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To examine RNA/protein synthesis of neutrophils and related dynamic changes during the inflammatory process, we investigated mRNA expressions in neutrophils, by RNA blot hybridization analyses using 12 different rabbit gene probes. We first selected five candidate genes encoding inflammation-related proteins, i.e. tumor necrosis factor (TNF-alpha) IL-1 alpha, IL-1 beta, neutrophil activating peptide-1/IL-8 (NAP-1/IL-8) and monocyte chemoattractant protein (MCP)-1. We further selected several genes on basis of the results from gene subtraction between cDNA libraries from neutrophils at an early (5 h) and at a late (24 h) stage of casein-induced acute peritonitis in rabbits, i.e. immune activation gene-2 (Act-2), migration inhibitory factor-related protein-8 (MRP-8), MRP-14, gamma-actin, and formyl-methionyl-leucyl-phenylalanine receptor (fMLP-R), and ferritin light (L) chain. In addition to these genes we used ferritin heavy (H) chain gene, another component of the ferritin molecule. We examined mRNA expressions by cytoplasmic slot blot analysis of the above 12 genes in neutrophils obtained from blood and from various stages of casein-induced inflammation in rabbits. The observed patterns of mRNA expression kinetics were classified into three. Pattern 1: mRNAs of MRP-8, MRP-14, and gamma-actin were constitutively expressed in blood neutrophils, and increased rapidly after emigration into inflammatory sites. Pattern 2: mRNAs of IL-1 beta, NAP-1/IL-8, Act-2, and fMLP-R were undetectable in blood neutrophils, and were induced rapidly after the onset of inflammation. Pattern 3 mRNAs of ferritin L and H chain were induced slowly, and increased with progression of the inflammatory process.(ABSTRACT TRUNCATED AT 250 WORDS)

Rabbits with casein‐induced peritonitis were febrile only during the early stage (1.5 to 12 hours) of inflammation. At that stage, peritoneal exudate cells (PEC) had preformed endogenous pyrogen (EP), while blood leukocytes and leukocytes obtained at the later stage of the inflammatory process did not. Early PEC consisted of 99% polymorphonuclear leukocytes (PMN). As early PEC and highly purified PMN (almost 100% pure) released almost identical amounts of EP, we concluded that PMN were the EP‐producing cells in the early PEC. EP from PMN of rabbits was composed of three factors with similar molecular weight (8,000 to 18,000) but different pI values (pI 7.2, 5.4, and 4.5). EP in the three fractions could not be separated from thymocyte comitogenic factor (TMF) in terms of m.w. and pI values. About 60% of EP activity and 92% of TMF activity in the culture supernatant were present in the pi 7.2 fraction. The pi 7.2 factor was weaker in EP activity and stronger in TMF activity than the other acidic fractions (pI 5.4 and 4.5). After high purification, 42 ng of the pI 7.2 factor had one unit of EP activity: one unit of TMF activity was present in 126 pg of highly purified pi 7.2 factor.