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Dendritic cells enhance growth and differentiation of CD40-activated B lymphocytes.

Dubois B, Vanbervliet B, Fayette J, Massacrier C, Van Kooten C, Brière F, Banchereau J, Caux C - J. Exp. Med. (1997)

Bottom Line: Furthermore, after CD40 ligation, DC enhanced by 30-300-fold the secretion of IgG and IgA by sIgD- B cells (essentially memory B cells).In the presence of DC, naive sIgD+ B cells produced, in response to interleukin-2, large amounts of IgM.Thus, in addition to activating naive T cells in the extrafollicular areas of secondary lymphoid organs, DC may directly modulate B cell growth and differentiation.

View Article: PubMed Central - PubMed

Affiliation: Schering Plough, Laboratory for Immunological Research, Dardilly, France.

ABSTRACT
After antigen capture, dendritic cells (DC) migrate into T cell-rich areas of secondary lymphoid organs, where they induce T cell activation, that subsequently drives B cell activation. Here, we investigate whether DC, generated in vitro, can directly modulate B cell responses, using CD40L-transfected L cells as surrogate activated T cells. DC, through the production of soluble mediators, stimulated by 3- to 6-fold the proliferation and subsequent recovery of B cells. Furthermore, after CD40 ligation, DC enhanced by 30-300-fold the secretion of IgG and IgA by sIgD- B cells (essentially memory B cells). In the presence of DC, naive sIgD+ B cells produced, in response to interleukin-2, large amounts of IgM. Thus, in addition to activating naive T cells in the extrafollicular areas of secondary lymphoid organs, DC may directly modulate B cell growth and differentiation.

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D–Lc strongly enhance Ig production by CD40-activated  memory B cells. B cells were cultured over CD40L L cells in the presence  or absence of D–Lc and their supernatants were harvested after 15 d and  assayed for presence of (A) IgG, IgA, and IgM. (B) Increasing numbers of  D–Lc were added to 104 CD40-activated B cells and total Ig production  was measured after 15 d of coculture. Igs levels are expressed as mean ±  SD of triplicate cultures (results from 1 experiment representative of 10).  (C) B cells were purified into IgD+ B cells and IgD− B cells using the  MACS system (purification detailed in Materials and Methods). IgD− B  cells were further separated into CD38+CD39− germinal center B cells  (GC) and CD38−CD39+ memory B cells using mAbs and bead depletion  as detailed in Materials and Methods. Increasing numbers of either total B  cells, MACS purified sIgD+ B cells, GC cells, or memory B cells were  cultured over 2.5 × 103 irradiated CD40L L cells in medium alone or in  the presence of 104 irradiated D–Lc. (SD ⩽ 10%). (results from 1 of   3 experiments). No significant B cell proliferation and differentiation was observed in absence of CD40L L cells.
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Figure 2: D–Lc strongly enhance Ig production by CD40-activated memory B cells. B cells were cultured over CD40L L cells in the presence or absence of D–Lc and their supernatants were harvested after 15 d and assayed for presence of (A) IgG, IgA, and IgM. (B) Increasing numbers of D–Lc were added to 104 CD40-activated B cells and total Ig production was measured after 15 d of coculture. Igs levels are expressed as mean ± SD of triplicate cultures (results from 1 experiment representative of 10). (C) B cells were purified into IgD+ B cells and IgD− B cells using the MACS system (purification detailed in Materials and Methods). IgD− B cells were further separated into CD38+CD39− germinal center B cells (GC) and CD38−CD39+ memory B cells using mAbs and bead depletion as detailed in Materials and Methods. Increasing numbers of either total B cells, MACS purified sIgD+ B cells, GC cells, or memory B cells were cultured over 2.5 × 103 irradiated CD40L L cells in medium alone or in the presence of 104 irradiated D–Lc. (SD ⩽ 10%). (results from 1 of   3 experiments). No significant B cell proliferation and differentiation was observed in absence of CD40L L cells.

Mentions: To investigate the effect of D–Lc on the differentiation of CD40-activated B cells into Ig-secreting cells, tonsillar B cells were cultured over CD40L L cells with or without D–Lc. In the absence of D–Lc, 104 B cells produced only marginal amounts of Igs (<0.2 μg/ml; Fig. 2 A). Addition of 104 D–Lc to CD40-activated B cells results in the production of microgram amounts of IgG (range, 4–70 μg/ml; mean increase, 125; n = 12) and IgA (range, 0.8–6 μg/ml; mean increase, 33; n = 12). In contrast, IgM secretion was increased only 5- to 15-fold (range, 0.5–2 μg/ml; mean increase, 10; n = 12). As few as 330 D–Lc induced an eightfold increase in total Ig secretion, maximal effect being obtained with 104 D–Lc per well (40-fold increase) (Fig. 2 B).


