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Germinal center formation and local immunoglobulin E (IgE) production in the lung after an airway antigenic challenge.

Chvatchko Y, Kosco-Vilbois MH, Herren S, Lefort J, Bonnefoy JY - J. Exp. Med. (1996)

Bottom Line: These assays confirmed that the isotypes observed in situ were a secreted product.These data demonstrate that antigen-driven differentiation of B cells via induction of an FDC network and germinal centers occurs in the parenchyma of inflamed lungs.These germinal centers would then provide a local source of IgE-secreting plasma cells that contribute to the release of factors mediating inflammatory processes in the lung.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Geneva Biomedical Research Institute, Glaxo Wellcome Research and Development S.A., Switzerland.

ABSTRACT
Airway inflammation plays a central role in the pathogenesis of asthma. However, the precise contribution of all cell types in the development and maintenance of airway hyperreactivity and histopathology during allergic inflammation remains unclear. After sensitization of mice in the periphery, challenge by multiple intratracheal (i.t.) instillations of ovalbumin (OVA) results in eosinophilia, mononuclear cell infiltration, and airway epithelial changes analogous to that seen in asthma (Blyth, D.I., M.S. Pedrick, T.J. Savage, E.M. Hessel, and D. Fattah. 1996. Am. J. Respir. Cell Mol. Biol. 14:425-438). To investigate further the nature of the cellular infiltrate, lungs from OVA-versus saline-treated mice were processed for histology and immunohistochemistry. One of the most striking features observed was the formation of germinal centers within the parenchyma of the inflamed lungs. In addition, follicular dendritic cells (FDCs) bearing OVA on their plasma membranes appeared and, adjacent to these sites, OVA-specific IgG1-, IgE-, and IgA-producing plasma cells emerged. To confirm that antigen-specific immunoglobulins (Ig) were being produced within the parenchyma, plasma cell number and antibody production were quantitated in vitro after isolation of cells from the lung. These assays confirmed that the isotypes observed in situ were a secreted product. As IgE-dependent mechanisms have been implicated as being central to the pathogenesis of bronchial asthma, airway hyperresponsiveness was evaluated. The mice undergoing lung inflammation were hyperresponsive, while the control group remained at baseline. These data demonstrate that antigen-driven differentiation of B cells via induction of an FDC network and germinal centers occurs in the parenchyma of inflamed lungs. These germinal centers would then provide a local source of IgE-secreting plasma cells that contribute to the release of factors mediating inflammatory processes in the lung.

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Quantification of the number of anti-OVA plasma cells isolated from lung. The low buoyant density cells prepared from lungs of  control mice, NaCl/NaCl, OVAip/NaCl, and OVAsc/NaCl (squares)  and of experimental mice, OVAip/OVA, and OVAsc/OVA (diamonds)  were incubated on OVA-coated wells before identification of the Ig isotype. Results are plotted as the number of OVA-specific antibody-forming cells (AFC) identified per 105 low buoyant density cells. Each data point  represents the mean of triplicate wells for cells from an individual mouse.
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Figure 4: Quantification of the number of anti-OVA plasma cells isolated from lung. The low buoyant density cells prepared from lungs of control mice, NaCl/NaCl, OVAip/NaCl, and OVAsc/NaCl (squares) and of experimental mice, OVAip/OVA, and OVAsc/OVA (diamonds) were incubated on OVA-coated wells before identification of the Ig isotype. Results are plotted as the number of OVA-specific antibody-forming cells (AFC) identified per 105 low buoyant density cells. Each data point represents the mean of triplicate wells for cells from an individual mouse.

Mentions: To determine the number of anti-OVA-producing cells present within the lung, an ELISPOT assay was utilized. The cells were obtained as outlined in Materials and Methods from enzymedigested lungs that had been prelavaged with saline. An enrichment step was introduced in order to concentrate the frequency of low buoyant density cells using sedimentation gradients. As can be seen in Fig. 4, only the lungs of mice that had received OVA in the trachea contained anti-OVAforming cells (AFCs). Their presence could not be attributed to a contaminating contribution from the residual blood of the lung, as no AFCs were detected in the OVAprimed but NaCl-challenged mice (OVA/NaCl groups). The isotypes again were restricted, as only IgM, IgG1, and IgA were detected and not IgG2a and IgG2b.


