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© 1985 All rights reserved

Isolation and Characterization of Human Interleukin-1 mRNA by Molecular Cloning

A. C. Webb, P. E. Auron, L. J. Rosenwasser, S. F. Mucci, A. Rich, S. M. Wolff and C. A. Dinarello


  Abstract

mRNA was isolated from LPS (hpopolysacchande)-stimulated monocytes and cDNA clones were propagated in bacteria Stimulation-specific cDNAs were characterized by injection of hybnd-selected mRNA into Xenopus laevis oocytes, followed by a bioassay for T-cell proliferation By this criterion, several cDNA clones appear to exhibit T-cell stimulating bioactivity, which could be inhibited by anti-IL-1 serum The cDNA clones were used as probes to characterize and isolate monocyle mRNA Northern blots reveal IL-1 mRNA to be approximately 1600 nucleotidcs long and represents about 5% of the total pA RNA in stimulated monocytes. IL-1 mRNA isolated by hybrid selection to cDNA is translated in rcticulocyte lysatc to yield a 35 kD polypeptide and in oocytes to produce biological activity. Similarly reticulocytc translation and immunoprecipitation of monocyte pA RNA fractionated on a sucrose gradient suggest that the IL-1 biological activity synthesized by oocytes is associated with translation of a precursor with a molecular weight around 35 kD.

KEY WORDS: Intrerleukin-1, mRNA, cDNA


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