6、5×106、10×106個(gè)CAR-T,每只一次性尾iv 100 μL細(xì)胞懸液)。采用活體成像技術(shù)檢測(cè)CAR-T治療腫瘤的消退效果;酶聯(lián)免疫吸附實(shí)驗(yàn)檢測(cè)血漿中γ-干擾素(IFN-γ)水平;觀察小鼠一般狀態(tài)、體溫、體質(zhì)量;治療后14 d對(duì)小鼠進(jìn)行解剖和采樣,利用RNAscope技術(shù)檢測(cè)CAR-T在小鼠組織的特異性分布。結(jié)果 成功建立了多發(fā)性骨髓瘤NSG小鼠移植腫瘤模型;在輸注BCMA-CAR-T后3、7、12 d,生物發(fā)光成像顯示,CAR-T組小鼠平均光子強(qiáng)度顯著低于細(xì)胞外液、MOCK-T組(P<0.05),中、高劑量組的治療效果明顯高于低劑量組(P<0.05);CAR-T組的IFN-γ分泌水平顯著高于細(xì)胞外液、MOCK-T組(P<0.05);記錄CAR-T治療組小鼠的一般情況,未見(jiàn)明顯不良反應(yīng);RNAscope檢測(cè)到CAR-T在輸注14 d后少量特異性分布于骨髓和脾臟。結(jié)論 本研究驗(yàn)證了BCMA-CAR-T抗多發(fā)性骨髓瘤藥效明顯,安全可靠,RNAscop技術(shù)可應(yīng)用于CAR-T分布檢測(cè)。;Objective Choose the appropriate mouse model, evaluate BCMA-CAR-T anti-tumor effects, and explore the application of RNAscope in CAR-T cell therapy. Methods MM. 1s-luc multiple myeloma mouse model was established in NSG mice with severe immunodeficiency, they were randomly divided into extracellular fluid group (100 μL PBS buffer per mouse), MOCK-T group (100 μL untransfected T cells), low, medium and high dose groups of CAR-T (1×106, 5×106, 10×106 CAR-T per mouse, and 100 μL cell suspension of one-off tail iv). In vivo imaging was performed to detect the tumor regression effect of CAR-T treatment. The level of interferon-gamma (IFN-gamma) in plasma was detected by ELISA. General clinical observation, body temperature, body weight monitoring were performed after CAR-T was given once. Mice were dissected and sampled 14 days after infusion. CAR-T distribution in the spleen and bone marrow of mice was detected by RNAscope technology, and the specific distribution of CAR-T in various tissues was detected. Results Successfully established many found myeloma NOD/SCID mice transplanted tumor model, At 3, 7 and 12 days after BCMA-CAR-T infusion, bioluminescence imaging showed that the average photon intensity of mice in CAR-T group was significantly lower than that in extracellular fluid and MOCK-T group (P<0.05), and the therapeutic effect in medium and high dose groups was significantly higher than that in low dose groups (P<0.05); the level of IFN-gamma secretion in CAR-T group was significantly higher than that in extracellular fluid and MOCK-T group (P<0.05); the general situation of mice in CAR-T group was recorded, but no significant difference was observed. Adverse reactions:RNA scope detected that CAR-T was localized in bone marrow and spleen 14 days after infusion. Conclusion The BCMA-CAR-T has been proven to be safe and reliable by some non-clinical evaluation methods. RNAscop technology can be applied to CAR-T cell distribution detection."/>