Studies On Immune Factor And Activity Of Phagocytes In Coelomic Fluid Of Strongylocentrotus Intermedius

1In order to study the immune mechanism of coelomic fluid of Strongylocentrotus intermedius, weresearched the phagocytosis activity in different medium and different treatment of yeast cell; Thenwe analysed the no cell coelomic fluid and ingredient which was isolated from regulating yeast cellsand no regulating yeast cells with SDS-PAGE, and no cell coelomic fluid was dialysis,SephadexG-200gel chromatography and SDS-PAGE; In isotonic buffer, cell free coelomic fluid by yeast cellabsorption, we determined the phagocytosis effected by opsonin-like of different concention anddifferent time. The result showed that isotonic buffer,cell free coelomic fluid by yeast cellsabsorption,and no cell coelomic fluid all can regulate phagocytosis,and the ratio is1.00:1.11:1.94,aswell as,the phagocytosis rate of regulating yeast cells was1.61times of no regulating yeast cells inisotonic buffer;There was a new opsonin-like molecular band from ingredient which was isolatedfrom regulating yeast cells, and the molecular weight is250.62ku,as well as,we also found themolecular weight is250.62ku after the no cell coelomic fluid was dialysised, SephadexG-200gelchromatography; in tested concentration range, the greater the opsonin concentration, the higher thephagocytosis, including5.0μg/ml,10.0μg/ml were higher than control group in each test timegroup. In isotonic buffer,10.0μg/ml and5.0μg/ml groups had highly significant differencecomparing to control group in30min after reaction.In cell free coelomic fluid by yeast cellabsorption,10.0μg/ml group had significant difference comparing to control group in30min afterreaction.These results indicted that,there is a kind of new opsonin-like molecular in coelomic fluidof Strongylocentrotus intermedius and the molecular weight is250.62ku which can promotephagocytosis.2In this part,the method of mammals hemolytic serum for complement factor B test to detect thecomplement B factor of Strongylocentrotus intermedius. We tested the coelomic fluid activity ofcomplement factor B after the heat treatment and without heat treatment. Results showed that thereis a complement factor B in the coelomic fluid of Strongylocentrotus intermedius,and the hemolyticactivity of complement factor B in coelomic fluid after heat treatment was lower than that withoutheat treatment.That is there is a complement factor B in the coelomic fluid,as well as it is not stableto high tempreture,and it is same to the character of mammal’s factor B.3Our experiment examined the phagocytic activity in different times and different temperatures,aswell as the injection of undaria pinnatifida lectin,we want to examine whether these factors canpromote phagocytic activity. Results showed that phagocytic activity had the highest rate at30minin30℃. We injected undaria pinnatifida to Strongylocentrotus intermedius, and injection time wasevery24h, continuely5days, and we found that after the injection of different time, threeconcentrations of lectin:0.57mg/ml,0.28mg/ml,1.13mg/ml can significantly or extremely significantly improve phagocytic activity,and the group of0.28mg/ml had the best effect in provingthe phagocytic rate,the group of0.75mg/ml had the best effect in proving the phagocytic count.