[Administration of a single chain variable fragments chimeric protein (SD) of ovalbumin epitopes internalizing receptor DEC-205 antibody inhibits food allergy in mice].

Chong Wan, Meiying Wu, Yuqing Zhang, Junwei Shao, Qingqing Luo, Jiyu Ju, Lingzhi Xu
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Abstract

Objective To investigate the preventive therapeutic effect and possible mechanism of single chain variable fragments chimeric protein (SD) of ovalbumin epitopes internalizing receptor DEC-205 antibody on food allergy in mice. Methods Mice were randomly divided to five groups (control, PBS, scFv DEC 100 μg, SD 50 μg, SD 100 μg) and treated for 24 hours before OVA administration. After challenge, the serum level of OVA-specific IgE, IgG1, IgG2a and IL-4 were detected by ELISA. Infiltration of eosinophils and mast cells in the jejunum was observed by HE staining and toluidine blue staining respectively. The bone marrow of tibia and femur was isolated and cultured to obtain immature dendritic cells(BMDCs), which were further treated with LPS (10 ng/mL), TSLP (50 ng/mL), scFv DEC protein (1000 ng/mL) and SD protein (10,100,1000)ng/mL for 24 hours, and the IL-10 level of supernatant was assayed by ELISA. Results Compared with PBS group, the number of SD-treated mice with diarrhea was markedly reduced. The difference in rectal temperature and the levels of serum OVA-specific IgE, IgG1, IgG2a and IL-4 decreased significantly after prophylactic administration of SD; The number of eosinophils and mast cells in jejunum also decreased significantly while the IL-10 level in the supernatant of BMDCs increased significantly after SD intervention. Conclusion SD mitigates experimental FA response by fosters the immune tolerance property of dendritic cells.

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[给药卵清蛋白表位的单链可变片段嵌合蛋白(SD)内化受体DEC-205抗体抑制小鼠食物过敏]。
目的探讨卵清蛋白表位内化受体DEC-205抗体单链可变片段嵌合蛋白(SD)对小鼠食物过敏的预防治疗作用及其可能机制。方法将小鼠随机分为5组(对照组、PBS组、scFv组,DEC 100 μg、SD 50 μg、SD 100 μg),治疗24 h后给药。攻毒后,采用ELISA法检测ova特异性IgE、IgG1、IgG2a和IL-4水平。HE染色和甲苯胺蓝染色分别观察空肠嗜酸性粒细胞和肥大细胞的浸润情况。分离培养胫骨、股骨骨髓,获得未成熟树突状细胞(BMDCs), LPS (10 ng/mL)、TSLP (50 ng/mL)、scFv DEC蛋白(1000 ng/mL)、SD蛋白(10,100、1000)ng/mL)处理24h, ELISA检测上清液中IL-10水平。结果与PBS组比较,sd组腹泻小鼠数量明显减少。预防给药SD后,小鼠直肠温度差异及血清ova特异性IgE、IgG1、IgG2a、IL-4水平均显著降低;SD干预后,空肠嗜酸性粒细胞和肥大细胞数量也显著减少,BMDCs上清液中IL-10水平显著升高。结论SD通过增强树突状细胞的免疫耐受特性来减轻FA的实验反应。
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