Lopez M, Amorim L, Gane P, Cristoph A, Bardinet D, Abina A M, Minty A, Bernard J
Faculté de Médecine, Inserm U76, Paris, France.
J Immunol Methods. 1997 Oct 27;208(2):117-29. doi: 10.1016/s0022-1759(97)00133-6.
Dendritic cells (DCs), which are antigen presenting cells of potential use in human antitumor vaccination trials, are presently the subject of intense investigation. Many recent studies have reported the possibility of generating ex vivo large numbers of DCs with high antigen presenting capacity by the culture of bone marrow or blood progenitors. In this study, we examined the differentiation into DCs of CD34+ progenitors isolated from the G-CSF mobilized blood of 3 healthy donors and 5 patients with breast cancer and cultured in the presence of GM-CSF + IL-13. The characteristics of the cells were compared to those of cells obtained in the presence of GM-CSF + TNF alpha. By day 15, one third of the bulk cells cultured with IL-13 were CD1a+/CD14- and strongly expressed CD1c, CD40, CD80 and HLA-DR. In contrast, cells obtained with TNF alpha expressed CD1a on one in three cells but with a considerably lower fluorescence intensity than on IL-13-cultured cells and strongly expressed CD14 on more than 50% of cells. CD1a+/CD14- cells emerged in IL-13 cultures at day 5, while in TNF alpha cultures CD14+ cells appeared before CD1a+ cells. Cells grown in the presence of IL-13 had an increased capacity to present antigens to autologous lymphocytes and to stimulate allogeneic T-lymphocytes. This effect was greater than that of cells grown in the presence of TNF alpha. These cells should therefore have greater effector potential in any therapeutic applications in humans.
树突状细胞(DCs)是在人类抗肿瘤疫苗试验中具有潜在应用价值的抗原呈递细胞,目前是深入研究的对象。最近许多研究报道了通过培养骨髓或血液祖细胞在体外大量生成具有高抗原呈递能力的DCs的可能性。在本研究中,我们检测了从3名健康供体和5名乳腺癌患者经粒细胞集落刺激因子(G-CSF)动员的血液中分离出的CD34+祖细胞在粒细胞-巨噬细胞集落刺激因子(GM-CSF)+白细胞介素-13(IL-13)存在下向DCs的分化情况。将这些细胞的特征与在GM-CSF+肿瘤坏死因子α(TNFα)存在下获得的细胞的特征进行了比较。到第15天,用IL-13培养的大部分细胞中有三分之一为CD1a+/CD14-,并强烈表达CD1c、CD40、CD80和人类白细胞抗原DR(HLA-DR)。相比之下,用TNFα获得的细胞中三分之一的细胞表达CD1a,但荧光强度明显低于用IL-13培养的细胞,且超过50%的细胞强烈表达CD14。在IL-13培养物中,CD1a+/CD14-细胞在第5天出现,而在TNFα培养物中,CD14+细胞在CD1a+细胞之前出现。在IL-13存在下生长的细胞向自体淋巴细胞呈递抗原以及刺激同种异体T淋巴细胞的能力增强。这种效应大于在TNFα存在下生长的细胞。因此,这些细胞在人类的任何治疗应用中应该具有更大的效应潜力。