closure. The IL-1β exposure did not significantly affect ECFC adhesion, DiI-Ac-LDL uptake, or ECFC proliferation.

CONCLUSIONS AND CLINICAL RELEVANCE

These results suggested a role for IL-1β in the inhibition of ECFC function in vitro. Functional changes in ECFCs following challenge with IL-1β did not appear to be due to changes in cell proliferative capacity. These findings have implications for designing microenvironments for and optimizing therapeutic effects of ECFCs used to treat ischemic diseases in horses.

Abstract

OBJECTIVE

To investigate the effects of recombinant equine IL-1β on function of equine endothelial colony-forming cells (ECFCs) in vitro.

SAMPLE

ECFCs derived from peripheral blood samples of 3 healthy adult geldings.

PROCEDURES

Function testing was performed to assess in vitro wound healing, tubule formation, cell adhesion, and uptake of 1,1′-dioctadecyl-3,3,3′,3′ tetramethylindocarbocyanine perchlorate–labeled acetylated low-density lipoprotein (DiI-Ac-LDL) by cultured ECFCs. Cell proliferation was determined by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide assay. Effects on function test results of different concentrations and exposure times of recombinant equine IL-1β were assessed.

RESULTS

Challenge of cultured ECFCs with IL-1β for 48 hours inhibited tubule formation. Continuous challenge (54 hours) with IL-1β in the wound healing assay reduced gap closure. The IL-1β exposure did not significantly affect ECFC adhesion, DiI-Ac-LDL uptake, or ECFC proliferation.

CONCLUSIONS AND CLINICAL RELEVANCE

These results suggested a role for IL-1β in the inhibition of ECFC function in vitro. Functional changes in ECFCs following challenge with IL-1β did not appear to be due to changes in cell proliferative capacity. These findings have implications for designing microenvironments for and optimizing therapeutic effects of ECFCs used to treat ischemic diseases in horses.

Contributor Notes

Dr. Winter's present address is the Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210.

Address correspondence to Dr. Wooldridge (aaw0002@auburn.edu).
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