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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.

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Allostimulatory and  processing capacities of D1 cells.  (a) Mixed lymphocyte reaction  (MLR) by sorted I-Abright and  I-Aint D1 cells shows that the  mature I-Abright cells are the ones  that have the highest allostimulatory capacity. Presentation of the  OVA protein is downregulated  by TNFα treatment. Antigenspecific presentation of exogenous OVA protein (b) or OVApeptide327-339 (c) by D1 cells  treated or not with TNFα for 1  or 7 d: presentation of the OVA  peptide is not affected by maturation but presentation of the  OVA protein is downregulated  by TNFα.
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Figure 8: Allostimulatory and processing capacities of D1 cells. (a) Mixed lymphocyte reaction (MLR) by sorted I-Abright and I-Aint D1 cells shows that the mature I-Abright cells are the ones that have the highest allostimulatory capacity. Presentation of the OVA protein is downregulated by TNFα treatment. Antigenspecific presentation of exogenous OVA protein (b) or OVApeptide327-339 (c) by D1 cells treated or not with TNFα for 1 or 7 d: presentation of the OVA peptide is not affected by maturation but presentation of the OVA protein is downregulated by TNFα.

Mentions: 1 d after sorting, I-Abright and I-Aint cells were used as stimulators of unprimed BALB/c lymphnode T lymphocytes. I-Abright cells resulted much more potent activators of allogeneic T cells as compared to I-Aint cells (Fig. 8 a), in agreement with the notion that mature DC acquire a stronger stimulatory activity. However the situation may be different for nominal class II–restricted protein antigens. Immature murine DC were in fact reported to present native protein antigens better than shortterm cultured DC (19). To address this issue, I-Abright and I-Aint sorted cells were compared for their capacity to process and present native OVA or OVA peptides to the specific T hybridoma BO97.10. Sorted I-Aint cells were more efficient in the presentation of native OVA whereas both I-Abrightand I-Aintcells were very efficient in the presentation of the peptide to the same hybridoma (data not shown).


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)

Allostimulatory and  processing capacities of D1 cells.  (a) Mixed lymphocyte reaction  (MLR) by sorted I-Abright and  I-Aint D1 cells shows that the  mature I-Abright cells are the ones  that have the highest allostimulatory capacity. Presentation of the  OVA protein is downregulated  by TNFα treatment. Antigenspecific presentation of exogenous OVA protein (b) or OVApeptide327-339 (c) by D1 cells  treated or not with TNFα for 1  or 7 d: presentation of the OVA  peptide is not affected by maturation but presentation of the  OVA protein is downregulated  by TNFα.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 8: Allostimulatory and processing capacities of D1 cells. (a) Mixed lymphocyte reaction (MLR) by sorted I-Abright and I-Aint D1 cells shows that the mature I-Abright cells are the ones that have the highest allostimulatory capacity. Presentation of the OVA protein is downregulated by TNFα treatment. Antigenspecific presentation of exogenous OVA protein (b) or OVApeptide327-339 (c) by D1 cells treated or not with TNFα for 1 or 7 d: presentation of the OVA peptide is not affected by maturation but presentation of the OVA protein is downregulated by TNFα.
Mentions: 1 d after sorting, I-Abright and I-Aint cells were used as stimulators of unprimed BALB/c lymphnode T lymphocytes. I-Abright cells resulted much more potent activators of allogeneic T cells as compared to I-Aint cells (Fig. 8 a), in agreement with the notion that mature DC acquire a stronger stimulatory activity. However the situation may be different for nominal class II–restricted protein antigens. Immature murine DC were in fact reported to present native protein antigens better than shortterm cultured DC (19). To address this issue, I-Abright and I-Aint sorted cells were compared for their capacity to process and present native OVA or OVA peptides to the specific T hybridoma BO97.10. Sorted I-Aint cells were more efficient in the presentation of native OVA whereas both I-Abrightand I-Aintcells were very efficient in the presentation of the peptide to the same hybridoma (data not shown).

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