诱导C57鼠获得抗β淀粉样蛋白抗体的新途径

New Way to Getting Anti-amyloid beta Antibody in C57 Mice

目的探讨诱导C57鼠产生β淀粉样蛋白抗体的新途径。方法收获小鼠骨髓细胞,定向培养为树突状细胞(DCs),培养期间加用突变型β淀粉样蛋白1-42多肽刺激致敏。流式细胞仪测定收获的DCs成熟度。将致敏DCs作为疫苗接种至C57鼠,以β淀粉样蛋白+氟氏佐剂接种作为阳性对照,用间接ELISA法测定小鼠的抗β淀粉样蛋白抗体滴度。结果用突变型β淀粉样蛋白刺激DCs可促进其表面CD11c、CD86、MHC-Ⅱ及CD80分子表达从54.69%、76.68%、44.77%和58.65%分别提高到77.49%、92.76%、85.52%和75.57%。接种DCs后14d抗β淀粉样蛋白抗体滴度开始逐渐升高,一直持续到免疫后35d仍未见降低。相对于佐剂疫苗7d显著升高、21d达峰的变化曲线有所平缓和滞后。DCs疫苗免疫后平均抗体滴度最高达到2425.17倍,但比佐剂疫苗组要低(平均最高3200倍)。结论采用DCs疫苗能有效地诱导C57鼠产生足够的抗β淀粉样蛋白抗体。

Objective To avoid severe adverse effects brought by amyloid beta vaccine with adju-vant,a new Alzheimer vaccine was developed from dendritic cells stimulated with mutanted amyloid beta 1-42 peptide and then inoculated to C57 mice. The efficacy of inducing anti-amyloid beta antibody was observed. Methods Bone marrow cells harvested from mouse femur was cultured committed to dendritic cells（DCs）. During the culture,mutanted amyloid beta 1-42 peptide was added to stimulate and sensitize DCs. Maturity of harvested DCs was assessed by means of flow cytometry. Then the sensitized DCs was inoculated into C57 mice as amyloid beta vaccine. Amyloid beta with Freund＇s adjuvant was inoculated at the same time to act as positive control. The anti-amyloid beta titre of mouse was measured by indirective ELISA. Results Expression of CD11c,CD86,MHC-Ⅱand CD80 on the surface of amyloid beta stimulating DCs was increased respectively from 54.69%,76.68%,44.77% and 58.65% up to 77.49%,92.76%,85.52% and 75.57%. Anti-amyloid beta antibody titre elevated significantly from 14 days after DCs vaccine inoculation,and the elevated antibody titre retained to 35 days after inoculation without reduction. This curve was gentle in a hysterotely way than that of adjuvant vaccine,which was elevated significantly 7 days and peaked 21 days after inoculation. The peak of geographic mean anti-body titre was 2 425.17,which was lower than that of vaccine with adjuvant （the peak value was 3200）. Conculsion DCs vaccine can induce adequate anti-amyloid beta antibodies in C57 mice efficiently.