巨噬细胞源性趋化因子对NTHiP6蛋白疫苗免疫效果的影响  

The immunological effect of MDC protein on P6 protein vaccine of nontypeable Haemophilus Influenzae

作　　者：韩小艳[1] 张彦霞[1] 杨新玲[1] 李小俊[1] 王晨红[1] 张玉妥[1] 

机构地区：[1]河北北方学院医学检验学院,张家口075000

出　　处：《现代免疫学》2015年第1期32-35,共4页Current Immunology

基　　金：河北省教育厅重点课题(Z2011208);张家口市科技项目(1321062D);河北北方学院青年基金(Q2010028)

摘　　要：观察巨噬细胞源性趋化因子(MDC)佐剂对NTHiP6蛋白疫苗免疫效果的影响。将原核表达质粒PGEX-6P2/P6转入E.coli XL1-Blue,IPTG诱导P6蛋白的表达并进行纯化。将BALB/c小鼠随机分为A-D四组,分别为PBS对照组、MDC对照组、P6蛋白组、P6蛋白联合MDC组。分别于0、14、28d经黏膜免疫,末次免疫后14d,每组12只小鼠取血和肺泡灌洗液,ELISA检测血清中IgG抗体和肺泡灌洗液中IgA抗体水平。每组取3只小鼠,制备脾淋巴细胞,ELISA检测IL-4、IL-17和IFN-γ水平。用10LD50NTHi攻击每组剩余15只小鼠,观察免疫保护作用。在大肠杆菌中成功表达P6蛋白。第三次免疫后,D组诱导的IgG抗体、IgA抗体、IL-4、IL-17和IFN-γ水平显著高于其他各组(P<0.05)。经NTHi攻击后,D组生存率达80%,与A组、B组相比差异有统计学意义(P<0.05),但C组、D组之间无显著性差异(P>0.05)。MDC作为佐剂可以使NTHiP6疫苗获得较好的免疫效果。The purpose of this study was to observe immune effect of macrophage-derived chemokine （MDC） on nontypeable Hae- mophilus Influenzae （NTHi）-P6 protein vaccine. The recombinant plasmid PGEX-6P2/P6 was transformed into E. coli XL1- Blue. The P6 protein was induced and expressed in the expression system. The P6 protein was purified by Glutathione Sepha- roseTM 413 affinity chromatography. P, ALB/e mice were randomized into four groups that were respectively inoculated intranasally with PBS, MDC,P6 protein and P6 protein ＋ MDC at 0, 14 and 28 days. At day 14, sera and bronchoalveolar lavage fluids were collected from 12 mice of each group, The levels of IgG and IgA antibodies were detected by ELISA. Splenocytes were i- solated from 3 mice of each group for determining the levels of IL 4, IL 17 and IFN-~/ by ELISA. Furthermore, the other 15 mice of each group were challenged with 10LD50 of Haemophilus Influenzae ATCC49247 to observe the protective effect of the vaccine. The P6 protein was successfully expressed and purified from E. coli XL1-Blue cells. After the third immunization, P6 protein ＋ MDC induced high levels of IgG and IgA antibodies as well as IL-4, IL-17 and IFN-γ （P〈0.05）. After chal- lenged with 10LD50 of NTHi ATCC49247, the survival rate of mice in P6 protein ＋ MDC group was 80%, significantly higher than those in the control groups（P〈0.05）, but no statistical difference was noted when compared with the P6 protein group. So, MDC protein could enhance the immune protective effect of NTHi-P6 protein vaccine.

关 键 词：不可分型流感嗜血杆菌 P6蛋白 巨噬细胞源性趋化因子 黏膜免疫 

分 类 号：R392.11[医药卫生—免疫学]