生物技术通讯
生物技術通訊
생물기술통신
LETTERS IN BIOTECHNOLOGY
2013年
3期
337-341
,共5页
任皎%赵莉%高孟%姜云水%郝明强%谭文杰%田厚文%阮力
任皎%趙莉%高孟%薑雲水%郝明彊%譚文傑%田厚文%阮力
임교%조리%고맹%강운수%학명강%담문걸%전후문%원력
人乳头瘤病毒%佐剂%治疗性疫苗
人乳頭瘤病毒%佐劑%治療性疫苗
인유두류병독%좌제%치료성역묘
human papillomavirus%adjuvant%therapeutic vaccines
目的：研究CpG佐剂、弗氏佐剂、聚肌胞苷酸佐剂及左旋咪唑、西米替丁作为佐剂对人乳头瘤病毒16型L2E7E6融合蛋白在小鼠体内产生的免疫效果的影响。方法：以单独蛋白组、蛋白加各佐剂组分别肌肉注射免疫C57BL/6小鼠，检测不同佐剂诱发小鼠产生的体液免疫和细胞免疫应答水平，并观察其对小鼠肿瘤生长的抑制作用。结果：各免疫组均能检测到高滴度的抗L2、E7、E6蛋白IgG抗体（以IgG1为主），其中弗氏佐剂能显著提高E6蛋白的IgG和IgG1抗体水平和E7蛋白的IgG1抗体水平（P<0.05），CpG佐剂明显提高了E7蛋白的IgG2a抗体水平（P<0.01）；而西米替丁佐剂则降低了E7抗原的IgG抗体水平（P<0.05）；同时可以检测到CpG佐剂组能诱发小鼠产生针对E7、E6较强的细胞免疫反应，且能抑制70%的荷瘤小鼠肿瘤生长；此外弗氏佐剂与聚肌胞苷酸佐剂可产生较弱的针对E7肽的细胞免疫反应，能延缓荷瘤小鼠肿瘤形成时间，与单纯蛋白组相比差异显著（P<0.05）。结论：CpG佐剂、弗氏佐剂和聚肌胞苷酸佐剂都能提高人乳头瘤病毒16型L2E7E6融合蛋白的细胞免疫反应水平和抑制肿瘤生长能力，其中CpG佐剂效果较好，为促进该蛋白作为疫苗的研发提供了实验依据。
目的：研究CpG佐劑、弗氏佐劑、聚肌胞苷痠佐劑及左鏇咪唑、西米替丁作為佐劑對人乳頭瘤病毒16型L2E7E6融閤蛋白在小鼠體內產生的免疫效果的影響。方法：以單獨蛋白組、蛋白加各佐劑組分彆肌肉註射免疫C57BL/6小鼠，檢測不同佐劑誘髮小鼠產生的體液免疫和細胞免疫應答水平，併觀察其對小鼠腫瘤生長的抑製作用。結果：各免疫組均能檢測到高滴度的抗L2、E7、E6蛋白IgG抗體（以IgG1為主），其中弗氏佐劑能顯著提高E6蛋白的IgG和IgG1抗體水平和E7蛋白的IgG1抗體水平（P<0.05），CpG佐劑明顯提高瞭E7蛋白的IgG2a抗體水平（P<0.01）；而西米替丁佐劑則降低瞭E7抗原的IgG抗體水平（P<0.05）；同時可以檢測到CpG佐劑組能誘髮小鼠產生針對E7、E6較彊的細胞免疫反應，且能抑製70%的荷瘤小鼠腫瘤生長；此外弗氏佐劑與聚肌胞苷痠佐劑可產生較弱的針對E7肽的細胞免疫反應，能延緩荷瘤小鼠腫瘤形成時間，與單純蛋白組相比差異顯著（P<0.05）。結論：CpG佐劑、弗氏佐劑和聚肌胞苷痠佐劑都能提高人乳頭瘤病毒16型L2E7E6融閤蛋白的細胞免疫反應水平和抑製腫瘤生長能力，其中CpG佐劑效果較好，為促進該蛋白作為疫苗的研髮提供瞭實驗依據。
목적：연구CpG좌제、불씨좌제、취기포감산좌제급좌선미서、서미체정작위좌제대인유두류병독16형L2E7E6융합단백재소서체내산생적면역효과적영향。방법：이단독단백조、단백가각좌제조분별기육주사면역C57BL/6소서，검측불동좌제유발소서산생적체액면역화세포면역응답수평，병관찰기대소서종류생장적억제작용。결과：각면역조균능검측도고적도적항L2、E7、E6단백IgG항체（이IgG1위주），기중불씨좌제능현저제고E6단백적IgG화IgG1항체수평화E7단백적IgG1항체수평（P<0.05），CpG좌제명현제고료E7단백적IgG2a항체수평（P<0.01）；이서미체정좌제칙강저료E7항원적IgG항체수평（P<0.05）；동시가이검측도CpG좌제조능유발소서산생침대E7、E6교강적세포면역반응，차능억제70%적하류소서종류생장；차외불씨좌제여취기포감산좌제가산생교약적침대E7태적세포면역반응，능연완하류소서종류형성시간，여단순단백조상비차이현저（P<0.05）。결론：CpG좌제、불씨좌제화취기포감산좌제도능제고인유두류병독16형L2E7E6융합단백적세포면역반응수평화억제종류생장능력，기중CpG좌제효과교호，위촉진해단백작위역묘적연발제공료실험의거。
Objective: To investigate the effects of CpG, freund adjuvant, poly I:C, levamisole and cimetidine on the immune responses to recombinant protein L2E7E6 of human papillomavirus type 16 in mice. Methods: C57BL/6 mice were immunized with recombinant protein L2E7E6 combined with or without CpG, freund adjuvant, poly I:C, levamisole or cimetidine through intramuscular. Then the humoral and cellular immune responses were detected by ELISA and ELISPOT respectively, and the effects of tumor growth regression were measured by using the TC-1 tumor bearing mice model. Results: All of groups mice immunized with the recombinant protein L2E7E6 combined with or without adjuvant could elicit high specific IgG antibodies titer against L2, E7 and E6 proteins (IgG1 antibodies were predominant), among which those with freund adjuvant could secret higher IgG and IgG1 antibodies against E6 antigen and higher IgG1 antibody against E7 antigen respectively(P<0.05), and those with CpG could secret higher Th1-associated IgG2a antibodies against E7 antigen(P<0.05), however those with cimeti?dine could secret lower IgG antibody against E7 antigen(P<0.05). Furthermore, the mice vaccinated with the re?combinant protein combined with CpG could elicit stronger T cell immune responses to peptides E7 and E6 re?spectively, and eliminate the established TC-1 tumors in 70% vaccinated mice. In addition, those mice vaccinated with freund adjuvant and poly I:C were able to elicit moderate cellular immune responses to peptide E7 and de?layed tumor appearance in tumor-bearing mice. Conclusion: Accompanied with CpG, freund adjuvant or poly I:C, the recombinant protein could be enhanced on the cellular immune response and the effects of tumor growth re?gression in immunized mice, and it could have better effects with the CpG, which will provide the basis to devel? op the L2E7E6 protein as one promising candidate vaccine.
