Anopheles Dirus Black Coated Yoelii Immune Mechanism

An. dirus refractory to P.yoe/ii is manifested by melanotic encapsulation of the oocyst after complete its passage through the mosquito's midgut. It is deduced that the PPO cascade may be involved in the melanotic encapsulation reaction of An.dirus. However, few evidences are directly support this opinion. It is known that PPO system consists of recognition/binding protein, PPO and PPAE. Of the system, P0 is one of the important components; it has two activities, MPO and DPO, and exists as proenzyme PPO in the insect. To explore the role of PPO cascade in melanization of Pyoelii, an An.dirus-Pyoelii system was used to observe alteration of P0 activity in An.dirus haemolymph following infection of P yoelii and the induced haemolymph proteins during P yoelii oocyst melanization. PPAE is a kind of serine proteinase about 30}CD. Serine proteinase is the key enzyme of serine proteinase cascade. We postulate that serine protinase of An.dirus may be also important in the regulation of melanotic encapsulation against Pyoelii. The aim of this study is to clone the serine proteinase from An. dirus to establish foundation to further analysis its role in regulation of melanotic encapsulation against Plasmodium. In addition, An.dirus S7 is to also be cloned to use as internal control to follow the transcriptional activity of serine proteinase during Plasmodium infection. Adult An.dirus females (3-5 days post enclosion) were divided into three groups: non-blood-fed (N), normal-blood-fed (B) and infected-blood-fed (I). Firstly, both MPO and DPO activities in all three groups were identified and measured by zymography following native PAGE at 5,7,11 and 15 days, respectively. Our data show MPO and o-DPO activity in group I is significantly higher than those in groups N and B (p