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Collection of lymph-borne dendritic cells in the rat

Nature Protocols volume 1, pages 2263–2270 (2006)Cite this article

Abstract

Dendritic cells (DCs) are crucial in immune induction. Not only do they collect antigens in peripheral tissues, and transport and process them for presentation to lymphocytes in draining lymph nodes, but they also regulate the immune response by modulating T-cell differentiation. Intestinal and hepatic DCs migrating in lymph can be collected from rats under near-physiological conditions. Initially, the mesenteric or celiac lymph nodes are removed from young rats (30 min). The afferent and efferent lymph vessels subsequently heal, permitting DCs to enter the thoracic duct. After at least 6 wk, the duct is cannulated (40 min). Lymph can be collected for up to 48 h. DCs can subsequently be identified, enriched and sorted to high degrees of purity. This two-stage technique generates large numbers of immunologically relevant DCs under near-physiological conditions. Lymph collection requires 2–3 h per animal over 6 wk.

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Figure 1: Specialist equipment for thoracic duct cannulation.
Figure 2: Animals shaved and arranged as if for surgery.
Figure 3: Mesenteric lymphadenectomy.
Figure 4: Thoracic duct cannulation.
Figure 5: Flow cytometry of DCs from lymph.

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Acknowledgements

The authors thank the staff of the animal facility for care of the animals. We gratefully acknowledge A. Milling and M. Nassar for critical reading of the manuscript. The work was funded by the Biotechnology and Biological Sciences Research Council, UK.

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  1. Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford, OX1 3RE, UK

    Simon WF Milling, Christopher Jenkins & Gordon MacPherson

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Correspondence to Simon WF Milling.

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Milling, S., Jenkins, C. & MacPherson, G. Collection of lymph-borne dendritic cells in the rat. Nat Protoc 1, 2263–2270 (2006). https://doi.org/10.1038/nprot.2006.315

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