Phosphatidylserine-containing liposomes: modulators of rhinovirus induced inflammatory responses

C.A. Stokes, N. Glanville, R. Kaur, R.D. Hume, D. Robinson, Y. Perrie, M.R. Edwards, V. O'Donnell, J. Harwood, L.C. Parker, I. Sabroe

Research output: Contribution to journalMeeting abstract

Abstract

Background: Human rhinoviral infections are major contributors to the healthcare burden associated with acute exacerbations of asthma. We, and others have recently demonstrated that rhinovirus (RV)-induced inflammatory responses are mediated by multiple signalling mechanisms, such as IL-1/MyD88 (1) and TLR3/RIGI (2). We have also previously published work showing that TLR signalling is effectively inhibited by phosphatidylserine-containing liposomes (SAPS), through the disruption of membrane microdomains (3). Evidence has also suggested that membrane microdomains may influence infections with RV. In this study, we explored the ability of SAPS to modulate responses to the natural viral pathogens, RV-1B and RV-16.
Method: The immortalized bronchial epithelial cell line, BEAS-2B or primary bronchial epithelial cells were infected with RV-1B or RV-16 at a TCID50/ml of 19107 for 1 h. Immediately following infection, various concentrations of SAPS were added and changes in cytokine release were measured at 24 h. SAPS remained present throughout. Type I and III interferon (IFN) expression and rates of viral replication were measured by quantitative PCR. Virus quantification was also performed using a viral CPE assay, and IFN signalling was measured by western blot. Liposome stability was characterised and intracellular trafficking of fluorescently labelled SAPS in BEAS-2B cells was investigated using confocal microscopy. For in vivo studies, female wt Balb/c mice were pre-treated with SAPS for 2 h prior to infection with RV as previously described and changes in BAL cell number, BAL cytokine production and viral replication were quantified (4).
Results: Characterisation of SAPS liposomes by mass spectrometry showed no obvious signs of oxidation over the time period tested, and liposome size remained constant. Preliminary confocal studies revealed that SAPS was rapidly internalised within the cell and was found to associate with intracellular compartments such as the early endosome and golgi. Viral infected BEAS-2B cells co-incubated with SAPS, showed notably impaired responses to RV as assessed by release of CXCL8 and CCL5. SAPS also reduced RV-induced IFNb production and STAT-1 phosphorylation, without significantly influencing viral replication rates. Modest increases in viral particle production were only observed at 48 and 72 h time points. Suppression of viral-induced cytokine production was also observed in primary bronchial epithelial cells and pilot in vivo studies showed that SAPS results in reduced KC production at 24 h post viral infection, and this was associated with reduced neutrophil numbers within the BAL fluid.
Conclusion: Our data demonstrates a potential means of modulating inflammatory responses induced by human rhinovirus.
LanguageEnglish
Article number287
Pages108
Number of pages1
JournalImmunology
Volume140
Issue numberS1
Early online date29 Nov 2013
DOIs
Publication statusPublished - Dec 2013
EventAnnual congress of the British Society for Immunology - Liverpool, United Kingdom
Duration: 2 Feb 20135 Feb 2013

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Rhinovirus
Phosphatidylserines
Liposomes
Dimercaprol
Membrane Microdomains
Epithelial Cells
Cytokines
Infection
Interferon Type I
Endosomes
Virus Diseases
Interleukin-1
Confocal Microscopy
Virion
Interferons
Mass Spectrometry
Neutrophils
Asthma
Cell Count
Western Blotting

Bibliographical note

Poster. Abstracts BSI Congress 2013
Special Issue: Abstracts of the Annual Congress of the British Society for Immunology, 2–5 December 2013, Liverpool, UK

