Comparison of several maturation inducing factors in dendritic cell differentiation.

BACKGROUND Dendritic cells (DCs) are professional antigen presenting cells that have an important role in the initiation of immune response. The use of maturation factors in dendritic cell differentiation provides a promising approach in immunotherapy. OBJECTIVE In this study, we compared tumor necrosis factor-α, polyribocytidylic acid, lipopolysacharide and CpG oligonucleotides in inducing dendritic cell maturation. METHODS We generated immature dendritic cells with GM-CSF in combination with IL-4 from peripheral blood mononuclear adherent cells and used tumor necrosis factor-α, polyribocytidylic acid, lipopolysacharide and CpG for the induction of dendritic cell maturation. CD83 maturation marker on the dendritic cells was analyzed by flowcytometry after 7 days. In addition, mixed leukocyte reaction between dendritic cells and T cells was performed by MTT proliferation assay. RESULTS Flow cytometry results demonstrated a comparable high level of CD83 expression on the mature dendritic cells generated by TNF-α, CpG, Poly I:C, and LPS treatment of the immature dendritic cells. However, a significantly poorer proliferation of lymphocytes cocultured with the Poly I:C-treated DCs was observed compared to the CpG-treated DCs in mixed leukocyte reaction (p=0.026). Conversely, a significantly stronger proliferation of lymphocytes was observed when cocultured with TNF-α-treated DCs compared to the LPS-treated DCs (p=0.025). CONCLUSION Our results indicated that all of studied maturation inducing factors can be used in DC maturation but TNF-α and CpG were the preferred in vitro maturation factors. It is concluded that maturation of dendritic cells by CpG motif and TNF-α can be used to regulate immune responses.

[1]  T. Ichinohe,et al.  Zymosan enhances the mucosal adjuvant activity of poly(I:C) in a nasal influenza vaccine , 2010, Journal of medical virology.

[2]  G. Rabinovich,et al.  Tolerogenic Dendritic Cells in the Control of Autoimmune Neuroinflammation: An Emerging Role of Protein-Glycan Interactions , 2010, Neuroimmunomodulation.

[3]  M. Rescigno Functional specialization of antigen presenting cells in the gastrointestinal tract. , 2010, Current opinion in immunology.

[4]  C. Norbury,et al.  Dendritic Cell Migration Limits the Duration of CD8+ T-Cell Priming to Peripheral Viral Antigen , 2010, Journal of Virology.

[5]  J. Banchereau,et al.  Ribosomal protein mRNAs are translationally-regulated during human dendritic cells activation by LPS , 2009, Immunome research.

[6]  S. Agrawal,et al.  Activation of human monocyte-derived dendritic cells in vitro by Thymax, a gross thymic extract. , 2009, Anticancer research.

[7]  Xiaoxi Li,et al.  Down-Regulation of TLR9 Expression Affects the Maturation and Function of Murine Bone Marrow-Derived Dendritic Cells Induced by CpG , 2009, Cellular and Molecular Immunology.

[8]  S. Abediankenari,et al.  Generation of immune inhibitory dendritic cells and CD4+T regulatory cells inducing by TGF-beta. , 2009, Iranian journal of allergy, asthma, and immunology.

[9]  E. Read,et al.  Effect of ex vivo culture duration on phenotype and cytokine production by mature dendritic cells derived from peripheral blood monocytes , 2009, Transfusion.

[10]  J. Peters,et al.  Poly(I:C) coated PLGA microparticles induce dendritic cell maturation. , 2009, International journal of pharmaceutics.

[11]  B. Kelsall Recent progress in understanding the phenotype and function of intestinal dendritic cells and macrophages , 2008, Mucosal Immunology.

[12]  N. Kadowaki,et al.  Natural Interferon α/β–Producing Cells Link Innate and Adaptive Immunity , 2000, The Journal of experimental medicine.

[13]  R. Verdijk,et al.  Polyriboinosinic polyribocytidylic acid (poly(I:C)) induces stable maturation of functionally active human dendritic cells. , 1999, Journal of immunology.

[14]  K. Kim CD4+T Cells , 1993 .

[15]  T. Mosmann Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[16]  M. Noguchi,et al.  Dendritic cell-derived TNF-alpha is responsible for development of IL-10-producing CD4+ T cells. , 2010, Cellular immunology.

[17]  C. Reis e Sousa,et al.  Activation of dendritic cells: translating innate into adaptive immunity. , 2004, Current opinion in immunology.

[18]  E. Borden,et al.  Polyriboinosinic:polyribocytidylic acid (poly i:c) as an antitumor compound: factors influencing in vivo activity. Abstr. , 1979 .