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IgE enhances mouse mast cell Fc(epsilon)RI expression in vitro and in vivo: evidence for a novel amplification mechanism in IgE-dependent reactions.

Yamaguchi M, Lantz CS, Oettgen HC, Katona IM, Fleming T, Miyajima I, Kinet JP, Galli SJ - J. Exp. Med. (1997)

Bottom Line: This IgE- and antigen-specific mast cell activation and mediator production is thought to be critical to the pathogenesis of allergic disorders, such as anaphylaxis and asthma, and also contributes to host defense against parasites.In turn, IgE-dependent upregulation of Fc(epsilon)RI expression significantly enhances the ability of mouse mast cells to release serotonin, interleukin-6 (IL-6), and IL-4 in response to challenge with IgE and specific antigen.The demonstration that IgE-dependent enhancement of mast cell Fc(epsilon)RI expression permits mast cells to respond to antigen challenge with increased production of proinflammatory and immunoregulatory mediators provides new insights into both the pathogenesis of allergic diseases and the regulation of protective host responses to parasites.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02215, USA.

ABSTRACT
The binding of immunoglobulin E (IgE) to high affinity IgE receptors (Fc(epsilon)RI) expressed on the surface of mast cells primes these cells to secrete, upon subsequent exposure to specific antigen, a panel of proinflammatory mediators, which includes cytokines that can also have immunoregulatory activities. This IgE- and antigen-specific mast cell activation and mediator production is thought to be critical to the pathogenesis of allergic disorders, such as anaphylaxis and asthma, and also contributes to host defense against parasites. We now report that exposure to IgE results in a striking (up to 32-fold) upregulation of surface expression of Fc(epsilon)RI on mouse mast cells in vitro or in vivo. Moreover, baseline levels of Fc(epsilon)RI expression on peritoneal mast cells from genetically IgE-deficient (IgE -/-) mice are dramatically reduced (by approximately 83%) compared with those on cells from the corresponding normal mice. In vitro studies indicate that the IgE-dependent upregulation of mouse mast cell Fc(epsilon)RI expression has two components: an early cycloheximide-insensitive phase, followed by a later and more sustained component that is highly sensitive to inhibition by cycloheximide. In turn, IgE-dependent upregulation of Fc(epsilon)RI expression significantly enhances the ability of mouse mast cells to release serotonin, interleukin-6 (IL-6), and IL-4 in response to challenge with IgE and specific antigen. The demonstration that IgE-dependent enhancement of mast cell Fc(epsilon)RI expression permits mast cells to respond to antigen challenge with increased production of proinflammatory and immunoregulatory mediators provides new insights into both the pathogenesis of allergic diseases and the regulation of protective host responses to parasites.

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IgE enhances mouse peritoneal mast cell FcεRI expression in  vitro in a time- and dose-dependent manner. (A) Identification of peritoneal mast cells (gated, in box) by flow cytometry. Freshly isolated peritoneal cells of retired breeder BALB/c mice were stained for the expression  of surface IgE receptor and c-kit. The gated cells were sorted and confirmed microscopically as 100% mast cells. (B–D) Regulation of surface  FcεRI expression on peritoneal mast cells by ascites IgE ex vivo. (B) Peritoneal cells isolated from BALB/c mice were cultured for 4 d without IgE  or with IgE at 0.005, 0.05, or 5.0 μg/ml, incubated with excess IgE at  4°C for 50 min to saturate mast cell FcεRI, and then mast cell surface  FcεRI expression was analyzed by flow cytometry. The background fluorescence intensity of mast cells stained only for c-kit is also shown (no  anti-IgE). (C) Dose-dependent effect of IgE on surface FcεRI expression  on peritoneal mast cells. Peritoneal cells from 5 BALB/c mice were combined, cultured with or without IgE for 4 d, and analyzed by flow cytometry for surface FcεRI. MESF, molecules of equivalent soluble fluorochrome units (see Materials and Methods). All data are mean ± SEM  (n = 3); ‡P = 0.076, ***P <0.0001 versus values for cells cultured without IgE for 4 d. (D) Time course of changes in mast cell surface FcεRI  expression in peritoneal cells (combined from 8 BALB/c mice) cultured  with or without IgE (5.0 μg/ml). All data are mean ± SEM (n = 3); **P  <0.001, ***P <0.0001 versus values at the same time point for cells cultured without IgE; †P <0.05, ††P <0.001, †††P <0.0001 versus baseline  (day 0) value.
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Figure 1: IgE enhances mouse peritoneal mast cell FcεRI expression in vitro in a time- and dose-dependent manner. (A) Identification of peritoneal mast cells (gated, in box) by flow cytometry. Freshly isolated peritoneal cells of retired breeder BALB/c mice were stained for the expression of surface IgE receptor and c-kit. The gated cells were sorted and confirmed microscopically as 100% mast cells. (B–D) Regulation of surface FcεRI expression on peritoneal mast cells by ascites IgE ex vivo. (B) Peritoneal cells isolated from BALB/c mice were cultured for 4 d without IgE or with IgE at 0.005, 0.05, or 5.0 μg/ml, incubated with excess IgE at 4°C for 50 min to saturate mast cell FcεRI, and then mast cell surface FcεRI expression was analyzed by flow cytometry. The background fluorescence intensity of mast cells stained only for c-kit is also shown (no anti-IgE). (C) Dose-dependent effect of IgE on surface FcεRI expression on peritoneal mast cells. Peritoneal cells from 5 BALB/c mice were combined, cultured with or without IgE for 4 d, and analyzed by flow cytometry for surface FcεRI. MESF, molecules of equivalent soluble fluorochrome units (see Materials and Methods). All data are mean ± SEM (n = 3); ‡P = 0.076, ***P <0.0001 versus values for cells cultured without IgE for 4 d. (D) Time course of changes in mast cell surface FcεRI expression in peritoneal cells (combined from 8 BALB/c mice) cultured with or without IgE (5.0 μg/ml). All data are mean ± SEM (n = 3); **P <0.001, ***P <0.0001 versus values at the same time point for cells cultured without IgE; †P <0.05, ††P <0.001, †††P <0.0001 versus baseline (day 0) value.

