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作 者:孙薏[1] 陈健[1] 蔡鹏[2] 呼永河[1] 钟国成[1] 冯怀志[1] 闵敏[1] 李硕[1] 张聪[1]
机构地区:[1]解放军第452医院肿瘤血液中心,四川成都610021 [2]北京市道培医院,北京100049
出 处:《中国实验血液学杂志》2010年第1期219-223,共5页Journal of Experimental Hematology
基 金:四川省卫生厅科学研究项目;编号050287;050288
摘 要:本研究探讨负载自身肿瘤裂解物的树突状细胞(dendritic cells,DC)与细胞因子诱导的杀伤细胞(cytokine-induced killers,CIK)共培养后,治疗复发难治性非霍奇金淋巴瘤患者的临床免疫学疗效、毒副反应。选择9例复发难治性非霍奇金氏淋巴瘤患者,分离外周血单个核细胞,贴壁细胞体外经GM-CSF和IL-4诱导产生DC,并负载自体肿瘤细胞裂解物,悬浮淋巴细胞经IFN-γ、IL-2、抗CD3单克隆抗体、IL-1α体外诱导产生CIK细胞。7天后将负载抗原的DC与CIK共培养,12天后回输体内。利用流式细胞术、T淋巴细胞核仁形成区嗜银蛋白(Ag-NORs)的检测及ELISA双抗夹心法检测过继免疫生物治疗前后各项免疫指标的变化,并观察临床疗效。结果表明:经细胞治疗后患者外周淋巴细胞Ag-NOR的IS值明显增高,呈上升趋势;CIK细胞培养后可见CD3+CD8+和CD3+CD56+细胞较培养前显著增加(p<0.01),负载肿瘤抗原的DC致敏后CD3+CD8+和CD3+CD56+的表达较之单纯的CIK进一步明显提高(p<0.01);将负载自身肿瘤抗原的DC与CIK共培养后,上清液中IFN-γ、IL-12的水平显著高于单独CIK对照组(p<0.05)。5例患者通过影像学复查显示肿瘤较治疗前明显缩小;除一过性发热、畏寒外未见其它不良反应。结论:负载自身抗原的DC-CIK细胞的具有特异性杀伤淋巴瘤细胞的能力;外周淋巴细胞Ag-NOR的IS值明显增高,分泌细胞因子水平高于单纯CIK细胞,临床治疗效果令人满意。This study was aimed to investigate the killing activity of cytokine-induced killer (CIK) cells after being incubated with autologous tumor cell lysate-pulsed dendritic cells (DC) and to evaluate the clinical efficacy and side effect of autologous tumor cell lysate-loaded DC in combination with CIK on relapsed or refractory non-Hodgkin's lymphoma (NHL). Peripheral blood mononuclear cells (PBMNC) were isolated from 9 patients with NHL, and cultured with granulocyte- macrophage colony-stimulating factor(GM-CSF)and interleukin-4(IL-4) to produce DC. The DC were pulsed with autologous tumor cell lysate. T lymphocytes from PBMNC were cultured with interferon-'y(IFN--,/), IL-2, CD3-moAb, and IL-lc~ to prepare CIK. After receiving the immunotherapy of DC and CIK, immunologic and clinical responses were evaluated. The results showed that the AgNOR, CD3+ CD8+ and CD3+CD56+ ratio were markedly improved after the imrnunotberapy (p 〈 0.01) ; IFN-,,/and IL-12 levels in supernatant of DC-CIK group were higher than that in CIK group (p 〈0.01) ; Tumor size were significantly decreased after the imrnunotherapy (p 〈 0. 05). Except transient fever and chill, no remarkable adverse event happened during or after the treatment. It is concluded that the autologous tumor cell lysate-pulsed DC in combination with CIK show ability to specifically kill the lymphoma cells, obviously increases the IS value of Ag-NOR in peripheral lymphocytes, secretes cytokines higher than CIK cells alone. This combination displays the short-term satisfied efficacy on NHL through inducing specific antitumor immunity ,and can be used as an effective adjuvant measure for the routine therapy of NHL.
关 键 词:非霍奇金淋巴瘤 树突状细胞 细胞因子诱导杀伤细胞 肿瘤裂解物 免疫治疗
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