Anti-tumoural activity of peripheral blood mononuclear cells against melanoma cells: discrepant in-vitro and in-vivo effects
Anti-tumoural activity of peripheral blood mononuclear cells against melanoma cells: discrepant in-vitro and in-vivo effects
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DOI：
10.1097/01.cmr.0000205016.31235.a9
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发表时间：
2006-08-01
影响因子：
2.2
通讯作者：
Albert, Philippe
中科院分区：
文献类型：
--
作者：
Joseph-Pietras, Debora;Carlier, Annie;Albert, Philippe
由于肿瘤细胞通过多种机制逃避化疗药物，本研究使用小鼠黑色素瘤细胞亚线耐多柔比星（B16R）来检测小鼠外周血单核细胞的抗肿瘤活性。已知多细胞球体是最适合模拟体内实体肿瘤的体外模型，并用于研究肿瘤生物学的许多方面。体外实验中，将尾侧穿刺后经Ficoll梯度离心恢复的小鼠外周血单个核细胞与B16R多细胞肿瘤球细胞共培养。形态学观察表明，共培养14 h后，外周血单核细胞聚集并聚集在球体周围，32 h内建立了接触。共培养38 ~ 62 h，球体大小明显减小。外周血单核细胞发挥与B16R黑素瘤细胞球体细胞死亡诱导相关的细胞溶解作用。对发挥抗肿瘤作用的外周血单核细胞进行定位和鉴定的免疫学研究表明，球体被单核/巨噬细胞深度浸润，在此阶段观察到显著的细胞溶解活性和较高的细胞死亡率。在体内研究中，肿瘤内注射了同基因的原始外周血单个核细胞。观察到这些细胞有微弱的潜在体内抗肿瘤作用（抑制B16R黑色素瘤生长20-25%），但与未处理的对照小鼠相比，外周血单个核细胞处理小鼠的中位生存时间没有增加。因此，尽管外周血单核细胞在体外对低免疫原性和高转移性耐药B16黑色素瘤细胞具有抗肿瘤活性，但在体内潜在的抗黑色素瘤作用，如果存在的话，并没有增加B16R黑色素瘤小鼠的寿命。
As tumour cells use multiple mechanisms to escape from chemotherapeutic drugs, the anti-tumoural activity of naive mouse peripheral blood mononuclear cells was examined in this study, using a mouse melanoma cell subline resistant to doxorubicin (B16R). Multicellular spheroids are known to be the most adapted in-vitro model to mimic solid tumours in vivo and are used to investigate many aspects of tumour biology. For in-vitro studies, murine peripheral blood mononuclear cells recovered by Ficoll gradient centrifugation after caudal puncture were co-cultured with multicellular tumour spheroids of B16R cells. Morphological investigations show that peripheral blood mononuclear cells were gathered and focused around the spheroids after 14 h of co-culture and contacts were established within 32 h. Between 38 and 62 h of co-culture, the size of the spheroids decreased significantly. The peripheral blood mononuclear cells exerted cytolytic effects that correlated with the induction of cell death in spheroids of B16R melanoma cells. Immunological investigations to localize and identify peripheral blood mononuclear cells that exerted anti-tumoural effects have shown that spheroids were deeply infiltrated by monocytes/macrophages at a stage in which a significant cytolytic activity and a strong cell death rate were observed. For in-vivo studies, intratumoural injections of syngeneic naive peripheral blood mononuclear cells were administered. A weak potential in-vivo anti-tumoural effect of these cells was observed (inhibition of B16R melanoma growth by 20-25%) but the median survival time of mice treated with peripheral blood mononuclear cells did not increase compared with untreated control mice. Thus, despite anti-tumoural activities of peripheral blood mononuclear cells against the poorly immunogenic and highly metastatic chemoresistant B16 melanoma cells in vitro, a potential anti-melanoma effect in vivo, if present, did not increase the life span of B16R melanoma-bearing mice.