PMID: 8459105Apr 2, 1993Paper

Cytokine-specific ELISPOT assay. Single cell analysis of IL-2, IL-4 and IL-6 producing cells

Journal of Immunological Methods
K FujihashiH Kiyono

Abstract

In order to assess cytokine-producing cells at the single cell level, the cytokine-specific ELISPOT assay has proven to be an important and sensitive method. The purpose of this study was to adapt this method to elucidate individual cells producing murine IL-2, IL-4 or IL-6. In order to establish these cytokine-specific ELISPOT assays, IL-2-, IL-4- and IL-6-specific cDNA transfected myeloma cell lines, e.g., X63-Ag8-653 X2, X63-Ag8-653 X4 and X63-Ag8-653 X6, respectively, were used as specific cytokine-producing cells. In the IL-2 ELISPOT assay, the coating reagent, monoclonal antibody (mAb) rat IgG2a anti-mouse IL-2 (CR #40014) was used while rabbit IgG polyclonal anti-mouse IL-2 was employed for detection of IL-2 spot forming cells (SFC). The mAbs anti-mouse IL-4, BVD4-1D11 and BVD6-24G2 were selected as capture and detection antibodies for enumeration of IL-4 SFC. For the IL-6 ELISPOT assay, anti-mouse IL-6 (MP5-20F3) mAb was used for coating and MP5-32C11 mAb was used for detection of IL-6 SFC. When IL-2 producing X63-Ag8-653 X2 cells were subjected to these three different ELISPOT assays, IL-2-specific SFC were only noted with the IL-2 ELISPOT system. In the case of IL-4 SFC, only X63-Ag8-653 X4 cells formed specific spots...Continue Reading

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Citations

Apr 1, 1995·Current Opinion in Immunology·N Shastri
Dec 11, 2002·Cellular Immunology·Katsumitsu AraiGunther Dennert
Jun 16, 2000·Journal of Immunological Methods·F H RininslandM Tary-Lehmann
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Feb 5, 2008·Digestive and Liver Disease : Official Journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver·B MarottiG Nardone
Aug 25, 2004·Comparative Immunology, Microbiology and Infectious Diseases·A NuntaprasertC Kai
Jul 28, 2004·Immunology Letters·Senad Divanovic, Alexander C K Lai
Dec 6, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Jian ChenJohn D Mountz
Nov 5, 1999·International Immunology·F D Finkelman, S C Morris
Apr 25, 2008·Current Protocols in Immunology·Fred FinkelmanDavid Sehy
Jan 1, 1994·International Reviews of Immunology·K FujihashiH Kiyono

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