Germinal center formation and local immunoglobulin E (IgE) production in the lung after an airway antigenic challenge.

Chvatchko Y, Kosco-Vilbois MH, Herren S, Lefort J, Bonnefoy JY - J. Exp. Med. (1996)

Quantification of the number of anti-OVA plasma cells isolated from lung. The low buoyant density cells prepared from lungs of  control mice, NaCl/NaCl, OVAip/NaCl, and OVAsc/NaCl (squares)  and of experimental mice, OVAip/OVA, and OVAsc/OVA (diamonds)  were incubated on OVA-coated wells before identification of the Ig isotype. Results are plotted as the number of OVA-specific antibody-forming cells (AFC) identified per 105 low buoyant density cells. Each data point  represents the mean of triplicate wells for cells from an individual mouse.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196373&req=5

Figure 4: Quantification of the number of anti-OVA plasma cells isolated from lung. The low buoyant density cells prepared from lungs of control mice, NaCl/NaCl, OVAip/NaCl, and OVAsc/NaCl (squares) and of experimental mice, OVAip/OVA, and OVAsc/OVA (diamonds) were incubated on OVA-coated wells before identification of the Ig isotype. Results are plotted as the number of OVA-specific antibody-forming cells (AFC) identified per 105 low buoyant density cells. Each data point represents the mean of triplicate wells for cells from an individual mouse.
Mentions: To determine the number of anti-OVA-producing cells present within the lung, an ELISPOT assay was utilized. The cells were obtained as outlined in Materials and Methods from enzymedigested lungs that had been prelavaged with saline. An enrichment step was introduced in order to concentrate the frequency of low buoyant density cells using sedimentation gradients. As can be seen in Fig. 4, only the lungs of mice that had received OVA in the trachea contained anti-OVAforming cells (AFCs). Their presence could not be attributed to a contaminating contribution from the residual blood of the lung, as no AFCs were detected in the OVAprimed but NaCl-challenged mice (OVA/NaCl groups). The isotypes again were restricted, as only IgM, IgG1, and IgA were detected and not IgG2a and IgG2b.

Bottom Line: These assays confirmed that the isotypes observed in situ were a secreted product.These data demonstrate that antigen-driven differentiation of B cells via induction of an FDC network and germinal centers occurs in the parenchyma of inflamed lungs.These germinal centers would then provide a local source of IgE-secreting plasma cells that contribute to the release of factors mediating inflammatory processes in the lung.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, Geneva Biomedical Research Institute, Glaxo Wellcome Research and Development S.A., Switzerland.

ABSTRACT
Airway inflammation plays a central role in the pathogenesis of asthma. However, the precise contribution of all cell types in the development and maintenance of airway hyperreactivity and histopathology during allergic inflammation remains unclear. After sensitization of mice in the periphery, challenge by multiple intratracheal (i.t.) instillations of ovalbumin (OVA) results in eosinophilia, mononuclear cell infiltration, and airway epithelial changes analogous to that seen in asthma (Blyth, D.I., M.S. Pedrick, T.J. Savage, E.M. Hessel, and D. Fattah. 1996. Am. J. Respir. Cell Mol. Biol. 14:425-438). To investigate further the nature of the cellular infiltrate, lungs from OVA-versus saline-treated mice were processed for histology and immunohistochemistry. One of the most striking features observed was the formation of germinal centers within the parenchyma of the inflamed lungs. In addition, follicular dendritic cells (FDCs) bearing OVA on their plasma membranes appeared and, adjacent to these sites, OVA-specific IgG1-, IgE-, and IgA-producing plasma cells emerged. To confirm that antigen-specific immunoglobulins (Ig) were being produced within the parenchyma, plasma cell number and antibody production were quantitated in vitro after isolation of cells from the lung. These assays confirmed that the isotypes observed in situ were a secreted product. As IgE-dependent mechanisms have been implicated as being central to the pathogenesis of bronchial asthma, airway hyperresponsiveness was evaluated. The mice undergoing lung inflammation were hyperresponsive, while the control group remained at baseline. These data demonstrate that antigen-driven differentiation of B cells via induction of an FDC network and germinal centers occurs in the parenchyma of inflamed lungs. These germinal centers would then provide a local source of IgE-secreting plasma cells that contribute to the release of factors mediating inflammatory processes in the lung.

Show MeSH
Related in: MedlinePlus