Limits...
Pollen lipidomics: lipid profiling exposes a notable diversity in 22 allergenic pollen and potential biomarkers of the allergic immune response.

Bashir ME, Lui JH, Palnivelu R, Naclerio RM, Preuss D - PLoS ONE (2013)

Bottom Line: Lipid antigens have attracted attention for their potent immunoregulatory effects.Three experiments compared pollen lipid profiles.Pollen lipids vary greatly among allergenic species and contain many molecules that have stimulatory or regulatory effects on immune responses.

View Article: PubMed Central - PubMed

Affiliation: Section of Otolaryngology-Head and Neck Surgery, Department of Surgery, Division of the Biological Sciences, The University of Chicago, Illinois, USA. mbashir@surgery.bsd.uchicago.edu

ABSTRACT

Background/aim: Pollen grains are the male gametophytes that deliver sperm cells to female gametophytes during sexual reproduction of higher plants. Pollen is a major source of aeroallergens and environmental antigens. The pollen coat harbors a plethora of lipids that are required for pollen hydration, germination, and penetration of the stigma by pollen tubes. In addition to proteins, pollen displays a wide array of lipids that interact with the human immune system. Prior searches for pollen allergens have focused on the identification of intracellular allergenic proteins, but have largely overlooked much of the extracellular pollen matrix, a region where the majority of lipid molecules reside. Lipid antigens have attracted attention for their potent immunoregulatory effects. By being in close proximity to allergenic proteins on the pollen surface when they interact with host cells, lipids could modify the antigenic properties of proteins.

Methodology/principal findings: We performed a comparative pollen lipid profiling of 22 commonly allergenic plant species by the use of gas chromatography-mass spectroscopy, followed by detailed data mining and statistical analysis. Three experiments compared pollen lipid profiles. We built a database library of the pollen lipids by matching acquired pollen-lipid mass spectra and retention times with the NIST/EPA/NIH mass-spectral library. We detected, identified, and relatively quantified more than 106 lipid molecular species including fatty acids, n-alkanes, fatty alcohols, and sterols. Pollen-derived lipids stimulation up-regulate cytokines expression of dendritic and natural killer T cells co-culture.

Conclusions/significance: Here we report on a lipidomic analysis of pollen lipids that can serve as a database for identifying potential lipid antigens and/or novel candidate molecules involved in allergy. The database provides a resource that facilitates studies on the role of lipids in the immunopathogenesis of allergy. Pollen lipids vary greatly among allergenic species and contain many molecules that have stimulatory or regulatory effects on immune responses.

Show MeSH

Related in: MedlinePlus

Effect of different lipid stimuli on TNF-α response of T cells.For analysis of cytokines by intracellular staining, conventional T cells harvested after culture in vitro were stimulated with 1 µg/ml lipid molecules for 24 to 36 hours in 6-well plates. After treatment with 10 µg/mL GolgiPlug, a protein transport inhibitor containing brefeldin A (eBiosciences) during the final 6 to 12 hours of stimulation, the cells were stained for surface markers and anti-mouse TNF-α as described in Materials and Methods. Flow cytometry was performed on a FACSCanto II instrument (BD Biosciences) and analyzed using FlowJo software (Tree Star).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3585183&req=5

pone-0057566-g003: Effect of different lipid stimuli on TNF-α response of T cells.For analysis of cytokines by intracellular staining, conventional T cells harvested after culture in vitro were stimulated with 1 µg/ml lipid molecules for 24 to 36 hours in 6-well plates. After treatment with 10 µg/mL GolgiPlug, a protein transport inhibitor containing brefeldin A (eBiosciences) during the final 6 to 12 hours of stimulation, the cells were stained for surface markers and anti-mouse TNF-α as described in Materials and Methods. Flow cytometry was performed on a FACSCanto II instrument (BD Biosciences) and analyzed using FlowJo software (Tree Star).

Mentions: We have used intracellular staining with cytokines combined with flow cytometry to examine the frequencies of TNF-α-producing cells from conventional T cells. FACS analysis showed that in the absence of lipids, 0.5% of unstimulated cells expressed intracellular TNF-α. This increased to 12% with 1 uM C24∶0, 2.5% with 1 uM nC23, 5% with 1 uM nC25 or 1-tetradecanol and 3.5% with 1 uM ribitol. Flow cytometric analysis showed several lipids found in pollen stimulate conventional T cells to express TNF-α (Figure 3).


