Differentiation and Assessment of Monocyte-Derived Dendritic Cells in the Presence of Endothelial Cells Conditioned Media and Maturation Factors

Document Type : Original Article (s)

Authors

1 Department of Biology, School of Science, Urmia University, Urmia, Iran

2 Assistant Professor, Department of Biology, School of Science, Urmia University, Urmia, Iran

3 Assistant Professor, Department of Immunology, School of Veterinary Medicine, Urmia University, Urmia, Iran

Abstract

Background: Dendritic cells (DCs) migrate from the periphery to the T cell zone of secondary lymphoid organs. Thus, activated DCs are specialized cells that obtain information from the periphery and transfer it to lymphoid organs. During their migration from the blood into peripheral tissues and then into the lymph nodes, DCs interact with endothelial cells and their soluble mediators. Therefore, we studied the effects of the endothelial condition medium (ECM) on phenotypic and functional characteristics of monocyte- driven dendritic cells.Methods: DCs were generated by culture of monocyte cells for 5 days in RPMI medium (RPMI 1640) supplemented with 10% FCS (fetal calf serum), IL-4 (interlukin-4), and GM-CSF (granulocyte, macrophage-colony stimulating factor). They were then incubated for 48 hours in MCM (monocyte conditioned medium), TNF-α (tumor necrotic factor alpha), and polyinosinic-polycytidylic acid (polyIC; control). ECM was added on day 5 and the beginning of the culture (day-0) to the control. The maturation of harvested DCs on day 7 was evaluated via flow cytometry, beta-counter and ELISA kits.Findings: DCs generated from peripheral blood monocytes specifically interact with ECM via soluble mediators. This induced the phenotypic maturation of DC via increased expression of CD83, CD80, HLA-DR, down-regulation of CD14, decreased phagocytic activity, and producing stimulatory cytokines in comparison with maturation factors alone. Moreover, ECM-matured DCs potently induced T cell activation reflected by increased T cell proliferation.Conclusion: Our results demonstrated that human endothelial cells with which DCs can encounter during their trafficking from tissue to lymph nodes can act as potent regulators of DC differentiation and function.

Keywords


  1. Katz SI, Tamaki K, Sachs DH. Epidermal Langerhans cells are derived from cells originating in bone marrow. Nature 1979; 282(5736): 324-6.
  2. Thomas DB. Viruses and the Cellular Immune Response. New York: Marcel Dekker; 1993.
  3. Tan JK, O'Neill HC. Maturation requirements for dendritic cells in T cell stimulation leading to tolerance versus immunity. J Leukoc Biol 2005; 78(2): 319-24.
  4. Sallusto F, Palermo B, Lenig D, Miettinen M, Matikainen S, Julkunen I, et al. Distinct patterns and kinetics of chemokine production regulate dendritic cell function. Eur J Immunol 1999; 29(5): 1617-25.
  5. D'Amico G, Bianchi G, Bernasconi S, Bersani L, Piemonti L, Sozzani S, et al. Adhesion, transendothelial migration, and reverse transmigration of in vitro cultured dendritic cells. Blood 1998; 92(1): 207-14.
  6. Allport JR, Ding H, Collins T, Gerritsen ME, Luscinskas FW. Endothelial-dependent mechanisms regulate leukocyte transmigration: a process involving the proteasome and disruption of the vascular endothelial-cadherin complex at endothelial cell-to-cell junctions. J Exp Med 1997; 186(4): 517-27.
  7. Thornhill MH, Kyan-Aung U, Haskard DO. IL-4 increases human endothelial cell adhesiveness for T cells but not for neutrophils. J Immunol 1990; 144(8): 3060-5.
  8. Randolph GJ, Luther T, Albrecht S, Magdolen V, Muller WA. Role of tissue factor in adhesion of mononuclear phagocytes to and trafficking through endothelium in vitro. Blood 1998; 92(11): 4167-77.
  9. Randolph GJ, Beaulieu S, Lebecque S, Steinman RM, Muller WA. Differentiation of monocytes into dendritic cells in a model of transendothelial trafficking. Science 1998; 282(5388): 480-3.
  10. Rafii S, Shapiro F, Rimarachin J, Nachman RL, Ferris B, Weksler B, et al. Isolation and characterization of human bone marrow microvascular endothelial cells: hematopoietic progenitor cell adhesion. Blood 1994; 84(1): 10-9.
  11. Tian F, Grimaldo S, Fujita M, Cutts J, Vujanovic NL, Li LY. The endothelial cell-produced antiangiogenic cytokine vascular endothelial growth inhibitor induces dendritic cell maturation. J Immunol 2007; 179(6): 3742-51.
  12. Methe H, Hess S, Edelman ER. Endothelial cell-matrix interactions determine maturation of dendritic cells. Eur J Immunol 2007; 37(7): 1773-84.
  13. Calvi LM, Adams GB, Weibrecht KW, Weber JM, Olson DP, Knight MC, et al. Osteoblastic cells regulate the haematopoietic stem cell niche. Nature 2003; 425(6960): 841-6.
  14. Pestka S, Krause CD, Sarkar D, Walter MR, Shi Y, Fisher PB. Interleukin-10 and related cytokines and receptors. Annu Rev Immunol 2004; 22: 929-79.
  15. Lan YY, Wang Z, Raimondi G, Wu W, Colvin BL, de CA, et al. "Alternatively activated" dendritic cells preferentially secrete IL-10, expand Foxp3+CD4+ T cells, and induce long-term organ allograft survival in combination with CTLA4-Ig. J Immunol 2006; 177(9): 5868-77.
  16. Brombacher F, Kastelein RA, Alber G. Novel IL-12 family members shed light on the orchestration of Th1 responses. Trends Immunol 2003; 24(4): 207-12.
  17. Alexander WS, Hilton DJ. The role of suppressors of cytokine signaling (SOCS) proteins in regulation of the immune response. Annu Rev Immunol 2004; 22: 503-29.
  18. Stark GR, Kerr IM, Williams BR, Silverman RH, Schreiber RD. How cells respond to interferons. Annu Rev Biochem 1998; 67: 227-64.
  19. Paglia P, Chiodoni C, Rodolfo M, Colombo MP. Murine dendritic cells loaded in vitro with soluble protein prime cytotoxic T lymphocytes against tumor antigen in vivo. J Exp Med 1996; 183(1): 317-22.
  20. Macatonia SE, Hosken NA, Litton M, Vieira P, Hsieh CS, Culpepper JA, et al. Dendritic cells produce IL-12 and direct the development of Th1 cells from naive CD4+ T cells. J Immunol 1995; 154(10): 5071-9.
  21. Delirezh N, Moazzeni SM, Shokri F, Shokrgozar MA, Atri M, Kokhaei P. Autologous dendritic cells loaded with apoptotic tumor cells induce T cell-mediated immune responses against breast cancer in vitro. Cell Immunol 2009; 257(1-2): 23-31.