用于研究外周血单个核细胞中IFNγ和IL-4分泌的马ELISPOT检测法。

Equine ELISPOT assay to study secretion of IFNγ and IL-4 from peripheral blood mononuclear cells.

作者信息

Hagen Jodi, Hartnett Chris, Houchins Jeffrey P, Giguère Steeve, Kalyuzhny Alexander E

机构信息

R&D Systems, Inc., Minneapolis, MN, USA.

出版信息

Methods Mol Biol. 2012;792:39-45. doi: 10.1007/978-1-61779-325-7_3.

DOI:10.1007/978-1-61779-325-7_3

PMID:21956499

Abstract

Human and mouse immune system cells are the most frequently used specimens in ELISPOT assays. In an effect to expand the application of ELISPOT assay to other species, we developed matched antibody pairs for ready-to-use kits designed for studying the frequency of equine IFNγ- and IL-4-secreting peripheral blood mononuclear cells (PBMCs). Equine PBMCs were stimulated with either concanavalin A (Con A) or calcium ionomycin mixed with phorbol 12-myristate 13-acetate (CaI + PMA). We found that Con A, in general, had a more profound stimulating effect than CaI + PMA on IL-4 secretion, whereas both stimulatory and inhibitory effects were observed on IFNγ secretion. Our data demonstrate a large dynamic range in IFNγ and IL-4 secretion among different donors, which may reflect animal health and serve as a valuable diagnostic marker.

摘要

人和小鼠免疫系统细胞是ELISPOT分析中最常用的样本。为了将ELISPOT分析的应用扩展到其他物种，我们开发了配对抗体对，用于即即用型试剂盒，以研究马外周血单个核细胞（PBMC）分泌IFNγ和IL-4的频率。用伴刀豆球蛋白A（Con A）或与佛波醇12-肉豆蔻酸酯13-乙酸酯混合的钙离子载体（CaI + PMA）刺激马PBMC。我们发现，一般来说，Con A对IL-4分泌的刺激作用比CaI + PMA更显著，而对IFNγ分泌则观察到刺激和抑制两种作用。我们的数据表明，不同供体之间IFNγ和IL-4分泌存在很大的动态范围，这可能反映动物健康状况，并可作为有价值的诊断标志物。