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© 1985 All rights reserved

Purified, 35S-Met, 3H-Leu-Labelled Human Monocyte Interleukin-1 (IL-1) with Endogenous Pyrogen Activity

C. A. Dinarello, H. A. Bernheim, J. G. Cannon, G. Lopreste, S. J. C. Warner, A. C. Webb and P. E. Auron


  Abstract

Adherent human blood monocytes were stimulated with heat-killed Staphylococcus albus in the presence of3i S-methionine and 3H-leucine After 36 hours of incubation, the supernatant medium was purified over an anti-human endogenous pyrogen immunoabsorbent followed by gel-filtration; each fraction was assayed for lymphocyte activating factor (LAF) and active fractions were pooled for chromatofocusing. Chromatofocusing confirmed the three isoelectnc focusing points of IL-1 Each fraction was also precipitated with trichloroacetic acid and run on SDS-PAGE. Fluorography revealed that the intensity of the single radioactive polypeptide in each fraction correlated with LAF activity. Each band differed by about 2000 daltons Pooled fractions were incubated with polymyxin B for 2 hours and then injected into rabbits. Only those fractions with LAF activity produced characteristic endogenous pyrogen fevers while fractions not containing LAF were non-pyrogenic. These studies suggest that the charge heterogeneity of IL-1 is associated with molecular weight differences and confirm previous studies that LAF and endogenous pyrogen appear to be identical molecules.

KEY WORDS: Interleukin-1, Endogenous pyrogen, Fever


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