| Summary: | 碩士 === 長榮大學 === 醫學研究所碩士班醫學組 === 93 === Allergen specific Immunotherapy (IT) induced immune tolerance with induction of TH2 cells anergy remain to be clarified. The aim of this study was to evaluate whether the mite allergen Dermatophagoides pteronyssinus (Der p) specific IT serially decreased IL-4+/CD4+ (TH2) lymphocytes and induced apoptosis of TH2 lymphocytes in asthmatic children. Sixty Der p-sensitive asthmatic children were randomly assigned to a received IT and an untreated group. Der p specific IT treated patients were examined at three time points: before IT, after 6 months of an increased dose phase and with maximum tolerated doses after 1 year. Peripheral blood mononuclear cells (PBMC) were isolated and cultured for 48 hours for cellular staining with CD4+, CD45RO, CD25+ cell phenotypes and interleukin (IL)-4 and interferon-γ expression by fluorescence monoclonal antibodies. Apoptosis was measured using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL) method. A simultaneous flow cytometric study using the same permeabilized cell was examined to determine whether apoptosis occurred preferentially in TH1 or TH2 lymphocytes and whether CD4+CD25+ regulatory T cells would be elevated after Der p specific IT. We also used Enzyme-linked immunosorbent assay (ELISA) to detect whether immuno-modulators such as Interleukin-10, Transforming growth factor-β (TGF-β) and soluble CD14 (sCD14) molecule will be increased after Der p specific IT.
The data demonstrated that Der p specific IT decreased Dpt-specific IgE levels (P < 0.01) after 1 year of treatment. In addition, decreased CD4+IL-4+ TH2 cells with increased CD4+IFN-γ+ TH1 cells were observed at 6 months and 1 year after IT treatment (P < 0.05). At the same time, apoptosis of CD4+IL-4+ TH2 lymphocytes in the IT group had increased after 1 year’s of treatment when compared with the results before treatment and after 6 months of treatment (P < 0.05). In addition, CD45RO cells apoptosis mainly occurred after 6 months of IT treatment and after 1-year period of IT treatment (P < 0.05). After 1-year period of IT, we found that increased CD4+CD25+ regulatory T cells with induction of IL-10, TGF-β and sCD14 levels (P < 0.05). Besides, increased sCD14 levels correlated with elevated CD4+CD25+ regulatory T cells were found (P < 0.05).
All above data suggested that Der p specific IT could decrease Der p specific IgE and CD4+IL-4+ TH2 lymphocytes with induction apoptosis of CD4+IL-4+ TH2 lymphocytes subsets serially. Besides, Increased CD4+CD25+ regulatory T cells with induction of IL-10, TGF-β, sCD14 levels may partially explain the mechanism of Der p specific IT.
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