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Dendritic cells pulsed with intact Streptococcus pneumoniae elicit both protein- and polysaccharide-specific immunoglobulin isotype responses in vivo through distinct mechanisms.

Colino J, Shen Y, Snapper CM - J. Exp. Med. (2002)

Bottom Line: Immature bone marrow-derived myeloid dendritic cells (BMDCs) are induced to undergo phenotypic maturation and secretion of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-12, and IL-10 when pulsed in vitro with intact Streptococcus pneumoniae.After transfer to naive mice, pulsed BMDCs induce immunoglobulin (Ig) isotype responses specific for both protein and polysaccharide pneumococcal antigens, having in common the requirement for viable BMDCs, T cells, and B7-dependent costimulation in the recipient mice.Whereas primary Ig isotype responses to bacterial proteins uniformly require BMDC expression of major histocompatibility complex class II, CD40, and B7, and the secretion of IL-6, but not IL-12, similar requirements for antipolysaccharide Ig responses were only observed for the IgG1 isotype.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USA.

ABSTRACT
Immature bone marrow-derived myeloid dendritic cells (BMDCs) are induced to undergo phenotypic maturation and secretion of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-12, and IL-10 when pulsed in vitro with intact Streptococcus pneumoniae. After transfer to naive mice, pulsed BMDCs induce immunoglobulin (Ig) isotype responses specific for both protein and polysaccharide pneumococcal antigens, having in common the requirement for viable BMDCs, T cells, and B7-dependent costimulation in the recipient mice. Whereas primary Ig isotype responses to bacterial proteins uniformly require BMDC expression of major histocompatibility complex class II, CD40, and B7, and the secretion of IL-6, but not IL-12, similar requirements for antipolysaccharide Ig responses were only observed for the IgG1 isotype.

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Kinetics of the in vivo antiprotein and antipolysaccharide Ig isotype induced by in vivo transfer of BMDCs pulsed with S. pneumoniae type 14. Groups of C57BL/6 mice (n = 6) received an intravenous injection of 106 BMDCs pulsed with S. pneumoniae type 14 (black symbols) or 106 unpulsed BMDCs as control (white symbols) at day 0. At day 21, both groups of mice received an intraperitoneal immunization with 5 × 106 free S. pneumoniae type 14 (indicated by arrows). The data shows the serum titers of Ig isotypes specific for PspA (top), PC (middle), and Cps14 (bottom) on different days after immunization. The data shown are representative of two independent experiments.
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fig2: Kinetics of the in vivo antiprotein and antipolysaccharide Ig isotype induced by in vivo transfer of BMDCs pulsed with S. pneumoniae type 14. Groups of C57BL/6 mice (n = 6) received an intravenous injection of 106 BMDCs pulsed with S. pneumoniae type 14 (black symbols) or 106 unpulsed BMDCs as control (white symbols) at day 0. At day 21, both groups of mice received an intraperitoneal immunization with 5 × 106 free S. pneumoniae type 14 (indicated by arrows). The data shows the serum titers of Ig isotypes specific for PspA (top), PC (middle), and Cps14 (bottom) on different days after immunization. The data shown are representative of two independent experiments.

Mentions: In preliminary experiments we established a set of conditions for inducing optimal in vivo protein- and polysaccharide-specific Ig responses upon transfer of S. pneumoniae-pulsed DCs (“pulsed BMDCs”) into naive mice (data not shown). On the basis of these studies, we chose to inject each mouse intravenously with 106 viable BMDCs that were pulsed with a ratio of 800 bacteria per individual DC for a 4–5 h period. This resulted in each DC internalizing an average of ≤10 bacteria (see Fig. 1 A). As illustrated above, these conditions for BMDCs pulsing further allowed for in vitro autocrine/paracrine exposure of BMDCs to stimulatory cytokines, but without significant exposure to immunosuppressive IL-10, and for continued cytokine secretion, with the exception of TNF-α, by BMDCs upon transfer in vivo. As shown in Fig. 2 , BMDCs pulsed with heat-killed S. pneumoniae and transferred into naive mice stimulated a primary Ig response specific for PspA, Cps14, and PC (hapten component of the cell wall C-polysaccharide). BMDCs pulsed with formalinized bacteria induced comparable primary antigen-specific Ig responses (data not shown). A primary Ig response specific for PsaA, a membrane-bound lipoprotein, was also induced (data not shown). In contrast, unpulsed BMDCs failed to induce any specific primary Ig response.


