Direct comparison of ELISPOT and ELISA-based assays for detection of individual cytokine-secreting cells.

Lymphokine and cytokine research Pub Date : 1994-08-01
S Tanguay, J J Killion
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引用次数: 0

Abstract

A direct comparison was made between the insoluble ELISPOT, solubilized ELISPOT, and ELISA assays, to detect cytokine secretion by cells, using sterile ELISA plates and commercially available monoclonal antibodies. We evaluated the IL-6 secretion by resident peritoneal macrophages of BALB/c mice and the secretion of IL-2, IL-4, IL-5, and IL-6 by the murine T helper clone, D10.G4.1 cells. Our results demonstrated that ELISPOT can detect cytokine secretion at the single cell level in either adherent or nonadherent cells. The level of detection by ELISPOT was 10 to 200 times more sensitive than ELISA performed on culture supernatants. We also demonstrated that the solubilized ELISPOT can detect cytokine secretion by cells with greater sensitivity than conventional ELISA. These ELISPOT assays can be used to characterize the cytokine secretion pattern of different cell populations in a simple, reproducible, and reliable manner.

直接比较ELISPOT和elisa法检测单个细胞因子分泌细胞。
使用无菌ELISA板和市售单克隆抗体,直接比较不溶性ELISPOT、溶解性ELISPOT和ELISA检测细胞分泌细胞因子的方法。我们评估了BALB/c小鼠常驻腹腔巨噬细胞分泌IL-6和小鼠T辅助克隆D10.G4.1细胞分泌IL-2、IL-4、IL-5和IL-6的情况。我们的结果表明,ELISPOT可以在贴壁和非贴壁细胞中检测单细胞水平的细胞因子分泌。ELISA检测水平比ELISA检测培养上清灵敏10 ~ 200倍。我们还证明,与常规ELISA相比,溶解ELISPOT可以检测细胞分泌的细胞因子,灵敏度更高。这些ELISPOT检测可用于以简单、可重复和可靠的方式表征不同细胞群的细胞因子分泌模式。
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