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Prostaglandin D2 and leukotriene E4 synergize to stimulate diverse TH2 functions and TH2 cell/neutrophil crosstalk.

Xue L, Fergusson J, Salimi M, Panse I, Ussher JE, Hegazy AN, Vinall SL, Jackson DG, Hunter MG, Pettipher R, Ogg G, Klenerman P - J. Allergy Clin. Immunol. (2014)

Bottom Line: The receptors for these mediators are drug targets in the treatment of allergic diseases, and hence understanding their interaction is likely to have clinical implications.The effects of PGD2, LTE4, and their combination on human TH2 cell gene expression were defined by using a microarray, and changes in specific inflammatory pathways were confirmed by means of PCR array, quantitative RT-PCR, ELISA, Luminex, flow cytometry, and functional assays, including analysis of downstream neutrophil activation.The combination of these lipids synergistically or additively enhanced TH2 responses and, strikingly, induced marked production of diverse nonclassical TH2 inflammatory mediators, including IL-22, IL-8, and GM-CSF, at concentrations sufficient to affect neutrophil activation.

View Article: PubMed Central - PubMed

Affiliation: Oxford NIHR Biomedical Research Centre, Translational Immunology Laboratory, Nuffield Department of Medicine, John Radcliffe Hospital, University of Oxford, Oxford, United Kingdom; Respiratory Medicine Unit, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom. Electronic address: luzheng.xue@ndm.ox.ac.uk.

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Involvement of integrins in CAIA of TH2 cells induced by PGD2 and LTE4. Cell aggregation after incubation with indicated treatments for 2 hours (A), for 2 or 9 hours in the presence of EDTA or MnCl2(B), or for 1 hour in the presence of the indicated antibodies (C). Scale bar = 0.5 mm. *P < .05 between PGD2 plus LTE4 and other treatments or between indicated conditions (n = 2-5).
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fig3: Involvement of integrins in CAIA of TH2 cells induced by PGD2 and LTE4. Cell aggregation after incubation with indicated treatments for 2 hours (A), for 2 or 9 hours in the presence of EDTA or MnCl2(B), or for 1 hour in the presence of the indicated antibodies (C). Scale bar = 0.5 mm. *P < .05 between PGD2 plus LTE4 and other treatments or between indicated conditions (n = 2-5).

Mentions: To explore the consequences of such increased integrin expression on TH2 cell-cell adhesion, we used an in vitro cell activation–induced aggregation (CAIA) assay.23 Stimulation with either lipid alone caused marked CAIA that formed within 0.5 and 1 hours and persisted for 4 to 6 hours (Fig 3, A). The combination of both mediators further enhanced the aggregation. To verify the involvement of integrins, which are critically dependent on Ca2+ ions, we tested the effect of EDTA, an inhibitory chelating agent, and MnCl2, an activator of integrin function, on PGD2/LTE4-induced CAIA. As shown in Fig 3, B, EDTA (5 mmol/L) inhibited and MnCl2 (1 mmol/L) prolonged the CAIA. Blocking antibodies to CD54 (ICAM1) and CD31 (PECAM1) were used to further confirm the contribution of integrins to the CAIA (Fig 3, C). Both antibodies partially reduced the intensity of CAIA in a concentration-dependent manner. The inhibitory effect of anti-CD54 was slightly more potent than that of anti-CD31, and the combination showed a marginal additive effect.


Prostaglandin D2 and leukotriene E4 synergize to stimulate diverse TH2 functions and TH2 cell/neutrophil crosstalk.

Xue L, Fergusson J, Salimi M, Panse I, Ussher JE, Hegazy AN, Vinall SL, Jackson DG, Hunter MG, Pettipher R, Ogg G, Klenerman P - J. Allergy Clin. Immunol. (2014)

Involvement of integrins in CAIA of TH2 cells induced by PGD2 and LTE4. Cell aggregation after incubation with indicated treatments for 2 hours (A), for 2 or 9 hours in the presence of EDTA or MnCl2(B), or for 1 hour in the presence of the indicated antibodies (C). Scale bar = 0.5 mm. *P < .05 between PGD2 plus LTE4 and other treatments or between indicated conditions (n = 2-5).
© Copyright Policy - CC BY
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4418751&req=5

fig3: Involvement of integrins in CAIA of TH2 cells induced by PGD2 and LTE4. Cell aggregation after incubation with indicated treatments for 2 hours (A), for 2 or 9 hours in the presence of EDTA or MnCl2(B), or for 1 hour in the presence of the indicated antibodies (C). Scale bar = 0.5 mm. *P < .05 between PGD2 plus LTE4 and other treatments or between indicated conditions (n = 2-5).
Mentions: To explore the consequences of such increased integrin expression on TH2 cell-cell adhesion, we used an in vitro cell activation–induced aggregation (CAIA) assay.23 Stimulation with either lipid alone caused marked CAIA that formed within 0.5 and 1 hours and persisted for 4 to 6 hours (Fig 3, A). The combination of both mediators further enhanced the aggregation. To verify the involvement of integrins, which are critically dependent on Ca2+ ions, we tested the effect of EDTA, an inhibitory chelating agent, and MnCl2, an activator of integrin function, on PGD2/LTE4-induced CAIA. As shown in Fig 3, B, EDTA (5 mmol/L) inhibited and MnCl2 (1 mmol/L) prolonged the CAIA. Blocking antibodies to CD54 (ICAM1) and CD31 (PECAM1) were used to further confirm the contribution of integrins to the CAIA (Fig 3, C). Both antibodies partially reduced the intensity of CAIA in a concentration-dependent manner. The inhibitory effect of anti-CD54 was slightly more potent than that of anti-CD31, and the combination showed a marginal additive effect.

Bottom Line: The receptors for these mediators are drug targets in the treatment of allergic diseases, and hence understanding their interaction is likely to have clinical implications.The effects of PGD2, LTE4, and their combination on human TH2 cell gene expression were defined by using a microarray, and changes in specific inflammatory pathways were confirmed by means of PCR array, quantitative RT-PCR, ELISA, Luminex, flow cytometry, and functional assays, including analysis of downstream neutrophil activation.The combination of these lipids synergistically or additively enhanced TH2 responses and, strikingly, induced marked production of diverse nonclassical TH2 inflammatory mediators, including IL-22, IL-8, and GM-CSF, at concentrations sufficient to affect neutrophil activation.

View Article: PubMed Central - PubMed

Affiliation: Oxford NIHR Biomedical Research Centre, Translational Immunology Laboratory, Nuffield Department of Medicine, John Radcliffe Hospital, University of Oxford, Oxford, United Kingdom; Respiratory Medicine Unit, Nuffield Department of Medicine, University of Oxford, Oxford, United Kingdom. Electronic address: luzheng.xue@ndm.ox.ac.uk.

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Related in: MedlinePlus