Abstract
Dendritic cells (DC) are widely considered to be the major antigen-presenting cell (APC) type in immune responses. These cells are obtained from adherent cells or are purified CD14+ monocytes from peripheral blood mononuclear cells (PBMC) by in vitro stimulation with granulocyte, macrophage-colony-stimulating factor (GM-CSF) plus interleukin (IL)-4. They express high levels of MHC class II and costimulatory molecules, internalize Ag rapidly via Fc receptors and mannose receptors, and, by macropinocytosis, produce large amounts of IL-12 on CD40 ligation, and are potent in presenting soluble Ag and in stimulating allogeneic mixed-leukocyte reactions.
To study primary T-cell responses and cytokine production in allergy patients, we have developed an in vitro system by using highly purified T cells as responder cells and monocyte-derived DC (MDC) as the APC. MDC provide a convenient and potent APC source for T-cell response studies.
Key Words
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Banchereau, J., Briere, F., and Caux, C., et al. (2000) Immunobiology of dendritic cells. Annu. Rev. Immunol. 18, 767–811.
Rossi, M. and Young, J. W. (2005) Human dendritic cells: potent antigen-presenting cells at the crossroads of innate and adaptive immunity. J. Immunol. 175, 1373–1381.
O’Neill, D. W., Adams, S., and Bhardwaj, N. (2004) Manipulating dendritic cell biology for the active immunotherapy of cancer. Blood 104, 2235–2246.
Sallusto, F., Cella, M., Danieli, C., and Lanzavecchia, A. (1995) Dendritic cells use macropinocytosis and the mannose receptor to concentrate macromolecules in the major histocompatibility complex class II compartment: downregulation by cytokines and bacterial products. J. Exp. Med. 182, 389–400.
Cella, M., Scheidegger, D., Palmer-Lehmann, K., Lane, P., Lanzavecchia, A., and Alber, G. (1996) Ligation of CD40 on dendritic cells triggers production of high levels of interleukin-12 and enhances T cell stimulatory capacity: T-T help via APC activation. J. Exp. Med. 184, 747–752.
Cella, M., Engering, A., Pinet, V., Pieters, J., and Lanzavecchia, A. (1997) Inflammatory stimuli induce accumulation of MHC class II complexes on dendritic cells. Nature 388, 782–787.
Dhodapkar, M. V., Krasovsky, J., Steinman, R. M., and Bhardwaj, N. (2000) Mature dendritic cells boost functionally superior CD8(+) T-cell in humans without foreign helper epitopes. J. Clin. Invest. 105, R9–R14.
Thurner, B., Haendle, I., Roder, C., et al. (1999) Vaccination with mage-3A1 peptidepulsed mature, monocyte-derived dendritic cells expands specific cytotoxic T cells and induces regression of some metastases in advanced stage IV melanoma. J. Exp. Med. 190, 1669–1678.
Sung, S. J., Taketomi, E. A., Smith, A. M., Platts-Mills, T. A., and Fu, S. M. (1999) Efficient presentation of house dust mite allergen Der p 2 by monocyte-derived dendritic cells and the role of beta 2 integrins. Scand. J. Immunol. 49, 96–105.
Sung, S. S., Nelson, R. S., and Silverstein, S. C. (1985) Mouse peritoneal macrophages plated on mannan-and horseradish peroxidase-coated substrates lose the ability to phagocytose by their Fc receptors. J. Immunol. 134, 3712–3717.
Wright, S. and Silverstein, S. (1982) Tumor-promoting phorbol esters stimulate C3b and C3b′ receptor-mediated phagocytosis in cultured human monocytes. J. Exp. Med. 156, 1149–1164.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2008 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Sung, Ss.J. (2008). Monocyte-Derived Dendritic Cells as Antigen-Presenting Cells in T-Cell Proliferation and Cytokine Production. In: Jones, M.G., Lympany, P. (eds) Allergy Methods and Protocols. Methods in Molecular Medicine, vol 138. Humana Press. https://doi.org/10.1007/978-1-59745-366-0_9
Download citation
DOI: https://doi.org/10.1007/978-1-59745-366-0_9
Publisher Name: Humana Press
Print ISBN: 978-0-89603-896-7
Online ISBN: 978-1-59745-366-0
eBook Packages: Springer Protocols