He Tianpei, Tang Chaoke, Xu Shulin, Moyana Terence, Xiang Jim
Research Unit, Health Research Division, Saskatchewan Cancer Agency, Department of Oncology, University of Saskatchewan, Saskatoon, Saskatchewan S7N 4H4, Canada.
Cell Mol Immunol. 2007 Apr;4(2):105-11.
Dendritic cells (DCs) are the most potent antigen-presenting cells (APCs) for the initiation of antigen (Ag)-specific immune responses. In most studies, mature DCs are generated from bone marrow cells or peripheral monocytes; in either case, the harvested cells are then cultured in medium containing recombinant GM-CSF, IL-4 and TNF-alpha for 7-10 days and stimulated with lipopolysaccharide (LPS). However, this approach is time-consuming and expensive. There is another less cost approach of using immobilized DC cell lines, which can easily grow in the medium. A disadvantage with the immobilized DC cell lines, however, is that they are immature DCs and lack expression of MHC class II and costimulatory CD40 and CD80 molecules. This, therefore, limits their capacity for inducing efficient antitumor immunity. In the current study, we investigated the possible efficacy of various stimuli (IL-1beta,IFN-gamma, TNF-alpha, CpG and LPS) in converting the immature dendritic cell line DC2.4 to mature DCs. Our findings were quite interesting since we demonstrated for the first time that IFN-gamma was able to stimulate the maturation of DC2.4 cells. The IFN-gamma-activated ovalbumin (OVA)-pulsed DC2.4 cells have capacity to upregulate MHC class II, CD40, CD80 and CCR7, and to more efficiently stimulate in vitro and in vivo OVA-specific CD8+ T cell responses and antitumor immunity. Therefore, IFN-gamma-activated immortal DC2.4 cells may prove to be useful in the study of DC biology and antitumor immunity.
树突状细胞(DCs)是启动抗原(Ag)特异性免疫反应最有效的抗原呈递细胞(APCs)。在大多数研究中,成熟的DCs是由骨髓细胞或外周单核细胞产生的;在任何一种情况下,收获的细胞随后在含有重组GM-CSF、IL-4和TNF-α的培养基中培养7-10天,并用脂多糖(LPS)刺激。然而,这种方法既耗时又昂贵。还有另一种成本较低的方法,即使用固定化的DC细胞系,它们可以在培养基中轻松生长。然而,固定化DC细胞系的一个缺点是它们是未成熟的DCs,缺乏MHC II类分子以及共刺激分子CD40和CD80的表达。因此,这限制了它们诱导有效抗肿瘤免疫的能力。在本研究中,我们研究了各种刺激物(IL-1β、IFN-γ、TNF-α、CpG和LPS)将未成熟树突状细胞系DC2.4转化为成熟DCs的可能效果。我们的发现非常有趣,因为我们首次证明IFN-γ能够刺激DC2.4细胞的成熟。IFN-γ激活的卵清蛋白(OVA)脉冲DC2.4细胞有能力上调MHC II类分子、CD40、CD80和CCR7,并更有效地刺激体外和体内OVA特异性CD8 + T细胞反应及抗肿瘤免疫。因此,IFN-γ激活的永生化DC2.4细胞可能在DC生物学和抗肿瘤免疫研究中有用。