[摘 要] 目的：探讨构建靶向CD7抗原的嵌合共刺激受体（CCR）并制备该受体修饰的健康供者来源γδ T细胞，以评估其对急性T淋巴细胞白血病（T-ALL）的体内与体外杀伤作用。方法：构建携带CD7-DAP10-CCR的慢病毒载体，并转导健康人外周血γδ T细胞，制备靶向CD7抗原的CCR γδ T细胞（CD7-DAP10-CCR-γδ T），所获细胞借助表达CD64、CD86和CD137L的人工抗原提呈细胞（aAPC）进行体外扩增。采用Annexin Ⅴ/7-AAD方法检测CD7-DAP10-CCR-γδ T对T-ALL细胞（Jurkat）、CD7缺陷型Jurkat细胞（CD7⁻ Jurkat）及异体健康人原代αβ T细胞的体外杀伤作用，共设置了3组效靶比（E∶T = 1∶1、1∶3和1∶10），孵育时间为18~24 h。其中，Jurkat细胞作为CD7阳性靶细胞，CD7⁻ Jurkat作为CD7阴性靶细胞以验证杀伤特异性，而异体健康人原代αβ T细胞则作为CD7阳性正常细胞对照，用于评估CD7-DAP10-CCR-γδ T细胞的脱靶效应。此外，在T-ALL荷瘤免疫缺陷小鼠体内验证药效，通过定期对荷瘤免疫缺陷小鼠进行活体成像、体质量检测及生存期观察，评估CD7-DAP10-CCR-γδ T细胞在荷瘤免疫缺陷小鼠体内的药效。结果：成功利用aAPC体外制备出CD7-DAP10-CCR-γδ T细胞，其平均扩增倍数超过10 000倍。体外杀伤实验表明，该细胞对T-ALL细胞具有较高的杀伤能力（P < 0.01），对Jurkat 细胞具有较高的毒性（P < 0.01），对CD7⁻ Jurkat细胞杀伤作用有限，而对高表达CD7的正常原代αβ T细胞基本无杀伤作用；荷瘤免疫缺陷小鼠体内药效试验结果显示，相对于对照PBS组，经CD7-DAP10-CCR-γδ T细胞治疗后，荷瘤免疫缺陷小鼠的生存期显著延长（P < 0.01）。结论：体外借助aAPC能成功制备CD7-DAP10-CCR-γδ T细胞，并且CD7-DAP10-CCR-γδ T细胞在体外、小鼠体内均对T-ALL细胞具有较强的杀伤作用，表明CD7-DAP10-CCR-γδ T细胞具备对T-ALL的治疗潜力。

Objective : To investigate the effect of overexpression of tyrosine kinase receptor 1 (Tie-1) in cervical cancer cells on the malignant biological behavior of tumor-related endothelial cells (TRECs) ． Methods : Immuno- histochemical method was used to detect the expression of Tie-1 in cervical cancer cells ( CCCs) and TRECs of 96 patients with cervical cancer，and to analyze the correlation between the expression of Tie-1 in TRECs and clinico- pathological features and prognosis of patients．HeLa cells overexpressing Tie-1 (Hela-Tie1OE) were constructed， and HeLa-Tie1OE was co-cultured with human umbilical vein endothelial cells ( HUVECs) by Transwell cell co- culture method to obtain cervical cancer TRECs．Western blot was used to analyze Tie-1 protein expression．The migration，invasion and tubulogenesis of TRECs were detected by cell scratch assay，Transwell invasion and migra- tion assay and tubulogenesis assay. Results : The expression of Tie-1 was positively correlated with CCCs and TRECs in 96 cervical cancer patients．The positive expression rate of Tie-1 in TRECs of patients with stage(FIGO) Ⅰ B2-ⅡA，tumor diameter ≥4 cm ，cervical muscle invasion depth ≥ 1 /2 full layer ，adenocarcinoma ，medium and low differentiation cervical cancer was higher than that of patients with Ⅰ A 1-Ⅰ B 1，tumor diameter＜4 cm， cervical muscle invasion depth ＜1 /2 full layer，squamous carcinoma，and high differentiation cervical cancer，re- spectively．The differences were statistically significant (P＜0. 05) ．The expression of Tie-1 in TRECs of cervical cancer patients had no significant correlation with age，lymph node metastasis and lymphatic space invasion．The positive expression of Tie-1 in cervical cancer TRECs was negatively correlated with 5-year progression-free survival time and overall survival time (P＜0. 05) ．The Tie-1 expression of TRECs obtained by co-culture of Hela-Tie1OE and HUVECs was up-regulated ，and the invasion ，migration and tubulogenesis of TRECs cells were enhanced. Conclusion The high expression of Tie-1 in TRECs of cervical cancer is related to FIGO stage，tumor diameter， degree of differentiation，depth of cervical muscular invasion，type of cervical cancer，and poor prognosis of pa- tients．Overexpression of Tie-1 can promote TRECs invasion，migration and tubulogenesis.