MIP-1α基因佐剂对NTHi P6 DNA疫苗免疫效果的影响  

Effect of a mouse MIP-1α gene adjuvant on enhancement of the immunogenicity of a DNA vaccine encoding the NTHi P6 protein

作　　者：韩小艳[1] 王晨红[1] 张玉妥[1] 

机构地区：[1]河北北方学院微生物教研室,河北张家口075000

出　　处：《中国病原生物学杂志》2015年第6期512-515,共4页Journal of Pathogen Biology

基　　金：张家口市科技项目(No.1321062D)

摘　　要：目的观察小鼠巨噬细胞炎性蛋白1α(macrophage inflammatory protein-1α,MIP-1α)基因佐剂对不可分型流感嗜血杆菌(nontypeable Haemophilus influenzae,NTHi)外膜蛋白P6(outer membrane protein P6,P6)DNA疫苗免疫效果的影响。方法将92只BALB/c小鼠随机分为4组,分别用pcDNA3.1、pcDNA3.1/MIP-1α、pcDNA3.1/P6和pcDNA3.1/MIP-1α+pcDNA3.1/P6,肌肉注射免疫,每2周免疫1次,共3次,质粒每次每只接种100μg。末次免疫后14d,每组取10只小鼠,采血,分离血清,采用ELISA法检测特异性IgG抗体;每组取3只小鼠,制备脾淋巴细胞,采用CCK-8法检测脾淋巴细胞增殖活性,采用ELISA法检测小鼠脾淋巴细胞分泌IL-4和IFN-γ水平。用8LD50NTHi攻击每组剩余10只小鼠,观察免疫保护作用。结果末次免疫后,pcDNA3.1/MIP-1α+pcDNA3.1/P6组小鼠脾淋巴细胞分泌IFN-γ水平为(1726.45±184.48)ng/L,增殖指数2.06±0.04,与pcDNA3.1/P6组(1443.09±187.70)ng/L和1.90±0.04比较差异均有统计学意义(P<0.05);抗体滴度和IL-4水平两组间差异无统计学意义(P>0.05)。经NTHi攻击后,pcDNA3.1/MIP-1α+pcDNA3.1/P6免疫组小鼠14d生存率达70%,与pcDNA3.1组0%和pcDNA3.1/MIP-1α组0%相比与差异有统计学意义(P<0.05),与pcDNA3.1/P6组60%相比差异无统计学意义(P>0.05)。结论MIP-1α基因佐剂在一定程度上能增强NTHi P6基因疫苗的免疫保护作用。Objective To observe the effects of a genetic adjuvant, MIP-1α, on a DNA vaccine encoding the P6 outer membrane protein of nontypeable Haemophilus inf[uenzae （NTHi）. Methods Ninety-two BALB/e mice were randomly divided into four groups that were injected 3 times in the quadriceps with pcDNA3. 1, pcDNA3. 1/MIP-1α, pcDNA3. 1/ P6, or pcDNA3.1/MIP -1α＋pcDNA3, l/P6 at 2-week intervals. The dose of plasmid administered was 100μg. Sera were collected from 10 mice in each group 14 days after the final injection and IgG antibodies were detected with ELISA. Splenocytes from 3 mice in each group were separated and cultured. IFN-γ and IL-4 were detected with ELISA. The pro- liferation of spleen lymphocytes was analyzed using CCK-8. Another 10 mice in each group were challenged with 8 times the LDS0 of tlaemophilus influenzae ATCC49247 to observe the protective effect of the vaccine. Results After the final immunization, the mice vaccinated with pcDNA3.1/MIP la＋pcDNA3, 1/P6 had a higher spleen lymphocyte stimulation index （2.06±0.04） and higher level of IFN-γ （1726.45±184.48 ng/L） than did mice vaccinated with pcDNA3, l/P6 （1.90±0.04, 1443.09±187.70 ng/L） （P〈0.05）. There was no difference between the two groups in terms of IL-4 levels and IgG antibody titers. After the challenge with 8 times the LD50 of H. influenzae ATCC49247, mice vaccinated with peDNA3.1/MIP-1α＋pcDNA3, 1/P6 had a survival rate of 70%, which was significantly higher than that of mice vaccinated with pcDNA3.1 （0%） or pcDNA3.1/MIP-1α （0%） （P〈0.05）, but there was no statistical difference in the survival rate in comparison to mice vaccinated with pcDNA3. 1/P6 （60%） （P〈0. 05）. Conclusion MIP-la enhanced the immunity conierred by a DNA vaccine encoding the NTHi P6 protein.

关 键 词：不可分型流感嗜血杆菌 核酸疫苗 巨噬细胞炎性蛋白1Α 小鼠 

分 类 号：R378.4[医药卫生—病原生物学]