Dendritic cells pulsed with intact Streptococcus pneumoniae elicit both protein- and polysaccharide-specific immunoglobulin isotype responses in vivo through distinct mechanisms.

Colino J, Shen Y, Snapper CM - J. Exp. Med. (2002)

Kinetics of the in vivo antiprotein and antipolysaccharide Ig isotype induced by in vivo transfer of BMDCs pulsed with S. pneumoniae type 14. Groups of C57BL/6 mice (n = 6) received an intravenous injection of 106 BMDCs pulsed with S. pneumoniae type 14 (black symbols) or 106 unpulsed BMDCs as control (white symbols) at day 0. At day 21, both groups of mice received an intraperitoneal immunization with 5 × 106 free S. pneumoniae type 14 (indicated by arrows). The data shows the serum titers of Ig isotypes specific for PspA (top), PC (middle), and Cps14 (bottom) on different days after immunization. The data shown are representative of two independent experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196017&req=5

fig2: Kinetics of the in vivo antiprotein and antipolysaccharide Ig isotype induced by in vivo transfer of BMDCs pulsed with S. pneumoniae type 14. Groups of C57BL/6 mice (n = 6) received an intravenous injection of 106 BMDCs pulsed with S. pneumoniae type 14 (black symbols) or 106 unpulsed BMDCs as control (white symbols) at day 0. At day 21, both groups of mice received an intraperitoneal immunization with 5 × 106 free S. pneumoniae type 14 (indicated by arrows). The data shows the serum titers of Ig isotypes specific for PspA (top), PC (middle), and Cps14 (bottom) on different days after immunization. The data shown are representative of two independent experiments.
Mentions: In preliminary experiments we established a set of conditions for inducing optimal in vivo protein- and polysaccharide-specific Ig responses upon transfer of S. pneumoniae-pulsed DCs (“pulsed BMDCs”) into naive mice (data not shown). On the basis of these studies, we chose to inject each mouse intravenously with 106 viable BMDCs that were pulsed with a ratio of 800 bacteria per individual DC for a 4–5 h period. This resulted in each DC internalizing an average of ≤10 bacteria (see Fig. 1 A). As illustrated above, these conditions for BMDCs pulsing further allowed for in vitro autocrine/paracrine exposure of BMDCs to stimulatory cytokines, but without significant exposure to immunosuppressive IL-10, and for continued cytokine secretion, with the exception of TNF-α, by BMDCs upon transfer in vivo. As shown in Fig. 2 , BMDCs pulsed with heat-killed S. pneumoniae and transferred into naive mice stimulated a primary Ig response specific for PspA, Cps14, and PC (hapten component of the cell wall C-polysaccharide). BMDCs pulsed with formalinized bacteria induced comparable primary antigen-specific Ig responses (data not shown). A primary Ig response specific for PsaA, a membrane-bound lipoprotein, was also induced (data not shown). In contrast, unpulsed BMDCs failed to induce any specific primary Ig response.

Bottom Line: Immature bone marrow-derived myeloid dendritic cells (BMDCs) are induced to undergo phenotypic maturation and secretion of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-12, and IL-10 when pulsed in vitro with intact Streptococcus pneumoniae.After transfer to naive mice, pulsed BMDCs induce immunoglobulin (Ig) isotype responses specific for both protein and polysaccharide pneumococcal antigens, having in common the requirement for viable BMDCs, T cells, and B7-dependent costimulation in the recipient mice.Whereas primary Ig isotype responses to bacterial proteins uniformly require BMDC expression of major histocompatibility complex class II, CD40, and B7, and the secretion of IL-6, but not IL-12, similar requirements for antipolysaccharide Ig responses were only observed for the IgG1 isotype.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814, USA.

ABSTRACT
Immature bone marrow-derived myeloid dendritic cells (BMDCs) are induced to undergo phenotypic maturation and secretion of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-12, and IL-10 when pulsed in vitro with intact Streptococcus pneumoniae. After transfer to naive mice, pulsed BMDCs induce immunoglobulin (Ig) isotype responses specific for both protein and polysaccharide pneumococcal antigens, having in common the requirement for viable BMDCs, T cells, and B7-dependent costimulation in the recipient mice. Whereas primary Ig isotype responses to bacterial proteins uniformly require BMDC expression of major histocompatibility complex class II, CD40, and B7, and the secretion of IL-6, but not IL-12, similar requirements for antipolysaccharide Ig responses were only observed for the IgG1 isotype.

Show MeSH
Related in: MedlinePlus