Dendritic cells enhance growth and differentiation of CD40-activated B lymphocytes.

Dubois B, Vanbervliet B, Fayette J, Massacrier C, Van Kooten C, Brière F, Banchereau J, Caux C - J. Exp. Med. (1997)

D–Lc strongly enhance Ig production by CD40-activated  memory B cells. B cells were cultured over CD40L L cells in the presence  or absence of D–Lc and their supernatants were harvested after 15 d and  assayed for presence of (A) IgG, IgA, and IgM. (B) Increasing numbers of  D–Lc were added to 104 CD40-activated B cells and total Ig production  was measured after 15 d of coculture. Igs levels are expressed as mean ±  SD of triplicate cultures (results from 1 experiment representative of 10).  (C) B cells were purified into IgD+ B cells and IgD− B cells using the  MACS system (purification detailed in Materials and Methods). IgD− B  cells were further separated into CD38+CD39− germinal center B cells  (GC) and CD38−CD39+ memory B cells using mAbs and bead depletion  as detailed in Materials and Methods. Increasing numbers of either total B  cells, MACS purified sIgD+ B cells, GC cells, or memory B cells were  cultured over 2.5 × 103 irradiated CD40L L cells in medium alone or in  the presence of 104 irradiated D–Lc. (SD ⩽ 10%). (results from 1 of   3 experiments). No significant B cell proliferation and differentiation was observed in absence of CD40L L cells.
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Related In: Results  -  Collection

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Figure 2: D–Lc strongly enhance Ig production by CD40-activated memory B cells. B cells were cultured over CD40L L cells in the presence or absence of D–Lc and their supernatants were harvested after 15 d and assayed for presence of (A) IgG, IgA, and IgM. (B) Increasing numbers of D–Lc were added to 104 CD40-activated B cells and total Ig production was measured after 15 d of coculture. Igs levels are expressed as mean ± SD of triplicate cultures (results from 1 experiment representative of 10). (C) B cells were purified into IgD+ B cells and IgD− B cells using the MACS system (purification detailed in Materials and Methods). IgD− B cells were further separated into CD38+CD39− germinal center B cells (GC) and CD38−CD39+ memory B cells using mAbs and bead depletion as detailed in Materials and Methods. Increasing numbers of either total B cells, MACS purified sIgD+ B cells, GC cells, or memory B cells were cultured over 2.5 × 103 irradiated CD40L L cells in medium alone or in the presence of 104 irradiated D–Lc. (SD ⩽ 10%). (results from 1 of   3 experiments). No significant B cell proliferation and differentiation was observed in absence of CD40L L cells.
Mentions: To investigate the effect of D–Lc on the differentiation of CD40-activated B cells into Ig-secreting cells, tonsillar B cells were cultured over CD40L L cells with or without D–Lc. In the absence of D–Lc, 104 B cells produced only marginal amounts of Igs (<0.2 μg/ml; Fig. 2 A). Addition of 104 D–Lc to CD40-activated B cells results in the production of microgram amounts of IgG (range, 4–70 μg/ml; mean increase, 125; n = 12) and IgA (range, 0.8–6 μg/ml; mean increase, 33; n = 12). In contrast, IgM secretion was increased only 5- to 15-fold (range, 0.5–2 μg/ml; mean increase, 10; n = 12). As few as 330 D–Lc induced an eightfold increase in total Ig secretion, maximal effect being obtained with 104 D–Lc per well (40-fold increase) (Fig. 2 B).

Bottom Line: Furthermore, after CD40 ligation, DC enhanced by 30-300-fold the secretion of IgG and IgA by sIgD- B cells (essentially memory B cells).In the presence of DC, naive sIgD+ B cells produced, in response to interleukin-2, large amounts of IgM.Thus, in addition to activating naive T cells in the extrafollicular areas of secondary lymphoid organs, DC may directly modulate B cell growth and differentiation.

View Article: PubMed Central - PubMed

Affiliation: Schering Plough, Laboratory for Immunological Research, Dardilly, France.

ABSTRACT
After antigen capture, dendritic cells (DC) migrate into T cell-rich areas of secondary lymphoid organs, where they induce T cell activation, that subsequently drives B cell activation. Here, we investigate whether DC, generated in vitro, can directly modulate B cell responses, using CD40L-transfected L cells as surrogate activated T cells. DC, through the production of soluble mediators, stimulated by 3- to 6-fold the proliferation and subsequent recovery of B cells. Furthermore, after CD40 ligation, DC enhanced by 30-300-fold the secretion of IgG and IgA by sIgD- B cells (essentially memory B cells). In the presence of DC, naive sIgD+ B cells produced, in response to interleukin-2, large amounts of IgM. Thus, in addition to activating naive T cells in the extrafollicular areas of secondary lymphoid organs, DC may directly modulate B cell growth and differentiation.

Show MeSH
Related in: MedlinePlus