Equine PBMC phenotyping during equine hepacivirus infection

Humans infected with HCV show significant changes in PBMC populations during infection. Equine hepacivirus (EqHV) is the closest genetic relative of HCV known. We sought to examine the equine immune responses to EqHV infection in order to determine whether horses might serve as a good model for HCV research, particularly in vaccine development and understanding of protective immunity.
Conclusion:
No significant changes were observed in population proportions over the course of infection in 7 horses. Numbers might have been too small to detect subtle changes. Validated flow cytometry panel by sorting major populations and performing PCR to confirm identity prior to this work. Also compared cell phenotype assignment by flow versus single cell RNA Seq for 2 horses in this dataset. Positive and negative selection markers used for most cell phenotypes.

感染丙型肝炎病毒（HCV）的人类在感染期间，其外周血单个核细胞（PBMC）群体存在显著变化。马肝炎病毒（EqHV）是目前已知与HCV亲缘关系最近的遗传近亲。本研究旨在探究马匹感染EqHV后的免疫应答，以验证马匹能否作为HCV研究的理想动物模型，尤其在疫苗开发与保护性免疫机制解析领域。

结论：
本研究纳入的7匹受试马匹在感染全过程中，其外周血单个核细胞群体比例未出现显著变化。该结果可能源于样本量不足，无法检测到细微的群体变动。本研究前期已通过分选主要细胞群体并结合聚合酶链式反应（PCR）验证细胞身份，完成了流式细胞术检测panel的方法学验证。此外，本数据集还针对其中2匹受试马匹，对比了流式细胞术与单细胞RNA测序（single cell RNA Seq）的细胞表型分型结果。多数细胞表型均通过阳性与阴性选择标记完成鉴定。