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The completion of sequencing human genome creates a new era of biological science and technology. Although the sequence of the human genome has been known, it is still hard to rapidly explore the whole functional genes, especially, their interaction with each other and the meaning to the body. However, the "reverse biology" which comes into being in the recent years provides us a series of novel ideas and technologies for discovering new functional gene, among which the immune-related genes have attracted more attentions, clarifying how functional gene works and their potential value in application.
RESUMEN
Abstract Objective:To observe and compare the effects of immune responses of HBV DNA caccine by insertion IL-2 gene.Try to ex-Plore the efficient way to enhance the immune effect of DNA vaccines. Methods:The recombinant expression plasmid vector peDNA3.1-S/IL-2with fusion gene of HBsAg and IL-2 cloned was constructed by PCR method and DNA recombinant technique.Then, it was transfered into Cos-7 cells by cation lyposomes and was detected its transient expressing product in vitro.At last,observed and compared the cellular and humoralimmune responses to HBsAg in C57BL/6 mice by intramuscular injection.Results:The pcDNA3. 1-S/IL-2 was constructed successfully by themethod of restriction endonucleases digestion, RCR and DNA sequencing. It can increase the titers of anti-HBs antibody. The bioactivity of IL-2and lymphocyte proliferation responses to HBsAg induced by the mouse spleen immunized with psDNA3.1-S/IL-2 were improved also. Conclu-sion:These results showed the expression of HBsAg-IL-2 fused gene can enhance the immune responses and suggested the insertion of IL-2 maybe a practicable way to enhance the DNA vaccine efficacy.
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Objective:To study the immune responses of HBV based immunization co injecting CpG oligodeoxynucleotides in HBV transgenic mice.Methods:The HBV transgenic mice were immunized by multiple sites intramuscular injection with V HBs and CpG ODN or V HBs alone or V 1012 controls.The anti HBs Ab and HBsAg in serum from immunized mice was measured by ELISA.The expression of HBsAg in liver tissue was detected by immunohistochemistry method(SP).The histological activity index was calculated with Knodell's method.Results:The anti HBs Ab in serum could be detected and HBsAg in serum and liver tissues could not be measured in 2 out of 6 HBV transgenic mice immunized with V HBs+CpG ODN.There were not significantly changes in the level of anti HBs Ab and HBsAg in serum from HBV transgenic mice immunized with V HBs or V 1012 alone.The histological activity indexes in liver tissues from mice immunized with V HBs+CpG ODN were higher than that in mice inoculated with V HBs or V 1012 alone.The large number of lymphocytes could be found in liver tissues in V HBs+CpG ODN immunization group by microscopy.Conclusion:The immuno response,which seems to be responsible for the disappearance of HBsAg,might be elicited in HBV transgenic mice immunize with V HBs+CpG ODN.