Nedumpun Teerawut, Ritprajak Patcharee, Suradhat Sanipa
Inter-department of Medical Microbiology, Graduate School, Chulalongkorn University, Pathumwan, Bangkok 10330, Thailand.
RU in Oral Microbiology and Immunology, Department of Microbiology, Faculty of Dentistry, Chulalongkorn University, Pathumwan, Bangkok 10330, Thailand; Oral Biology Research Center, Faculty of Dentistry, Chulalongkorn University, Pathumwan, Bangkok 10330, Thailand.
Vet Immunol Immunopathol. 2016 Dec;182:63-68. doi: 10.1016/j.vetimm.2016.10.002. Epub 2016 Oct 13.
In vitro derivation of dendritic cells (DCs) is an alternative approach to overcome the low frequency of primary DCs and the difficulty of isolation techniques for studying DC immunobiology. To date, the conventional culture protocol of porcine monocyte-derived DCs (MoDCs) has been widely used. However, this protocol is not practical due to the requirement of a substantial number of blood monocytes, and the process often interferes with DC maturation. To improve in vitro porcine MoDC generation, we modified the previous conventional DC generation protocol, based on the human and mouse primary DC culture system, and compared phenotypic and functional features of MoDCs derived from the modified protocol to the conventional protocol. The modified protocol consumed fewer monocytes but generated higher CD1 cells with DC-like morphology and the ability of maturation. In addition, MoDCs from the modified protocol exhibited increased antigen uptake and IFN-γ production in response to LPS stimulation. Our findings indicate that the modified protocol is expedient and reliable for generating potent MoDCs that substitute for primary DCs. This will be a valuable platform for future research in antigen delivery, vaccines and immunotherapy in pigs, as well as relevant veterinary species.
体外衍生树突状细胞（DCs）是一种替代方法，可克服原代DCs频率低以及研究DC免疫生物学时分离技术困难的问题。迄今为止，猪单核细胞衍生DCs（MoDCs）的传统培养方案已被广泛使用。然而，由于需要大量血液单核细胞，该方案并不实用，而且该过程常常干扰DC成熟。为了改进体外猪MoDC的生成，我们基于人和小鼠原代DC培养系统修改了先前的传统DC生成方案，并将修改方案衍生的MoDCs与传统方案衍生的MoDCs的表型和功能特征进行了比较。修改后的方案消耗的单核细胞较少，但产生了更多具有DC样形态和成熟能力的CD1细胞。此外，修改方案衍生的MoDCs在LPS刺激下表现出抗原摄取增加和IFN-γ产生增加。我们的研究结果表明，修改后的方案对于生成替代原代DCs的有效MoDCs是方便且可靠的。这将成为未来猪以及相关兽医物种的抗原递送、疫苗和免疫治疗研究的宝贵平台。
