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Immune Responses to HBV Surface Protein Using DNA Prime and Prome and Protein Boost Stuategy in Balb/c Mice

Author: ZhaoMingLi
Tutor: XingYiPing
School: Nanjing Medical University
Course: Internal Medicine
Keywords: DNA vaccine Hepatitis B Surface antigen The beginning of heterogeneity Free - strengthen immunization strategy
CLC: R392
Type: Master's thesis
Year: 2009
Downloads: 11
Quote: 0
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Abstract


Background Hepatitis B virus infection is a global health concern, hepatitis B virus infection can lead to chronic hepatitis, cirrhosis, and even hepatocellular carcinoma. Currently, effective in preventing hepatitis B vaccine is based on HBsAg as a main component, a recombinant subunit vaccine, can make the 90% to 95% of vaccinated produce protective antibodies after vaccination, but still 5% to 10% of people Even if vaccination over three HBsAg vaccine can not produce a protective immune. Chronic HBV carriers, due to its specific cellular immune function is low, the body of HBsAg tolerated, can not produce a humoral and cellular immunity after vaccination. Since the advent of DNA vaccine, its advantages gradually show up and be recognized in the prevention of infection by pathogens such as viruses, intracellular bacteria and parasites have a greater advantage. Some studies have shown that HBsAg DNA vaccine can stimulate the body to produce specific immune response of a certain intensity, and has a potential therapeutic effect. Present an obvious problem is the lower animals, the greater the effectiveness of DNA vaccine. A variety of ways to enhance the efficiency of the nucleic acid vaccine, the beginning of heterogeneity Free - strengthen immunization strategy (Heterologous Prime-Boost regimen) is one of the immunization strategy to study more. This strategy the first immunization, and strengthen the immune using different immunogen or immunological methods, can induce the immune system to produce a strong specific immune response. Studies show heterogeneity Free early - to strengthen immunization strategy, a better role in the induction of cellular immune responses, and shows a good prospect of application. The previous build of the study group the novel nucleic acid vaccine pSW3891/MHBs/adr of (abbreviated ADR), the nucleic acid vaccine induced high titers of antibody in mice, showing good immunogenicity. The purpose of this experiment is to observe the beginning of of adr nucleic acid vaccines Free protein vaccine booster immunization strategy of immune responses in mice. Objective To observe the DNA vaccine primary immunization of Balb / c mice immune response, enhanced by the hepatitis B virus HBsAg protein vaccine immunization strategy. The liposomal transfection method TransfectionTM Law hepatitis B virus surface antigen protein (MHBs) nucleic acid the vaccine pSW3891/MHBs/adr, and the empty vector pSW3891 (vector) in vitro transfection of 293T cells. Immunoblotting (Western blot) detection adr vector in vitro expression; animal study selected 18 Balb / c mice (n = 6), the number and then randomly divided into three groups, namely the empty vector plasmid group (vector vector group ), adr DNA vaccine combined HBsAg protein vaccine (adr protein group) HBsAg protein vaccine combined with HBsAg protein vaccine (protein protein group); intramuscular injection at week 0, respectively, to the vector, adr, and HBsAg protein vaccine immunized mice at 4 weeks intramuscular injection Vector, HBsAg protein vaccine, and HBsAg protein vaccine immunized mice. Detect serum HBsAg specific antibodies by enzyme-linked immunosorbent assay (ELISA), enzyme-linked immunosorbent spot (ELISPOT) assay mouse splenocytes HBsAg polypeptide specific IFN-γ-secreting cells. Results of the adr in vitro transfection of 293T cells, able to express the surface antigen of hepatitis B virus the protein (MHBs); vivo study results show that: In addition to the negative control vector vector group the adr protein group, protein protein group mice could be detected in serum anti-HBs, dynamic changes: immune two weeks after the start to produce antibodies reached a peak, the sixth week of anti-HBs, the the protein protein group anti-HBs endpoint titer (1:72900) and the adr protein group (1:24300 ) was significantly higher, with statistical significance (P lt; 0, 05); application ELISPOT assay specificity INF-γ secretion of spleen cells specific number of spots for, adr protein group 713.94 ± 8.029/10 ~ 5 cells ( X ± SD) protein protein group specific spots for 379.22 ± 43.06/10 ~ 5 cells (X ± SD), vector vector group specific number of spots is only of 9 ± 3.42/10 ~ 5 cells. adr protein group was significantly higher than that of protein protein group (p lt; 0.001), with a statistically significant (p lt; 0.05). Conclusion of hepatitis B virus surface antigen protein (MHBs) nucleic acid vaccine pre-sensitized Balb / c mice significantly increased the hepatitis B HBsAg protein vaccine cellular immune response level.

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