日本血吸虫虫源性抗原诱导效应性B细胞分化的研究

目的 目的 观察日本血吸虫可溶性虫卵抗原 （SEA） 和可溶性成虫抗原 （SWAP） 诱导小鼠效应性B细胞分化情况。方 方 法 法 SEA、 SWAP和脂多糖 （LPS） 分别体外刺激小鼠脾脏单个核细胞和脾脏CD19+ B细胞， 72 h后采用流式细胞术检测 CD19+ IL?6+ 及CD19+ IFN?γ+ 细胞比例； 用酶联免疫吸附试验 （ELISA） 检测抗原刺激后脾脏B细胞培养上清中IL?6和IFN?γ 水平。用SEA、 SWAP、 PBS分别和不完全弗氏佐剂混合免疫小鼠， 每周1次， 共3次， 末次免疫后7 d取小鼠脾脏， 流式细胞 术检测CD19+ IL?6+ 及CD19+ IFN?γ+ 细胞比例， 用ELISA法检测培养上清中IL?6和IFN?γ水平。 结果 结果 SEA与LPS可以体 外诱导脾脏单个核细胞和脾脏B细胞分化为CD19+ IL?6+ B细胞 （F = 5.099， P < 0.05； F = 6.951， P < 0.05）， 并刺激脾脏B细 胞分泌IL?6 （F = 55.184， P < 0.01）； SEA免疫小鼠后， 可以在体内刺激小鼠脾脏B细胞分泌IL?6 （F = 19.244， P < 0.01）， 并诱 导小鼠脾脏单个核细胞分化为CD19+ IL?6+ B细胞 （F = 14.904， P < 0.05）。SWAP可以体外诱导脾脏单个核细胞分化为 CD19+ IFN?γ+ B细胞 （F = 3.277， P < 0.05）， 但不能单独诱导脾脏B细胞分化为IFN?γ+ B细胞， 也不能刺激脾脏B细胞分泌 IFN?γ； SWAP免疫小鼠后， 可以在体内刺激小鼠脾脏B细胞分泌IFN?γ （F = 31.886， P < 0.01）， 并诱导其分化为CD19+ IFN? γ+ B细胞 （F = 49.873， P < 0.01）。 结论 结论 日本血吸虫SEA可以优势诱导小鼠脾脏单个核细胞分化为CD19+ IL?6+ B细胞， 而 SWAP可以优势诱导小鼠脾脏单个核细胞分化为CD19+ IFN?γ+ B细胞， 但依赖于其他免疫细胞的参与。

Effects of soluble egg antigen and adult worm antigen of Schistosoma japoni- cum on differentiation of effector B cells of mice

Objective To observe the differentiation ability of effector B cells induced by soluble egg antigen （SEA） and sol- uble adult worm antigen （SWAP） of Schistosoma japonicum. Methods The mouse spleen mononuclear cells and CD19＋ B cells sorted by beads were stimulated by SEA, SWAP and lipopolysaccharide （LPS）, respectively. The ratios of CD19＋ IL-6＋ cells and CD19＋IFN-＇,/＋ cells were analyzed by flow cytometry after 72 hours. At the same time, the cytokines IL-6 and IFN-＋ levels in the cultured supernatants were detected by ELISA. The mouse was immunized with the mixture of SEA or SWAP or LPS and the incom- plete Freund＇ s adjuvant for three times, respectively. The mouse spleen mononuclear cells were isolated at the seventh day after the last immunization. The ratios of CD 19＋IL-6＋ cells and CD 19＋ IFN-y＋cells were analyzed, and the cytokines IL-6 and IFN-＋ lev- els in the culture supernatants were detected. Results The ratio of CD19＋ IL-6＋ cells in spleen mononuclear cells and splenic B cells was significantly increased in the groups stimulated by SEA and LPS （P 〈 0.05）, and the cytokines IL-6 level in the CD19y＋B cells culture supernatants were significantly increased （P 〈 0.01 ）. Furthermore, the ratio of CD 19＋ IL-6＋ cells and the cytokines IL -6 level were significantly increased in the SEA immunized group （P 〈 0.01）. SWAP could induce a significantly higher ratio of the CD19＋ IFN-y＋ ceils in spleen cells, instead of in splenic CD19＋ B cells （P 〈 0.05）. The CD19＋ IFN-y＋ cells and the cytokine IFN-y＋level in the culture supernatants in the SWAP immunized group were significantly higher than those in the SEA and PBS im- munized groups （P 〈 0.01）. Conclusions SEA preferentially induces the increase of CD19＋ IL-6＋ ceils in mouse spleen cells; while SWAP preferentially induces the CD19 ＋IFN-y＋ cells＇ production of mouse spleen cells, depending on the effects of other im-mune cells.