为探明蝶蛹金小蜂Pteromalus puparum毒液对其寄主菜粉蝶Pieris rapae蛹颗粒血细胞和浆血细胞的包囊与吞噬能力的影响,本研究分别采用Na2-EDTA处理和尼龙毛法对菜粉蝶蛹颗粒血细胞与浆血细胞进行分离纯化;再采用离体细胞培养方法,研究了菜粉蝶蛹颗粒血细胞、浆血细胞各自在包囊和吞噬反应中的作用,以及蝶蛹金小蜂毒液对其所产生的影响。结果表明:颗粒血细胞和浆血细胞均参与了包囊反应,其中前者包囊作用明显,后者作用微弱,但两者同时存在时包囊作用最为明显;血淋巴浆质对颗粒血细胞和浆血细胞包囊反应均无显著影响。毒液对颗粒血细胞和浆血细胞的包囊能力均存在显著的抑制作用,且抑制作用具有明显的剂量效应特征。此外,菜粉蝶蛹颗粒血细胞和浆血细胞均具吞噬能力,其中前者吞噬能力显著强于后者;毒液对颗粒血细胞与浆血细胞的吞噬能力亦均存在显著的抑制作用,且该抑制作用亦具有显著的剂量效应特征。结果说明,菜粉蝶蛹颗粒血细胞及浆血细胞均参与寄主的细胞免疫反应,蝶蛹金小蜂毒液对其寄主颗粒血细胞和浆血细胞的包囊与吞噬能力均存在显著的抑制作用。
In order to investigate the effects of Pteromalus puparum venom on the encapsulation and phagocytotic capacities of granulocytes(GRs) and plasmatocytes(PLs) in its host,Pieris rapae,we first purified these two types of host hemocytes using Na2-EDTA treatment and nylon wool method,respectively.Then,with the method of cell culture in vitro we also studied the different roles of GRs and PLs in host encapsulation and phagocytosis,and the effects of parasitoid venom targeting to these two cellular responses.The results indicated that both GRs and PLs took part in the host encapsulation.The encapsulation performed by GRs was stronger than by PLs.The highest rate was presented in encapsulation reaction mediated by the mixture of GRs and PLs.The host cell-free plasma did not significantly influence the encapsulation performed by GRs and PLs,respectively.The encapsulation capacities of GRs and PLs were both markedly inhibited by the parasitoid venom in a dose-dependent manner.Additionally,in the host,both GRs and PLs possessed the phagocytotic capacity.The phagocytotic capacity of GRs was much stronger than that of PLs.Parasitoid venom also noticeably suppressed the phagocytosis performed by GRs and PLs,respectively,also in a dose-dependent manner.The results suggest that both GRs and PLs in the host,P.rapae,play key roles in the host cellular responses,and P.puparum venom can significantly decrease host cellular responses including encapsulation and phagocytosis performed by GRs and PLs.