| Objective: This study was designed to get know of Th subsets function statius of asthmatic children in remission period, and to observe the effects of aminophylline on Th subsets CKs from PBMC of children with chronic asthma during remission in vitro, at the same time, to analysis the anti-inflammation mechanisms of aminophylline and provide dosage reference for prophylaxis of asthma with aminophylline during remission period. Methods: PBMCs were separated from vein blood of 20 asthmatic children, devided into four groups: ACG,ATG1,ATG2,ATG3,according to aminophylline concentration (0,5,10,and 20 mg/L). At the same time, NCG including 11 nomal children＇PBMCs were determined. The concentration of serum total IgE and levels of IL-4,IFN-γ,IL-10 in culture supernatant of PBMC with PMA- ionomycin-stimulated and cultivated for 24h were measured with Enzyme-linked immunoadsordent assay (ELISA). IL-4+ and IFN-γ+ cell percentage of PBMC with PMA-ionomycin-monesin-stimulated and cultivated for 5h were detected with Enzyme-linked immunospot assay (ELISPOT). Meanwhile peripheral blood EOS numbers of every subject were counted. Results: ①EOS and IgE in ACG were significantly lower than NCG (t= 2.402 and 2.197, P<0.01 and 0.05); ② IL-4,IL-10 levels in ACG were significantly higher than NCG (P<0.01and 0.05), while IFN-γ level was significantly lower than NCG (t=-4.257,P<0.001); ③ IL-4 +cell percentage in ACG were higher than NCG (t=2.174,P <0.05), while IFN-γ+cell percentagewas lower than NCG(t=-2.437,P<0.05); ④ IL-4/IFN-γ ratio and IL-4+ cell/IFN-γ+ cell ratio in ACG were significantly higher than NCG (P<0.01); ⑤ Aminophylline mediated a slight inhibition to IL-4 and IL-4+ cell percentage, and this inhibition only was obvious at the concentrion of 20mg/L(P<0.01 and 0.05); ⑥ IFN-γ levels were significantly increased only in ATG3 (P<0.05), and IFN-γ+cell percentages weren't changed by aminophylline; ⑦ Aminophylline reduced IL-4/IFN-γ ratio and IL-4+ cell%/IFN-γ+ cell % ratio significantly in ATG3 (P<0.01 and 0.05);⑧ Aminophylline supressed IL-10 synthesis of PBMCs significantly in ATG3 (P<0.05)；⑨ Significant positive correlations were found between serum levels of IgE and IL-4(r= 0.845,P<0.01), IL-4+ cell %(r = 0.660, P<0.01), IL-4/IFN-γ(r= 0.822, P<0.01), IL-4+ cell /IFN-γ+ cell (r= 0.475 ,P<0.05).There were negative correlations between IL-4 and IFN-γ(r=-0.492,P<0.05),IL-4+ cell % and IFN-γ+cell %(r=-0.475,P<0.05)in ACG,while positive correlations were found between IL-4 level and IL-4+cell percentage(r=0.874,P<0.01),IL-4 level and IL-10 level(r=0.568, P<0.01), IL-10 level and IL-4/IFN-γ ratio(r=0.558,P <0.05),IFN-γ and IFN-γ+cell percentage(r=0.789, P<0.01),IL-4/IFN-γ ratio and IL-4+cell%/IFN-γ+cell% ratio(r=0.639, P<0.01).There were significant positive correlations between EOS and IL-4/IFN-γ ratio(r=0.704,P<0.01),EOS and IL-4+ cell%/IFN-γ+ cell% ratio(r=0.667, P<0.01).Conclusions: ①There is Th2 subsets hyperfunction during asthma remission period, characterized by higher EOS in peripheral blood and higher serum IgE, increase of IL-10 ,higher IL-4/IFN-γ ratio and IL-4+ cell%/IFN-γ+cell% ratio; physician should pay attention to therapy of asthma in remission period; ②IL-10 levels in ACG were significantly higher than NCG in the study. We conclude that it may be due to the Th2 subset of asthmatic children in remission period has more powerful potential to secrete IL-10 than that in acute period. Meanwhile, significant positive correlationswere found between IL-10 level and IL-4/IFN-γ ratio, IFN-γ and IFN-γ+ cell percentage,which may hint that IL-10, one of the important adjustment cytokines to Th2/Th1 clones, strengthen Th2 subset function and mediate asthma episode; ③These data suggest that cytokines expressions of Th2 are inhibited at therapeutic concentration superior limit (20mg/L) of aminophylline, so we guess which Th2 cytokines were suppressed by aminophylline is likely to be an important mechanism of anti-inflammation at t...
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