CAJ | 학술논문

目的初步评价Al(OH)3对重组HPV16的L2E6E7融合蛋白(简称HPV16L2E6E7)的免疫佐剂作用.方法将108只雌性C57BL/6小鼠随机分成Al(OH)3组(85.7 μg/只,肌肉注射)、HPV16L2E6E7组(120.0μg/只,肌肉注射)、A1 (OH)3+HPV16L2E6E7组[每只 Al(OH)3 85.7μg+HPV16L2E6E7120.0μg,肌肉注射],每组36只,免疫程序为0、3、7d.ELISPOT法检测IFN-γ,间接ELISA法检测IgG抗体水平.另将40只皮下接种TC-1肿瘤细胞(1×104/只)的雌性C57BL/6小鼠随机分为PBS组(100.0 μL)、Al (OH)3组(85.7μg/只,肌肉注射)、HPV16L2E6E7组(120.0μg,/只,肌肉注射)、Al(OH)3+ HPV16L2E6E7组[每只Al(OH)385.7μg+HPV16L2E6E7 120.0μg,肌肉注射],每组10只.免疫程序为0、3、7d.观察肿瘤形成时间、体积变化及成瘤率,观察期为53 d.结果免疫后第10、14、21、28、35和42d,HPV16L2E6E7组与Al(OH)3+HPV16L2E6E7组IgG抗体滴度都高于Al(OH)3对照组,差异具有统计学意义(P均＜0.01),免疫后21、28、35、42 d Al(OH)3+ HPV16L2E6E7组特异性IgG抗体显著高于HPV16L2E6E7组(P均＜0.01);免疫后14、21、28d Al(OH)3+ HPV16L2E6E7组E7特异性IFN-γ水平显著高于HPV16L2E6E7组(P均＜0.01);小鼠抑瘤实验中,HPV16L2E6E7组抑瘤率为60％,Al(OH)3+ HPV16L2E6E7抑瘤率为40％,两组无明显差异(P＞0.05),但与对照组比较都有显著性差异(P＜ 0.01,P＜0.05).结论初步评价Al(OH)3能与HPV16L2E6E7较好吸附,可诱导C57BL/6雌性小鼠产生更强的特异性体液和细胞免疫应答,具有免疫增强作用.
목적 초보평개Al(OH)3대중조HPV16적L2E6E7융합단백(간칭HPV16L2E6E7)적면역좌제작용.방법 장108지자성C57BL/6소서수궤분성Al(OH)3조(85.7 μg/지,기육주사)、HPV16L2E6E7조(120.0μg/지,기육주사)、A1 (OH)3+HPV16L2E6E7조[매지 Al(OH)3 85.7μg+HPV16L2E6E7120.0μg,기육주사],매조36지,면역정서위0、3、7d.ELISPOT법검측IFN-γ,간접ELISA법검측IgG항체수평.령장40지피하접충TC-1종류세포(1×104/지)적자성C57BL/6소서수궤분위PBS조(100.0 μL)、Al (OH)3조(85.7μg/지,기육주사)、HPV16L2E6E7조(120.0μg,/지,기육주사)、Al(OH)3+ HPV16L2E6E7조[매지Al(OH)385.7μg+HPV16L2E6E7 120.0μg,기육주사],매조10지.면역정서위0、3、7d.관찰종류형성시간、체적변화급성류솔,관찰기위53 d.결과 면역후제10、14、21、28、35화42d,HPV16L2E6E7조여Al(OH)3+HPV16L2E6E7조IgG항체적도도고우Al(OH)3대조조,차이구유통계학의의(P균＜0.01),면역후21、28、35、42 d Al(OH)3+ HPV16L2E6E7조특이성IgG항체현저고우HPV16L2E6E7조(P균＜0.01);면역후14、21、28d Al(OH)3+ HPV16L2E6E7조E7특이성IFN-γ수평현저고우HPV16L2E6E7조(P균＜0.01);소서억류실험중,HPV16L2E6E7조억류솔위60％,Al(OH)3+ HPV16L2E6E7억류솔위40％,량조무명현차이(P＞0.05),단여대조조비교도유현저성차이(P＜ 0.01,P＜0.05).결론 초보평개Al(OH)3능여HPV16L2E6E7교호흡부,가유도C57BL/6자성소서산생경강적특이성체액화세포면역응답,구유면역증강작용.
Objective To evaluate the adjuvant effects of Al(OH)3 on a recombinant human papillomavirus 16L2E6E7 fusion protein (HPV16L2E6E7).Methods There were 108 female C57BL/6 mice randomly divided into Al(OH)3 group [85.7 μg for each mouse,intramuscular injection (im)],HPV16L2E6E7 group (120.0 μg for each mouse,im) and Al(OH)3 + HPV16L2E6E7 group (85.7μg Al(OH)3 and 120.0 μg HPV16L2E6E7 for each mouse,im),36 mice in each group,and the injection were given on 0,3,7 d.ELISA and ELISPOT were used to detect IgG and IFN-γ levels.Forty female C57BL/6 mice subcutaneously inoculated with 1 × 104 TC-1 tumor cells were randomly divided into PBS group (100.0 μL,im),Al(OH)3 group (85.7 μg for each mouse,im),HPV16L2E6E7 group (120.0μg for each mouse,im),Al(OH)3 + HPV16L2E6E7 (85.7 μg Al(OH)3 and 120.0μg HPV16L2E6E7 for each mouse,im),10 mice in each group,the injection were given on 0,3,7 d.The tumor formation time,tumor volume and antitumor rates were recorded throughout 53-day observation.Results The IgG titers of HPV16L2E6E7 and Al (OH)3 + HPV16L2E6E7 group were significantly higher than control group (P all ＜ 0.01) on 10,14,21,28,35 and 42 d.The specific IgG antibody titers of Al(OH)3 + HPV16L2E6E7 group were significantly higher than HPV16L2E6E7 group (P all ＜0.01) on 21,28,35,42 d.The IFN-γ levels of Al(OH)3 + HPV16L2E6E7 group were significantly higher than HPV16L2E6E7 group (P all ＜ 0.01) on 14,21,28 d.In TC-1 cell tumor model of female C57BL/6 mice,the antitumor rate of HPV16L2E6E7 was 60％ and it decreased to 40％ in Al(OH)3 + HPV16L2E6E7 group (P ＞ 0.05).Compared to the control group,there were no significant differences (P ＜ 0.01,P ＜ 0.05).Conclusions HPV16L2E6E7 fusion protein accompanied by Al(OH)3 adjuvant can induce stronger antigen-specific humoral and cellular immune responses which demonstrates that it can improve the immunogenicity of HPV16L2E6E7.