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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+IFNγ+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IFNγ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+IFNγ+细胞比例,用ELISA法检测培养上清中IL 6和IFNγ水平。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+IFNγ+B细胞(F=3.277,P<0.05),但不能单独诱导脾脏B细胞分化为IFNγ+B细胞,也不能刺激脾脏B细胞分泌IFNγ;SWAP免疫小鼠后,可以在体内刺激小鼠脾脏B细胞分泌IFNγ(F=31.886,P<0.01),并诱导其分化为CD19+IFNγ+B细胞(F=49.873,P<0.01)。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+IFNγ+B细胞,但依赖于其他免疫细胞的参与。
Objective To observe the differentiation of effector B cells induced by soluble antigen of swine schistosoma japonicum (SEA) and soluble adult worm antigen (SWAP). Methods The spleen mononuclear cells and spleen CD19 + B cells were stimulated with SEA, SWAP and lipopolysaccharide (LPS) in vitro respectively. The proportion of CD19 + IL 6+ and CD19 + IFNγ + cells was measured by flow cytometry 72 h later. The levels of IL-6 and IFN-γ in the supernatant of splenic B cells after antigen stimulation were detected by ELISA. The mice were immunized with SEA, SWAP, PBS and incomplete Freund’s adjuvant respectively once a week for 3 times, and the spleens of mice were taken 7 days after the last immunization. The levels of CD19 + IL 6+ and CD19 were detected by flow cytometry + IFNγ + cell ratio, IL 6 and IFNγ levels in culture supernatants were detected by ELISA. Results SEA and LPS induced spleen mononuclear cells and splenic B cells to differentiate into CD19 + IL 6 + B cells in vitro (F = 5.099, P <0.05; F = 6.951, (F = 55.184, P <0.01). The SEA-immunized mice could stimulate the splenic B cells to secrete IL-6 (F = 19.244, P <0.01) in vivo and induce the differentiation of mouse spleen mononuclear cells into CD19 + IL 6 + B cells (F = 14.904, P <0.05). SWAP could induce spleen mononuclear cells to differentiate into CD19 + IFNγ + B cells in vitro (F = 3.277, P <0.05), but could not induce splenic B cells to differentiate into IFNγ + B cells alone or stimulate splenic B cells to secrete IFNγ; SWAP After immunization, the spleen B cells secreting IFNγ (F = 31.886, P <0.01) were stimulated in vivo and induced to differentiate into CD19 + IFNγ + B cells (F = 49.873, P <0.01). Conclusions SEA of Schistosoma japonicum can induce spleen mononuclear cells to differentiate into CD19 + IL 6 + B cells predominantly, while SWAP can predominantly induce the differentiation of mouse spleen mononuclear cells into CD19 + IFNγ + B cells. However, depending on other immune cells participate.