Schnabel Christiane L, Babasyan Susanna, Freer Heather, Wagner Bettina
Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA.
Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA.
Vet Immunol Immunopathol. 2019 Jan;207:25-30. doi: 10.1016/j.vetimm.2018.11.016. Epub 2018 Nov 24.
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)14CD16 MHCII 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.
C-X-C基序配体10（CXCL10）是一种促炎趋化因子，已在人和小鼠中得到广泛评估。在马中，由于缺乏特异性抗体，CXCL10及其在宿主免疫中的作用很少被分析。我们利用杂交瘤技术制备了一种针对马趋化因子CXCL10的单克隆抗体（mAb）。通过对表达马重组CXCL10的中国仓鼠卵巢（CHO）细胞进行细胞内染色和流式细胞术分析，证实了抗体的特异性，而表达马重组CXCL9的CHO细胞未被检测到。体外刺激后，通过流式细胞术分析不同年龄组马外周血单个核细胞（PBMC）中天然CXCL10的表达。通过CXCL10与细胞表面标志物的三重染色来表征表达CXCL10的PBMC。用干扰素-γ刺激5小时同样诱导成年马和断奶幼驹分化簇（CD）14⁺CD16⁺MHCII单核细胞产生CXCL10。新制备的mAb能够通过流式细胞术对CXCL10进行细胞内定量分析，并为改善马炎症反应的评估提供了一种新的有价值的工具。
