Cytotoxic sinusoidal lymphocytes of liver transplant are effective against circulating tumor cells
Cytotoxic sinusoidal lymphocytes of liver transplant are effective against circulating tumor cells
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DOI：
10.1016/s0041-1345(98)01789-8
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发表时间：
1999-02-01
影响因子：
0.9
通讯作者：
Lukomska, B
中科院分区：
文献类型：
--
作者：
Durowicz, S;Olszewski, WL;Lukomska, B
材料与方法选用体重250 ~ 300 g的SWistar（W/Wag）（RT 1u）大鼠。通过将106个CC 531结肠癌细胞注射到门静脉中诱导肝肿瘤。CC 531是1，2-二甲基肼诱导的、中等分化的、弱免疫原性的结肠腺癌，与WAG/Rij大鼠同源。18细胞由荷兰鹿特丹大学医院-Dijkzigt的RL Marquet博士友情提供。3周后的总肝脏肿瘤质量平均为15.1 g（范围为5.2 - 20.7 g;平均占总肝脏重量的56.5%）。通过门静脉洗出正常W/Wag和具有肝CC 531转移的W/Wag的肝，并在Lymphoprep梯度（Nyegaard & Co A/S，Norway）上从灌注液中分离淋巴细胞。从正常大鼠中回收的淋巴细胞被命名为肝相关淋巴细胞1（LAL 1 n），从荷瘤大鼠中回收的淋巴细胞被命名为LAL 1 t。下一步，在37 ℃的温度下，在具有蠕动泵、氧合器和热交换器的体外肝灌注模型中，用来自正常大鼠的同系血液原位灌注肝脏1小时。灌注后再次冲洗肝脏血管，分离正常大鼠（LAL 2n）和荷瘤大鼠（LAL 2 t）积聚的淋巴细胞，测量转移瘤和正常肝窦中冲洗出的血细胞数量。此外，使用针对W3/13（CD 43）-（白唾液酸蛋白抗原）、W3/25（CD 4）-辅助/诱导细胞、OX 8（CD 8）-抑制/细胞毒性细胞、OX 19（CD 5）-T细胞的单克隆抗体，3.2进行洗出细胞的表型特征。3-（CD 56）NK细胞（Serotec，UK）。用铬酸钠（阿默舍姆，UK）标记的靶细胞（3 × 106）进行洗出的细胞对CC 531和K562（红白血病系）肿瘤细胞的细胞毒性，以使效应物与靶物的比率为80：1至5：1。数据表示为每个样品的51 Cr释放百分比。
MATERIALS AND METHODSWistar (W/Wag)(RT1u) rats with body weight 250 to 300 g were used. Liver tumors were induced by injection of 106 CC531 colon cancer cells into the portal vein. CC531 is a 1, 2-dimethylhydrazineinduced, moderately differentiated, and weakly immunogenic adenocarcinoma of the colon, syngeneic with WAG/Rij rats. 18 The cells were kindly provided by Dr RL Marquet, University Hospital-Dijkzigt, Rotterdam, The Netherlands. The total liver tumor mass after 3 weeks was on the average 15.1 g (range from 5.2 to 20.7 g; mean 56.5% of total liver weight). Livers of normal W/Wag and W/Wag with liver CC531 metastases were washed out through the portal vein, and lymphocytes were isolated from the perfusion ffuid on Lymphoprep gradient (Nyegaard & Co A/S, Norway). Lymphocytes retrieved from normal rats were named liver-associated-lymphocytes 1 (LAL1n) and those from tumor bearing rats (LAL1t). As the next step, livers were perfused in situ for 1 hour with syngeneic blood from normal rats in an extracorporal liver perfusion model with peristaltic pump, oxygenator, and heat exchanger at the temperature of 37 C. After perfusion, liver vasculature was washed out again, and the accumulating lymphocytes from normal rats (LAL2n) and tumor bearing rats (LAL2t) were isolated.The number of blood cells washed out from metastatic and normal liver sinusoids was measured. Furthermore, a phenotypical characteristic of washed out cells was performed using monoclonal antibodies against W3/13 (CD43)-(leucosialin antigen), W3/25 (CD4)-helper/inducer cells, OX8 (CD8)-suppressor/cytotoxic cells, OX19 (CD5)-T cells, 3.2. 3-(CD56) NK cells (Serotec, UK). Cytotoxicity of washed-out cells against CC531 and K562 (erythroleukemic line) tumor cells was carried out with target cells (3 106) labelled with sodium chromate (Amersham, UK) to make the effector to target ratio from 80: 1 to 5: 1. The data were expressed as the percentage of 51Cr-release for each sample.