Limits...
Maturation stages of mouse dendritic cells in growth factor-dependent long-term cultures.

Winzler C, Rovere P, Rescigno M, Granucci F, Penna G, Adorini L, Zimmermann VS, Davoust J, Ricciardi-Castagnoli P - J. Exp. Med. (1997)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: CNR Centre of Cellular and Molecular Pharmacology, Milan, Italy.

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.

Show MeSH

Related in: MedlinePlus

Phenotypical maturation of D1 cells.  Surface markers by FACS® analysis after cytokine  or living bacteria treatment. MHC class II (I-A),  B7.2, CD40, and ICAM-1 molecules of D1 bulk  culture growing in R1 medium or in medium supplemented with TNFα, LPS, IL-1β, or IL-6 (A) or  after treatment with living E. coli or S. aureus (B).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2196118&req=5

Figure 4: Phenotypical maturation of D1 cells. Surface markers by FACS® analysis after cytokine or living bacteria treatment. MHC class II (I-A), B7.2, CD40, and ICAM-1 molecules of D1 bulk culture growing in R1 medium or in medium supplemented with TNFα, LPS, IL-1β, or IL-6 (A) or after treatment with living E. coli or S. aureus (B).

Mentions: TNFα, LPS, IL-1β, and IL-6 were tested for their ability to modulate the expression of markers involved in APC/T cell interactions and differentially expressed in immature versus mature DC. Cells were stimulated for 24 h with either 100 U/ml TNFα, 1 μg/ml LPS or 10 ng/ml IL-1β, stained, and then analyzed by flow cytometry. I-A, B7.2, CD40, and ICAM-1 molecules were strongly upregulated by all stimuli, whereas expression of the same surface molecules did not change after treatment with 10 ng/ml IL-6 (Fig. 4 A).


Maturation stages of mouse dendritic cells in growth factor-dependent long-term cultures.

Winzler C, Rovere P, Rescigno M, Granucci F, Penna G, Adorini L, Zimmermann VS, Davoust J, Ricciardi-Castagnoli P - J. Exp. Med. (1997)

Phenotypical maturation of D1 cells.  Surface markers by FACS® analysis after cytokine  or living bacteria treatment. MHC class II (I-A),  B7.2, CD40, and ICAM-1 molecules of D1 bulk  culture growing in R1 medium or in medium supplemented with TNFα, LPS, IL-1β, or IL-6 (A) or  after treatment with living E. coli or S. aureus (B).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196118&req=5

Figure 4: Phenotypical maturation of D1 cells. Surface markers by FACS® analysis after cytokine or living bacteria treatment. MHC class II (I-A), B7.2, CD40, and ICAM-1 molecules of D1 bulk culture growing in R1 medium or in medium supplemented with TNFα, LPS, IL-1β, or IL-6 (A) or after treatment with living E. coli or S. aureus (B).
Mentions: TNFα, LPS, IL-1β, and IL-6 were tested for their ability to modulate the expression of markers involved in APC/T cell interactions and differentially expressed in immature versus mature DC. Cells were stimulated for 24 h with either 100 U/ml TNFα, 1 μg/ml LPS or 10 ng/ml IL-1β, stained, and then analyzed by flow cytometry. I-A, B7.2, CD40, and ICAM-1 molecules were strongly upregulated by all stimuli, whereas expression of the same surface molecules did not change after treatment with 10 ng/ml IL-6 (Fig. 4 A).

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: CNR Centre of Cellular and Molecular Pharmacology, Milan, Italy.

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.

Show MeSH
Related in: MedlinePlus