日本血吸虫可溶性虫卵抗原诱导BIO细胞产生的研究

目的观察日本血吸虫可溶性虫卵抗原（SEA）与成虫抗原（SwAP）是否能够诱导小鼠调节性B细胞的产生。方法SEA、SWAP和脂多糖（LPS）分别体外刺激小鼠脾脏单个核细胞和磁珠分选后的脾脏CD19＋B细胞，72h后用流式细胞术分别检测CD19＋IL-10＋细胞比例，和细胞表面CD80、CD86及CD40的表达，用ELISA法检测培养上清中IL-10的水平。体内实验用SEA、SWAP、PBS分别和不完全弗氏佐剂混合免疫小鼠，1次／N，共3次，末次免疫后7d取小鼠脾脏，流式细胞术检测CD19＋IL-10＋细胞比例，用ELIsA法检测培养上清中IL-10的水平。结果LPS与SEA体外均能显著诱导小鼠脾脏单个核细胞分化为CD19＋IL-10＋细胞，两者无显著差异，而swAP组不能诱导小鼠脾脏单个核细胞分化为CD19＋II-10＋。细胞；SEA与LPS能够诱导脾脏CD19＋B细胞表面高表达CD80、CD86和CD40，而SwAP不能诱导脾脏CD19＋B细胞活化；SEA与LPS诱导脾脏CDt9＋B细胞分化为IL-10＋细胞的比例也显著升高，同时能刺激脾脏CD19＋B细胞分泌高水平的IL-10，两者无显著差异；而SwAP不能诱导小鼠脾脏CD19＋。B细胞分化为IL—10＋细胞并分泌IL-10。SEA体内免疫小鼠后，小鼠脾脏单个核细胞分化为CD19＋IL-10＋细胞比例显著升高，并且能够分泌高水平的IL-10，而swAP和PBS免疫组均不能诱导小鼠脾脏单个核细胞分化为CD-9＋IL-10＋细胞并分泌1L-10。结论SEA体外体内均能诱导小鼠产生分泌IL-10的B10细胞，并且在体外不依赖于脾脏其他免疫细胞的参与，而SwAP体外体内均不能诱导B10细胞的产生。

BIO cells production induced by Schistosoma japonicum soluble egg antigens

Induction of the production of regulatory B cells by the soluble egg antigen （SEA） and adult worm antigen （SWAP） was studied in the research. Mouse spleen mononuclear cells and CD19＋ B cells sorted by beads were stimulated by SEA, SWAP and LPS, respectively. And the CD19＋ IL-10＋ cells ratio and expression of CD80, CD86 and CD40 were analyzed by flow cytometry after 72 hour. At the same time, the cytokines IL-10 lever in cultured supernatants was detected by ELISA. Mouse was immunized with the mixture of SEA or SWAP or PBS and incomplete Freund＇s adjuvant for three times, respective- ly. The mouse spleen mononuclear cells were isolated, and the CD19＋ IL-10＋ cells ratio and cytokines IL-10 lever in culture su- pernatants were analyzed on day 7 after last immunization. Results showed that the CD19＋ IL-10＋ cells ratio in spleen mononu clear cells was significantly increased in groups stimulated by SEA and LPS, comparing to the SWAP stimulated group （P〈 0.01）. Furthermore, IL-10＋ B cells ratio and expression of CD80, CD86, and CD40 in spleen CD19＋ B cells stimulated by SEA and LPS also significantly increased, instead of SWAP stimulation （P〈0.01）. The cytokines IL-10 levers in CD19＋ B cells cul- ture supernatants stimulated with SEA and LPS were significantly higher than those in SWAP stimulated group. The CD19＋ IL-t0＋ cells ratio and the cytokines IL-10 levers were significantly increased in SEA immunized group compared with the groups of SWAP and PBS immunization （P〈0.05）. In conclusion, instead of SWAP, SEA could induce the production and activation of B10 cells secreted IL-10 in vivo and in vitro, being independent of other immune cells effect in vitro.