Bonnefont-Rebeix Catherine, de Carvalho Camila Miranda, Bernaud Janine, Chabanne Luc, Marchal Thierry, Rigal Dominique
Immunology Department, Etablissement Français du Sang de Lyon,1-3 Rue du Vercors, 69007 Lyon, France.
Vet Immunol Immunopathol. 2006 Jan 15;109(1-2):167-76. doi: 10.1016/j.vetimm.2005.08.027. Epub 2005 Sep 30.
In this study, canine monocyte-derived dendritic cells (cMo-DC) were produced in presence of canine GM-CSF (cGM-CSF) and canine IL-4 (cIL-4), and they were characterized by their dendritic morphology, MLR functionality and phenotype. We noticed that cMo-DC were labelled with three anti-human CD86 (FUN-1, BU63 and IT2.2 clones), whereas resting and activated lymphocytes or monocytes were not stained. CD86 expression was induced by cIL-4 and was up-regulated during the differentiation of the cMo-DC, with a maximum at day 7. Furthermore, cMo-DC were very potent even in low numbers as stimulator cells in allogeneic MLR, and BU63 mAb was able to completely block the cMo-DC-induced proliferation in MLR. We also observed that cMo-DC highly expressed MHC Class II and CD32, but we failed to determine their maturation state since the lack of commercially available canine markers. Moreover, cMo-DC contained cytoplasmic periodic microstructures, potentially new ultrastructural markers of canine DC recently described. In conclusion, this work demonstrates that the CD86 costimulatory marker is now usable for a better characterization of in vitro canine DC.
在本研究中,犬单核细胞衍生树突状细胞(cMo-DC)在犬粒细胞-巨噬细胞集落刺激因子(cGM-CSF)和犬白细胞介素-4(cIL-4)存在的情况下产生,并通过其树突形态、混合淋巴细胞反应(MLR)功能和表型进行表征。我们注意到cMo-DC被三种抗人CD86(FUN-1、BU63和IT2.2克隆)标记,而静息和活化的淋巴细胞或单核细胞未被染色。CD86表达由cIL-4诱导,并在cMo-DC分化过程中上调,在第7天达到最大值。此外,cMo-DC即使数量很少,作为异基因MLR中的刺激细胞也非常有效,并且BU63单克隆抗体能够完全阻断cMo-DC在MLR中诱导的增殖。我们还观察到cMo-DC高度表达MHC II类分子和CD32,但由于缺乏市售的犬标记物,我们未能确定它们的成熟状态。此外,cMo-DC含有细胞质周期性微观结构,这可能是最近描述的犬DC的新超微结构标记物。总之,这项工作表明,CD86共刺激标记物现在可用于更好地表征体外犬DC。