| OBJACTIVE:To investigate the therapic mechanism of DF sublingual immunotherapy by studying the change law of 33D1 dendritic cells and NF-κB before and after the DF sublingual immunotherapy for a asthma mouse model and further comparing the therapic effect of DF vaccine of high dose and low dose.METHODS:1. Twenty eight SPF-grade Bal/c mice aged 4 to 6 weeks were divided into four groups: the control group of normal mouse; the group of asthmatic model; the group treated with high dose of Dermatophagoides farinae(DF) vaccine and the group treated with low dose of DF vaccine. The control group was without any treatment, and the other three groups were all allergized with 50μl crude solution of DF allergen and 2mg Al(OH)3 on the 1st, 8th and 15th day and conducted with sublingual vaccine (for 2 min) under the condition of anesthesia on 28th day, and then was respectively treated with PBS, high dose of DF vaccine (1mg) and low dose of DF vaccine (100μg) every 2 days. The three groups, a week after the immunotherapy, were all challenged by dropping nose with 50μg Der f1 every day continuously for 7days.2. The day next to the last challenge, airway hyperactivity of the mouse of these four groups was detected, first with PBS for a basic Penh value, and then chanllenged for observing the change of Penh by turns with methacholine of 1mg/ml, 10mg/ml, 30mg/ml, 50mg/ml and 100mg/ml.3. After two days of the last challenge, BAL was conducted upon each mouse under anesthesia and with 500ul PBS which was absorbed three times to take back the BALF to record its volume and Liu's staining was used to classify and count of 200 cells in BALF.4. Three days after the last challenge, a double staining immunohistochemical method was used to locate the 33D1 and NF-κB in lung tissues and submaxillary lymph nodes which were taken out from every mouse under anesthesia, and stereological method were used further to caculate their volume density(Vv), number density (Nv) , surface area density (Sv) under light microscope.RESULTS:1. Mice of the model group showed the symptoms as cough, gasp,fast and deep breathing and restlessness and the group treated with high dose of vaccine showed a clearly decreased symptoms.2. The number density (Nv), the volume (Vv), surface area density (Sv) of 33D1 in submaxillary lymph nodes of model group mice were(2.03±0.52)×10-3μm-3 , (2.75±1.33)×10-3μm0 and (2.32±1.20)×10-3μm-1 respectively. Those parameters of asthmatic mouse model group were (14.88±3.04)×10-3μm-3,(23.99±5.09)×10-3μm0, (22.06±1.67)×10-3μm-1 respectively and those of the group treated high dose of vaccine were(2.52±0.90)×10-3μm-3,(4.32±1.34)×10-3μm0(,3.17±0.85)×10-3μm-1 respectively. There was significant difference of the detective parameters of 33D1 between the group treated with high dose of vaccine and the group of model group(P<0.01), so there was between the model group and the control group(P<0.05).3. The number density (Nv) , the volume (Vv) ,surface area density (Sv) of NF-κB in lung tissues of normal mouse was 3.02±0.26)×10-3μm-3,(4.88±1.02)×10-3μm0 and (3.86±0.95)10-3μm-1 respectively ;Those parameters of mice asthma of model group wa(s70.46±2.37)×10-3μm-3(,88.26±1.73)×10-3μm0 and(86.03±1.48)×10-3μm-1 respectively ;and those of the group treated high dose of vaccine were(10.68±1.39)×10-3μm-3,(14.30±1.09)×10-3μm0 and(14.02±0.51)10-3μm-1 respectively .There was significant difference of the detective parameters of NF-κB between the group treated with high dose of vaccine and the group of model group(P<0.01),so was there between the model group and the control group(P<0.01), but wasn't between the model group and the group treated with lose dose of vaccine(P﹥0.05).4. The number of total cells and eosinophile granulocyte in BALF of model was (79.05±11.58)and(17.63±4.98) respectively, and that of the group treated with high dose of vaccine was (37.99±5.27) and(3.80±0.67)respectively, but that of the control group still remained was (63.06±11.66) and(17.43±2.87). The difference in the number of total cells and eosinophile granulocytes in BALF was significant between the group treated with high dose of vaccine and model group, but wasn't so between the model group and the group treated with low dose of vaccine.5.The airway hyperactivity of the group treated with dose of vaccine was obviously lower than that of the model group, but difference of the airway hyperactivity between the group of low dose vaccine and the model group was not significant.CONCLUSION:1. Sublingual DF vaccine immunotherapy for mice asthma is effective,and the accumulating dose of vaccine was900μg Der f1/18mg,which means that sublingual immunotherapy with total protein functions is an effective therapy for mice asthma.2. The mechanisms of the therapeutic effect of DF vaccine sublingual immunotherapy on mouse asthma involves the immunotolerance of 33D1 and the mechanisms of the down-regulation of NF-κB.
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