The liposome-incorporating cell wall skeleton of Mycobacterium bovis bacillus Calmette-Guein can directly enhance the susceptibility of cancer cells to lymphokine-activated killer cells through up-regulation of natural-killer group 2, member D ligands
The liposome-incorporating cell wall skeleton of Mycobacterium bovis bacillus Calmette-Guein can directly enhance the susceptibility of cancer cells to lymphokine-activated killer cells through up-regulation of natural-killer group 2, member D ligands
复制标题
DOI：
10.1111/j.1464-410x.2010.10056.x
复制
发表时间：
2011-11-01
影响因子：
4.5
通讯作者：
Akaza, Hideyuki
中科院分区：
文献类型：
--
作者：
Miyazaki, Jun;Kawai, Koji;Akaza, Hideyuki
目的：对自然杀伤细胞裂解膀胱癌细胞的能力进行临床前评价，所述膀胱癌细胞经R8-脂质体-卡介苗（BCG）-细胞壁骨架（CWS）处理而被修饰以显示增强的自然杀伤组2，成员D（NKG 2D）配体表达。用常规制备试剂盒获得外周血单个核细胞，将纯化的外周血单个核细胞经白细胞介素2（IL-2）刺激诱导产生淋巴因子激活的杀伤（LAK）细胞，观察LAK细胞对未经处理的小鼠和R8-脂质体-BCG-CWS共转染小鼠的抗肿瘤作用。使用细胞毒性WST-8测定方法，在培养4小时时，在各种效应物/靶（E：在T24细胞和RT-112细胞上，用BCG处理后，主要组织相容性复合物I类相关链B（MIC B）的表达增加了约1.5倍。在T24细胞和RT-112细胞上，用BCG也使结合蛋白（ULBP）1表达增加约1.5倍。R8-脂质体-BCG-CWS使MICB在T24细胞上的表面表达增加2.2倍，但在RT-112细胞上没有显著增加。ULBP 1在RT-112细胞上的表达增加约2.2倍，尽管在用R8-脂质体-BCG-CWS共培养的T24细胞中，在ULBP 2和3的表达之间没有观察到差异，CWS显示在E：T比为4时，对LAK细胞的细胞溶解敏感性增加约1.3倍，RT-112细胞显示在E：T比为4时，对LAK细胞的细胞溶解敏感性增加约1.4倍。结论在本研究中，R8-脂质体-BCG-CWS对表面NKG 2D配体的诱导使癌细胞对LAK细胞的细胞溶解更敏感，T24细胞和RT-112细胞，即使在没有免疫细胞的情况下单独培养时，R8-脂质体-BCG-CWS也能直接应答。因此，本研究中获得的结果可能表明一种新的针对膀胱癌的过继免疫疗法。
OBJECTIVETo conduct a preclinical evaluation of the ability of natural killer cells to cytolyze bladder cancer cells that were modified to show enhanced expression of natural-killer group 2, member D (NKG2D) ligands by R8-liposome-bacillus Calmette-Guein (BCG)-cell wall skeleton (CWS) treatment.MATERIALS AND METHODSThe T24 cells and RT-112 cells were co-cultured with R8-liposome-BCG-CWS and BCG for 2, 4, or 6 h, and then the surface expression of NKG2D ligands was analyzed using TaqMan real-time quantitative RT-PCR.Peripheral blood mononuclear cells were obtained with a conventional preparation kit, and then lymphokine-activated killer (LAK) cells were generated from these purified peripheral blood mononuclear cells via interleukin-2 stimulation.The anti-tumour effect of LAK cells against untreated and R8-liposome-BCG-CWS co-cultured with cells of the human bladder cancer cell lines T24 and RT-112 was analyzed using the cytotoxic WST-8 assay method at 4 h of culture at various effector/target (E : T) ratios.RESULTSMajor histocompatibility complex class I-related chain B (MICB) expression was increased approximate to 1.5-fold on T24 cells and RT-112 cells with BCG.UL-16-binding protein (ULBP) 1 expression was also increased approximate to 1.5-fold on T24 cells and RT-112 cells with BCG. R8-liposome-BCG-CWS increased the surface expression of MICB 2.2-fold on T24 cells but did not increase it significantly on RT-112 cells.ULBP1 expression was increased approximate to 2.2-fold on RT-112 cells, although no differences were observed between the expression of ULBP2 and 3 with R8-liposome-BCG-CWS.T24 cells that were co-cultured with R8-liposome-BCG-CWS showed an approximate to 1.3-fold increase in sensitivity to cytolysis by LAK cells at an E : T ratio of 4 and RT-112 cells showed an approximate to 1.4-fold increase at an E : T ratio of 2.CONCLUSIONSIn the present study, the induction of surface NKG2D ligands by R8-liposome-BCG-CWS rendered cancer cells more susceptible to cytolysis by LAK cells.T24 cells and RT-112 cells, even when cultured singly in the absence of immune cells, can directly respond to R8-liposome-BCG-CWS.The results obtained in the present study may therefore indicate a novel adoptive immunotherapy against bladder cancers.