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

Research output: Contribution to journalArticlepeer-review

24 Scopus citations

Abstract

Adherent human blood monocytes were stimulated with heat-killed Staphylococcus albus in the presence of 35S-methionine and 3H-leucine. After 36 hours of incubation, the supernatant medium was purified over an anti-human endogenous pyrogen immunosorbent followed by gel-filtration, each fraction was assayed for lymphocyte activating factor (LAF) and active fractions were pooled for chromatofocusing. Chromatofocusing confirmed the three isoelectric 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.

Original languageEnglish (US)
Pages (from-to)59-64
Number of pages6
JournalBritish Journal of Rheumatology
Volume24
Issue numberSUPPL. 1
StatePublished - 1985

ASJC Scopus subject areas

  • Rheumatology
  • Neuroscience(all)

Fingerprint Dive into the research topics of 'Purified, <sup>35</sup>S-Met, <sup>3</sup>H-Leu-labelled human monocyte interleukin-1 (IL-1) with endogenous pyrogen activity'. Together they form a unique fingerprint.

Cite this