© The Rockefeller University Press, 0022-1007/1997/1/317/ $5.00
The Journal of Experimental Medicine, Volume 185, Number 2, January 20, 1997 317-328
Maturation Stages of Mouse Dendritic Cells in Growth Factor–dependent Long-Term Cultures
Claudia Winzler*,
Patrizia Rovere
,
Maria Rescigno*,
Francesca Granucci*,
Giuseppe Penna
,
Luciano Adorini
,
Valerie S. Zimmermann
,
Jean Davoust
, and
Paola Ricciardi-Castagnoli*
From the * CNR Centre of Cellular and Molecular Pharmacology, Milan 20129, Italy;
Roche Milano Ricerche, Milan 20132, Italy;
Centre d'Immunologie INSERM-CNRS de Marseille Luminy, Parc Scientifique de Luminy, Case 906, Marseille 13288, France
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.
Address correspondence to Paola Ricciardi-Castagnoli, CNR (National Council Research) Centre of Cellular and Molecular Pharmacology, Via Vanvitelli 32, 20129 Milano, Italy.
This work was supported by the Italian Ministry of Public Health (Istituto Superiore di Sanità: AIDS grant 9403-92 and MS grant 52), by the National Council Research (CNR), by Biotop, and by the French Association against Cancer, grant 6058 (ARC).
1 Abbreviations used in this paper: DC, dendritic cells; LC, Langerhans cells; SN, supernatant.

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