Dendritic cell (DC) maturation approved to be a pivotal process for initiating immunity. Many protocols were established in order the isolated peripheral blood mononuclear cells (PBMCs) from healthy donors to mature into dendritic cells (DCs). The purpose of this study was to present an effective and reliable DC maturation procedure by comparing three different protocols (Interleukin-4/Tumor Necrosis Factor-appha (IL-4/TNFa) DC protocol, Interferon alpha (IFNa) DC protocol and FAST DC protocol). Whole blood was collected from six healthy donors and PBMCs were isolated by Ficoll gradient centrifugation. The counted cells were incubated with the addition of three different cocktails of supplements for appropriate time period. The final mature DC population was examined either by its phenotypic characteristics under light microscope or by measuring the expression of antigen presenting molecules such as CD80 and CD86 by flow cytometry. It was found that the mature DCs, generated from the IL-4/TNFa DC protocol, expressed higher levels of CD80 and CD86. Furthermore, they sharply exhibited their phenotypic hallmarks.
KeywordsDC MaturationTNFaIL-4IFNa
Cranmer, L.D., Trevor, K.T. and Hersh, E.M. (2004) Clinical Applications of Dendritic Cell Vaccination in the Treatment of Cancer. Cancer Immunology, Immunotherapy, 53, 275-306. http://dx.doi.org/10.1007/s00262-003-0432-5
Han, T.H., Jin, P., Ren, J., Slezak, S., Marincola, F.M. and Stroncek, D.F. (2009) Evaluation of 3 Clinical Dendritic Cell Maturation Protocols Containing Lipopolysaccharide and Interferon-Gamma. Journal of Immunotherapy, 32, 399-407. http://dx.doi.org/10.1097/CJI.0b013e31819e1773
De Costa, A.M., Justis, D.N., Schuyler, C.A. and Young, M.R. (2012) Administration of a Vaccine Composed of Dendritic Cells Pulsed with Premalignant Oral Lesion Lysate to Mice Bearing Carcinogen-Induced Premalignant Oral Lesions Stimulates a Protective Immune Response. International Immunopharmacology, 13, 322-330. http://dx.doi.org/10.1016/j.intimp.2012.05.004
Cella, M., Sallusto, F. and Lanzavecchia, A. (1997) Origin, Maturation and Antigen Presenting Function of Dendritic Cells. Current Opinion in Immunology, 9, 10-16. http://dx.doi.org/10.1016/S0952-7915(97)80153-7
Pulendran, B., Smith, J.L., Caspary, G., Brasel, K., Pettit, D., Maraskovsky, E. and Maliszewski, C.R. (1999) Distinct Dendritic Cell Subsets Differentially Regulate the Class of Immune Response in Vivo. Proceedings of the National Academy of Sciences of the USA, 96, 1036-1041. http://dx.doi.org/10.1073/pnas.96.3.1036
Pulendran, B. (2004) Modulating TH1/TH2 Responses with Microbes, Dendritic Cells, and Pathogen Recognition Receptors. Immunologic Research, 29, 187-196. http://dx.doi.org/10.1385/IR:29:1-3:187
Syme, R., Bajwa, R., Robertson, L., Stewart, D. and Gluck, S. (2005) Comparison of CD34 and Monocyte-Derived Dendritic Cells from Mobilized Peripheral Blood from Cancer Patients. Stem Cells, 23, 74-81. http://dx.doi.org/10.1634/stemcells.2004-0070
Zhou, L.J. and Tedder, T.F. (1996) CD14+ Blood Monocytes Can Differentiate into Functionally Mature CD83+ Dendritic Cells. Proceedings of the National Academy of Sciences of the USA, 93, 2588-2592. http://dx.doi.org/10.1073/pnas.93.6.2588
Harada, S., Kimura, T., Fujiki, H., Nakagawa, H., Ueda, Y., Itoh, T., Yamagishi, H. and Sonoda, Y. (2007) Flt3 Ligand Promotes Myeloid Dendritic Cell Differentiation of Human Hematopoietic Progenitor Cells: Possible Application for Cancer Immunotherapy. International Journal of Oncology, 30, 1461-1468. http://dx.doi.org/10.3892/ijo.30.6.1461
Caux, C., Vanbervliet, B., Massacrier, C., Azuma, M., Okumura, K., Lanier, L.L. and Banchereau, J. (1994) B70/B7-2 Is Identical to CD86 and Is the Major Functional Ligand for CD28 Expressed on Human Dendritic Cells. Journal of Experimental Medicine, 180, 1841-1847. http://dx.doi.org/10.1084/jem.180.5.1841
Palucka, K. and Banchereau, J. (2012) Cancer Immunotherapy via Dendritic Cells. Nature Reviews Cancer, 12, 265-277. http://dx.doi.org/10.1038/nrc3258
Matera, L. and Galetto, A. (2003) In Vitro Maturation of Dendritic Cells from Blood Progenitors. Methods in Molecular Biology, 215, 417-426.
