| DNA vaccine is derived from gene therapy research and developed a new field of study in recent years, it refers to a protein encoding the recombinant eukaryotic expression vectors of the immune body, so that foreign gene expression in vivo, thereby stimulating the immune system induced by a specific immune response. So, in this thesis, I am currently studying for dental caries DNA vaccine, and tuberculosis DNA vaccine which were preduced by magnifying technology in our institute.Experiment 1 Magnifying technology produced Anti-caries DNA vaccine immune efficacy experimentGnotobiotic rats, were immunized with targeted anti-caries plasmid pGJA- P/VAX1 which build by School of stomatology Wuhan University,study pGJA- P/VAX1 immune efficacy. 56 rats were randomly divided into 7 groups, infected with S. mutans at 20 days of age and given cariogenic diet, factorial experiment designed to immune rats by 3 factor (plasmid, protein, adjuvant) and 2 level (intramuscular injection(i.m.)or intranasal(i.n.)). Collection of serum and saliva samples at 0w,2w,4w,6w,8w,10w. An indirect ELISA method was established and vertify for serum IgG and salivary IgA detection, and determined the antibody responses induced by the vaccines. At the end of the experiment, Keys caries score was used to evaluate the anticaries effectiveness of the vaccines at the terminal study. SSPS software analyzed result show that:(1) As for the antibody reactions and caries scores, there were significantly different -ces (P<0.01) between rats immunized with DNA vaccine and immunized with PVAX1.(2) As the same immune pathway, rats immunized with pGJA-P/VAX and PAc protein had the significantly (P<0.01) highest level of specific serum anti-PAc IgG antibodies. There were no significant difference between the salivary anti-PAc IgA antibodies and their Keyes score of rats immunized with only plasmid.(3) The same immunization program, intranasal routes displayed the higher specific anti-PAc IgA antibody in salivare and lower Keyes score (P<0.05), serum antibo- dies was no difference. (4) In the mucosal immunity, plasmid immunitied with adjuvant and enhanced with PAc displayed the significantly high antibody reactions (P<0.01). Plasmid alone without adjuvant had much higher Keyes score than the other three groups (P <0.05).Conclude: The recombinant plasmid pGJA-P can effectively inhibit the occurrence and development of dental caries. Anticaries DNA vaccine pGJA-P/VAX1 by intrana- sal could prevent rats from dental caries more effectively than vaccine by intramus -cular route.Experiment 2 Initial Immune effects experimental of Tuberculosis DNA Vaccine produced by magnifying technologyMouse,were immunized with encoding Ag85A antigen tuberculosis DNA vaccine which build by the People's Liberation Army 309 hospital, study the effection of Immunity. 20 Balb / c mice,4-6weeks old of age,were randomly divided into 2 groups, immunized with TB DNA vaccine or PBS as control by intramuscular injection under design. The basis of the blood collected before immunization, a total of three immunization, 3 weeks after the third immunization mice were killed and collect whole blood, separation of serum for the detection of serum specific antibody and cytokine (Interferon-γ, IL-4); While detected the spleen cells Ag85A specific Inter- feron-γsecretion and CD4 +, CD8 + percentage. Results statistical analysis indicated that:(1) Mice with the TB DNA vaccine cann't stimulate a significant antibody.(2) But with higher Serum cytokines IFN-γand lower IL-4 than PBS control group.(3) ELISPOT experiment show TB group spleen cells IFN-γsecretion higher than PBS group (P <0.05).(4) TB group CD4 + cell percentage was higher than PBS control group (0.025 |
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