Role Of MiR-146a In Enforcing Effect Of Specific Immunotherapy On Allergic Rhinitis

ObjectThis study was to investigate the effect and mechanism of miR-146a in specific immunotherapy of allergic rhinitis, and its expression was detected in the nasal mucosa of patients with allergic rhinitis. We have established a mouse model of allergic rhinitis, and to study the role of miR-146a in time by specific immunotherapy in the mouse model, aimed at obtaining an experimental basis for the search for new gene-specific immune efficiency and apply it clinical.Methods(1) Quantitative PCR was used to detect the expression of miR-146a mRNA in allergic patients and healthy people in the nasal mucosa.(2) We used 50 ?g ovalbumin (OVA),5 mg of aluminum hydroxide gel as adjuvant,1 mL of sterile saline formulated as a suspension, which was subsequently injected into BALB/c mice by intraperitoneal injection. Each injection time is every other day, a total of 7 injections; after the foundation sensitized mice, each group of mice were intranasally using 5% OVA sterile saline, each side in an amount of 10 ?L, frequency per day 1 times. We use the above method of allergic rhinitis (AR) model group were consecutive excitation model for 10 days,control group mice were injected with 0.9% saline by intraperitoneal injection, followed by the use of 0.9% saline for the group of mice intranasally. Enzyme-linked immunosorbent assay detected in the peripheral blood. Quantitative PCR 146a mRNA expression in (AR) nasal mice.(3) We were used OVA (NPVO), miR-146a (NPVm) and OVA+miR-146a (NPVom) vaccine nanoparticles act on the AR model mice, we observed changes in symptoms of AR and related indicators of the mice in the above operation.Results(1) The expression of miR-146a in the nasal mucosa of allergic rhinitis was significantly lower than its expression in normal nasal mucosa (P<0.01);(2) Mice with typical symptoms of allergic rhinitis in the AR model group. EOS count within the AR nasal mucosa of micep was higher than the control group; serum concentrations of OVA-sIgE in AR model mice was significantly higher than the control group (P<0.05) and with the extension of time is sensitized gradual upward trend; miR-146a expression in nasal mucosa in AR model mice was significantly lower than normal nasal mucosa (P<0.01).(3) Symptoms of allergic rhinitis was significantly worse in NPVo group mice; allergic rhinitis symptoms, no significant changes in NPVm treated mice; allergic rhinitis symptoms was significantly inhibited in NPVom treated mice; ELISA test results show, IgE and IL-4 in serum and nasal mucosa was significantly higher in the group of mice; flow cytometry results showed that proliferation of OVA-specific antigens Tregs significantly increased; Tregs proliferation activity in each group AR mice were used AR NPVo and NPVm processing, T cell proliferative activity did not change significantly, and when using NPVom mice, T cell proliferation in the group; model nasal mucosa and spleen of mice was significantly lower than the normal control group mice significantly increased; further analysis found that Tregs in the OVA-specific NPVom treated mice; after NPVm or NPVom mice, enabling DCs expressed TGF-? mRNA and the corresponding protein was significantly heighten.Co-cultured with naive CD4+T cells and NPV sensitized DCs, results showed NPVm/NPVom enables naive CD4+CD25-T cells into CD4+CD25+Foxp3+TGF-?+Tregs; with OVA effect, by the NPVom induction of Tregs proliferative activity was significantly increased, while the NPVm induced proliferation of Tregs did not change significantly; proliferation effect NPVom sensitized with BSA DCs were detected, the results showed that Tregs proliferative activity had no significant effect. The Tregs were adoptively transferred into the AR mice, the transfer of Tregs generated by NPVom significantly inhibited the AR-related symptoms in AR mice while the NPVm-induced Treg or naive CD4+T cells did not.ConclusionmiR-146a can enforce OVA specific immunotherapy via inducing antigen specific regulatory T cells. miR-146a may have therapeutic potential to be used in the immunotherapy of allergic diseases.