Mentions: As assessed by flow cytometry to detect binding of IgE (see Materials and Methods), we found that the baseline levels of FcεRI expression on the surface of BALB/c mouse peritoneal mast cells, identified as an IgE+, c-kit+ subpopulation of unfractionated peritoneal cells (Fig. 1 A), were greatly enhanced, in a concentration- (Fig. 1, B and C) and time- (Fig. 1 D) dependent manner, upon incubation of the cells in vitro with a mouse monoclonal IgE antibody. Moreover, FcεRI expression progressively diminished when peritoneal mast cells were incubated in vitro without added IgE (Fig. 1 D). As a result, levels of FcεRI expression in peritoneal mast cells incubated for 6 d with IgE at 5 μg/ml (i.e., in the range observed in the serum of parasite-infected mice, reference 28) were 20-fold higher than those in cells incubated for 6 d without IgE (Fig. 1 D). And mast cells incubated for 4 d with a concentration of IgE (0.05 μg/ml) comparable to that present in the serum of normal mice (28, 29) had levels of FcεRI expression that were 3.6-fold those in cells incubated for 4 d without IgE (Fig. 1 C).


IgE enhances mouse mast cell Fc(epsilon)RI expression in vitro and in vivo: evidence for a novel amplification mechanism in IgE-dependent reactions.

Yamaguchi M, Lantz CS, Oettgen HC, Katona IM, Fleming T, Miyajima I, Kinet JP, Galli SJ - J. Exp. Med. (1997)

IgE enhances mouse peritoneal mast cell FcεRI expression in  vitro in a time- and dose-dependent manner. (A) Identification of peritoneal mast cells (gated, in box) by flow cytometry. Freshly isolated peritoneal cells of retired breeder BALB/c mice were stained for the expression  of surface IgE receptor and c-kit. The gated cells were sorted and confirmed microscopically as 100% mast cells. (B–D) Regulation of surface  FcεRI expression on peritoneal mast cells by ascites IgE ex vivo. (B) Peritoneal cells isolated from BALB/c mice were cultured for 4 d without IgE  or with IgE at 0.005, 0.05, or 5.0 μg/ml, incubated with excess IgE at  4°C for 50 min to saturate mast cell FcεRI, and then mast cell surface  FcεRI expression was analyzed by flow cytometry. The background fluorescence intensity of mast cells stained only for c-kit is also shown (no  anti-IgE). (C) Dose-dependent effect of IgE on surface FcεRI expression  on peritoneal mast cells. Peritoneal cells from 5 BALB/c mice were combined, cultured with or without IgE for 4 d, and analyzed by flow cytometry for surface FcεRI. MESF, molecules of equivalent soluble fluorochrome units (see Materials and Methods). All data are mean ± SEM  (n = 3); ‡P = 0.076, ***P <0.0001 versus values for cells cultured without IgE for 4 d. (D) Time course of changes in mast cell surface FcεRI  expression in peritoneal cells (combined from 8 BALB/c mice) cultured  with or without IgE (5.0 μg/ml). All data are mean ± SEM (n = 3); **P  <0.001, ***P <0.0001 versus values at the same time point for cells cultured without IgE; †P <0.05, ††P <0.001, †††P <0.0001 versus baseline  (day 0) value.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2196143&req=5

