[Immunotherapeutic Effect of Dermatophagoides pteronyssinus Group 1 Allergen T Cell Epitope Peptide Against Allergic Asthma in Mice].
Chao-pin Li, Beibei Zhao, Xiao-dong Zhan
Abstract
Chao-pin Li, Beibei Zhao, Xiao-dong Zhan
Abstract
Objective: To study the specific immunotherapeutic effect of Dermatophagoides pteronyssinus group 1 major allergen T-cell fusion epitope peptide vaccine TAT-IhC-DPTCE against allergic asthma. Methods: One hundred and twenty SPF-grade BALB/c mice were randomized into PBS group (group A), asthma group (group B), and immune treatment groups respectively receiving intraperitoneal (i.p.) injections of ProDer p 1 allergen (group C), DPTCE (group D), TAT-DPTCE (group E) or TAT-IhC-DPTCE (group F) (n=20 in each group). In detail, PBS (group A) or allergen extract derived from Dermatophagoides pteronyssinus (groups B-F, 10 μg) was intraperitoneally injected on days 0, 7 and 14, and was continued by aerosol inhalation from day 21 for 7 consecutive days (0.5 μg/ml, once/day, 30 min each time). The mice in groups C-F received i.p. injections of 100 μg/ml ProDer p 1, DPTCE, TAT-DPTCE and TAT-IhC-DPTCE respectively 30 min prior to inhalation challenge on days 25-27 as a specific immunotherapy, while those in groups A and B received 200 μl PBS. Twenty-four hours after the last inhalation challenge, all the mice were sacrificed. The lung histopathological changes were examined by HE staining. The levels of IFN-γ, IL-13, IL-10 and TGF-β in the bronchoalveolar lavage fluid (BALF) was determined with ELISA, and eosinophils in the BALF were counted (n=20 mice in each group). The serum level of IgE, IgG1 and IgG2a in orbital blood was determined by ELISA(n=5 mice in each group). Results: HE staining revealed increased BALF eosinophils and decreased pulmonary inflammation in group F compared with group B. The IFN-γ level in group F [(298.75±26.09) pg/ml] was significantly higher than those in groups B[(158.71±20.89) pg/ml], C[(210.38±18.92) pg/ml], D [(229.44±13.00) pg/ml] and E[(233.24±20.39) pg/ml] (all P<0.01). Similar results were also found for IL-10 and TGF-β, while the IL-13 levels in groups C [(47.35±4.71) pg/ml], D [(41.90±4.28) pg/ml], E[(41.05±6.50) pg/ml] and F[(18.53±5.67) pg/ml] were all significantly lower than that in group B [(66.68±6.63) pg/ml](all P<0.01). The number of BALF eosinophils in group B [5.65±0.91]×105/ml] was significantly higher than that in group A [(0.45±0.39)×105/ml] (P<0.01), while the BALF eosinophils in groups C [(4.00±0.59)×105/ml], D [(3.39±0.63)×105/ml], E [(3.24±0.69)×105/ml] and F [(1.42±0.49)×105/ml] decreased after immune treatment (all P<0.01). ELISA results showed that the serum IgE level in group F [(5.26±1.72) ng/ml] was significantly lower than those in group B [(32.81±2.98) ng/ml] and the other 3 treatment groups [group C, (20.06±3.17) ng/ml; D, (17.06±3.18) ng/ml; E, (16.23±3.61) ng/ml]. Similar results were also obtained for IgG1. In contrast, the serum IgG2a level in group F[(43.10±1.34) ng/ml] was significantly higher than those in group B[(12.61±1.87) ng/ml] and the other 3 treatment groups [group C, (23.37±2.67) ng/ml; D, (25.60±2.10) ng/ml; E, (25.91±1.33) ng/ml] (all P<0.01). Conclusion: Immunotherapy with chimeric TAT-IhC-DPTCE can effectively ameliorate the allergic airway response and pulmonary inflammation in mice.
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Objective: To study the specific immunotherapeutic effect of Dermatophagoides pteronyssinus group 1 major allergen T-cell fusion epitope peptide vaccine TAT-IhC-DPTCE against allergic asthma. Methods: One hundred and twenty SPF-grade BALB/c mice were randomized into PBS group (group A), asthma group (group B), and immune treatment groups respectively receiving intraperitoneal (i.p.) injections of ProDer p 1 allergen (group C), DPTCE (group D), TAT-DPTCE (group E) or TAT-IhC-DPTCE (group F) (n=20 in each group). In detail, PBS (group A) or allergen extract derived from Dermatophagoides pteronyssinus (groups B-F, 10 μg) was intraperitoneally injected on days 0, 7 and 14, and was continued by aerosol inhalation from day 21 for 7 consecutive days (0.5 μg/ml, once/day, 30 min each time). The mice in groups C-F received i.p. injections of 100 μg/ml ProDer p 1, DPTCE, TAT-DPTCE and TAT-IhC-DPTCE respectively 30 min prior to inhalation challenge on days 25-27 as a specific immunotherapy, while those in groups A and B received 200 μl PBS. Twenty-four hours after the last inhalation challenge, all the mice were sacrificed. The lung histopathological changes were examined by HE staining. The levels of IFN-γ, IL-13, IL-10 and TGF-β in the bronchoalveolar lavage fluid (BALF) was determined with ELISA, and eosinophils in the BALF were counted (n=20 mice in each group). The serum level of IgE, IgG1 and IgG2a in orbital blood was determined by ELISA(n=5 mice in each group). Results: HE staining revealed increased BALF eosinophils and decreased pulmonary inflammation in group F compared with group B. The IFN-γ level in group F [(298.75±26.09) pg/ml] was significantly higher than those in groups B[(158.71±20.89) pg/ml], C[(210.38±18.92) pg/ml], D [(229.44±13.00) pg/ml] and E[(233.24±20.39) pg/ml] (all P<0.01). Similar results were also found for IL-10 and TGF-β, while the IL-13 levels in groups C [(47.35±4.71) pg/ml], D [(41.90±4.28) pg/ml], E[(41.05±6.50) pg/ml] and F[(18.53±5.67) pg/ml] were all significantly lower than that in group B [(66.68±6.63) pg/ml](all P<0.01). The number of BALF eosinophils in group B [5.65±0.91]×105/ml] was significantly higher than that in group A [(0.45±0.39)×105/ml] (P<0.01), while the BALF eosinophils in groups C [(4.00±0.59)×105/ml], D [(3.39±0.63)×105/ml], E [(3.24±0.69)×105/ml] and F [(1.42±0.49)×105/ml] decreased after immune treatment (all P<0.01). ELISA results showed that the serum IgE level in group F [(5.26±1.72) ng/ml] was significantly lower than those in group B [(32.81±2.98) ng/ml] and the other 3 treatment groups [group C, (20.06±3.17) ng/ml; D, (17.06±3.18) ng/ml; E, (16.23±3.61) ng/ml]. Similar results were also obtained for IgG1. In contrast, the serum IgG2a level in group F[(43.10±1.34) ng/ml] was significantly higher than those in group B[(12.61±1.87) ng/ml] and the other 3 treatment groups [group C, (23.37±2.67) ng/ml; D, (25.60±2.10) ng/ml; E, (25.91±1.33) ng/ml] (all P<0.01). Conclusion: Immunotherapy with chimeric TAT-IhC-DPTCE can effectively ameliorate the allergic airway response and pulmonary inflammation in mice.
Key concepts: Allergen, Immunology, Medicine, Bronchoalveolar lavage, Inhalation, Group A, Immunoglobulin E, Group B