Generation of canine dendritic cells from peripheral blood monocytes without using purified cytokines
Generation of canine dendritic cells from peripheral blood monocytes without using purified cytokines
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DOI：
10.1016/j.vetimm.2006.07.002
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发表时间：
2006-11-15
影响因子：
1.8
通讯作者：
Inaba, Toshio
中科院分区：
文献类型：
--
作者：
Wijewardana, Viskam;Sugiura, Kikuya;Inaba, Toshio
树突状细胞（Dendritic cells，DCs）是一种由外周血单核细胞（PBMO）或骨髓前体细胞在体外分化而来的细胞，是一种很有前途的肿瘤免疫治疗候选细胞。这种狗和人类一样沿着着常见的癌症，是癌症研究的理想大型动物模型。然而，由于尚未建立体外分化的适当条件，因此犬中单核细胞衍生的DC尚未得到很好的表征。为了解决这个问题，我们已经开发了一种条件培养基，通过培养T细胞与固定化的抗犬CD 3抗体，并试图诱导分化的DC从PBMO。当在25%T细胞条件培养基（TCCM）存在下培养纯化的CD 14（+）PBMO时，PBMO在培养的第12天增大尺寸并且具有延长的树突状突起。发现培养的PBMO增加了MHC 11类和CD I a分子的表达，并且在混合白细胞反应中显著增加了对同种异体T细胞的刺激活性。此外，当用聚肌胞苷酸（Poly（1：C））刺激时，细胞显著增加其IL-18和IFN-γ的表达。细胞具有降低的吞噬活性，这是成熟DC中的常见缺陷。从这些结果可以看出，TCCM确实诱导了来自PBMO的DC的分化。(c)2006 Elsevier B. V.保留所有权利。
Dendritic cells (DCs), which differentiate in vitro from peripheral blood monocytes (PBMOs) or bone marrow precursors, are a promising candidate for immunotherapy against cancer. The dog, which suffers common types of cancers along with humans, make an ideal large animal model for cancer studies. Monocyte-derived DCs in the dog have not been well characterized, however, since the appropriate condition for in vitro differentiation has not been established. To tackle this problem, we have developed a conditioned media by culturing T cells with immobilized anti-canine CD3 antibody, and sought to induce differentiation of DCs from PBMOs. When purified CD14(+) PBMOs were cultured in the presence of 25% T cell conditioned medium (TCCM), the PBMOs increased size and had extended dendritic processes by day 12 of the culture. The cultured PBMOs were found to increase the expression of MHC class 11 and CD I a molecules, and significantly increased stimulatory activity for allogeneic T cells in the mixed leukocyte reaction. Moreover, the cells significantly increased their expression of IL-18 and IFN-gamma when stimulated with polyinosinic-polycytidylic acid (Poly (1:C)). The cells have a reduced phagocytic activity, which is a common defect in mature DCs. It follows from these results that TCCM does induce the differentiation of DCs from PBMOs. (c) 2006 Elsevier B.V. All rights reserved.