Dynamic changes in mRNA expression of neutrophils during the course of acute inflammation in rabbits.
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)
Mori
S
S
Department of Pathology, Kumamoto University School of Medicine, Japan.
Goto
K
K
Goto
F
F
Murakami
K
K
Ohkawara
S
S
Yoshinaga
M
M
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D016207:Cytokines / Q000096:biosynthesis; D005260:Female; D015870:Gene Expression; D007249:Inflammation / Q000276:immunology*; D007930:Leucine / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000378:metabolism*; D010538:Peritonitis / Q000276:immunology; D012333:RNA, Messenger / Q000096:biosynthesis*; D011817:Rabbits; D014529:Uridine / Q000378:metabolism
D016207:Cytokines; D012333:RNA, Messenger; D007930:Leucine; D014529:Uridine