| Fasciolosis is a common disease of domestic water buffalos(Babalus bubalis).Better understanding of the characteristics of water buffalos immunocyte is to control this disease.In the first part of our study,We used enzyme linked immunosorbent assay (ELISA)to selecte 15 non-infected buffalos of 9,15 and 24 months.White blood cell(WBC)counts and differential WBC counts were calculated for each animal. Simultaneously,we isolated the PBMC of buffalos in differential age groups. These PBMC were stimulated with PHA and ConA at 0.125,0.25,0.5,1,2,3 and 4μg/well,respectively.MTT was applied to determine the PBMC proliferation and the stimulation index(SI)was calculated.The results showed that WBC counts of 9,15 and 24 months were 17.1×10~9 entries/ml,16.4×10~9 entries/ml and 16.5×10~9 entries/ml,respectively.Lymphocyte counts were 9.7×10~9 entries/ml,9.1×10~9 entries/ml and 7.9×10~9 entries/ml.Eosinophile granulocyte count of every group were 0.3×10~9 entries/ml,0.1×10~9 entries/ml and 0.3×10~9 entries/ml,respectively.Unremarkable changes were observed in WBC counts and differential counts.PBMC proliferation showed that the SI correlated positively with PHA and ConA concentration 0.125-4.000μg/well.In 9,15 and 24 months age groups,The SI of PHA-stimula ted PBMC were 1.3-3.3, 1.0-3.0 and 1.5-3.6,respectively.The SI of ConA-stimulated PBMC was 1.0-1.5, 1.1-1.6 and 1.0-1.6,respectively.The differences of SI in all age groups were insignificant.Further-research was facilated through above experiments.In the second part of our study,to know the modulation of IFN-γ,and IL-10 prouduction by F.gigantica excretory secretory products(FgESP)in water buffalos PBMC.FgESP was made from F.gigantica adults collected from gallbladders of buffalos.It was used as a stimulator along non-specific stimulators(ConA,PHA and LPS)to stimulate PBMC.We established FgESP, LPS,LPS+FgESP,ConA,ConA+FgESP,PHA and PHA+FgESP seven experimental groups and one control.The IFN-γ,and IL-10 production were estimated by ELISA at 4 time points(6,12,24 and 48h).The results showed that IFN-γlevels increased significantly at 6h in group of ConA,PHA and LPS. IFN-γproduction in PHA group was early and displayed a pesk at 6h(83.41 pg/mL).IFN-γproduction in ConA group was late displayed a pesk at 24h (87.76 pg/mL).Little IL-10 production In these groups was estimated.In groups of PHA,ConA and LPS,IFN-γlevels in the group of ConA was the highest (87.76 pg/mL)at 24h.IFN-γlevel in LPS group was the lowest at 6h(47.5 pg/mL).Little IL-10 and little IFN-γproduction was estimated in FgESP group in any time point.Through comparing ConA+FgESP,PHA+FgESP with ConA, PHA and conrol,we found the trends of little dropping on IFN-γand hight rising on IL-10(P＜0.05).The max upregulating index in ConA+FgESP and PHA+FgESP groups were 2.91 and 2.92 at 24h.The differents of IFN-γand IL-10 between groups FgESP+LPS and LPS was not significant.In this part of our study suggested that water buffalos PBMC was highly susceptible to ConA, it facilitated foundation of vaccine design in future.FgESP can induce regulation in immunity-escaped of Fasciola gigantica.
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