中华妇产科杂志
中華婦產科雜誌
중화부산과잡지
CHINESE JOUNAL OF OBSTETRICS AND GYNECOLOGY
2013年
7期
523-527
,共5页
乳头状瘤病毒科%癌基因蛋白质类,病毒性%衣壳蛋白质类%DNA疫苗%免疫活性%小鼠,近交系
乳頭狀瘤病毒科%癌基因蛋白質類,病毒性%衣殼蛋白質類%DNA疫苗%免疫活性%小鼠,近交繫
유두상류병독과%암기인단백질류,병독성%의각단백질류%DNA역묘%면역활성%소서,근교계
Papillomaviridae%Oncogene proteins,viral%Capsid proteins%Vaccines,DNA%Immunocompetence%Mice,inbred strains
目的 初步观察PVAX1-HPV58mE6E7FcGB复合DNA疫苗的抗肿瘤免疫效果.方法 在检测该复合疫苗的免疫效果前,自行构建了稳定表达HPV58E6E7融合基因的B16-HPV58E6E7成瘤细胞系,表达并纯化了HPV58E6E7-GST融合蛋白作为抗原.将该疫苗免疫C57BL/6小鼠(免疫组)后,通过抗肿瘤移植保护实验和肿瘤生长抑制实验观察该疫苗对小鼠负荷的B16-HPV58E6E7移植瘤的防治效果；通过特异性抗体检测、酶联免疫斑点检测、特异性杀伤实验观察该疫苗在被免疫小鼠体内诱发的体液和细胞免疫反应.以单纯抗原PVAX1-HPV58mE6E7 Fc免疫小鼠作为单纯抗原组.结果 抗肿瘤移植保护实验显示,经PVAX1-HPV58mE6E7FcGB疫苗免疫的小鼠成瘤率仅为9/15,但成瘤时间最长[为(13.6±1.7)d],且肿瘤生长最慢.在肿瘤生长抑制实验中,疫苗组小鼠的肿瘤生长抑制率达81.4％.体液免疫检测显示,该疫苗和单纯抗原均能在小鼠体内诱发特异性抗体,最高滴度分别为1∶25 600和1∶12 800,两组比较,差异无统计学意义(P＞0.05)；但在细胞免疫检测中,疫苗组激活的特异性T淋巴细胞数[(219±34)个/4×105个脾淋巴细胞]约为单纯抗原组[(55±25)个/4×105个脾淋巴细胞]的4倍,两组比较,差异有统计学意义(P＜0.05)；且该疫苗诱发的特异性针对B 16-HPV58 E6 E7细胞的杀伤率(最高为43.3％)也明显高于单纯抗原组(杀伤率最高为31.3％),两组比较,差异有统计学意义(P＜0.05).结论 PVAX1-HPV58mE6E7FcGB复合DNA疫苗能有效激发特异性的体液和细胞免疫反应,显著抑制B16-HPV58E6E7肿瘤细胞的生长,在细胞免疫上优于单纯抗原,可作为HPV58阳性相关肿瘤及其癌前病变免疫治疗的候选疫苗.
目的 初步觀察PVAX1-HPV58mE6E7FcGB複閤DNA疫苗的抗腫瘤免疫效果.方法 在檢測該複閤疫苗的免疫效果前,自行構建瞭穩定錶達HPV58E6E7融閤基因的B16-HPV58E6E7成瘤細胞繫,錶達併純化瞭HPV58E6E7-GST融閤蛋白作為抗原.將該疫苗免疫C57BL/6小鼠(免疫組)後,通過抗腫瘤移植保護實驗和腫瘤生長抑製實驗觀察該疫苗對小鼠負荷的B16-HPV58E6E7移植瘤的防治效果；通過特異性抗體檢測、酶聯免疫斑點檢測、特異性殺傷實驗觀察該疫苗在被免疫小鼠體內誘髮的體液和細胞免疫反應.以單純抗原PVAX1-HPV58mE6E7 Fc免疫小鼠作為單純抗原組.結果 抗腫瘤移植保護實驗顯示,經PVAX1-HPV58mE6E7FcGB疫苗免疫的小鼠成瘤率僅為9/15,但成瘤時間最長[為(13.6±1.7)d],且腫瘤生長最慢.在腫瘤生長抑製實驗中,疫苗組小鼠的腫瘤生長抑製率達81.4％.體液免疫檢測顯示,該疫苗和單純抗原均能在小鼠體內誘髮特異性抗體,最高滴度分彆為1∶25 600和1∶12 800,兩組比較,差異無統計學意義(P＞0.05)；但在細胞免疫檢測中,疫苗組激活的特異性T淋巴細胞數[(219±34)箇/4×105箇脾淋巴細胞]約為單純抗原組[(55±25)箇/4×105箇脾淋巴細胞]的4倍,兩組比較,差異有統計學意義(P＜0.05)；且該疫苗誘髮的特異性針對B 16-HPV58 E6 E7細胞的殺傷率(最高為43.3％)也明顯高于單純抗原組(殺傷率最高為31.3％),兩組比較,差異有統計學意義(P＜0.05).結論 PVAX1-HPV58mE6E7FcGB複閤DNA疫苗能有效激髮特異性的體液和細胞免疫反應,顯著抑製B16-HPV58E6E7腫瘤細胞的生長,在細胞免疫上優于單純抗原,可作為HPV58暘性相關腫瘤及其癌前病變免疫治療的候選疫苗.
