【摘要】 为探索高效安全的分子免疫增强剂,本实验设计合成含11个CpG基序的寡聚核苷酸(CpG ODN),制备壳聚糖纳米颗粒包裹CpG ODN,研究新型CpG ODN壳聚糖纳米颗粒(CpG-CNP)对人乙型肝炎疫苗的免疫佐剂效应。在肌注乙型肝炎疫苗免疫6周龄昆明小鼠的同时,肌注接种裸CpG ODN 30 pmol/只(A1组)和CpG-CNP 5pmol/只(A2组),口服CpG-CNP 30 pmol/只(B组),并设生理盐水空白疫苗对照组(C组)。在接种后每周采血,Sandwich ELISA检测实验小鼠IL-2、IL-4、IL-6、IgG、IgA和IgM水平以及特异性抗体(HbsAb)含量和免疫细胞数量的变化。结果发现各实验组小鼠的白细胞介素、血清免疫球蛋白、乙肝特异抗体和外周血免疫细胞均较对照组显著增多(P<0.05);CpG-CNP肌注组小鼠血液的免疫球蛋白、特异性乙肝表面抗体和白细胞介素含量、淋巴细胞和单核细胞数量较A1组和B组显著增加(P<0.05)。CpG-CNP口服组与裸CpG肌肉注射组无显著差异(P>0.05)。这些表明CpG-CNP不仅能高效诱导小鼠对乙肝疫苗的体液免疫应答,同时也显著增强其细胞免疫应答;CNP包裹可提高CpG的免疫刺激效应,减少CpG剂量,提示新CpG-CNP肌注或口服可用于HBV的免疫预防。更多还原

【Abstract】 In order to develop a safe and effective immunoadjuvant to enhance the immunity of human to resist HBV infection,an novel CpG Oligodeoxynucleotides(CpG ODN) containing 11 CpG motifs was synthesized and entrapped with chitosan nanoparticles(CNP) which prepared by ionic cross linkage method,and employed to intramuscular injection on 6 weeks old Kunming mice with 5 μg HBsAg vaccine/ capita.Meanwhile,A1 group received i.m.inoculation with CpG ODN at the dose of 30 pmol/ mouse,A2 with CpG-CNP 5 pmol/ mouse,B group were orally inoculated with CpGCNP 30 pmol/ mouse,and C group only with saline as the blank control.The blood from the mice was weekly collected to detect the changes of the number of immune cells,IL-2,IL-4,IL-6,IgG,IgA and IgM by Sandwich ELISA,and HBsAb and immune cells were also analyzed by indirect ELISA and routine method.The result indicated the level of IL-2,IL-6,IgG,IgA,IgM and specific HBs antibody were significantly increased in the treated mice compared with the control mice,so as the number of lymphocytes and monocytes in all the treated mice(P<0.05);especially the humoral and cellular immune responses of A2 group were remarkably stronger than those of A1 and B mice(P<0.05).The oral inoculation with CpG-CNP provoked the similar humoral and cellular immunity in B group as those of A1 mice group(P>0.05).These suggested that the novel CpG ODN could significantly enhance the cellular and humoral immune response induced by HBsAg in mice,and CNP-CpG was a potentially promising effective immune potentiator to promote the vaccination control of of HBV in the future.更多还原