CXCL10 production in equine monocytes is stimulated by interferon-gamma

Abstract

C-X-C motif ligand 10 (CXCL10) is a pro-inflammatory chemokine and has been extensively evaluated in people and mice. In horses, CXCL10 and its involvement in host immunity has rarely been analyzed due to the lack of specific antibodies. We generated a mAb specific for the equine chemokine CXCL10 using hybridoma technology. Antibody specificity was confirmed by intracellular staining and flow cytometric analysis of Chinese Hamster Ovary (CHO) cells expressing equine rCXCL10, while CHO cells expressing equine rCXCL9 were not detected. Native CXCL10 expression in PBMC from horses of different age groups was analyzed by flow cytometry after in vitro stimulation. CXCL10 expressing PBMC were characterized by triple staining of CXCL10 combined with cell-surface markers. Stimulation with IFN-γ for 5 h similarly induced CXCL10 production in cluster of differentiation (CD)14⁺CD16^- MHCII^high monocytes of adult horses and weanlings. The newly generated mAb enables the quantitative intracellular analysis of CXCL10 by flow cytometry and provides a new valuable tool to improve the evaluation of inflammatory responses in horses.

Keywords: C-X-C motif ligand 10; Chemokine; Flow cytometry; Horse; IP-10; mAb.