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KMID : 0350519940470031275
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Journal of Catholic Medical College
1994 Volume.47 No. 3 p.1275 ~ p.1283
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Effect of Interleukin-5 on Chemotaxis and Superoxide Anion Generation of Guinea Pig Eosinophils
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Abstract
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Eosinophils have been implicated as major effector cells in pathogenesis of allergic diseases. In order to observation of tissue injury in allergic diseases, the eosinophils have to migrate to target organ and generate inflammatory chemical
mediators,
granule-associate basic proteins and highly reactive oxygen metabolites. The potential role of IL-5 as eosinophil-proliferating, activating and/or recruiting mediator led us to evaluate the effects of IL-5, PAF, fMLP or PMA on superoxide anion
generation by eosinophils and chemotaxis by eosinophils with or without priming by IL-5. Eosinophil-rich peritoneal exudate was obtained in guinea pig by horse serum or polymyxin B sulfate as a stimulator and the eosinophils were purified by
Percoll
gradient centrifugation method. Chemotactic activity was evaluated on micro-Boyden chamber and results were expressed as the number of eosinophils in the presence of HBSS(Hnak's balanced salt solution), PAF(platelet activating factor), fMLP
(formyl-methionly-leucyl-phenylalanine) or IL-5(interleukin-5), Superoxide anion generation by eosinophils stimulated with PAF, PMA, fMLP or IL-5 was determined as the superoxide dismutase(20μg/ml)-inhibitable reduction of cytochrome C.
@ES The results were as follows;
@EN 1. The mean total cell count, the total eosinophil count, and the proportion of eosinophils in peritoneal exudate of guinea pig were 25.1±3.4(×10E6) and 34.3% after injection of polymxyxin B sulfate, and 26.8±5.3(×10E6), 8.4±2.9(×10E6),
31.4%
after injection of horse serum respectively.
2. Superoxide anion generation of eosinophils in response to PAF, fMLP, PMA or IL-5 was increased in time dependent manner. Superoxide anion generation by eosinophils was 12.45±4.26M, 36.12±14.37nM, 43.83*17.21nM, 42.29±16.52nM, at 20, 40, 60
and 90
minute after stimulation with PMA(phorbol myristate acetate). Superoxide anion generation by eosinophis was 8.62±2.5nM, 14.13±5.18M, 18.24±6.25M, 20.76±6.93nM, at 20, 40, 60 and 90 minute after stimulation with PAF. Superoxide anion
generation
by
eosinophils was 6.32±1.95nM, 14.52±3.92nM, 14.52±4.93nM, 17.26±5.37nM, at 20, 40, 60, 90 minute after stimulation with fMLP. Superoxide anion generation by eosinophils was 9.45±2.47nM. 23.32±6.37nM, 25.17±8.63nM, 26.92±9.43nM, at 20, 40,
60, 90
minute after stimulation with IL-5.
3. The mean numbers of migrating eosinophils after stimulation with HBSS, PAF, fMLP or IL-5 to eosinophils 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(400ng/ml) were 11±4.7, 98±30.1, 45±19.8, 106±42.8. Eosinophils with priming by IL-5
were
showed more chemotactic activity 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 upregulation of chemotactic response toward different agents. IL-5 as well as other stimulator
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 chemotaxis and superoxide anion generation.
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