Cite this

Stokes, C. A., Glanville, N., Kaur, R., Hume, R. D., Robinson, D., Perrie, Y., ... Sabroe, I. (2013). Phosphatidylserine-containing liposomes: modulators of rhinovirus induced inflammatory responses. Immunology, 140(S1), 108. [287]. https://doi.org/10.1111/imm.12184
Stokes, C.A. ; Glanville, N. ; Kaur, R. ; Hume, R.D. ; Robinson, D. ; Perrie, Y. ; Edwards, M.R. ; O'Donnell, V. ; Harwood, J. ; Parker, L.C. ; Sabroe, I. / Phosphatidylserine-containing liposomes : modulators of rhinovirus induced inflammatory responses. In: Immunology. 2013 ; Vol. 140, No. S1. pp. 108.
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abstract = "Background: Human rhinoviral infections are major contributors to the healthcare burden associated with acute exacerbations of asthma. We, and others have recently demonstrated that rhinovirus (RV)-induced inflammatory responses are mediated by multiple signalling mechanisms, such as IL-1/MyD88 (1) and TLR3/RIGI (2). We have also previously published work showing that TLR signalling is effectively inhibited by phosphatidylserine-containing liposomes (SAPS), through the disruption of membrane microdomains (3). Evidence has also suggested that membrane microdomains may influence infections with RV. In this study, we explored the ability of SAPS to modulate responses to the natural viral pathogens, RV-1B and RV-16.Method: The immortalized bronchial epithelial cell line, BEAS-2B or primary bronchial epithelial cells were infected with RV-1B or RV-16 at a TCID50/ml of 19107 for 1 h. Immediately following infection, various concentrations of SAPS were added and changes in cytokine release were measured at 24 h. SAPS remained present throughout. Type I and III interferon (IFN) expression and rates of viral replication were measured by quantitative PCR. Virus quantification was also performed using a viral CPE assay, and IFN signalling was measured by western blot. Liposome stability was characterised and intracellular trafficking of fluorescently labelled SAPS in BEAS-2B cells was investigated using confocal microscopy. For in vivo studies, female wt Balb/c mice were pre-treated with SAPS for 2 h prior to infection with RV as previously described and changes in BAL cell number, BAL cytokine production and viral replication were quantified (4).Results: Characterisation of SAPS liposomes by mass spectrometry showed no obvious signs of oxidation over the time period tested, and liposome size remained constant. Preliminary confocal studies revealed that SAPS was rapidly internalised within the cell and was found to associate with intracellular compartments such as the early endosome and golgi. Viral infected BEAS-2B cells co-incubated with SAPS, showed notably impaired responses to RV as assessed by release of CXCL8 and CCL5. SAPS also reduced RV-induced IFNb production and STAT-1 phosphorylation, without significantly influencing viral replication rates. Modest increases in viral particle production were only observed at 48 and 72 h time points. Suppression of viral-induced cytokine production was also observed in primary bronchial epithelial cells and pilot in vivo studies showed that SAPS results in reduced KC production at 24 h post viral infection, and this was associated with reduced neutrophil numbers within the BAL fluid.Conclusion: Our data demonstrates a potential means of modulating inflammatory responses induced by human rhinovirus.",
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Stokes, CA, Glanville, N, Kaur, R, Hume, RD, Robinson, D, Perrie, Y, Edwards, MR, O'Donnell, V, Harwood, J, Parker, LC & Sabroe, I 2013, 'Phosphatidylserine-containing liposomes: modulators of rhinovirus induced inflammatory responses' Immunology, vol. 140, no. S1, 287, pp. 108. https://doi.org/10.1111/imm.12184

Phosphatidylserine-containing liposomes : modulators of rhinovirus induced inflammatory responses. / Stokes, C.A.; Glanville, N.; Kaur, R.; Hume, R.D.; Robinson, D.; Perrie, Y.; Edwards, M.R.; O'Donnell, V.; Harwood, J.; Parker, L.C.; Sabroe, I.

In: Immunology, Vol. 140, No. S1, 287, 12.2013, p. 108.

Research output: Contribution to journalMeeting abstract

TY - JOUR

T1 - Phosphatidylserine-containing liposomes

T2 - Immunology

AU - Stokes, C.A.

AU - Glanville, N.

AU - Kaur, R.

AU - Hume, R.D.

AU - Robinson, D.

AU - Perrie, Y.

AU - Edwards, M.R.

AU - O'Donnell, V.

AU - Harwood, J.

AU - Parker, L.C.

AU - Sabroe, I.

N1 - Poster. Abstracts BSI Congress 2013 Special Issue: Abstracts of the Annual Congress of the British Society for Immunology, 2–5 December 2013, Liverpool, UK

