Dissertation > Excellent graduate degree dissertation topics show

Echinococcus Granulosus Recombinant GST Protective Immunity and Its Immune Mechanism in Mice

Author: XiongYing
Tutor: ZhaoWei
School: Ningxia Medical University
Course: Genetics
Keywords: Echinococcus granulosus Glutathione s- transferase Protective immunity Immune mechanism
CLC: R392
Type: Master's thesis
Year: 2010
Downloads: 30
Quote: 0
Read: Download Dissertation

Abstract


The purpose of Echinococcus granulosus (Echinococcus granulosus, Eg) have been cloned and successfully constructed reorganization the prokaryotic expression plasmid pET-28a/GST of the foundation on expression purified recombinant protein Eg.GST, animal experiments and immune mechanisms of protective immunity study to identify the potential of its vaccine. Recombinant expression plasmid of the method of (1) inducing expression and purification: and IPTG was induction expression of recombinant proteins EgGST (rEgGST), and purified by His-bind resin column containing Ni2 rEgGST. (2) animal grouping and immunization programs: select 132 female 6-week-old ICR mice were randomly divided into three groups namely experimental group, adjuvant PBS and PBS group, n = 44; using subcutaneously in a multi-point injection, continuous immune three times, at intervals of two weeks; immunized mice after 10 weeks, each mouse with 1500 living former head larva intraperitoneal injection attack infection. 0 weeks before immunization and after the attack, two weeks, four weeks, six weeks, 10 weeks, 18 weeks, 30 weeks, were killed and mice, and the orbital blood serum was separated. (3) rEgGST protective immunity in mice induced force calculation: according to the Dempster the formula: immune protection force (%) = (1 - Immunohistochemical average number of cysts / control group, the average number of cysts) × 100% calculation immune protection force (4) rEgGST induced protective immunity in mice and its mechanism: ① by ELISA method rEgGST induced the mouse humoral immunity indicators change research; (2) through streaming technology on rEgGST induced changes in indicators of immune cells in research ; ③ rEgGST induced mouse cytokine changes and MTT assay mouse spleen lymphocyte proliferation. Results (1): The genetic engineering have constructed the strains Eg.GST/pET-28a/BL21 (DE3) plysS., Expression and purification of a molecular weight of about 28kD the reorganization EgGST of. (2): According to the formula rEgGST can induce protective immunity of 89.39%. (3) ① rEgGST mice after antigen after immunization and challenge infection, produce high levels of IgG, low levels of IgG1 and IgE, IgG2a, IgG2b and IgG3 antibody levels before immunization compared is elevated, and prompt IgG IgG3 , IgG2a and IgG2b may be involved in the mechanism of the protective immune response. The ② rEgGST mice after challenge infection not only increase in CD4 T cells, CD8 T cells have slightly increased CD4 / CD8 ratio with PBS control group decreased; CD25 cells and adjuvant challenge infection after the 30 weeks rEgGST group with compared to the control group significant difference between the experimental group and the control group had no significant difference. ③ rEgGST mice after immunization generates a high level of IL-2, IFN-γ and TNF-α, a low level of IL-4 and IL-10; late attack infected IL-2, IFN-γ and TNF-α reducing IL-4 and IL-10 increased. Reaction prompted the recombinant antigen to induce Th1-type immune response mainly; attacks infected spleen lymphocyte proliferation test, MTT assay confirmed that the immunized mice rEgGST in rEgGST stimulated spleen lymphocyte proliferation was significantly higher than the PBS control group (P lt; 0.01). Conclusion (1): successfully expressed and purified rEgGST. (2): rEgGST can induce protective immunity of mice to produce 89.39%, suggesting rEgGST protein can induce strong protection force, is a highly the potential anti packages coccidiosis vaccine candidate molecules. (3): rEgGST protective immunity by inducing host humoral immune response and Th1-type immune response the show that rEgGST candidate vaccine is a promising anti-echinococcosis.

Related Dissertations

  1. Identification of B.melitensis 16M Protein Immunodominant Antigens,S852.61
  2. The Development of PCR Diagnosis Kit for Detecting the Echinococus Granulosus in Canine Feces,S858.292
  3. Anti- hydatid drugs mebendazole soft capsules in the development and bioavailability studies,R96
  4. Effects of Biliary Fistula on Life Period of Hepatic Cystic Echinococcosis,R532.32
  5. Echinococcus granulosus ( Chinese mainland strain ) recombinant antigen Eg14-3-3 immune protection and immune mechanisms,R392
  6. Studies of Immunoportection and Its Mechanism of Recombinant Myophilin Against Echinococcus Granulosus in Mice,R392
  7. Echinococcus granulosus cDNA library screening and diagnostic target antigen,R532.32
  8. Turbot important bacterial diseases pentavalent vaccine and immune adjuvant effect of,S943
  9. Proteomics of Echinococcus Granulosus and Cyst Wall,R392
  10. Echinococcus Granulosus:study on the Mechanism of Immunological Protective Differences between Recombinant Ferritin and Recombinant mMDH,R392
  11. Preliminary Research on Biological Features of Recombinant EG95-(C3d)3 Protein,S852.5
  12. Immunoadjuvant Effect of the Extracellular Domain of Sheep CTLA-4 on the EG95 Antigen of Echinococus Granulosus,S852.5
  13. Sequence Analyzing and Its Expression of the egG1Y162 Gene from Echinococcus Granulosus,R392
  14. Construction and Expression in E.coli of the Recombinant Plasimid pET30a-EgA31-Eg95 of Echinococcus Granulosus and Its Serological Evaluation,R392
  15. Molecular Cloning and Characterization of EgERK1 from Protoscolex of Echinococcus Granulosus in Xinjiang,R346
  16. Echinococcus Granulosus (the Chinese Mainland Stain): Expression of Recombinant Myophilin Vaccine & Its Immune Protection and Mechanism in Mice,R392
  17. Echinococcus Granulosus: Cloning, Expression of EgP-29 and Its Protective Immunity and Mechanism in Mice,R392
  18. Recombinant Expression, Identification Immunological Characters and Immunological Protection of Ferritin Gene pET-28a from Echinococcus Granulosus,R392
  19. Echinococcus Granulosus: Dynamic Study on ICR Mice Immunized with Recombinant Protein EgP-29,R392
  20. Research on Motif Discovery Method Based on Immune Genetic Algorithm,Q75

CLC: > Medicine, health > Basic Medical > Medical Immunology
© 2012 www.DissertationTopic.Net  Mobile