| Objective: Hepatitis is an infectious disease and badly harm to humanhealth. In the world, there are about 5 hundred millions HBV and HCVinfector and hepatitis is correlated with the occurrence of hepatocirrhosisand liver cell cancer. Although interferon and anti-virus nucleotide drugshave new evolvement, there are no effective drugs to treat hepatitis.Recently, injected hepatitis B vaccine is the main way that can prevent andtherapy hepatitis B, but it should inject big dose and many times. Nowmany studies showed that prokaryotic DNA containing unmethylated CpGmotifs could stimulate the immune system and induce the secretion ofimmunoglobulins and several kinds of cytokines. At the same time,synthesized CpG oligodeoxynucleotides (ODN) could potently stimulatemost types of immune cells. It could directly stimulate B lymphocytes,macrophages and dendritic cells (DC) through interaction with Toll likereceptor 9. As an effective adjuvant, CpG-ODN is capable of promotingantigen specific immune responses. At the same time, HBV transgenic micesimulate the state of human HBV infection, so they can be regarded as ananimal model to study human chronic HBV infection. The aim of thisarticle is to investigate the effects of CpG-ODN as an adjuvant on cellularimmune responses in HBV Transgenic Mice immunized with recombinanthepatitis B vaccine. Methods: HBV Transgenic Mice were divided into four groups(n=10), and were partly immunized with N.S. , rIFNα-2b(20000IU/time),recombinant hepatitis B vaccine(4μg/time)and vaccine(4μg/0.5ml/time)+CpG-ODN(15μg/0.1ml/time)into the left leg muscles.After immunized, blood was collected and the subsets of T lymphocytewere investigated through FACSVantageTMSE, IL-2,IL-12(p70)and IFN-γwere examined by ELISA, the proliferation of spleen cells was measuredby MTT and cytotoxic T lymphocyte (CTL) response was tested by lacticacid dehydrogenase (LDH) releasing method. Results: CD3+%,CD4+%,CD8+% of recombinant hepatitis B vaccinegroup were(25.008±2.852)%,(13.640±1.616)%,(9.989±1.123)%;the amount of IL-2,IL-12(p70),IFN-γwere(1304.288±572.212)pg/mL,(203.810±91.807)pg/mL,(860.736±336.888)pg/mL . Compared withN.S., recombinant hepatitis B vaccine could distinctly raise CD3+%,CD4+%,CD8+% and the amount of IL-2,IL-12(p70),IFN-γ, and inducestrong lymphocyte proliferation reaction and cytotoxic T lymphocyteactivities, (P<0.05). CD3+%,CD4+%,CD8+% of vaccine + CpG groupwere (28.952±4.057)%,(15.413±1.307)%,(11.109±1.159)%, theamount of IL-2,IL-12(p70),IFN-γwere (1930.719±623.924)pg/mL,(329.090±162.155)pg/mL,(1 554.827±425.243)pg/mL. The results ofvaccine + CpG group had large difference with vaccine group, (P<0.05).Immunized HBV Transgenic Mice with CpG-ODN showed higher specificcellular responses to HBV than those without CpG-ODN (P<0.05).Because of times and doses of rIFNα-2b, immunized with rIFNα-2b andN.S. have no obvious difference. Conclusion: 1.After HBV Tg mice were immunized with CpG-ODNand recombinant Hepatitis B vaccine, the percent of CD4+ T cell(Th cell)was more higher than vaccine group. 2. The amount of IL-2,IL-12(p70),IFN-γ,the proliferation ofspleen cells and cytotoxic T lymphocyte (CTL) response were largelyhigher than vaccine group. 3.CpG-ODN and Recombinant Hepatitis B Vaccine can induce morestrong HBV specific cellular immune response, specially Th1 cellularimmune responses. 4.CpG-ODN can act as adjuvants to enhance cellular immuneresponses in HBV Transgenic Mice immunized with recombinant hepatitisB vaccine.
|
PDF Full Text Request