Endothelial protein C receptor-dependent antichemotactic effects of canine protein C
| Autor: | Olivier Côté, R. Darren Wood, M. Anthony Hayes, Valerie M. Wong, Dorothee Bienzle |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Chemokine Neutrophils Receptors Cell Surface Biology Polymerase Chain Reaction Flow cytometry 03 medical and health sciences Dogs Zymogen medicine Animals Dog Diseases Endothelial protein C receptor General Veterinary medicine.diagnostic_test Chemotaxis General Medicine Flow Cytometry Molecular biology Blood Coagulation Factors Reverse transcription polymerase chain reaction Chemotaxis Leukocyte 030104 developmental biology biology.protein Blood Coagulation Tests Antibody Protein C medicine.drug |
| Zdroj: | American journal of veterinary research. 78(2) |
| ISSN: | 1943-5681 |
| Popis: | OBJECTIVE To determine whether canine protein C (CnPC) had antichemotactic effects on canine neutrophils, whether endothelial protein C receptor (EPCR) was expressed on canine neutrophils, and the role of EPCR in neutrophil chemotaxis. SAMPLE Neutrophils isolated from blood samples from healthy dogs (n = 6) and sick dogs with (2) or without (3) an inflammatory leukogram. PROCEDURES Neutrophils were analyzed by reverse transcriptase PCR assay and flow cytometry for detection of EPCR mRNA and protein expression, respectively. Neutrophils were incubated with CnPC zymogen or canine activated protein C (CnAPC), with or without RCR-379 (an anti–human EPCR antibody). Neutrophils were then allowed to migrate through a filter membrane toward a chemokine. Untreated neutrophils served as positive control samples. Migration was quantified by fluorescence measurement, and chemotaxis index (Chx) values (fluorescence of test sample/fluorescence of positive control sample) were computed. RESULTS The cDNA for EPCR was amplified, and EPCR expression was detected on neutrophil surfaces. Obtained Chx values were significantly higher in cells treated with RCR-379 than in cells treated with CnPC or CnAPC alone. The Chx values for neutrophils treated with RCR-379 were not significantly different from 1, whereas those for neutrophils treated without RCR-379 were significantly less than 1. The effects of RCR-379 on neutrophil migration were independent of concentration or activation status of protein C. CONCLUSIONS AND CLINICAL RELEVANCE Canine neutrophils expressed EPCR, and inhibition of neutrophil chemotaxis by CnPC and CnAPC depended on EPCR. Interventions with EPCR signaling may have therapeutic application in dogs. |
| Databáze: | OpenAIRE |
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