虾类原肌球蛋白的Balb/c小鼠致敏动物模型构建研究  

Establishment of a sensitization animal model based on Balb/c mice for Shrimp Tropomyosin

作　　者：孙佳益[1] 王锡昌[1] 李振兴[2] 卢瑛[1] 

机构地区：[1]上海海洋大学食品学院上海水产品加工及贮藏工程技术研究中心,201306 [2]中国海洋大学食品科学与工程学院,青岛266003

出　　处：《免疫学杂志》2013年第2期161-164,共4页Immunological Journal

基　　金：上海市科技兴农重点攻关项目(沪农科攻字(2009)第6-1号和(2010)第2-6号);上海市教育委员会优秀青年教师基金资助项目(B-8101-09-0036);上海水产品加工及贮藏工程技术研究中心(11DZ2280300)

摘　　要：目的虾类主要过敏原原肌球蛋白(Tropomyosin,Tm)的Balb/c小鼠致敏动物模型的构建及Tm的低过敏性处理效果评价。方法本研究选用Balb/c小鼠,设立常见致敏蛋白卵清蛋白(OVA)组、虾主要过敏原Tm组、经无花果蛋白酶消减处理的Tm(Tm-E)和阴性对照PBS组,将各组样品与氢氧化铝佐剂(3∶1)混合后采用腹腔注射方式以构建Tm致敏动物模型,每次注射时间间隔为1周。第4天采集小鼠血清,通过ELISA评价血清中IgE和IgG抗体的效价变化。结果 OVA和Tm刺激38 d后产生大量IgE和IgG抗体;阴性对照组在整个刺激过程中其血清中不含有IgE和IgG抗体;Tm-E组在致敏24 d、38 d时其IgE的OD值分别较Tm组低77.83%和64.06%(P<0.01)。结论本研究成功构建了虾主要过敏原Tm的致敏动物模型,此外发现无花果蛋白酶解处理方式(酶活与底物比13.6 U/g,35℃酶解10 min)可有效降低Tm的过敏原性。Object of this study is to establish an animal model based on Balb/c mice,sensitization to shrimp major allergen tropomyosin(Tm),and to assess the efficacy of desensitized treatment to Tm.Ovalbumin(OVA),Tm,ficin enzymolyed Tm(Tm-E),and a negative control of PBS were respectively applied as immunogens.Then Balb/c mice were immunized by intraperitoneal injection of emulsions prepared by mixing these immunogens respectively with aluminum hydroxide adjuvant(v/v,3∶1),and the injection was repeated once every other week.Blood samples were collected at the fourth day after every exposure,and specific IgE and IgG antibodies in antiserum were detected using enzyme-linked immunosorbent assay(ELISA).The results showed that OVA and Tm immunogens induced vigorous IgE and IgG antibody responses at the 38th day.In contrast,PBS failed to induce vigorous IgE and IgG production,with no detectable IgE and IgG.The specific IgE of Tm-E group was gradually increased at the 24th and 38th day,however,compared with the Tm group,it was decreased by 77.83% and 64.06%(P < 0.01),respectively.In conclusion,one approach of sensitization animal model for shrimp allergen tropomyosin has been established successfully.Furthermore,it is obviouse that allergenicity of Tm could be reduced by ficin(enzyme activity/ substrate at 13.6 U/g,enzymolysis in 35 ℃,10 min).

关 键 词：虾 过敏原 原肌球蛋白 动物模型 无花果蛋白酶 

分 类 号：R392.8[医药卫生—免疫学]