怀牛膝果寡糖对H1N1流感疫苗佐剂活性及免疫细胞功能的影响  

Adjuvant effect of fructooligosaccharide from Achyranthes bidentata on H1N1 influenza vaccine and immunocyte function

作　　者：汪艳艳[1,2] 李海霞[2] 巫亚俊 赵修南[2] 麻浩[2] 刘坤璐[2] 武军华[2] 单俊杰[2] 王玉霞[2] 王海南[1,3] 

机构地区：[1]贵州大学药学院,贵州贵阳550025 [2]军事医学科学院毒物药物研究所,北京100850 [3]国家食品药品监督管理总局,北京100053

出　　处：《中国药理学与毒理学杂志》2016年第2期113-121,共9页Chinese Journal of Pharmacology and Toxicology

基　　金：国家科技支撑计划课题(2011BAD26B02-3)~~

摘　　要：目的研究牛膝果寡糖ABP-50-FOS对H1N1流感疫苗的佐剂活性及免疫细胞功能的影响。方法采用凝胶渗透色谱、毛细管电泳、红外光谱和核磁共振等方法研究ABP-50-FOS的理化性质;BALB/c小鼠肌肉注射H1N1流感疫苗3μg和ABP-50-FOS 200μg初次免疫,28 d后加强免疫。分别于初次和再次免疫后14 d用ELISA法检测小鼠血清特异性Ig G,Ig G1,Ig G2a,Ig G2b,Ig G3和Ig M抗体水平;流式细胞仪测定脾细胞中CD3+,CD19+,CD4+和CD8+淋巴细胞亚群百分率;MTT法测定小鼠脾细胞增殖活性;ELISA法测定脾细胞培养上清干扰素γ(IFN-γ)、胸腺细胞培养上清白细胞介素4(IL-4)、腹腔巨噬细胞培养上清肿瘤坏死因子α(TNF-α)和一氧化氮及小鼠骨髓细胞和DC2.4树突状细胞培养上清IL-12p70含量。结果ABP-50-FOS是一种果寡糖,峰高分子质量为1885 u,骨架结构为1,2-连接和1,6-连接的果糖。该寡糖能明显升高H1N1流感疫苗免疫小鼠血清特异性Ig G,Ig G2a和Ig M抗体水平(P<0.01),提高免疫小鼠脾CD3+,CD4+和CD8+T淋巴细胞亚群的百分率。体外实验表明,ABP-50-FOS能促进小鼠脾细胞增殖反应和IFN-γ分泌(P<0.01),促进胸腺细胞IL-4分泌(P<0.01),刺激腹腔巨噬细胞分泌TNF-α(P<0.01)和DC2.4树突状细胞分泌IL-12p70(P<0.01),对巨噬细胞一氧化氮的分泌有抑制作用。结论牛膝果寡糖ABP-50-FOS对H1N1流感疫苗具有良好的佐剂活性,体外对小鼠免疫细胞功能具有促进作用。OBJECTIVE To investigate chemical properties of a fructooligosaccharide （ABP-50-FOS） separated from Achyranthes bidentata and immune response in mice immunized H1N1 influenza vaccine.METHODS The methods of GPC,CE,IR and NMR were used to study chemical properties of ABP-50-FOS.BALB/c mice were immunized intramuscularly twice with H1N1 influenza vaccine （3 μg） plus ABP-50-FOS （200 μg） each mouse.The serum total antibody titer and its isotypes titers were analyzed by ELISA.The populations of CD4^＋,CD8^＋,CD3^＋and CD19^＋lymphocytes were deter-mined by flow cytometry.The proliferation activities of spleen T and B lymphocytes were determined with MTT method.The levels of cytokines interferon-γ（IFN-γ）,tumor necrosis factor-α（TNF-α）,inter-leukin-4（IL-4）,IL-12 and NO were measured by ELISA kits.RESULTS ABP-50-FOS was a fructooli-gosaccharide with moleculer mass 1885 u.Its bone linkages contained 1,2-and 1,6-fructose residues.ABP-50-FOS could induce high specific-IgG,IgG1,IgG2a,IgG2b and IgM titers after immunization with H1N1 influenza antigen twice （P〈0.01）.ABP-50-FOS significantly elevated the percentage of CD3^＋,CD4^＋and CD8^＋spleen lymphocytes and IFN-γ secretions （P〈0.01） in vitro.It also stimulated peritoneal macrophage of mice and DC2.4 dendritic cells to produce TNF-α and IL-12p70 respectively （P〈0.01）.However,ABP-50-FOS inhibited secretions of NO in macrophage.CONCLUSION The fruc-tooligosaccharide ABP-50-FOS separated from A.bidentata can exhibit strong adjuvant activity for H1N1 influenza vaccine.

关 键 词：牛膝 多糖 果寡糖 疫苗 佐剂 

分 类 号：R392[医药卫生—免疫学]