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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.
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Dinarello et al. (1985) studied this question.
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