Della Bella, S., Gennaro, M., Vaccari, M., Ferraris, C., Nicola, S., Riva, A., Clerici, M., Greco, M. and Villa, M.L. (2003) Altered Maturation of Peripheral Blood Dendritic Cells in Patients with Breast Cancer. British Journal of Cancer, 89, 1463-1472. http://dx.doi.org/10.1038/sj.bjc.6601243
Korthals, M., Safaian, N., Kronenwett, R., Maihofer, D., Schott, M., Papewalis, C., Diaz blanco, E., Winter, M., Czibere, A., Haas, R., Kobbe, G. and Fenk, R. (2007) Monocyte Derived Dendritic Cells Generated by IFN-Alpha Acquire Mature Dendritic and Natural Killer Cell Properties as Shown by Gene Expression Analysis. Journal of Translational Medicine, 5, 46. http://dx.doi.org/10.1186/1479-5876-5-46
Trevejo, J.M., Marino, M.W., Philpott, N., Josien, R., Richards, E.C., Elkon, K.B. and Falck-Pedersen, E. (2011) TNF-Alpha-Dependent Maturation of Local Dendritic Cells Is Critical for Activating the Adaptive Immune Response to Virus Infection. Proceedings of the National Academy of Sciences of the United States of America, 98, 12162-12167. http://dx.doi.org/10.1073/pnas.211423598
Van de Laar, L., Coffer, P.J. and Woltman, A.M. (2012) Regulation of Dendritic Cell Development by GM-CSF: Molecular Control and Implications for Immune Homeostasis and Therapy. Blood, 119, 3383-3393. http://dx.doi.org/10.1182/blood-2011-11-370130
Hiasa, M., Abe, M., Nakano, A., Oda, A., Amou, H., Kido, S., Takeuchi, K., Kagawa, K., Yata, K., Hashimoto, T., Ozaki, S., Asaoka, K., Tanaka, E., Moriyama, K. and Matsumoto, T. (2009) GM-CSF and IL-4 Induce Dendritic Cell Differentiation and Disrupt Osteoclastogenesis through M-CSF Receptor Shedding by Up-Regulation of TNF-Alpha Converting Enzyme (TACE). Blood, 114, 4517-4526. http://dx.doi.org/10.1182/blood-2009-04-215020
Conti, L., Cardone, M., Varano, B., Puddu, P., Belardelli, F. and Gessani, S. (2008) Role of the Cytokine Environment and Cytokine Receptor Expression on the Generation of Functionally Distinct Dendritic Cells from Human Monocytes. European Journal of Immunology, 38, 750-762. http://dx.doi.org/10.1002/eji.200737395
Mariotti, S., Sargentini, V., Marcantonio, C., Todero, E., Teloni, R., Gagliardi, M.C., Ciccaglione, A.R. and Nisini, R. (2008) T-Cell-Mediated and Antigen-Dependent Differentiation of Human Monocyte into Different Dendritic Cell Subsets: A Feedback Control of Th1/Th2 Responses. The FASEB Journal, 22, 3370-3379. http://dx.doi.org/10.1096/fj.08-108209
Zhang, A.L., Colmenero, P., Purath, U., Teixeira De Matos, C., Hueber, W., Klareskog, L., Tarner, I.H., Engleman, E.G. and Soderstrom, K. (2007) Natural Killer Cells Trigger Differentiation of Monocytes into Dendritic Cells. Blood, 110, 2484-2493. http://dx.doi.org/10.1182/blood-2007-02-076364
