CAJ | 학술논문

目的应用活体动物体内光学成像系统观察抗体药物Rituximab在荷淋巴瘤裸鼠体内的生物分布.方法制备FITC标记的Rituximab(FITC-Rituximab),用激光扫描共聚焦显微镜和流式细胞仪体外分析FITC-Rituximab与人淋巴瘤Raji细胞的亲和力;建立裸鼠淋巴瘤皮下移植瘤模型,应用活体动物体内光学成像系统观察FITC-Rituximab在荷瘤小鼠体内的分布.结果流式细胞仪和激光扫描共聚焦显微镜检测证明FITC-Rituximab与淋巴瘤Raji细胞有较好的亲合活性,主要结合在细胞膜表面.体内活体动物光学成像分析表明,FITC-Rituximab在1 h内即可特异性地在肿瘤部位富集,3～4 h即可进入肿瘤组织内部并达到最大浓度富集,8～10 h后在肿瘤组织仍可观察到FITCRituximab的特异性的富集,而在其他组织器官没有可见的荧光存在;双侧皮下移植瘤模型再次证明FITC-Rituximab具有对CD20抗原过表达的淋巴瘤特异性结合能力.结论动物活体成像系统能够准确地监测FITC标记Rituximab在荷瘤裸鼠体内的动态分布情况,该技术对实时分析抗体药物在荷瘤小鼠体内的靶向效果具有参考价值和指导意义.
목적 응용활체동물체내광학성상계통관찰항체약물Rituximab재하림파류라서체내적생물분포.방법 제비FITC표기적Rituximab(FITC-Rituximab),용격광소묘공취초현미경화류식세포의체외분석FITC-Rituximab여인림파류Raji세포적친화력;건립라서림파류피하이식류모형,응용활체동물체내광학성상계통관찰FITC-Rituximab재하류소서체내적분포.결과 류식세포의화격광소묘공취초현미경검측증명FITC-Rituximab여림파류Raji세포유교호적친합활성,주요결합재세포막표면.체내활체동물광학성상분석표명,FITC-Rituximab재1 h내즉가특이성지재종류부위부집,3～4 h즉가진입종류조직내부병체도최대농도부집,8～10 h후재종류조직잉가관찰도FITCRituximab적특이성적부집,이재기타조직기관몰유가견적형광존재;쌍측피하이식류모형재차증명FITC-Rituximab구유대CD20항원과표체적림파류특이성결합능력.결론 동물활체성상계통능구준학지감측FITC표기Rituximab재하류라서체내적동태분포정황,해기술대실시분석항체약물재하류소서체내적파향효과구유삼고개치화지도의의.
Objective To conduct an in vivo optical imaging analysis of the biodistribution of antibody Rituximab in lymphoma tumor-bearing nude mice. Methods Laser scanning confocal microscope and flow cytometry were employed to determine the affinity of FITC-labeled Rituximab (FITC-Rituximab)with human lymphoma Raji cells. And the in vivo optical imaging system was used to analyze the biodistribution of FITC-Rituximab in lymphoma-transplanted xenograft nude mice. Results The results of flow cytometry and laser scanning confocal microscope demonstrated that FITC-Rituximab had remarkable affinity with lymphoma Raji cells and was mainly bound at cell membrane. The results of in vivo imaging analysis suggested that FITC-Rituximab could specifically accumulated at peritumor tissue less than 1 h, then penetrated into the interior of tumor and concentrated in 3-4 h. And the specific concentration of FITCRituximab could still been observed more than 8-10 h whereas there was no apparent fluorescence at other tissues. Furthermore, the results observed from a two-flank tumor xenograft model showed that FITCRituximab possessed specific binding affinity for CD20-overexpressed lymphoma. Conclusion The in vivo optical imaging system can accurately monitor the distribution of FITC-Rituximab in tumor-bearing nude mice. And this technique has a reference value and significance for a real-time analysis of tumor-targeting capability of antibody drugs.