Maturation Stages of Mouse Dendritic Cells in Growth Factor–dependent Long-Term Cultures

Abstract

The signals controlling the checkpoints of dendritic cells (DC) maturation and the correlation between phenotypical and functional maturational stages were investigated in a defined model system of growth factor–dependent immature mouse DC. Three sequential stages of DC maturation (immature, mature, and apoptotic) were defined and characterized. Immature DC (stage 1) had low expression of costimulatory molecules, highly organized cytoskeleton, focal adhesion plaques, and slow motility; accordingly, they were very efficient in antigen uptake and processing of soluble proteins. Further, at this stage most of major histocompatibility complex class II molecules were within cytoplasmic compartments consistent with a poor allostimulatory capacity. Bacteria or cytokines were very efficient in inducing progression from stage 1 towards stage 2 (mature). Morphological changes were observed by confocal analysis including depolymerization of F-actin and loss of vinculin containing adhesive structures which correlates with acquisition of high motility. Antigen uptake and presentation of native protein antigen was reduced. In contrast, presentation of immunogenic peptides and allostimulatory activity became very efficient and secretion of IL-12 p75 was detectable after antigen presentation. This functional DC maturation ended by apoptotic cell death, and no reversion to the immature phenotype was observed.

Targeting dendritic cells with antigen via dendritic cell-associated promoters

“…The DC cell line, D1 (kindly provided by Dr Paola Ricciardi-Castagnoli) was maintained in culture as previously described. 25 The B3Z T-cell hybridoma 26 and the OVA-transfected, murine melanoma cell line, B16OVA, 27 (derived from B16F10, clone MO5) was kindly provided by Dr Kenneth Rock. The medium used for the in vitro T-cell stimulation was RPMI 1640 supplemented with 5% FCS, glutamine, 50 mM b-Mercaptoethanol and antibiotics/antimycotics.…”

A recombinant E. coli vaccine to promote MHC class I-dependent antigen presentation: application to cancer immunotherapy

“…[13][14][15] Recombinant E. coli have been used as a standard molecular biology tool for over three decades and are widely used for the production of recombinant proteins. Therefore, such bacteria could be promising candidates for the delivery of tumour antigens to DCs.…”

Saturated and Polyunsaturated Fatty Acids Reciprocally Modulate Dendritic Cell Functions Mediated through TLR4

“…Thus, we determined whether the expression of cytokines is similarly modulated by the fatty acids during the maturation of DCs. IL-12p70 is responsible for driving CD4 ϩ T cells to a Th1 response, and its production by DCs correlates with the functional maturation of DCs (23). Unlike IL-12p70, IL-6 does not promote DC maturation, but induces acute phase responses (23).…”