Kitawaki, T., Kadowaki, N., Sugimoto, N., Kambe, N., Hori, T., Miyachi, Y., Nakahata, T. and Uchiyama, T. (2006) IgE-Activated Mast Cells in Combination with Pro-Inflammatory Factors Induce Th2-Promoting Dendritic Cells. International Immunology, 18, 1789-1799. http://dx.doi.org/10.1093/intimm/dxl113
Yu, Y., Tang, L., Wang, J., Liu, S., Wang, W., An, H., Qi, R., Zhang, M. and Cao, X. (2002) Psoriatic Lesional Keratinocytes Promote the Maturation of Human Monocyte-Derived Langerhans Cells. Dermatology, 204, 94-99. http://dx.doi.org/10.1159/000051824
Van Lieshout, A.W., Barrera, P., Smeets, R.L., Pesman, G.J., Van Riel, P.L., Van Den Berg, W.B. and Radstake, T.R. (2005) Inhibition of TNF Alpha during Maturation of Dendritic Cells Results in the Development of Semi-Mature Cells: A Potential Mechanism for the Beneficial Effects of TNF Alpha Blockade in Rheumatoid Arthritis. Annals of the Rheumatic Diseases, 64, 408-414. http://dx.doi.org/10.1136/ard.2004.023259
Castiello, L., Sabatino, M., Jin, P., Clayberger, C., Marincola, F.M., Krensky, A.M. and Stroncek, D.F. (2011) Monocyte-Derived DC Maturation Strategies and Related Pathways: A Transcriptional View. Cancer Immunology, Immunotherapy, 60, 457-466. http://dx.doi.org/10.1007/s00262-010-0954-6
Severa, M., Remoli, M.E., Giacomini, E., Ragimbeau, J., Lande, R., Uze, G., Pellegrini, S. and Coccia, E.M. (2006) Differential Responsiveness to IFN-Alpha and IFN-Beta of Human Mature DC through Modulation of IFNAR Expression. Journal of Leukocyte Biology, 79, 1286-1294. http://dx.doi.org/10.1189/jlb.1205742
Blanco, P., Palucka, A.K., Pascual, V. and Banchereau, J. (2008) Dendritic Cells and Cytokines in Human Inflammatory and Autoimmune Diseases. Cytokine & Growth Factor Reviews, 19, 41-52. http://dx.doi.org/10.1016/j.cytogfr.2007.10.004
Simmons, D.P., Wearsch, P.A., Canaday, D.H., Meyerson, H.J., Liu, Y.C., Wang, Y., Boom, W.H. and Harding, C.V. (2012) Type I IFN Drives a Distinctive Dendritic Cell Maturation Phenotype That Allows Continued Class II MHC Synthesis and Antigen Processing. The Journal of Immunology, 188, 3116-3126. http://dx.doi.org/10.4049/jimmunol.1101313
Swetman, C.A., Leverrier, Y., Garg, R., Gan, C.H., Ridley, A.J., Katz, D.R. and Chain, B.M. (2002) Extension, Retraction and Contraction in the Formation of a Dendritic Cell Dendrite: Distinct Roles for Rho GTPases. European Journal of Immunology, 32, 2074-2083. http://dx.doi.org/10.1002/1521-4141(200207)32:7 3.0.CO;2-S
Zheng, Y., Manzotti, C.N., Liu, M., Burke, F., Mead, K.I. and Sansom, D.M. (2004) CD86 and CD80 Differentially Modulate the Suppressive Function of Human Regulatory T Cells. The Journal of Immunology, 172, 2778-2784. http://dx.doi.org/10.4049/jimmunol.172.5.2778