Pollen lipidomics: lipid profiling exposes a notable diversity in 22 allergenic pollen and potential biomarkers of the allergic immune response.

Bashir ME, Lui JH, Palnivelu R, Naclerio RM, Preuss D - PLoS ONE (2013)

Effect of different lipid stimuli on TNF-α response of T cells.For analysis of cytokines by intracellular staining, conventional T cells harvested after culture in vitro were stimulated with 1 µg/ml lipid molecules for 24 to 36 hours in 6-well plates. After treatment with 10 µg/mL GolgiPlug, a protein transport inhibitor containing brefeldin A (eBiosciences) during the final 6 to 12 hours of stimulation, the cells were stained for surface markers and anti-mouse TNF-α as described in Materials and Methods. Flow cytometry was performed on a FACSCanto II instrument (BD Biosciences) and analyzed using FlowJo software (Tree Star).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585183&req=5

pone-0057566-g003: Effect of different lipid stimuli on TNF-α response of T cells.For analysis of cytokines by intracellular staining, conventional T cells harvested after culture in vitro were stimulated with 1 µg/ml lipid molecules for 24 to 36 hours in 6-well plates. After treatment with 10 µg/mL GolgiPlug, a protein transport inhibitor containing brefeldin A (eBiosciences) during the final 6 to 12 hours of stimulation, the cells were stained for surface markers and anti-mouse TNF-α as described in Materials and Methods. Flow cytometry was performed on a FACSCanto II instrument (BD Biosciences) and analyzed using FlowJo software (Tree Star).
Mentions: We have used intracellular staining with cytokines combined with flow cytometry to examine the frequencies of TNF-α-producing cells from conventional T cells. FACS analysis showed that in the absence of lipids, 0.5% of unstimulated cells expressed intracellular TNF-α. This increased to 12% with 1 uM C24∶0, 2.5% with 1 uM nC23, 5% with 1 uM nC25 or 1-tetradecanol and 3.5% with 1 uM ribitol. Flow cytometric analysis showed several lipids found in pollen stimulate conventional T cells to express TNF-α (Figure 3).

Bottom Line: Lipid antigens have attracted attention for their potent immunoregulatory effects.Three experiments compared pollen lipid profiles.Pollen lipids vary greatly among allergenic species and contain many molecules that have stimulatory or regulatory effects on immune responses.

View Article: PubMed Central - PubMed

Affiliation: Section of Otolaryngology-Head and Neck Surgery, Department of Surgery, Division of the Biological Sciences, The University of Chicago, Illinois, USA. mbashir@surgery.bsd.uchicago.edu

ABSTRACT

Background/aim: Pollen grains are the male gametophytes that deliver sperm cells to female gametophytes during sexual reproduction of higher plants. Pollen is a major source of aeroallergens and environmental antigens. The pollen coat harbors a plethora of lipids that are required for pollen hydration, germination, and penetration of the stigma by pollen tubes. In addition to proteins, pollen displays a wide array of lipids that interact with the human immune system. Prior searches for pollen allergens have focused on the identification of intracellular allergenic proteins, but have largely overlooked much of the extracellular pollen matrix, a region where the majority of lipid molecules reside. Lipid antigens have attracted attention for their potent immunoregulatory effects. By being in close proximity to allergenic proteins on the pollen surface when they interact with host cells, lipids could modify the antigenic properties of proteins.

Methodology/principal findings: We performed a comparative pollen lipid profiling of 22 commonly allergenic plant species by the use of gas chromatography-mass spectroscopy, followed by detailed data mining and statistical analysis. Three experiments compared pollen lipid profiles. We built a database library of the pollen lipids by matching acquired pollen-lipid mass spectra and retention times with the NIST/EPA/NIH mass-spectral library. We detected, identified, and relatively quantified more than 106 lipid molecular species including fatty acids, n-alkanes, fatty alcohols, and sterols. Pollen-derived lipids stimulation up-regulate cytokines expression of dendritic and natural killer T cells co-culture.

Conclusions/significance: Here we report on a lipidomic analysis of pollen lipids that can serve as a database for identifying potential lipid antigens and/or novel candidate molecules involved in allergy. The database provides a resource that facilitates studies on the role of lipids in the immunopathogenesis of allergy. Pollen lipids vary greatly among allergenic species and contain many molecules that have stimulatory or regulatory effects on immune responses.

Show MeSH
Related in: MedlinePlus