Generation of functional monocyte-derived fast dendritic cells suitable for clinical application in the absence of interleukin-6

Abstract

To develop dendritic cells (DCs)-based immunotherapy for cancer patients, it is necessary to have a standardized, reproducible, fast, and easy to use protocol for in vitro generation of fully functional DCs. Recently, a new strategy was described for differentiation and maturation of human monocyte (Mo)-derived fast-DCs with full T cell stimulatory capacity within only 48-72 h of in vitro culture. Interleukin (IL)-6 plus tumour necrosis factor (TNF)-a, IL-1b, and prostaglandin (PG)-E 2 were used in this strategy to induce maturation of the generated DCs. The present study further modifies this strategy by excluding IL-6 from the cytokines cocktail used for DCs maturation. The results showed that maturation of fast-DCs without IL-6 did not significantly alter the morphology, phenotype and the yield of mature DCs (P [ 0.05, compared with those generated with IL-6). Moreover, fast-DCs generated without IL-6 are functional antigen presenting cells, have the ability to induce tetanus toxoid-specific autologous T cell proliferation, and are suitable for gene delivery through adenoviral vector transduction as those generated with IL-6 (P [ 0.05). In conclusion, the present study proves that fully mature and functional Mo-derived fast-DCs can be generated in vitro without adding IL-6, which not only reduces the number of required recombinant cytokines, but may also resemble DCs development in vivo more closely.

PGE2 Is Crucial for the Generation of FAST Whole- Tumor-Antigens Loaded Dendritic Cells Suitable for Immunotherapy in Glioblastoma

“…Dauer and colleagues on 2003 described a strategy for differentiation and maturation of monocyte derived DC within only 48-72 h of in vitro culture, called "Fast" protocol, and several other groups have further elaborated on this pioneering work [13][14][15][16][17]. Here we described this method modified from our group to obtain Fast mature autologous DC, loaded with antigens obtained from whole tumor lysate, suitable for the immunotherapy of glioblastoma patients.…”

“…Fully mature DC are generally obtained in 7-10 days of culture: differentiation step take place in 5-7 days with GM-CSF and IL4; immature DC are subsequently induced to maturation for 1-3 days with pro-inflammatory stimuli to generate a population of immunogenic mature DC [12]. Several groups have shown that it is possible to obtain mature DC in 2-3 days cell culture [13][14][15][16][17]. The in vitro generation of DC with Fast protocol will shorten the time from the patients' recruiting to DC treatment and therefore be beneficial in the clinical setting: a Fast protocol would result in a simplified processing that minimizes inter-preparations variability and the risk of contamination, also resulting less expensive.…”

Advances and challenges: dendritic cell vaccination strategies for glioblastoma

“…‘FastDCs’ have been obtained after mere 2-day culturing periods and retain the essential ability to induce antigen-specific T-cell responses [33, 34]. Important differences of fastDCs compared to standard DCs include higher yields in culture, lower release of IL-12 p70, higher CCL19-induced chemokinesis, better intracellular antigen processing, and more effective priming of tumor-specific cytotoxic T-cells [33, 35, 36].…”

Specific immunotherapy of experimental myasthenia gravis in vitro and in vivo: The Guided Missile strategy

“…(Note that KLH antibody levels in our ELISA system are about 10-fold higher than AChR antibody levels). selective culture (Jones et al, 2012), (Ramadan, 2011), and newer methods of deriving large quantities of DCs from iPS cells are promising (Senju et al, 2010). Fortunately, mature human DCs are not vulnerable to killing by FasL (Ashany et al, 1999;Hoves et al, 2003), and should therefore be well suited for this purpose.…”