Figures & data
Figure 1. Two-dimensional PAGE of purification steps of 35S-labeled proteins in supernatants from heat-killed Staphylococcus epidermidis-stimulated human blood monocytes. The first-dimension pH measurements are derived from direct measurement of a parallel standard gel. Each panel indicates the purification step. Fractions eluting from the immunoaffinity step were chromatographed on Sephadex G-50 and the pyrogenic activity of fractions less than 20000 Daltons were pooled and subjected to chromatofocusing. The fractions eluting from the chromatofocusing step in the neutral pH range were then pooled and subjected to 2-dimensional PAGE. A single protein is visible by autoradiography with a molecular weight near 18000 Daltons and a pI between 6 and 7. Previous chromatofocusing in flat-beds revealed that the pI of leukocytic pyrogen was at 6.8.Citation21 The figure is reprinted from.Citation22
Figure 2. Co-elution of IL1 activities using radiolabeled proteins. A. SDS-PAGE of trichloracetic acid-precipitated fractions 24–36 of 35S-methionine-labeled monocyte supernatants during chromatofocusing. The pH of the chromatofocusing gradient is shown. Before chromatofocusing, 4 Ls of pooled monocyte supernatants were concentrated and subjected to sequential immunoadsorption and gel filtration.Citation23 B. Top. The same fractions shown in A were assayed for induction of PGE2 from dermal fibroblasts. Fever was assessed in trained rabbits and LAF activity was measured using in D10.G4.1 cells. Serum SAA was determined following intraperitoneal injection into mice.Citation30 Adapted from.Citation22
Figure 3. Multiple activities of human leukocytic pyrogen. A compilation of activities using recombinant human IL1β derived fromCitation2 and other published studies.Citation25,53
