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KMID : 0369319950150010024
Allergy
1995 Volume.15 No. 1 p.24 ~ p.35
The effects of interleukin-5 on chemotaxia sna superoxide anion generation by guinea pig eosinophils
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Abstract
Eosinophils have been implicated as major cells in the pathogenesis of allergic diseases. For tissue injury in allergic diseases, the eosinophils have to migrate to target organ and generate inflammatory chemical mediators, granuleassociat basic proteins and highly reactive oxygen metabolites. The potential role of IL-5 as eosinophil-proliferating, activating and/or recruiting mediator led us to evaIuate the effects of IL-5, PAF, fMLsP or PMA on superoxide anion generation by eosinophils and chemotaxis by eosinophils with or without priming by IL-5. Eosinophirich peritoneal exudate was obtained in guinea pig by horse serum or polymyx- in B sulfate as a stirnulator and the eosinophils were purified by percoll gradient centrifugation method. Chemotactic activity was evaluated on micro-Boyden chamber and results wrer expressed as the number of eosinophils in the presence of HBSS (Hanks balanced salt solution), PAF (platelet activating factor), fMLP (formyl- methionyl-leucyl-phenylalanine) or IL-5 (interl- eukin-5). Superoxide anion generation by eosinophils stirnulated with PAF, PMA, fMLP or IL-5 was determined as the superoxide dismutase (20 ug /ml)-inhibitable reduction of cytochrome C. The results were as follows', 1. The mean total cell count, the total eosino- phil count and the proportion of eosinophils in peritoneal exudate of guinea pig after injection of polymyxin B sulfate or horse serum into peritoneal cavity were 25.1+3.4(x10') or 26.8+5.3 (x10'), 8.6+2.5(x10') or 8.4x2.9(x10') and 34.3 % or 31.4 %. 2. Superoxide anion generation of eosinophils in response to PAF, fMLP, PMA or IL-5 was in- creased in time dependent manner. At 20, 40, 60, 90 minute after stimulation with PMA (phorbol muris-tate acetate), superoxide anion generation by eosinophils was 12.45+4.26 nM, 36.12+14.37nM, 43.83+17.21 nM, 42.29+-16.5 2 nM. At 20, 40, 60, 90 minute after stimulation with PAF, superoxide anion generation by eosinophils was 8.62+2.5 nM, 14.13+5.18 nM, 18. 24+6.25 nM, 20.76+6.93 nM. At 20, 40, 60, 90 mintute after stimulation with fMLP, superoxide anion generation by eosinophils was 6.32 +1.95nM, 14.52+3.92nM, 14.52+4.93 nM, 17. 26+5.37 nM. At 20, 40, 60, 90 minute after stimulation with IL-5, superoxide anion genera- tion by eosinophils was 9.45+2.47 nM, 23.32+ 6.37 nM, 25.17+ 8.63 nM, 26.92+9.43 nM. 3. The rnean numbers of migrating eosino- phils after stimulation with HBSS, PAF, fMLP or IL-5 to eosinophils without priming by IL-5 were 943.2, 64+21.5, 34+13.6, 73+-27.9. Chemotactic activity of eosinophils to PAF, fMLP or IL-5 was significantly higher than that of HBSS(p<0.05). The mean numbers of migrating eosinophils after stimulation with HBSS, PAF, fMLP or IL-5 to eosinophils with priming by IL-5(400 ng/ml) were ll+4.7, 98+ 30.1, 45+-19.8, 106+42.8. Eosinophils with priming by IL-5 were showed more chemotactic ctivity than eosinophils without priming when stimulated by PAF, or IL-5(P<0.05). In chemotactic activity to eosinophils, IL-5 or PAF was significantly increased compared with fMLP(P < 0.01). In conclusion, IL-5 together with PAF are important chemotactic agents for eosinophils and IL-5 is modulator of eosinophil chemotaxis causing selective up regulation of chemotactic response toward different agents. IL-5 like other stimulators induces superoxide anion generation by eosinophils in time dependent manner. So IL-5 may play a significant role in increasing the functional activities of eosinophils such as chomotaxis and superoxide anion generation in allergic diseases.
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