流感嗜血杆菌D蛋白黏膜免疫保护效果研究

发布时间：2018-04-05 17:31

本文选题：D蛋白　切入点：流感嗜血杆菌　出处：《免疫学杂志》2016年05期

【摘要】：目的评估D蛋白(protein D,PD)黏膜免疫对流感嗜血杆菌感染的保护效果,研究其免疫应答机制,为发展新疫苗提供依据。方法运用分子克隆技术构建PD蛋白表达质粒;用含有或无霍乱毒素(cholera toxin,CT)的PD蛋白黏膜免疫C57BL/6小鼠,ELISA方法检测血清Ig G、唾液Ig A及脾细胞的细胞因子分泌水平;建立定植和致死性感染模型,观察细菌清除及小鼠生存情况;以裸鼠和μMT小鼠来研究细胞和体液免疫应答在PD免疫保护效果中的作用。结果单独PD蛋白黏膜免疫野生鼠能诱导高滴度的抗PD Ig G和s Ig A,血清中Ig G1、Ig G2a和Ig G2b亚型水平相当,脾细胞分泌IL-17A增加,小鼠鼻咽部流感嗜血杆菌清除增加,致死性感染生存率为82%;裸鼠和μMT小鼠免疫PD后对流感嗜血杆菌的清除效应和致死性感染保护作用受损,PD抗血清被动免疫μMT小鼠后其抗感染能力恢复;PD+CT组抗体滴度及脾细胞分泌IL-4和IL-17A水平均高于PD组,细菌清除效应与PD组相似,但无致死性感染保护能力。结论无佐剂PD蛋白黏膜免疫能有效诱导黏膜应答,抵抗流感嗜血杆菌的定植与致死性感染,为黏膜疫苗的发展提供了可能的选择。
[Abstract]:Objective to evaluate the protective effect of D protein D PDD on Haemophilus influenzae infection, and to study the mechanism of its immune response, and to provide evidence for the development of new vaccine.Methods the expression plasmid of PD protein was constructed by molecular cloning technique, and the levels of serum IgG, saliva IgA and cytokine secretion of spleen cells were detected by Elisa method in C57BL/6 mice immunized with or without cholera toxin cholera toxin.The colonization and lethal infection models were established to observe the clearance of bacteria and the survival of mice, and the role of cellular and humoral immune responses in the immune protection of PD was studied in nude mice and 渭 MT mice.Results the wild mice immunized with PD protein could induce high titer of anti-PD IgG and sIg A, the levels of IgG G2a and IgG2b in serum were the same, the secretion of IL-17A from spleen cells was increased, and the clearance of Haemophilus influenzae in nasopharynx was increased.The survival rate of fatal infection was 82.The scavenging effect of PD on Haemophilus influenzae in nude mice and 渭 MT mice and the protective effect of lethal infection were impaired. The anti-infective ability of mice immunized with PD antiserum was recovered after immunization with 渭 MT mice.The levels of IL-4 and IL-17A secreted by splenocytes were higher in PD group than in PD group.The bacterial scavenging effect was similar to that of PD group, but had no protective effect on fatal infection.Conclusion Mucosal immunity of adjuvant free PD protein can effectively induce mucosal response and resist the colonization and fatal infection of Haemophilus influenzae, which provides a possible choice for the development of mucosal vaccine.
【作者单位】：重庆医科大学附属第一医院输血科;重庆医科大学附属永川医院检验科;重庆医科大学检验医学院临床检验诊断学教育部重点实验室;重庆医科大学附属第一医院检验科;重庆医科大学附属第一医院辅助生殖中心;
【基金】：重庆市医学科研计划(20142004) 重庆市科技计划(cstc2014jcyj A10042)
【分类号】：R392

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