Limits...
Surfactant Protein D modulates allergen particle uptake and inflammatory response in a human epithelial airway model.

Schleh C, Rothen-Rutishauser BM, Blank F, Lauenstein HD, Nassimi M, Krug N, Braun A, Erpenbeck VJ, Gehr P, Hohlfeld JM - Respir. Res. (2012)

Bottom Line: SPP were taken up by epithelial cells, macrophages, and dendritic cells.This uptake coincided with secretion of pro-inflammatory cytokines and chemokines.SP-D modulated the uptake of SPP in a cell type specific way (e.g. increased number of macrophages and epithelial cells, which participated in allergen particle uptake) and led to a decreased secretion of pro-inflammatory cytokines.

View Article: PubMed Central - HTML - PubMed

Affiliation: Fraunhofer Institute of Toxicology and Experimental Medicine, Nikolai-Fuch-Str 1, 30635 Hannover, Germany. carsten.schleh@web.de

ABSTRACT

Background: Allergen-containing subpollen particles (SPP) are released from whole plant pollen upon contact with water or even high humidity. Because of their size SPP can preferentially reach the lower airways where they come into contact with surfactant protein (SP)-D. The aim of the present study was to investigate the influence of SP-D in a complex three-dimensional human epithelial airway model, which simulates the most important barrier functions of the epithelial airway. The uptake of SPP as well as the secretion of pro-inflammatory cytokines was investigated.

Methods: SPP were isolated from timothy grass and subsequently fluorescently labeled. A human epithelial airway model was built by using human Type II-pneumocyte like cells (A549 cells), human monocyte derived macrophages as well as human monocyte derived dendritic cells. The epithelial cell model was incubated with SPP in the presence and absence of surfactant protein D. Particle uptake was evaluated by confocal microscopy and advanced computer-controlled analysis. Finally, human primary CD4+ T-Cells were added to the epithelial airway model and soluble mediators were measured by enzyme linked immunosorbent assay or bead array.

Results: SPP were taken up by epithelial cells, macrophages, and dendritic cells. This uptake coincided with secretion of pro-inflammatory cytokines and chemokines. SP-D modulated the uptake of SPP in a cell type specific way (e.g. increased number of macrophages and epithelial cells, which participated in allergen particle uptake) and led to a decreased secretion of pro-inflammatory cytokines.

Conclusion: These results display a possible mechanism of how SP-D can modulate the inflammatory response to inhaled allergen.

Show MeSH

Related in: MedlinePlus

Secretion of Interleukin (IL)-8 into supernatants of an epithelial airway model after 8 hours incubation with 10 million subpollen particles (SPP) ± surfactant protein D (SP-D) and further 72 h incubation with fresh medium without particles and proteins. IL-8 was measured by enzyme linked immunosorbent assay. Values are means ± SEM from of least 5 experiments. # p < 0.05; ## p < 0.01; Detection limit was 3.2 pg/ml; Limit of quantification was 64 pg/ml.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3295667&req=5

Figure 5: Secretion of Interleukin (IL)-8 into supernatants of an epithelial airway model after 8 hours incubation with 10 million subpollen particles (SPP) ± surfactant protein D (SP-D) and further 72 h incubation with fresh medium without particles and proteins. IL-8 was measured by enzyme linked immunosorbent assay. Values are means ± SEM from of least 5 experiments. # p < 0.05; ## p < 0.01; Detection limit was 3.2 pg/ml; Limit of quantification was 64 pg/ml.

Mentions: Incubation with 10 million SPP for 8 hours increased secretion of IL-8 when cells were incubated for additional 72 hours with fresh medium in the presence of CD4+-T-cells. The baseline value of 61.3 ± 11.3 ng/ml, measured in the supernatants of the cells alone, was increased to 103.3 ± 24.3 ng/ml. Importantly, co-incubation with SP-D during the 8 hour-particle exposure period decreased IL-8 secretion significantly (SP-D 1 μg/ml: 56.0 ± 9.8 ng/ml and SP-D 10 μg/ml: 50.5 ± 10.5 ng/ml/Figure 5).