목적 초보관찰PVAX1-HPV58mE6E7FcGB복합DNA역묘적항종류면역효과.방법 재검측해복합역묘적면역효과전,자행구건료은정표체HPV58E6E7융합기인적B16-HPV58E6E7성류세포계,표체병순화료HPV58E6E7-GST융합단백작위항원.장해역묘면역C57BL/6소서(면역조)후,통과항종류이식보호실험화종류생장억제실험관찰해역묘대소서부하적B16-HPV58E6E7이식류적방치효과；통과특이성항체검측、매련면역반점검측、특이성살상실험관찰해역묘재피면역소서체내유발적체액화세포면역반응.이단순항원PVAX1-HPV58mE6E7 Fc면역소서작위단순항원조.결과 항종류이식보호실험현시,경PVAX1-HPV58mE6E7FcGB역묘면역적소서성류솔부위9/15,단성류시간최장[위(13.6±1.7)d],차종류생장최만.재종류생장억제실험중,역묘조소서적종류생장억제솔체81.4％.체액면역검측현시,해역묘화단순항원균능재소서체내유발특이성항체,최고적도분별위1∶25 600화1∶12 800,량조비교,차이무통계학의의(P＞0.05)；단재세포면역검측중,역묘조격활적특이성T림파세포수[(219±34)개/4×105개비림파세포]약위단순항원조[(55±25)개/4×105개비림파세포]적4배,량조비교,차이유통계학의의(P＜0.05)；차해역묘유발적특이성침대B 16-HPV58 E6 E7세포적살상솔(최고위43.3％)야명현고우단순항원조(살상솔최고위31.3％),량조비교,차이유통계학의의(P＜0.05).결론 PVAX1-HPV58mE6E7FcGB복합DNA역묘능유효격발특이성적체액화세포면역반응,현저억제B16-HPV58E6E7종류세포적생장,재세포면역상우우단순항원,가작위HPV58양성상관종류급기암전병변면역치료적후선역묘.
Objective To initially observe the antitumor immune of PVAX1-HPV58mE6E7FcGB composite DNA vaccine.Methods Before detecting immune effect of the vaccine,the B16-HPV58E6E7 tumor cell line was built which could steadily express HPV58E6E7 fusion gene.Then,HPV58E6E7-GST fusion protein as an antigen was expressed and purified.Before or after immunized with the vaccine,the C57BL/6 mice were challenged by B16-HPV58E6E7 cells.Anti-tumor transplantation and tumor growth inhibition experiment were performed to observe prevention and treatment effects on the vaccine.Specific humoral and cellular immune responses in the immunized mice were detected by ELISA,enzyme linked immunospot assay (ELISPOT) and cytotoxic T lymphocyte (CTL) method.Results In the anti-tumor transplantation experiment,tumor formation rate was only 9/15 in the mice which were immunized by PVAX1-HPV58mE6E7FcGB vaccine,time before tumor formation was the longest [(13.6 ± 1.7) days] and tumor growth was the slowest in the vaccine group.In tumor growth inhibition experiment,inhibition rate reached 81.4％ in the vaccine group.Except tumor formation rate,all data in the vaccine group was superior to the pure antigen PVAX1-HPV58mE6E7Fc group (P ＜ 0.05).Humoral immune effect showed that both the vaccine and the pure antigen could induce specific antibody in the immunized mice,and the highest titer were 1 ∶ 25600 and 1 ∶ 12800,respectively.Although there was not significant difference of antibody titer between the vaccine and the pure antigen group (P ＞ 0.05),the number of activated T cells in the vaccine group was almost four times as that in the pure antigen group [(219 ±34)/4 × 105 spleen lymphocytes versus (55 ±25)/4 × 105 spleen lymphocytes,P ＜ 0.05],and the highest specific CTL that vaccine induced was significantly higher than that of pure antigen (43.3％ versus 31.3％,P ＜ 0.05).Conclusions Humoral and cellular immune response could be effectively stimulated by PVAX1-HPV58mE6E7FcGB composite DNA vaccine.Growth of B16-HPV58E6E7 cells was significantly inhibited in the immunized mice.The cellular immune effect on the vaccine was superior to the pure antigen.Therefore,PVAX1-HPV58mE6E7FcGB could be used as a candidate vaccine for immune therapy to the HPV58 positive tumors and precancerous lesions.
