Allergy.  1995 Mar;15(1):24-35.

The effects of interleukin5 on chemotaxis and superoxide anion generation by guinea pig eosinophils

• Cho SH
• Lee JS
• Kim HH

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 eosinophibproliferating, activa-ting and/or recruiting mediator led us to evaluate 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. Eosinophilrich peritoneal exudate was obtained in guinea pig by horse serum or polymyx in B sulfate as a stimulator and the eosinophils were purified by percoil gradient centrifugation method. Chemotactic activity was evaluated on micro-Boydenchamber and results wrer expressed as the number of eosinophils in the presence of HBSS (Hank's balanced salt solution), PAF (platelet activating factor), fMLP (formylmethiony-leucyl-phenylalanine) or IL-5 interl-eukin-5). Superoxide anion generation by eosmophils stimulated with PAF, PMA, fMLP or IL-5 was determined as the superoxide dismutase (20 /micro gram /ml)-inhibitable reduction of cytochrome C. The results were as follows; 1. The mean total cell count, the total eosinophil 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(x106) or 26.8+/-5.3 (x106), 8.6+/-2.5(x106) or 8.4+2.9(x106) and 34.3 % or 31.4 %. 2. Superoxide anion generation of eosmophils in response to PA?, fMLP, PMA or IL 5 was increased 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+/-14.37 nM, 43.83+/-17.21 nM, 42.29+/-6.5 2 nM. At 20, 40, 60, 90 minute after stimulation with PAF, superoxide anion generation by eosinophfis was 8.62+/-2.5 nM, 14.13+/-5.18 nM, 18. 24+/-6.25nM, 20.76+/-6.93nM. At 20, 40, 60, 90 mintute after stimulation with fMLP, super-oxide anion generation by eosinophils was 6.32+/-1.95 nM, 14.52+/-3.92 nM, 14.52+/-4.93 nM, 17. 26+/-5.37nM. At 20, 40, 60, 90minute after stimulation with IL-5, superoxide anion generation 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 mean numbers of migrating eosinophiIs after stimulation with HBSS, PAF, fMLP or IL-5 to eosmophils without priming by IL- 5 were 9+/-3.2, 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 11+/-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 chemotaetic agents for eosinophfis and IL-5 is modulator of eosmophil chemotaxis causing selective up regulation of chemotactic response toward different agents. IL-5 like other stimulators induces superoxide anion generation by eosmophils 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.

MeSH Terms

Animals
Cell Count
Centrifugation
Chemotaxis*
Cytochromes c
Eosinophils*
Exudates and Transudates
Guinea Pigs*
Guinea*
Horses
Interleukin-5
Oxygen
Peritoneal Cavity
Polymyxin B
Superoxide Dismutase
Superoxides*
Up-Regulation
Cytochromes c
Interleukin-5
Oxygen
Polymyxin B
Superoxide Dismutase
Superoxides