Murine aortic smooth muscle cells acquire, though fail to present exogenous protein antigens on major histocompatibility complex class II molecules

Marcella Maddaluno, Neil MacRitchie, Gianluca Grassia, Armando Ialenti, John P. Butcher, Paul Garside, James M. Brewer, Pasquale Maffia

Research output: Contribution to journalArticle

Abstract

In the present study aortic murine smooth muscle cell (SMC) antigen presentation capacity was evaluated using the E alpha-GFP/Y-Ae system to visualize antigen uptake through a GFP tag and tracking of E.. peptide/MHCII presentation using the Y-Ae Ab. Stimulation with IFN-gamma (100 ng/mL) for 72 h caused a significant (P < 0.01) increase in the percentage of MHC class II positive SMCs, compared with unstimulated cells. Treatment with E alpha-GFP (100 mu g/mL) for 48 h induced a significant (P < 0.05) increase in the percentage of GFP positive SMCs while it did not affect the percentage of Y-Ae positive cells, being indicative of antigen uptake without its presentation in the context ofMHC class II. After IFN-gamma-stimulation, ovalbumin-(OVA, 1 mg/mL) or OVA(323-339) peptide-(0.5 mu g/mL) treated SMCs failed to induce OT-IICD4(+) T cell activation/proliferation; this was also accompanied by a lack of expression of key costimulatory molecules (OX40L, CD40, CD70, and CD86) on SMCs. Finally, OVA-treated SMCs failed to induce DO11.10-GFP hybridoma activation, a process independent of costimulation. Our results demonstrate that while murine primary aortic SMCs express MHC class II and can acquire exogenous antigens, they fail to activate T cells through a failure in antigen presentation and a lack of costimulatory molecule expression.
Original languageEnglish
Article number949845
Number of pages10
JournalBiomed Research International
Volume2014
DOIs
Publication statusPublished - 2014
Externally publishedYes

Cite this

Maddaluno, Marcella ; MacRitchie, Neil ; Grassia, Gianluca ; Ialenti, Armando ; Butcher, John P. ; Garside, Paul ; Brewer, James M. ; Maffia, Pasquale. / Murine aortic smooth muscle cells acquire, though fail to present exogenous protein antigens on major histocompatibility complex class II molecules. In: Biomed Research International. 2014 ; Vol. 2014.
@article{b563f620baab4fb58d163957d90f8e4f,
title = "Murine aortic smooth muscle cells acquire, though fail to present exogenous protein antigens on major histocompatibility complex class II molecules",
abstract = "In the present study aortic murine smooth muscle cell (SMC) antigen presentation capacity was evaluated using the E alpha-GFP/Y-Ae system to visualize antigen uptake through a GFP tag and tracking of E.. peptide/MHCII presentation using the Y-Ae Ab. Stimulation with IFN-gamma (100 ng/mL) for 72 h caused a significant (P < 0.01) increase in the percentage of MHC class II positive SMCs, compared with unstimulated cells. Treatment with E alpha-GFP (100 mu g/mL) for 48 h induced a significant (P < 0.05) increase in the percentage of GFP positive SMCs while it did not affect the percentage of Y-Ae positive cells, being indicative of antigen uptake without its presentation in the context ofMHC class II. After IFN-gamma-stimulation, ovalbumin-(OVA, 1 mg/mL) or OVA(323-339) peptide-(0.5 mu g/mL) treated SMCs failed to induce OT-IICD4(+) T cell activation/proliferation; this was also accompanied by a lack of expression of key costimulatory molecules (OX40L, CD40, CD70, and CD86) on SMCs. Finally, OVA-treated SMCs failed to induce DO11.10-GFP hybridoma activation, a process independent of costimulation. Our results demonstrate that while murine primary aortic SMCs express MHC class II and can acquire exogenous antigens, they fail to activate T cells through a failure in antigen presentation and a lack of costimulatory molecule expression.",
author = "Marcella Maddaluno and Neil MacRitchie and Gianluca Grassia and Armando Ialenti and Butcher, {John P.} and Paul Garside and Brewer, {James M.} and Pasquale Maffia",
year = "2014",
doi = "10.1155/2014/949845",
language = "English",
volume = "2014",
journal = "Biomed Research International",
issn = "2314-6133",
publisher = "Hindawi Publishing Corporation",

}

Murine aortic smooth muscle cells acquire, though fail to present exogenous protein antigens on major histocompatibility complex class II molecules. / Maddaluno, Marcella; MacRitchie, Neil; Grassia, Gianluca; Ialenti, Armando; Butcher, John P.; Garside, Paul; Brewer, James M.; Maffia, Pasquale.

