Abstract
Dendritic cells (DC) are rare cells in peripheral tissues, and their isolation from tissues is fraught with problems. Thus, the proportion of DC within a tissue that is extracted is unknown, isolation procedures may select for subpopulations, and the isolation procedure itself may affect their properties. As part of their life history, DC migrate from peripheral tissues, via peripheral, afferent lymph to lymph nodes, even in the absence of exogenous antigenic stimulation. They are extracted within the node and very few, if any, appear in efferent lymph. These lymph DC (L-DC) represent a population that has matured in the periphery, that may have acquired antigen (Ag), and that may be engaged in active Ag transport to lymph nodes. As such they are a physiologically relevant DC population. In large animals such as sheep and cattle, L-DC can be isolated by direct cannulation of peripheral lymphatics, but yields are relatively low. In rodents, direct cannulation of some peripheral lymphatics is possible (1), but yields of cells are minuscule. To get around this problem we and others (1 -7), have utilized lymphadenectomy as a means of collecting pseudo-afferent lymph. When lymph nodes are removed, over a period of weeks, the afferent and efferent lymphatics join as part of the healing process, leaving cells in peripheral lymph free to enter central lymph. Central lymphat ics are relatively easy to cannulate, and cannulation can be maintained for considerable periods of time (see Note 1).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Pugh, C.W., MacPherson, G. G., and Steer, H. W. (1983) Characterization of nonlymphoid cells derived from rat peripheral lymph. J. Exp. Med. 157, 1758ā79.
Mayrhofer, G., Holt, P. G., and Papdimitriou, J. M. (1986) Functional characteristics of the veiled cells in afferent lymph from the rat intestine. Immunology 58, 379ā387.
Harkiss, G., Hopkins, J., and McConnell, I. (1990) Uptake of antigen by afferent lymph dendritic cells mediated by antibody. Eur. J. Immunol. 20, 2367ā2373.
Bujdoso, R., Hopkins, J., Dutia, B. M., Young, P., McConnell, I. (1989) Characterisation of sheep afferent lymph dendritic cells and their role in antigen carriage. J. Exp. Med. 170, 1285ā1302.
Bujdoso, R., Harkiss, G., Hopkins, J., and McConnell, I. (1990) Afferent lymph dendritic cells: a model for antigen capture and presentation in vivo. Int. Rev. Immunol. 6, 177ā86.
Howard, C. J., Sopp, P., Brownlie, J., Parsons, K. R., Kwong, L. S., and Collins, R. A. (1996) Afferent lymph veiled cells stimulate proliferative responses in allogeneic CD4+and CD8+T cells but not gamma delta TCR+T cells. Immunology 88, 558ā64.
Howard, C. J., Sopp, P., Brownlie, J., Kwong, L. S., Parsons, K. R., and Taylor, G. (1997) Identification of two distinct populations of dendritic cells in afferent lymph that vary in their ability to stimulate T cells. J. Immunol. 159, 5372ā5382.
Matsuno, K., Ezaki, T., Kudo, S., and Uehara, Y. (1996) A life stage of particleladen rat dendritic cells in vivo: their terminal division, active phagocytosis, and translocation from the liver to the draining lymph. J. Exp. Med. 183, 1865ā1878.
Kudo, S., Matsuno, K., Ezaki, T., and Ogawa, M. (1997) A novel migration pathway for rat dendritic cells from the blood: hepatic sinusoids-lymph translocation. J. Exp. Med. 185, 777ā784.
Gowans, J. L. (1959) The recirculation of lymphocytes from blood to lymph in the rat. J. Physiol. 146, 54.
Bollman, J. L., Cain, J. C., and Grindley, J. H. (1948) Techniques for the collection of lymph from the liver, small intestine or thoracic duct of the rat. J. Lab. Clin. Med. 33, 1349.
Steinman, R. M., Cohn, Z. A. (1973) Identification of a novel cell type in peripheral lymphoid organs of mice. I. Morphology, quantitation, tissue distribution. J. Exp. Med. 137, 1142ā1162.
Klinkert, W. E. F., LaBadie, J. H., and Bowers, W. E. (1982) Accessory and stimulating properites of dendritic cells and macrophages isolated from various rat tissues. J. Exp. Med. 156, 1ā19.
Metlay, J. P., Witmer-Pack, M. D., Agger, R., Crowley, M. T., Lawless, D., and Steinman, R. M. (1990) The distinct leukocyte integrins of mouse spleen dendritic cells as identified with new hamster monoclonal antibodies. J. Exp. Med. 171, 1753ā1771.
Inaba, K., Pack, M., Inaba, M., Sakuta, H., Isdell, F., and Steinman, R. M. (1997) High levels of amajorhistocompatibility complex II-self peptide complex on dendritic cells from the T cell areas of lymph nodes. J. Exp. Med. 186, 665ā672.
Schuler, G., and Steinman, R. M. (1985) Murine epidermal Langerhans cells mature into potent immunostimulatory dendritic cells in vitro. J. Exp. Med. 161, 526ā546.
Austyn, J. M., Hankins, D. F., Larsen, C. P., Morris, P. J., Rao, A. S., Roake, J. A. (1994) Isolation and characterization of dendritic cells from mouse heart and kidney. J. Immunol. 152, 2401ā2410.
MacPherson, G. G. (1989) Properties of lymph-borne (veiled) dendritic cells in culture. I. Modulation of phenotype, survival and function: partial dependence on GM-CSF. Immunology 68, 102ā107.
Leenen, P. J., Radosevic, K., Voerman, J. S., et al. (1998) Heterogeneity of mouse spleen dendritic cells: in vivo phagocytic activity, expression of macrophage markers, and subpopulation turnover. J. Immunol. 160, 2166ā2173.
Gyure, L. and Hall, J. (1990) A quick method for obtaining lymph borne dendritic macrophages from rats. J. Immunol. Methods. 131, 49ā53.
Liu, L. M., Zhang, M., Jenkins, C., and MacPherson, G. G. (1998) Dendritic cell heterogeneity in vivo: Two functionally different dendritic cell populations in rat intestinal lymph can be distinguished by CD4 expression. J. Immunol. 161, 1146ā1155.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
Ā© 2001 Humana Press Inc.
About this protocol
Cite this protocol
MacPherson, G., Wykes, M., Huang, FP., Jenkins, C. (2001). Isolation of Dendritic Cells from Rat Intestinal Lymph and Spleen. In: Robinson, S.P., Stagg, A.J., Knight, S.C. (eds) Dendritic Cell Protocols. Methods in Molecular Medicineā¢, vol 64. Humana Press. https://doi.org/10.1385/1-59259-150-7:29
Download citation
DOI: https://doi.org/10.1385/1-59259-150-7:29
Publisher Name: Humana Press
Print ISBN: 978-0-89603-584-3
Online ISBN: 978-1-59259-150-3
eBook Packages: Springer Protocols