中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2008年
9期
822-827
,共6页
赵枫姝%褚莉莉%窦骏%唐权%王永仿%吴昀%曹明刚%潘猛%顾宁
趙楓姝%褚莉莉%竇駿%唐權%王永倣%吳昀%曹明剛%潘猛%顧寧
조풍주%저리리%두준%당권%왕영방%오윤%조명강%반맹%고저
小鼠IL-21%糖基化磷脂酰肌醇%抗肿瘤免疫
小鼠IL-21%糖基化燐脂酰肌醇%抗腫瘤免疫
소서IL-21%당기화린지선기순%항종류면역
Murine IL-21%Glycosyl phosphatidylinositol%Anti-tumor immunity
目的 构建糖基化磷脂酰肌醇(glycosyl phosphafidylinositol,GPI)修饰的小鼠IL-21瘤苗,并对此瘤苗的抗肿瘤效应及其机制作初步探讨.方法 通过重叠PCR方法获得IL-21-GPI融合基因并将其插入空载体pcDNA3.1.将鉴定过的重组载体以脂质体法转染B16F10细胞制成瘤苗,细胞间接免疫荧光法及流式细胞仪检测转染瘤细胞膜表面IL-21的表达,通过对小鼠脾细胞的增殖作用鉴定表达的IL-21的生物学活性.将瘤苗接种小鼠后,通过观察小鼠肿瘤体积和生存率分析瘤苗的抗瘤性,并检测了瘤苗免疫鼠的细胞免疫活性.结果 正确构建了pcDNA3.1/IL-21-GPI重组载体,膜表达的IL-21有良好的生物学活性,制备的瘤苗能发挥抗肿瘤效应,其机制与免疫鼠细胞免疫活性增强有关.结论 成功构建了具有抗肿瘤活性的GPI修饰的IL-21瘤苗,为其进一步抗肿瘤免疫治疗研究奠定了基础.
目的 構建糖基化燐脂酰肌醇(glycosyl phosphafidylinositol,GPI)脩飾的小鼠IL-21瘤苗,併對此瘤苗的抗腫瘤效應及其機製作初步探討.方法 通過重疊PCR方法穫得IL-21-GPI融閤基因併將其插入空載體pcDNA3.1.將鑒定過的重組載體以脂質體法轉染B16F10細胞製成瘤苗,細胞間接免疫熒光法及流式細胞儀檢測轉染瘤細胞膜錶麵IL-21的錶達,通過對小鼠脾細胞的增殖作用鑒定錶達的IL-21的生物學活性.將瘤苗接種小鼠後,通過觀察小鼠腫瘤體積和生存率分析瘤苗的抗瘤性,併檢測瞭瘤苗免疫鼠的細胞免疫活性.結果 正確構建瞭pcDNA3.1/IL-21-GPI重組載體,膜錶達的IL-21有良好的生物學活性,製備的瘤苗能髮揮抗腫瘤效應,其機製與免疫鼠細胞免疫活性增彊有關.結論 成功構建瞭具有抗腫瘤活性的GPI脩飾的IL-21瘤苗,為其進一步抗腫瘤免疫治療研究奠定瞭基礎.
목적 구건당기화린지선기순(glycosyl phosphafidylinositol,GPI)수식적소서IL-21류묘,병대차류묘적항종류효응급기궤제작초보탐토.방법 통과중첩PCR방법획득IL-21-GPI융합기인병장기삽입공재체pcDNA3.1.장감정과적중조재체이지질체법전염B16F10세포제성류묘,세포간접면역형광법급류식세포의검측전염류세포막표면IL-21적표체,통과대소서비세포적증식작용감정표체적IL-21적생물학활성.장류묘접충소서후,통과관찰소서종류체적화생존솔분석류묘적항류성,병검측료류묘면역서적세포면역활성.결과 정학구건료pcDNA3.1/IL-21-GPI중조재체,막표체적IL-21유량호적생물학활성,제비적류묘능발휘항종류효응,기궤제여면역서세포면역활성증강유관.결론 성공구건료구유항종류활성적GPI수식적IL-21류묘,위기진일보항종류면역치료연구전정료기출.
Objective To construct the murine IL-21 (mIL-21) tumor vaccine modified by glyco-syl phosphafidylinositol(GPI), and to evaluate its anti-tumor effect and mechanisms. Methods The IL-21-GPI gene was acquired by overlap PCR and inserted into PeDNA3.1. The recombinant plnsmid pcDNA3.1/ IL-21-GPI was transformed into cell B16F10, and the expression of mIL-21 on cell membrane was deter-mined by cell indirect immumofluorescence and flow cytometry (FCM). The bioactivity of mIL-21 was iden-tiffed according to its effects on the proliferation of mouse spleen cells. The anti-tumor effect was evaluated depending on the tumor size and the survival of tumor-beating mice after the tumor vaccine was inoculated into C57BL/6 mice. And the activity of cell-mediated immunity in immunized mice was detected at the same time. Results The recombinant plasmid pcDNA3.1/IL-21-GPI was correctly constructed, which could ex-press mIL-21 binding the membrane with good bioactivity. The vaccine had good anti-tumor effect, and the cell-mediated immunity had been improved in immunized mice. Conclusion The GPI modified mIL-21 tumor vaccine with anti-tumor activity was constructed successfully, which provided a good foundation for studying anti-tumor immunity and therapy in future.
