SiO_(2)-Pickering乳液佐剂增强沙眼衣原体Pgp3蛋白免疫效果  

Effect of SiO_(2)-Pickering emulsion adjuvant on enhancing Pgp3 protein immunity of Chlamydia trachomatis

作　　者：史可靓 方春霞 赵兰华 周辉[2] 李忠玉 SHI Keliang;FANG Chunxia;ZHAO Lanhua;ZHOU Hui;LI Zhongyu(Pathogenic Biology Institute,Hunan Provincial Key Laboratory for Special Pathogens Prevention and Control,Hengyang Medical School,University of South China,Hengyang 421001,Hunan,China;The First Hospital of Hunan University of Chinese Medicine,Changsha 410007,Hunan,China)

机构地区：[1]南华大学衡阳医学院病原生物学研究所特殊病原体防控湖南省重点实验室,湖南衡阳421001 [2]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《中南医学科学杂志》2023年第3期319-324,共6页Medical Science Journal of Central South China

基　　金：国家自然科学基金(82272383、32070189);湖南省自然科学基金(2021JJ30594);湖南省教育厅项目(20A421、20C1390);湖南省卫健委项目(202111000475)。

摘　　要：目的优化二氧化硅(SiO_(2))Pickering(SPE)乳液佐剂,研究其对沙眼衣原体(Ct)Pgp3蛋白的免疫增强效应,为预防Ct感染性疾病提供实验依据。方法优化SiO_(2)质量浓度、水油比及超声功率,超声乳化制备SPE佐剂。30只小鼠均分为SPE+Pgp3组(Pgp3蛋白和SPE混合免疫)、Pgp3组(Pgp3蛋白单独免疫)和PBS组。ELISA检测各组小鼠血清抗体水平和脾细胞上清细胞因子水平,流式细胞术检测脾细胞γ干扰素(IFN-γ)、白细胞介素-4(IL-4)水平。结果SPE最优制备条件为超声功率337 W、水油比10∶2、SiO_(2)质量浓度1 g/L,最优条件下SPE直径为(300.12±44.26)nm,聚合物分散性指数为0.33±0.12。除PBS组外,其他组均检测到了特异性抗体,SPE+Pgp3组总抗体IgG及其亚类IgG1、IgG2a抗体水平高于Pgp3组,且以IgG2a升高为主(P<0.05)。SPE+Pgp3组、Pgp3组脾细胞IFN-γ和刺激指数高于PBS组,且SPE+Pgp3组高于Pgp3组(P<0.05);IFN-γ主要由CD4^(+)T细胞和CD8^(+)T细胞分泌。各组IL-4含量均较低。结论SPE乳液佐剂具有良好的佐剂效应,能有效增强Pgp3蛋白体液免疫和细胞免疫,且可优势诱导Th1型免疫反应。Aim To optimize the emulsion adjuvant of silica(SiO_(2))Pickering(SPE)and study its immune-enhancing effect on Pgp3 protein of Chlamydia trachomatis(Ct),providing experimental basis for the prevention of Ct infectious diseases.Methods SPE adjuvant was prepared by phacoemulsification with SiO_(2) concentration,water-oil ratio and ultrasonic power optimized.All 30 mice were divided into SPE+Pgp3 group(Pgp3 protein and SPE mixed immunization),Pgp3 group(Pgp3 protein alone immunization)and PBS group.Serum antibody levels and splenocyte supernatant cytokine levels were detected by ELISA,and spleen cells interferon-γ(IFN-γ)and interleukin-4(IL-4)levels were detected by flow cytometry.Results The optimal preparation conditions of SPE were ultrasonic power 337 W,water-oil ratio 10∶2,SiO_(2) concentration 1 g/L,SPE size diameter(300.12±44.26)nm,polymer dispersion index 0.33±0.12.Specific antibodies were detected in all groups except PBS group.The levels of total IgG and its subclasses IgG1 and IgG2a in SPE+Pgp3 group were higher than those in Pgp3 group,and IgG2a was mainly increased(P0.05).The IFN-γand stimulation indices of spleen cells in SPE+Pgp3 and Pgp3 groups were higher than those in PBS group,and SPE+Pgp3 group was higher than those in Pgp3 group(P0.05);IFN-γis mainly secreted by CD4^(+)T cells and CD8^(+)T cells.The content of IL-4 in each group was low.Conclusion SPE emulsion adjuvant had a good adjuvant effect,which effectively enhanced the humoral and cellular immunity induced by Pgp3 protein in mice,and the immune response induced by SPE+Pgp3 vaccine preparation tended to be Th1 type.

关 键 词：SPE乳液佐剂 沙眼衣原体 Pgp3蛋白 免疫反应 

分 类 号：R374.1[医药卫生—病原生物学]