In: Biomed Research International, Vol. 2014, 949845, 2014.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Murine aortic smooth muscle cells acquire, though fail to present exogenous protein antigens on major histocompatibility complex class II molecules

AU - Maddaluno, Marcella

AU - MacRitchie, Neil

AU - Grassia, Gianluca

AU - Ialenti, Armando

AU - Butcher, John P.

AU - Garside, Paul

AU - Brewer, James M.

AU - Maffia, Pasquale

PY - 2014

Y1 - 2014

N2 - In the present study aortic murine smooth muscle cell (SMC) antigen presentation capacity was evaluated using the E alpha-GFP/Y-Ae system to visualize antigen uptake through a GFP tag and tracking of E.. peptide/MHCII presentation using the Y-Ae Ab. Stimulation with IFN-gamma (100 ng/mL) for 72 h caused a significant (P < 0.01) increase in the percentage of MHC class II positive SMCs, compared with unstimulated cells. Treatment with E alpha-GFP (100 mu g/mL) for 48 h induced a significant (P < 0.05) increase in the percentage of GFP positive SMCs while it did not affect the percentage of Y-Ae positive cells, being indicative of antigen uptake without its presentation in the context ofMHC class II. After IFN-gamma-stimulation, ovalbumin-(OVA, 1 mg/mL) or OVA(323-339) peptide-(0.5 mu g/mL) treated SMCs failed to induce OT-IICD4(+) T cell activation/proliferation; this was also accompanied by a lack of expression of key costimulatory molecules (OX40L, CD40, CD70, and CD86) on SMCs. Finally, OVA-treated SMCs failed to induce DO11.10-GFP hybridoma activation, a process independent of costimulation. Our results demonstrate that while murine primary aortic SMCs express MHC class II and can acquire exogenous antigens, they fail to activate T cells through a failure in antigen presentation and a lack of costimulatory molecule expression.

AB - In the present study aortic murine smooth muscle cell (SMC) antigen presentation capacity was evaluated using the E alpha-GFP/Y-Ae system to visualize antigen uptake through a GFP tag and tracking of E.. peptide/MHCII presentation using the Y-Ae Ab. Stimulation with IFN-gamma (100 ng/mL) for 72 h caused a significant (P < 0.01) increase in the percentage of MHC class II positive SMCs, compared with unstimulated cells. Treatment with E alpha-GFP (100 mu g/mL) for 48 h induced a significant (P < 0.05) increase in the percentage of GFP positive SMCs while it did not affect the percentage of Y-Ae positive cells, being indicative of antigen uptake without its presentation in the context ofMHC class II. After IFN-gamma-stimulation, ovalbumin-(OVA, 1 mg/mL) or OVA(323-339) peptide-(0.5 mu g/mL) treated SMCs failed to induce OT-IICD4(+) T cell activation/proliferation; this was also accompanied by a lack of expression of key costimulatory molecules (OX40L, CD40, CD70, and CD86) on SMCs. Finally, OVA-treated SMCs failed to induce DO11.10-GFP hybridoma activation, a process independent of costimulation. Our results demonstrate that while murine primary aortic SMCs express MHC class II and can acquire exogenous antigens, they fail to activate T cells through a failure in antigen presentation and a lack of costimulatory molecule expression.

U2 - 10.1155/2014/949845

DO - 10.1155/2014/949845

M3 - Article

VL - 2014

JO - Biomed Research International

JF - Biomed Research International

SN - 2314-6133

M1 - 949845

ER -