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

• Published in: Rheumatology (Oxford, England) Vol. XXIV; no. suppl-1; pp. 59 - 64
• Main Authors: Dinarello, C. A., Bernheim, H. A., Cannon, J. G., Lopreste, G., Warner, S. J. C., Webb, A. C., Auron, P. E.
• Format: Journal Article
• Language: English
• Published: Oxford University Press 01.01.1985
• Subjects: Endogenous pyrogen; Fever; Interleukin-1
• ISSN: 1462-0324; 1462-0332
• DOI: 10.1093/rheumatology/XXIV.suppl_1.59

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.