| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 1999: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1998: ¥2,600,000 (Direct Cost: ¥2,600,000)
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| Research Abstract |
1. Purification of a novel neutrophil chemotactic factor from rat granulation tissue :
A novel neutrophil chemotactic factor has been purified from the granulation tissue of air-pouch/carrageenin-induced inflammation in rats. The purified chemoattractant was a basic protein with heparin-binding site, and gave a single band corresponding to a molecular mass of 16-kDa on SDS-PAGE under reducing conditions. The chemoattractant was treated with lysylendopeptidase and the resulting peptides were isolated by reversed-phase HPLC. Amino acid sequences of the peptides were almost identical with the sequence of N-terminal fibronectin type III domain of human collagen type XIV, suggesting that the purified chemoattractant consists mainly of N-terminal fibronectin type III domain and the adjacent heparin-binding site of rat collagen type XIV. The 16-kDa fragment of collagen type XIV dose-dependently attracted rat neutrophils and transiently increased the intracellular free CaィイD12+ィエD1 concentratio
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n of neutrophils. The results suggest that the novel chemoattractant play a role in neutrophil recruitment in rat inflammation.
2. Roles of chemokines in neutrophil infiltration in rat inflammation models :
In the present study, by use of ELISA specific for each chemokine, we have determined the levels of the chemokines (CINC-1, -2, -3 and rat MIP-1α) in the pouch fluids of two types of inflammation models (FITC-ovalbumin-induced allergic inflammation and LPS-induced inflammation in rats). Effects of anti-chemokine antibodies on neutrophil chemotaxis were determined in vivo and in vitro. Anti-CINCs-antibodies, which inhibited all the CINCs, suppressed both neutrophil infiltration in vivo and neutrophil chemotactic activity of the 8-hr pouch fluid in vitro, whereas anti-MIP-1α antibody slightly suppressed the chemotaxis in vivo and in vitro. Our results suggest that CINC-1/-2 for the allergic inflammation and CINC-2/-3 for the LPS-induced inflammation, respectively, play an important role in infiltration of neutrophils into inflammatory sites of rat inflammation models. Less
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