Lebbé C, de Crémoux P, Millot G, Podgorniak M P, Verola O, Berger R, Morel P, Calvo F
Laboratory of Pharmacology, Hôpital Saint-Louis, Paris, France.
Arch Dermatol Res. 1997 Jun;289(7):421-8. doi: 10.1007/s004030050215.
We established long-term cultures from skin tumors of nine patients suffering from classical Kaposi's sarcoma (KS). Spindle cells obtained after enzymatic digestion were cultured on gelatin- or fibronectin-coated flasks in DMEM with 15% fetal calf serum, aFGF and heparin. Immunohistochemical staining was positive for MHC class I, laminin, type IV collagen, vimentin, alpha smooth muscle actin (20-40% of cells), caldesmon (20%), calponin (20-40%) and smooth muscle myosin (20-40%), and was negative for common leukocyte antigen, CD4, LFA1, CD34 and cytokeratin. Around 20% of cells up to the third passage in culture expressed the endothelial markers CD36, BMA 120 but were negative for UEA and Fc von Willebrand. Smooth muscle proteins were detected with immunoblotting. Using the polymerase chain reaction, human herpes virus 8 (HHV8) sequences were detected in primary cultures of three out of seven cell lines but were rapidly lost during in vitro passaging. KS-derived cells did not proliferate in serum-free medium, had a normal karyotype and did not grow in soft agar medium. Tumors formed in nude mice injected with KS-derived cells. The tumors were composed of mouse cells and were highly vascularized. Our results suggest that KS-derived cells are heterogeneous: the majority of cells have either a smooth muscle cell or a fibroblastic phenotype. Another minor cell compartment was composed of endothelium-derived cells. KS cells do not possess the characteristics of transformed cells in vitro and may be composed of polyclonal activated cells. Recombinant alpha interferon (rIFN) slightly inhibited the growth of KS-derived cells and increased the expression of MHC class I antigens. While cells were resistant to natural killer (NK) cell-mediated cytotoxicity, they became sensitive to rIFN-primed NK cells. Thus, the antitumor potential of rIFN against KS in vivo could result from immunomodulatory rather than from direct antiproliferative effects.
我们从9例经典型卡波西肉瘤（KS）患者的皮肤肿瘤中建立了长期培养体系。酶消化后获得的梭形细胞在涂有明胶或纤连蛋白的培养瓶中，于含有15%胎牛血清、碱性成纤维细胞生长因子（aFGF）和肝素的DMEM培养基中培养。免疫组织化学染色显示，MHC I类分子、层粘连蛋白、IV型胶原、波形蛋白、α平滑肌肌动蛋白（20 - 40%的细胞）、钙调蛋白（20%）、钙结合蛋白（20 - 40%）和平滑肌肌球蛋白（20 - 40%）呈阳性，而普通白细胞抗原、CD4、淋巴细胞功能相关抗原1（LFA1）、CD34和细胞角蛋白呈阴性。在培养至第三代的细胞中，约20%的细胞表达内皮细胞标志物CD36、BMA 120，但对荆豆凝集素（UEA）和血管性血友病因子（Fc von Willebrand）呈阴性。通过免疫印迹法检测到了平滑肌蛋白。利用聚合酶链反应，在7个细胞系中的3个原代培养物中检测到了人类疱疹病毒8（HHV8）序列，但在体外传代过程中迅速丢失。KS来源的细胞在无血清培养基中不增殖，具有正常的核型，且在软琼脂培养基中不生长。将KS来源的细胞注射到裸鼠体内可形成肿瘤。这些肿瘤由小鼠细胞组成，且血管高度丰富。我们的结果表明，KS来源的细胞具有异质性：大多数细胞具有平滑肌细胞或成纤维细胞表型。另一个较小的细胞部分由内皮来源的细胞组成。KS细胞在体外不具备转化细胞的特征，可能由多克隆活化细胞组成。重组α干扰素（rIFN）对KS来源细胞的生长有轻微抑制作用，并增加了MHC I类抗原的表达。虽然细胞对自然杀伤（NK）细胞介导的细胞毒性具有抗性，但它们对经rIFN预处理的NK细胞变得敏感。因此，rIFN在体内对KS的抗肿瘤潜力可能源于免疫调节作用而非直接的抗增殖作用。