PY - 2013/12

Y1 - 2013/12

N2 - Background: Human rhinoviral infections are major contributors to the healthcare burden associated with acute exacerbations of asthma. We, and others have recently demonstrated that rhinovirus (RV)-induced inflammatory responses are mediated by multiple signalling mechanisms, such as IL-1/MyD88 (1) and TLR3/RIGI (2). We have also previously published work showing that TLR signalling is effectively inhibited by phosphatidylserine-containing liposomes (SAPS), through the disruption of membrane microdomains (3). Evidence has also suggested that membrane microdomains may influence infections with RV. In this study, we explored the ability of SAPS to modulate responses to the natural viral pathogens, RV-1B and RV-16.Method: The immortalized bronchial epithelial cell line, BEAS-2B or primary bronchial epithelial cells were infected with RV-1B or RV-16 at a TCID50/ml of 19107 for 1 h. Immediately following infection, various concentrations of SAPS were added and changes in cytokine release were measured at 24 h. SAPS remained present throughout. Type I and III interferon (IFN) expression and rates of viral replication were measured by quantitative PCR. Virus quantification was also performed using a viral CPE assay, and IFN signalling was measured by western blot. Liposome stability was characterised and intracellular trafficking of fluorescently labelled SAPS in BEAS-2B cells was investigated using confocal microscopy. For in vivo studies, female wt Balb/c mice were pre-treated with SAPS for 2 h prior to infection with RV as previously described and changes in BAL cell number, BAL cytokine production and viral replication were quantified (4).Results: Characterisation of SAPS liposomes by mass spectrometry showed no obvious signs of oxidation over the time period tested, and liposome size remained constant. Preliminary confocal studies revealed that SAPS was rapidly internalised within the cell and was found to associate with intracellular compartments such as the early endosome and golgi. Viral infected BEAS-2B cells co-incubated with SAPS, showed notably impaired responses to RV as assessed by release of CXCL8 and CCL5. SAPS also reduced RV-induced IFNb production and STAT-1 phosphorylation, without significantly influencing viral replication rates. Modest increases in viral particle production were only observed at 48 and 72 h time points. Suppression of viral-induced cytokine production was also observed in primary bronchial epithelial cells and pilot in vivo studies showed that SAPS results in reduced KC production at 24 h post viral infection, and this was associated with reduced neutrophil numbers within the BAL fluid.Conclusion: Our data demonstrates a potential means of modulating inflammatory responses induced by human rhinovirus.

AB - Background: Human rhinoviral infections are major contributors to the healthcare burden associated with acute exacerbations of asthma. We, and others have recently demonstrated that rhinovirus (RV)-induced inflammatory responses are mediated by multiple signalling mechanisms, such as IL-1/MyD88 (1) and TLR3/RIGI (2). We have also previously published work showing that TLR signalling is effectively inhibited by phosphatidylserine-containing liposomes (SAPS), through the disruption of membrane microdomains (3). Evidence has also suggested that membrane microdomains may influence infections with RV. In this study, we explored the ability of SAPS to modulate responses to the natural viral pathogens, RV-1B and RV-16.Method: The immortalized bronchial epithelial cell line, BEAS-2B or primary bronchial epithelial cells were infected with RV-1B or RV-16 at a TCID50/ml of 19107 for 1 h. Immediately following infection, various concentrations of SAPS were added and changes in cytokine release were measured at 24 h. SAPS remained present throughout. Type I and III interferon (IFN) expression and rates of viral replication were measured by quantitative PCR. Virus quantification was also performed using a viral CPE assay, and IFN signalling was measured by western blot. Liposome stability was characterised and intracellular trafficking of fluorescently labelled SAPS in BEAS-2B cells was investigated using confocal microscopy. For in vivo studies, female wt Balb/c mice were pre-treated with SAPS for 2 h prior to infection with RV as previously described and changes in BAL cell number, BAL cytokine production and viral replication were quantified (4).Results: Characterisation of SAPS liposomes by mass spectrometry showed no obvious signs of oxidation over the time period tested, and liposome size remained constant. Preliminary confocal studies revealed that SAPS was rapidly internalised within the cell and was found to associate with intracellular compartments such as the early endosome and golgi. Viral infected BEAS-2B cells co-incubated with SAPS, showed notably impaired responses to RV as assessed by release of CXCL8 and CCL5. SAPS also reduced RV-induced IFNb production and STAT-1 phosphorylation, without significantly influencing viral replication rates. Modest increases in viral particle production were only observed at 48 and 72 h time points. Suppression of viral-induced cytokine production was also observed in primary bronchial epithelial cells and pilot in vivo studies showed that SAPS results in reduced KC production at 24 h post viral infection, and this was associated with reduced neutrophil numbers within the BAL fluid.Conclusion: Our data demonstrates a potential means of modulating inflammatory responses induced by human rhinovirus.

UR - http://onlinelibrary.wiley.com/doi/10.1111/imm.12184/abstract

U2 - 10.1111/imm.12184

DO - 10.1111/imm.12184

M3 - Meeting abstract

VL - 140

SP - 108

JO - Immunology

JF - Immunology

SN - 0019-2805

IS - S1

M1 - 287

ER -

Stokes CA, Glanville N, Kaur R, Hume RD, Robinson D, Perrie Y et al. Phosphatidylserine-containing liposomes: modulators of rhinovirus induced inflammatory responses. Immunology. 2013 Dec;140(S1):108. 287. https://doi.org/10.1111/imm.12184