Echinococcus Granulosus:study On The Mechanism Of Immunological Protective Differences Between Recombinant Ferritin And Recombinant MMDH

Objective Through recombinant ferritin on immune protection and do not have protective immunity of recombinant mMDH comparative analysis of proteins, To find out, mechanism of immune protection caused by vaccine immune and mechanism of Ec hinococcus granulosus protoscolex in mice, provide theoretical basis for wider use of vaccines. Methods (1) ICR mice(female)were divided into three groups with random process;(2) All the mice of experimental group were vaccinated subcutaneously on the back with 10μl antigen dissolved in phosphate-buffered saline(PBS)and emulsified with 50ul Freund's complete adjuvant(FCA)respectively in the first time,two weeks after the first vaccination with the same preparation except that FCA was replaced by Freund's incomplete adjvant(FIA),all mice were vaccinated thirdly, the same as the second time with an interval week,PBS group did merely with PBS 100ul each time,the process was as before. (3) All the mice were challenged with 1500 protoscolices after vaccinating thirdly about 2 weeks; (4) Attack after 20 weeks of infection after cesarean section to kill mice, statistical number of cysts in mice peritoneal and diameter, calculation of immune protection;(5) Profile control and kill the mice, aseptic preparation of spleen, separation of spleen cells;(6) Spleen cells isolated, Application of FCM for detecting percentage of T-cell CD4~+, and CD8~+ change;(7) MTT assay for detection of splenic lymphocyte proliferation;(8) Obtain data on immune protection and do not have protective immunity of two group comparisons, analysis of inherent mechanism.Result (1)Using Dempster theory: rEg.ferritin can induce 86.37% immune protective efficiency in mice against the challenge infection, However rEg.mMDH Antigen immune ICR mice did not get immune protection,it indicated that rEg.ferritin has the potential of vaccines, rEg.mMDH is not.(2)①Immunity of mice with recombinant ferritin before the generation kill, CD4~+T cell increase(P﹤0.01), no obvious changes in CD8~+T cells(P﹥0.05), spleen lymphocyte subsets of mice with rEg.mMDH compared with control group, no significant change,it indicated that protective mechanism relate with participation of CD4~+T cells, role of CD8~+T cells are not significant;②As demonstrated by MTT assay,the proliferation activity of splenic T lymphocyte in rEg.ferritin group was higher than that in the control group(P﹤0.01), rEg.mMDH is not(P﹥0.05).It suggested that mechanisms of rEg.ferritin-induced mice relate with proliferation of splenic lymphocytes.Conclusions Cell immunity protective mechanism of rEg.ferritin induced mice may be associated with activated CD4~+T cell and stimulated proliferation of splenic lymphocytes.rEg.mMDH antigen non- immune protective mechanism may be associated with non-activated CD4~+T cell and non-stimulated proliferation of splenic lymphocytes.