Isolation and functional analysis of lamina propria dendritic cells from the mouse small intestine

Abstract

Dendritic cells (DCs) are the most professional antigen-presenting cells that are indispensable for the initiation of adaptive immune responses. DCs are heterogeneous in terms of their origin, anatomical location, cell-surface markers, and functions. Previous studies have demonstrated that there exist several groups of DCs in the lamina propria (LPDC) of gastrointestinal tract, which collectively contribute to the maintenance of gut homeostasis through the regulation of the balance between active immunity and tolerance. However, although intestinal LPDCs are attractive research target for understanding the immunological mechanisms in the gut, isolation of the LPDCs is complicated and technically difficult for unskilled people. Therefore, establishment of the method to isolate intestinal LPDCs is a major obstacle in this research. Here, we describe the methods that we have established for the isolation of primary DCs from the LP of mouse small intestine. Our isolation method provides high yield of viable LP leukocytes (LPLs) including DCs. Combination with FACS sorting allows for the selective isolation of CD103+ CD8α+ DCs and CD103+ CD8α− DCs from the LPLs. Furthermore, isolated LPDCs can be subjected to immunological assays, such as measurement of cytokine productions following stimulation of Toll-like receptors. Thus, our methods would be useful for studying the functions of LPDCs of mouse small intestine.