Abstract: Objective To study the effect of non-methylated CpG oligodeoxynucleotide (cytosine-phosphate-guanosine-oligodeoxynucleotides, CpG ODN) combined with heat shock protein 70 (heat shock protein 70, HSP70)/CD80DNA vaccine on pulmonary inflammation, serum levels of IL-4, IL-13, IFN-γ and the changes of IL-4, IL-25 and IL-33 in bronchoalveolar lavage fluid(BALF) of asthmatic mice, so as to provide experimental basis for the treatment of asthma by CpG ODN-HSP70/CD80DNA vaccine. Methods Ovalbumin (OVA) and aluminum hydroxide [Al (OH)₃] adjuvant were used to sensitize mice with asthma model.From June to September 2019,totally 32 BALB/c mice aged 6—8 weeks were selected and were randomly divided into four groups, including control group, asthma group, HSP70/CD80DNA vaccine group and combined vaccine group (CpG ODN combined with HSP70/CD80DNA vaccine group).HE staining was used to observe the pathological and morphological changes of lung tissue.The changes of serum levels of IL-4, IL-13, IFN-γ in and IL-4, IL-25, IL-33 in BALF were detected by ELISA. Results HE staining of lung tissue showed that the inflammatory response of lung tissue in combined vaccine group was significantly less than that in the asthma group, and the infiltration of inflammatory cells around the airway was significantly reduced.ELISA results showed that compared with the control group, serum levels of IL-4 and IL-13 in asthma group were significantly higher (P<0.05), and serum IFN-γ level was significantly lower (P<0.05).Compared with the asthma group, serum levels of IL-4 and IL-13 in the Hsp70/CD80DNA vaccine group and the combined vaccine group were significantly lower (P<0.05), and the serum IFN-γ level was significantly higher (P<0.05).Compared with the Hsp70/CD80DNA vaccine group, the serum levels of IL-4 and IL-13 in combined vaccine group were significantly decreased (P<0.05), and the serum IFN-γ were significantly increased (P<0.05).The results of BALF showed that compared with the control group, the levels of BALF IL-4, IL-25 and IL-33 in the asthma group were significantly increased (P<0.05).Compared with the asthma group, the levels of IL-4, IL-25 and IL-33 in BALF in the Hsp70/CD80 DNA vaccine group and the combined vaccine group were significantly decreased (P<0.05).And the levels of IL-33 in BALF in the combined vaccine group were significantly lower than those in the Hsp70/CD80 DNA vaccine group (P<0.05). Conclusion CpG ODN combined with Hsp70/CD80DNA vaccine can inhibit the production of IL-4, IL-13, IL-25 and IL-33 in mice, enhance the production of IFN-γ in mice, and reduce airway inflammation in asthmatic mice.

Key words: HSP70/CD80DNA vaccine, CpG ODN, airway inflammation, asthmatic mice

摘要： 目的 研究非甲基化CpG基序的寡聚脱氧核苷酸 (CpG ODN)联合热休克蛋白70(HSP70)/CD80DNA疫苗对哮喘小鼠肺部炎症、血清IL-4、IL-13、IFN-γ及支气管肺泡灌洗液(BALF)中IL-4、IL-25、IL-33水平变化的影响,为联合 CpG ODN -HSP70/CD80 DNA疫苗治疗哮喘提供实验依据。方法 2019年6-9月选用 6~8周龄BALB/c小鼠32只, 卵清蛋白(OVA)加氢氧化铝[Al(OH)₃]佐剂致敏的方法制备小鼠哮喘模型, 随机分成4组,分别为对照组、哮喘组、HSP70/CD80DNA疫苗组及联合疫苗组(CpG ODN联合HSP70/CD80DNA疫苗组),HE染色观察肺组织病理形态学改变,ELISA法检测血清IL-4、IL-13、IFN-γ和BALF中IL-4、IL-25、IL-33水平变化。结果 肺组织HE染色结果显示HSP70/CD80疫苗联合CpG ODN治疗组小鼠肺组织炎症反应较哮喘组明显减轻,气道周围炎症细胞浸润显著减少。ELISA结果显示与对照组比较,哮喘组血清IL-4及IL-13水平显著增高(P<0.05),血清IFN-γ水平明显下降(P<0.05);与哮喘组比较,HSP70/CD80DNA疫苗组和联合疫苗组血清IL-4及IL-13水平显著降低(P<0.05),血清IFN-γ水平明显升高(P<0.05);与HSP70/CD80DNA疫苗组比较,联合疫苗组血清IL-4及IL-13水平显著降低(P<0.05),血清IFN-γ水平明显升高(P<0.05);支气管肺泡灌洗液(BALF)结果显示与对照组比较,哮喘组BALF IL-4、IL-25、IL-33水平显著增高(P<0.05);与哮喘组比较,HSP70/CD80DNA疫苗组和联合疫苗组BALF IL-4、IL-25、IL-33水平显著下降(P<0.05);与HSP70/CD80DNA疫苗组比较,联合疫苗组BALF IL-4、IL-25、IL-33水平显著降低(P<0.05)。结论 CpG ODN联合HSP70/CD80DNA疫苗能抑制小鼠体内产生IL-4、IL-13、IL-25、IL-33, 增强小鼠体内产生IFN-γ,减轻哮喘小鼠气道炎症。

关键词: HSP70/CD80DNA疫苗, CpG ODN, 气道炎症, 哮喘小鼠