| IntroductionThe dysfunction of T helper cell and disorder of the percent of Th1/Th2 are one of the pathogenesis that asthma happened. The function of Th2, in contrast with Th1, is enhanced. This results the cytokines pruduction of Th2 type are increased, including interleukin-4 interleukin- 5 and interleukin-13. these cytokines will lead to immunol cascade reaction in asthma, for example: the contraction of airway smooth musual, the infiltration of eosinophil, the increased secrete of immunoglobulin E and bronchial hyperresponsiveness.The safty and effectivity dencity of theophylline lies 10-20mg/L, 5-10mg/L be regarded as low dose. Anti-CD86mAb may inhibit activation of CD4+T cell by blocking signal conduction between allergen-presenting cell and CD4+T, as the result, the secret of inflammatorycytokines is decreased. We cocultured monocytes or alveolar macrophages and CD4+T in vitro,in order to investigate the effects of the low-dose theophylline and anti-CD86mAb on the inflammatory cytokines production of T helper cells and explore its pathogenesis.Material & MethodsMaterial Derp allergen; theophylline; Anti-CD86 mono- anti body; PE-anti-human IL-4 & IFN-r antibody; CD4+T immunomagnetic bead isolate system; foetal calf serum;RPM! Medium 1640 culture fluid; incubator; IL-4 IL-5 IFN-renzyme link immunosorbent assay kits; enzyme immnoassay apparatus; flow cytometry.Experiment I 10 subjects with Derp allergic asthma. Monocytes and CD4+T were isolated from peripheric venous blood which were obtained from 10 subjects with mild atopic asthma. Monocytes and CD4+T were cocultrued at the condition of 37CC.5%CO2 with or without theophylline. The experiment was divided into two groups: 5mg/ml and 10mg/ml. The supernatants were harvested after 72 hours coculture. The concentration of interferon-r(INF-r), interleukin -4(IL-4) and interleukin-5 (IL-5) were determined by ELISA. The percents of Th1 and Th2 that were marked by fluorescent antibody (IL-4Ab& IFN-rAb) were examined through FCM.Experiment II The specimens of brochoalveolar lavage fluid (BALF) and peripheric blood were derived from 10 house dust mite Dermatophagoides pteronyssinus (Der p)-allergic asthma patients. The alveolar macro- phages(AMs) and CD4+T cells were isolated from BALF and peripheric blood. AMs was stimulated and activated by allergen which were abstracted fromhouse dust-mite. Then AMs and CD4+T cells were co-cultured with or without anti-CD86mAb at 37癈.5%(v/v)CO2 condition. The experiment group was subdivided into two groups. One was 0.1mg/L,the other was 1mg/LThe super- natants were harvested after 72 hours coculture. The concentra- tion of INF-gamma IL-4 and IL-5 were determined by ELISA. The CD86 molecule expression on AMs was detected by flow cytometry.ResultsExperiment I In contrast with the controls, the production of IL-4 and IL-5 were lower .while that of INF-r was higher in experiment group. This variations ware significantly. (P<0.01) There were no difference in two experimemtal groups(P>.05). By contrast with the controls, the percentage of Th1/Th2 was increased in experiment group.Experiment II The expression of CD86 molecule on AMs was significantly higher than that in controls (P<0.01). The production of IL-4 and IL-5 of the control were increased than that of the anti-CD86mAb groups. While the secrets of IFN-gamma has no difference between two groups(P>0.05). IL-4 and IL-5 secretion in high-dose group were higher than that in low-dose one among experiment groups(P<0.01).Conclusion1. More and more studies and evidences show that theophlline can counteracts airway inflammation, attenuates plasma infiltration and inflammatory edema of bronchial mucous, inhibits airway hyperresponsiveness reaction. Fome our experiment, in contrast with the controls, the production of IL-4 and IL-5 were lower .while that of INF-r was higher in experimentgroup. This variations ware significantly. (P<0.01).There were no significant difference between two-dose experimental groups(P>0.05). These indicate that theophlline may effec...
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