PMID: 8946016Nov 13, 1996Paper

A nitric oxide production bioassay for interferon-gamma

Journal of Immunological Methods
Y M Kim, K Son

Abstract

Interferon-gamma (IFN-gamma), produced by stimulated T lymphocytes and natural killer cells, has an antitumor and antiviral activity by inhibition of cell growth, immunomodulation, and/or activation of macrophages. Although several IFN-gamma assays have been used, there are no simple, inexpensive, and specific assays. We have developed a new bioassay for IFN-gamma which measures the concentration of nitric oxide (NO) generated by a macrophage cell line RAW264.7 stimulated with IFN-gamma. NO production, determined by nitrite (NO2-) accumulation in culture medium, was linear at IFN-gamma concentrations between 0 and 10 U/ml and logarithmically linear between 2 and 100 U/ml, when RAW cells (1 x 10(5) cells/200 microliters/well in 96 well plate) were incubated with murine recombinant IFN-gamma for 24 h. The new assay has a high sensitivity with the detection limit of 0.1-0.2 ng/ml IFN-gamma, which is similar to that of the enzyme-linked immunosorbent assay (ELISA) and antiviral assays. Other cytokines such as IFN-alpha, IFN-beta, tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1 beta, IL-2, and IL-6, either alone or in combination did not produce NO from RAW264.7. Exogenous NO2-, NO3- or NO2- plus NO3- did not interfere w...Continue Reading

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Citations

Nov 19, 2003·Nitric Oxide : Biology and Chemistry·A PanossianG Wikman
Oct 3, 1999·Veterinary Immunology and Immunopathology·B LambrechtT P van den Berg
Jul 6, 2000·Immunology Letters·J GołabM Jakóbisiak
Aug 8, 2002·Parasite Immunology·Jolanta M DzikElzbieta Wałajtys-Rode
Jan 21, 2012·Evidence-based Complementary and Alternative Medicine : ECAM·Tao LiuDeqi Su
Sep 16, 2004·Parasite Immunology·J M DzikE Wałajtys-Rode
Jul 9, 2009·Journal of Biomedical Materials Research. Part B, Applied Biomaterials·Joyce C BregerNam Sun Wang
May 4, 2011·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Ruchi BansalKlaas Poelstra
Feb 15, 2001·American Journal of Physiology. Cell Physiology·E D Chan, D W Riches

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