摘要
目的研究茯苓总多糖（PCP）对人用H1N1流感疫苗抗原和乙肝疫苗抗原的佐剂作用。方法在50℃条件下,采用水提、醇沉、透析和冷冻干燥方法制备PCP。苯酚-硫酸法测定总多糖含量,凝胶渗透色谱法测定多糖相对分子质量分布,毛细管电泳法测定单糖组成。H1N1流感病毒裂解液为抗原（每鼠3μg）,与PCP（每鼠0.2或1.0 mg）联用,肌内注射免疫小鼠1次,免疫后14 d采用ELISA法测定小鼠血清特异性抗体Ig G滴度。乙肝表面抗原（HBs Ag）蛋白为抗原（每鼠2μg）,与PCP（每鼠1.0 mg）联用,肌内注射免疫小鼠2次,于第2次免疫后14,21,28和35 d,采用ELISA法测定小鼠血清特异性抗体Ig G滴度。结果PCP中多糖含量为40.8%,由岩藻糖、甘露糖、葡萄糖和半乳糖组成,摩尔比为1.00∶1.36∶0.48∶2.67。PCP与H1N1流感抗原联用免疫小鼠1次,能明显提高小鼠抗原特异性抗体Ig G滴度（P〈0.05）,效果与铝佐剂相当。与单用HBs Ag抗原组相比,PCP与HBs Ag联用初次免疫即能显著提高小鼠血清抗原特异性抗体滴度（P〈0.01）,且与铝佐剂组无显著性差异;二次免疫后14~21 d,PCP与HBs Ag联用组小鼠抗体水平进一步升高,且高于铝佐剂组（P〈0.05）,28~35 d PCP与HBs Ag联用组小鼠抗体滴度与铝佐剂组相当。结论PCP对人用H1N1流感疫苗和HBs Ag疫苗具有良好的佐剂活性,该多糖能显著提高小鼠体液免疫功能。
OBJECTIVE To investigate the adjuvant activity of total polysaccharides from Poria cocos（ PCP） with human H1N1 influenza antigen and HBs Ag in mice. METHODS PCP was obtained successively via extraction with water at 50℃,ethanol precipitation and dialysis. The sugar content was determined by phenol-sulfuric acid method,monosaccharide compositions were analyzed with the alditol acetates by gas chromatography,and molecular mass of PCP was determined with gel filtration chromatography. BALB / c mice were co-immunized intramuscularly one time with PCP（ 0. 2 or 1. 0 mg per mouse） plus H1N1 influenza antigen（ 3 μg per mouse）. HBs Ag（ 2 μg per mouse） plus PCP（ 1. 0 mg per mouse） was injected intramuscularly to immunize mice twice,and the serum specific antibody titers were determined at 14,21,28 and 35 d after the second immunization. The titers of serum antigenspecific antibody of mice were assayed using ELISA method. RESULTS The sugar content of PCP was40. 8%. PCP was composed of fucose,mannose,glucose and galactose,the molar ratio of which was1. 00∶ 1. 36∶ 0. 48∶ 2. 67. After immunization with H1N1 influenza antigen combined with PCP one time for two weeks,PCP could rapidly induce the production of serum antibody Ig G against H1N1 influenza antigen in mice compared with H1N1 influenza antigen alone group（ P 0. 05）. After the first immunization with HBs Ag combined with PCP for 14 d,PCP could increase the production of serum antibody Ig G against HBs Ag compared with HBs Ag alone group（ P 0. 05）. After the second immunization with HBs Ag combined with PCP for 14- 21 d,the adjuvant effect of PCP was stronger than that of alum adjuvant（ P 0. 01）. CONCLUSION PCP exhibits good adjuvant effect on human H1N1 influenza and HBs Ag.PCP significantly increases the humoral immunity response of mice.
出处
《中国药理学与毒理学杂志》
CAS
CSCD
北大核心
2015年第1期60-64,共5页
Chinese Journal of Pharmacology and Toxicology
基金
国家科技重大专项(2012ZX09301-001-003)
国家科技重大专项(2013ZX09402103)
国家科技支撑计划课题(2011BAD26B02-3)~~
关键词
茯苓
多糖
佐剂
疫苗
抗原
Poria cocos
polysaccharides
adjuvant
vaccine
antigen
