PMID: 2506615Jun 1, 1989Paper

Improvement of an ELISA bioassay for the routine titration of murine interferon-gamma

Research in Immunology
M A Le MoalP Truffa-Bachi

Abstract

We improved an ELISA bioassay for murine IFN-gamma (MuIFN-gamma) based on measurement of Ia antigen on P388D1, a mouse macrophagic tumour line. Cells were cultured in microtitre plates in medium containing dilutions of IFN-gamma source. They were then washed and stained with a rat anti-mouse I amAb followed by mouse anti-rat peroxidase-labelled antibody. After incubation with substrate, the OD was read directly from microtitre plates. Standard curves obtained with reference NIH MuIFN-gamma showed that this assay allowed for the definition of unit values (giving 50% of the maximal effect) comparable to NIH international units (IU). It detected as low as 0.2 IU/ml of MuIFN-gamma and, in contrast to antiviral assays, was insensitive to IFN-alpha/beta. We used a concanavalin A-conditioned supernatant, which is a mixed source of lymphokines, to assess the specificity of our assay. Indeed, Ia expression induced by ConA-conditioned supernatant was fully inhibited by preincubation with anti-MuIFN-gamma antibodies. Using a stable indicator cell line, the present cell surface assay is easier to perform than other ELISA using bone-marrow-derived macrophages, and does not require cell fixation; its high sensitivity and specificity are comp...Continue Reading

References

Dec 1, 1988·Immunology Today·J A Langer, S Pestka
Mar 17, 1988·Nature·M B Sporn, A B Roberts
Apr 1, 1988·European Journal of Immunology·S LandolfoG Cavallo
Nov 23, 1987·Journal of Immunological Methods·R C CurryG L Spitalny
Feb 1, 1988·Immunology Today·A O'GarraJ Christiansen
Apr 1, 1988·Immunology Today·D Cosman
Jan 1, 1987·European Journal of Immunology·E SeverinsonT Honjo
Dec 17, 1985·Journal of Immunological Methods·R L Fairchild, J W Moorhead
Aug 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·T W ChangJ Le
Dec 1, 1980·The Journal of Experimental Medicine·P S SteegJ J Oppenheim
Dec 1, 1982·The Journal of Experimental Medicine·P S SteegJ J Oppenheim
Apr 1, 1985·Immunology Today·G Trinchieri, B Perussia

❮ Previous
Next ❯

Citations

Jul 20, 1990·Journal of Immunological Methods·P H NibberingR Van Furth
Dec 1, 1995·Molecular and Biochemical Parasitology·A C La FlammeW C Van Voorhis
Nov 1, 1990·Infection and Immunity·S J MedingJ Langhorne

❮ Previous
Next ❯

Related Concepts

Related Feeds

Antivirals

Antivirals are medications that are used specifically for treating viral infections. Discover the latest research on antivirals here.

Antivirals (ASM)

Antivirals are medications that are used specifically for treating viral infections. Discover the latest research on antivirals here.