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 journalArticlepeer-review

Abstract

In the present study aortic murine smooth muscle cell (SMC) antigen presentation capacity was evaluated using the Eα-GFP/Y-Ae system to visualize antigen uptake through a GFP tag and tracking of Eα peptide/MHCII presentation using the Y-Ae Ab. Stimulation with IFN-γ (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 Eα-GFP (100 μg/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 of MHC class II. After IFN-γ-stimulation, ovalbumin- (OVA, 1 mg/mL) or OVA323-339 peptide-(0.5 μg/mL) treated SMCs failed to induce OT-II CD4(+) 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
Pages (from-to)1-10
Number of pages10
JournalBioMed Research International
Volume2014
DOIs
Publication statusPublished - 2014

Keywords

  • Animals
  • Antigen Presentation
  • Aorta/cytology
  • Flow Cytometry
  • Histocompatibility Antigens Class I/immunology
  • Histocompatibility Antigens Class II/immunology
  • Lymphocyte Activation/immunology
  • Mice
  • Myocytes, Smooth Muscle/immunology
  • Peptides/immunology
  • T-Lymphocytes/immunology

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