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 journalArticle

24 Citations (Scopus)

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

Fingerprint

Interleukin-1
Monocytes
Photofluorography
Polymyxin B
Immunosorbents
Trichloroacetic Acid
Isoelectric Point
leukocyte endogenous mediator
Staphylococcus
Leucine
Methionine
Gel Chromatography
Polyacrylamide Gel Electrophoresis
Fever
Hot Temperature
Molecular Weight
Rabbits
Peptides

ASJC Scopus subject areas

  • Rheumatology
  • Neuroscience(all)

Cite this

Purified, 35S-Met, 3H-Leu-labelled human monocyte interleukin-1 (IL-1) with endogenous pyrogen activity. / Dinarello, C. A.; Bernheim, H. A.; Cannon, J. G.

In: British Journal of Rheumatology, Vol. 24, No. SUPPL. 1, 1985, p. 59-64.

Research output: Contribution to journalArticle

Dinarello, C. A. ; Bernheim, H. A. ; Cannon, J. G. / Purified, 35S-Met, 3H-Leu-labelled human monocyte interleukin-1 (IL-1) with endogenous pyrogen activity. In: British Journal of Rheumatology. 1985 ; Vol. 24, No. SUPPL. 1. pp. 59-64.
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