Non-specificity Immune Vaccine Regulates Asthma Via Th1/Th2/Treg/Th17Cell Balance In An Acute Mouse Model

Objective：Asthma is a common respiratory disease which impairs patientsphysically and mentally. Immune deregulation plays an important role inasthma, within which the Th1/Th2/Treg/Th17imbalance contributessignificantly to the pathogenesis of asthma. Intervention against abnormalimmunity shows an effective path to regulate asthma. Heat shock protein70(Hsp70) and immune stimulator CD80regulate cellular and humoral immunebalances. Here we utilize a non-specificity immune vaccine (Hsp70/CD80DNA vaccine) to cure mouse acute asthma, and explore the mechanism ofTh1/Th2/Treg/Th17rebalancing during the treatment with this novel vaccine.Methods: pVXA1plasmid vector was used to construct a recombinantHsp70/CD80fusion gene vaccine.40BALB/c mice were delivered into fourgroups (for control, asthma-model, pVXA1-vector and vaccine treatment), eachgroup contained10animals. OVA-induced acute asthma mice were treated withvaccine by intramscular injection. After latest OVA stimulation, the airwayhyperresponsiveness (AHR) was evaluated by inhaling methacholine; serumIgE was measured by ELISA; pathological changes was evaluated by HE andPAS staining; expression of cytokines (IFN-γ,IL-4, TGF-βand IL-17) inBALF was examined by ELISA; expression of transcription factors (T-bet,GATA-3, Foxp3and RORγt) in lung tissue was assayed by Real-time PCR.Results: Compared with control group the asthma-model group changed asfollows:(1) AHR increased about1~2folds, P<0.05;(2) serum IgE increasedabout30%, P<0.05;(3) infiltration of eosinophils and monocytes increased inlung tissue, proliferation and hypertrophy of goblet cells increased, bronchialmucus deposited. Compared with asthma-model group, vaccine group changedas follows:(1) AHR decreased about1~1.5folds, resembling with the control group, P<0.05;(2) serum IgE decreased about20%, P<0.05;(3) inflammatorychanges decreased in lung tissues, proliferation and hypertrophy of goblet cellsand bronchial mucus were repressed. These findings indicated that wesuccessfully builded OVA-induced acute asthma-model, and the Hsp70/CD80DNA vaccine contributed in the treatment of acute asthma. Furthermore, weexplored the molecule mechanism of this vaccine treatment, focusing on theTh1/Th2/Treg/Th17rebalancing. We found expression of cytokines (IFN-γ,IL-4, TGF-βand IL-17) and transcription factors (T-bet, GATA-3, Foxp3andRORγt) changed through the vaccine treatment: IFN-γ/IL-4and TGF-β/IL-17decreased about50%and70%individually in asthma group compared withcontrol group, P<0.05; and increased about80%and250%in vaccine groupcompared with asthma-model group, P<0.05. T-bet/GATA-3and Foxp3/RORγt decreased about40%and60%individually in asthma group compared withcontrol group, P<0.05; and increased about three folds in vaccine groupcompared with asthma-model group, P<0.05. These findings suggested theHsp70/CD80DNA vaccine regulated asthma, depending on cytokines (IFN-γand TGF-β) and transcription factors (T-bet, Foxp3). Conclusion: TheHsp70/CD80DNA vaccine was effective to asthma, which up-regulated theexpression level of cytokines (IFN-γ and TGF-β) and transcription factors(T-bet and Foxp3), resulted changes of the ratio (IFN-γ/IL-4, TGF-β/IL-17,T-bet/GATA-3, Foxp3/RORγt) to restore Th1/Th2/Treg/Th17cell balancing.