【摘要】 目的 :探讨Aβ4 2 及其亚单位肽疫苗接种小鼠后特异性抗Aβ4 2抗体的产生情况。方法 :75只 6wk龄雄性BALB/c小鼠 ,随机分为 5组 :即对照组、Aβ4 2 组 ,Aβ3 6 - 42 组、Aβ1- 15 组和FAβ1- 15组。分别用PBS +MF59佐剂 ,Aβ4 2 +MF59,Aβ36～ 4 2 +七聚赖氨酸 (MAP) +MF59,Aβ1- 15 +MAP +MF59,Aβ1- 15 +MAP +福氏佐剂 ,免疫BALB/c小鼠 4次。用间接ELISA法 ,检测各组免疫小鼠血清和脑组织匀浆上清液中特异性抗体的滴度。将Aβ42 、Aβ3 6 - 42 和Aβ1- 15 与培养的PC12细胞共同培养 7d ,用MTT比色法检测 3种抗原肽对PC12细胞的毒性。将各组免疫小鼠的血清加入到含 2 0mg/LAβ4 2 的培养基中 ,再与培养的PC12细胞一起培养 7d ,用MTT比色法测定PC12细胞的存活率。结果 :第 2次免疫后 ,各实验组小鼠的免疫血清均有抗Aβ42 抗体产生 ,且抗体滴度随接种次数的增多而增高。同时 ,在脑组织匀浆上清液中也可检测出低滴度的抗Aβ4 2 抗体。Aβ42 可降低PC12细胞的存活率 ;而不同浓度的Aβ3 6 - 42 和Aβ1- 15 则对PC12细胞的存活率无显著影响。将 4组疫苗免疫血清和 2 0mg/LAβ42 同时加入培养基中培养PC12细胞时 ,可显著提高其存活率。结论 :Aβ42 及其亚单位疫苗 (Aβ1- 15 及Aβ36～ 4 2 )结合MF59佐剂免疫B更多还原

【Abstract】 AIM: To explore the production of anti-Aβ 42 antibody after immun ization with Aβ 42 and its subunit peptide vaccines. METHODS: Seventy five male BALB/c with the age of 6 weeks were randomly divided into 5 groups, namely, control group, Aβ 42 group, Aβ 36～42 group, Aβ 1-15 group and F Aβ 1-15 group. The BALB/c mice were immunized four t imes with PBS+MF59 adjuvant, Aβ 42 +MF59, Aβ 36-42 +heptalysine (MAP)+ MF59, Aβ 1-15 +MAP+MF59 and Aβ 1-15 +MAP+Freud’s adjuvant, respective ly. The titers of specific antibodies in sera and supernatants of brain tissue h omogenates from the immunized mice in every group were detected by indirect ELIS A. After Aβ 42 , Aβ 36-42 , Aβ 1-15 and F Aβ 1-15 were co -cultured together with cultured PC12 cells for 7 days, the cytotoxicity of th e 3 antigen peptides to PC12 cells were determined by MTT colorimetry. In additi on, after immune sera from each group were added to culture medium containing 2 0 mg/L Aβ 42 and co-cultured with PC12 cells for 7 days, the survival rat e of PC12 cells were examined by MTT assay. RESULTS: The product ion of anti-Aβ 42 antibodies was detected in sera of each experimental gr oup after the second time immunization, and the titer of antibody rose with the increase of immunizing times. In addition, the anti-Aβ 42 antibody with low titer was also detected in supernatants of brain tissue homogenates. The Aβ 42 could reduce the survival rate of PC12 cells, whereas Aβ 36～42 a n d Aβ 1-15 had no obvious effect on survival rate of PC12 cells. After immu ne sera from 4 experimental groups and Aβ 42 were co-cultured with PC 12 cells, their survival rate was found improved. CONCLUSION: C ombination of Aβ 42 and its subunit (Aβ 36～42 and Aβ 1-15 ) v a ccines with MF59 adjuvant can induce BALB/c mice to produce anti-Aβ 42 an tibody. The antibody may neutralize the cytotoxicity of Aβ 42 .更多还原