Li Na, Jiang Yu-xin, Diao Ji-dong, Zhao Bei-bei, Li Chao-pin
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2014 Aug;32(4):280-4.
To explore the effect of specific immunotherapy with major 3 group recombinant allergen rDer f 3 of Dermatophagoides farinae in murine asthma model.
Forty BALB/c mice were randomly divided into 4 groups, namely PBS group (negative control), ovalbumin(0VA) group (positive control), rDerf3 allergen sensitization group (asthma group), and rDerf3 specific immunotherapy group(SIT group). The mice in asthma group and SIT group were injected intraperitoneally with purified rDer f 3 protein on days 0, 7 and 14, respectively, and rDer f 3 solution was inhalated from day 21 for 7 days. During the 25th-27th day, mice in SIT group were injected subcutaneously with 100 jg rDer f 3 allergen for 30 min before nasal inhalation. Mice in groups of PBS and OVA were treated with PBS and OVA, respectively. Twenty-four hours after the final challenge, all mice were sacrificed, the bronchoalveolar lavage fluid (BALF) was collected and the total number of white blood cells and the number of eosinophils were recorded. The levels of IL-5 and IFN-gamma in BALF and supernatant of cultured splenocytes were detected by ELISA, as well as the serum levels of specific IgE and IgG2, antibodies. Lung tissues were stained with haematoxylin and eosin for histological analysis.
Compared with the asthma group, the rDer f 3-induced lung inflammation was significantly alleviated in SIT group. The total number of white blood cells [(7.03 +/- 1.38) x 10(8)/ml] in SIT group was considerably lower than that of OVA group [(22.11 +/- 3.70) x 10(8)/ml] and asthma group [(22.75 +/- 3.24) x 10(8)/ml] (P<0.01). The change trend of eosinophil leukocytes was similar with that of white blood cells. IL-5 levels in BALF [(108.20 +/- 11.02) pg/ml] and splenocyte culture supernatant [(98.34 +/- 13.06) pg/ml] in SIT group were significantly lower than that of OVA group [(182.04 +/- 13.94) pg/ml, (208.26 +/- 10.63) pg/ml] and asthma group [(195.33 +/- 15.33) pg/mL, (179.54 +/- 13.65) pg/ml] (P<0.01). Whereas, the level of IFN-gamma in BALF [(107.98 +/- 12.64) pg/ml] and supernatant of cultured splenocytes [(105.51 +/- 1.62) pg/ml] in SIT group was significantly higher than those of OVA group and asthma group (P<0.01). Compared with OVA group [(26.87 +/- 4.30) IU/ml] and asthma group [(35.25 +/- 8.84) IU/ml], a lower level of allergen-specific IgE [(9.12 +/- 3.78) IU/ml] and higher level of allergen-specific IgG2, [(38.52 +/- 6.33) microg/ml] were observed in SIT group (P<0.01).
rDer f 3 allergen can reverse allergen-induced airway and lung inflammation in murine asthma model.
探讨粉尘螨主要重组变应原rDer f 3特异性免疫疗法对小鼠哮喘模型的影响。
将40只BALB/c小鼠随机分为4组，即PBS组（阴性对照组）、卵清蛋白（OVA）组（阳性对照组）、rDerf3变应原致敏组（哮喘组）和rDerf3特异性免疫疗法组（SIT组）。哮喘组和SIT组小鼠分别于第0、7和14天腹腔注射纯化的rDer f 3蛋白，并从第21天开始吸入rDer f 3溶液，持续7天。在第25 - 27天，SIT组小鼠在鼻腔吸入前30分钟皮下注射100μg rDer f 3变应原。PBS组和OVA组小鼠分别用PBS和OVA处理。末次激发后24小时，处死所有小鼠，收集支气管肺泡灌洗液（BALF），记录白细胞总数和嗜酸性粒细胞数。采用酶联免疫吸附测定法（ELISA）检测BALF和脾细胞培养上清液中IL - 5和IFN - γ水平以及血清特异性IgE和IgG2抗体水平。肺组织用苏木精和伊红染色进行组织学分析。
与哮喘组相比，SIT组rDer f 3诱导的肺部炎症明显减轻。SIT组白细胞总数[（7.03±1.38）×10⁸/ml]显著低于OVA组[（22.11±3.70）×10⁸/ml]和哮喘组[（22.75±3.24）×10⁸/ml]（P<0.01）。嗜酸性粒细胞的变化趋势与白细胞相似。SIT组BALF中IL - 5水平[（108.20±11.02）pg/ml]和脾细胞培养上清液中IL - 5水平[（98.34±13.06）pg/ml]显著低于OVA组[（182.04±13.94）pg/ml，（208.26±10.63）pg/ml]和哮喘组[（195.33±15.33）pg/mL，（179.54±13.65）pg/ml]（P<0.01）。然而，SIT组BALF中IFN - γ水平[（107.98±12.64）pg/ml]和脾细胞培养上清液中IFN - γ水平[（105.51±1.62）pg/ml]显著高于OVA组和哮喘组（P<0.01）。与OVA组[（26.87±4.30）IU/ml]和哮喘组[（35.25±8.84）IU/ml]相比，SIT组变应原特异性IgE水平较低[（9.12±3.78）IU/ml]，变应原特异性IgG2水平较高[（38.52±6.33）μg/ml]（P<0.01）。
rDer f 3变应原可逆转小鼠哮喘模型中变应原诱导的气道和肺部炎症。
