Cunha Patricia, Gilbert Florence B, Bodin Jennifer, Godry Lise, Germon Pierre, Holbert Sebastien, Martins Rodrigo Prado
ISP, INRAE, Université de Tours, UMR1282, Nouzilly, France.
Front Vet Sci. 2022 Jun 9;9:891893. doi: 10.3389/fvets.2022.891893. eCollection 2022.
Dendritic cells are sentinels of the immune system responsible for the initiation of adaptive immune mechanisms. In that respect, the study of these cells is essential for a full understanding of host response to infectious agents and vaccines. In ruminants, the large blood volume facilitates the isolation of abundant monocytes and their derivation to other antigen-presenting cells such as dendritic cells and macrophages. However, the available protocols for the production of bovine monocyte-derived dendritic cells (moDCs) rely mostly on time-consuming and costly techniques such as density gradient centrifugation and magnetic sorting of cells. In this study, we describe a simplified protocol for the production of bovine moDC using conventional and serum-free media. We also employ moDC produced by this approach to carry out a flow cytometry-based antigen presentation assay adapted to blood fresh or frozen cells. The experimental strategies described here might enable the setup of studies involving a large number of individuals, requiring a large number of dendritic cells, or relying on the utilization of cryopreserved blood cells. These simplified protocols might contribute to the elucidation of cell-mediated immune responses in bovine.
树突状细胞是免疫系统的哨兵，负责启动适应性免疫机制。在这方面，对这些细胞的研究对于全面了解宿主对感染因子和疫苗的反应至关重要。在反刍动物中，大量的血容量便于分离出丰富的单核细胞，并将其衍生为其他抗原呈递细胞，如树突状细胞和巨噬细胞。然而，现有的用于生产牛单核细胞衍生树突状细胞（moDCs）的方案大多依赖于耗时且昂贵的技术，如密度梯度离心和细胞磁性分选。在本研究中，我们描述了一种使用常规培养基和无血清培养基生产牛moDC的简化方案。我们还采用通过这种方法产生的moDC进行基于流式细胞术的抗原呈递试验，该试验适用于新鲜或冷冻的血液细胞。这里描述的实验策略可能有助于开展涉及大量个体、需要大量树突状细胞或依赖于使用冷冻保存血细胞的研究。这些简化方案可能有助于阐明牛的细胞介导免疫反应。