Surfactant Protein D modulates allergen particle uptake and inflammatory response in a human epithelial airway model.

Schleh C, Rothen-Rutishauser BM, Blank F, Lauenstein HD, Nassimi M, Krug N, Braun A, Erpenbeck VJ, Gehr P, Hohlfeld JM - Respir. Res. (2012)

Secretion of Interleukin (IL)-8 into supernatants of an epithelial airway model after 8 hours incubation with 10 million subpollen particles (SPP) ± surfactant protein D (SP-D) and further 72 h incubation with fresh medium without particles and proteins. IL-8 was measured by enzyme linked immunosorbent assay. Values are means ± SEM from of least 5 experiments. # p < 0.05; ## p < 0.01; Detection limit was 3.2 pg/ml; Limit of quantification was 64 pg/ml.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3295667&req=5

Figure 5: Secretion of Interleukin (IL)-8 into supernatants of an epithelial airway model after 8 hours incubation with 10 million subpollen particles (SPP) ± surfactant protein D (SP-D) and further 72 h incubation with fresh medium without particles and proteins. IL-8 was measured by enzyme linked immunosorbent assay. Values are means ± SEM from of least 5 experiments. # p < 0.05; ## p < 0.01; Detection limit was 3.2 pg/ml; Limit of quantification was 64 pg/ml.
Mentions: Incubation with 10 million SPP for 8 hours increased secretion of IL-8 when cells were incubated for additional 72 hours with fresh medium in the presence of CD4+-T-cells. The baseline value of 61.3 ± 11.3 ng/ml, measured in the supernatants of the cells alone, was increased to 103.3 ± 24.3 ng/ml. Importantly, co-incubation with SP-D during the 8 hour-particle exposure period decreased IL-8 secretion significantly (SP-D 1 μg/ml: 56.0 ± 9.8 ng/ml and SP-D 10 μg/ml: 50.5 ± 10.5 ng/ml/Figure 5).

Bottom Line: SPP were taken up by epithelial cells, macrophages, and dendritic cells.This uptake coincided with secretion of pro-inflammatory cytokines and chemokines.SP-D modulated the uptake of SPP in a cell type specific way (e.g. increased number of macrophages and epithelial cells, which participated in allergen particle uptake) and led to a decreased secretion of pro-inflammatory cytokines.

View Article: PubMed Central - HTML - PubMed

Affiliation: Fraunhofer Institute of Toxicology and Experimental Medicine, Nikolai-Fuch-Str 1, 30635 Hannover, Germany. carsten.schleh@web.de

ABSTRACT

Background: Allergen-containing subpollen particles (SPP) are released from whole plant pollen upon contact with water or even high humidity. Because of their size SPP can preferentially reach the lower airways where they come into contact with surfactant protein (SP)-D. The aim of the present study was to investigate the influence of SP-D in a complex three-dimensional human epithelial airway model, which simulates the most important barrier functions of the epithelial airway. The uptake of SPP as well as the secretion of pro-inflammatory cytokines was investigated.

Methods: SPP were isolated from timothy grass and subsequently fluorescently labeled. A human epithelial airway model was built by using human Type II-pneumocyte like cells (A549 cells), human monocyte derived macrophages as well as human monocyte derived dendritic cells. The epithelial cell model was incubated with SPP in the presence and absence of surfactant protein D. Particle uptake was evaluated by confocal microscopy and advanced computer-controlled analysis. Finally, human primary CD4+ T-Cells were added to the epithelial airway model and soluble mediators were measured by enzyme linked immunosorbent assay or bead array.

Results: SPP were taken up by epithelial cells, macrophages, and dendritic cells. This uptake coincided with secretion of pro-inflammatory cytokines and chemokines. SP-D modulated the uptake of SPP in a cell type specific way (e.g. increased number of macrophages and epithelial cells, which participated in allergen particle uptake) and led to a decreased secretion of pro-inflammatory cytokines.

Conclusion: These results display a possible mechanism of how SP-D can modulate the inflammatory response to inhaled allergen.

Show MeSH
Related in: MedlinePlus