Figure 1: IgE enhances mouse peritoneal mast cell FcεRI expression in vitro in a time- and dose-dependent manner. (A) Identification of peritoneal mast cells (gated, in box) by flow cytometry. Freshly isolated peritoneal cells of retired breeder BALB/c mice were stained for the expression of surface IgE receptor and c-kit. The gated cells were sorted and confirmed microscopically as 100% mast cells. (B–D) Regulation of surface FcεRI expression on peritoneal mast cells by ascites IgE ex vivo. (B) Peritoneal cells isolated from BALB/c mice were cultured for 4 d without IgE or with IgE at 0.005, 0.05, or 5.0 μg/ml, incubated with excess IgE at 4°C for 50 min to saturate mast cell FcεRI, and then mast cell surface FcεRI expression was analyzed by flow cytometry. The background fluorescence intensity of mast cells stained only for c-kit is also shown (no anti-IgE). (C) Dose-dependent effect of IgE on surface FcεRI expression on peritoneal mast cells. Peritoneal cells from 5 BALB/c mice were combined, cultured with or without IgE for 4 d, and analyzed by flow cytometry for surface FcεRI. MESF, molecules of equivalent soluble fluorochrome units (see Materials and Methods). All data are mean ± SEM (n = 3); ‡P = 0.076, ***P <0.0001 versus values for cells cultured without IgE for 4 d. (D) Time course of changes in mast cell surface FcεRI expression in peritoneal cells (combined from 8 BALB/c mice) cultured with or without IgE (5.0 μg/ml). All data are mean ± SEM (n = 3); **P <0.001, ***P <0.0001 versus values at the same time point for cells cultured without IgE; †P <0.05, ††P <0.001, †††P <0.0001 versus baseline (day 0) value.
Mentions: As assessed by flow cytometry to detect binding of IgE (see Materials and Methods), we found that the baseline levels of FcεRI expression on the surface of BALB/c mouse peritoneal mast cells, identified as an IgE+, c-kit+ subpopulation of unfractionated peritoneal cells (Fig. 1 A), were greatly enhanced, in a concentration- (Fig. 1, B and C) and time- (Fig. 1 D) dependent manner, upon incubation of the cells in vitro with a mouse monoclonal IgE antibody. Moreover, FcεRI expression progressively diminished when peritoneal mast cells were incubated in vitro without added IgE (Fig. 1 D). As a result, levels of FcεRI expression in peritoneal mast cells incubated for 6 d with IgE at 5 μg/ml (i.e., in the range observed in the serum of parasite-infected mice, reference 28) were 20-fold higher than those in cells incubated for 6 d without IgE (Fig. 1 D). And mast cells incubated for 4 d with a concentration of IgE (0.05 μg/ml) comparable to that present in the serum of normal mice (28, 29) had levels of FcεRI expression that were 3.6-fold those in cells incubated for 4 d without IgE (Fig. 1 C).

Bottom Line: This IgE- and antigen-specific mast cell activation and mediator production is thought to be critical to the pathogenesis of allergic disorders, such as anaphylaxis and asthma, and also contributes to host defense against parasites.In turn, IgE-dependent upregulation of Fc(epsilon)RI expression significantly enhances the ability of mouse mast cells to release serotonin, interleukin-6 (IL-6), and IL-4 in response to challenge with IgE and specific antigen.The demonstration that IgE-dependent enhancement of mast cell Fc(epsilon)RI expression permits mast cells to respond to antigen challenge with increased production of proinflammatory and immunoregulatory mediators provides new insights into both the pathogenesis of allergic diseases and the regulation of protective host responses to parasites.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, Massachusetts 02215, USA.

ABSTRACT
The binding of immunoglobulin E (IgE) to high affinity IgE receptors (Fc(epsilon)RI) expressed on the surface of mast cells primes these cells to secrete, upon subsequent exposure to specific antigen, a panel of proinflammatory mediators, which includes cytokines that can also have immunoregulatory activities. This IgE- and antigen-specific mast cell activation and mediator production is thought to be critical to the pathogenesis of allergic disorders, such as anaphylaxis and asthma, and also contributes to host defense against parasites. We now report that exposure to IgE results in a striking (up to 32-fold) upregulation of surface expression of Fc(epsilon)RI on mouse mast cells in vitro or in vivo. Moreover, baseline levels of Fc(epsilon)RI expression on peritoneal mast cells from genetically IgE-deficient (IgE -/-) mice are dramatically reduced (by approximately 83%) compared with those on cells from the corresponding normal mice. In vitro studies indicate that the IgE-dependent upregulation of mouse mast cell Fc(epsilon)RI expression has two components: an early cycloheximide-insensitive phase, followed by a later and more sustained component that is highly sensitive to inhibition by cycloheximide. In turn, IgE-dependent upregulation of Fc(epsilon)RI expression significantly enhances the ability of mouse mast cells to release serotonin, interleukin-6 (IL-6), and IL-4 in response to challenge with IgE and specific antigen. The demonstration that IgE-dependent enhancement of mast cell Fc(epsilon)RI expression permits mast cells to respond to antigen challenge with increased production of proinflammatory and immunoregulatory mediators provides new insights into both the pathogenesis of allergic diseases and the regulation of protective host responses to parasites.

Show MeSH
Related in: MedlinePlus