AKAPs in lipid rafts are required for optimal antigen presentation by dendritic cells

Robynn V. Schillace, Casey L. Miller, Daniel Carr

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Dendritic cell (DC) maturation and antigen presentation are regulated by activation of protein kinase A (PKA) signaling pathways, through unknown mechanisms. We have recently shown that interfering with PKA signaling through the use of anchoring inhibitor peptides hinders antigen presentation and DC maturation. These experiments provide evidence that DC maturation and antigen presentation are regulated by A-kinase anchoring proteins (AKAPs). Herein, we determine that the presence of AKAPs and PKA in lipid rafts regulates antigen presentation. Using a combination of western blotting and immuno-cytochemistry, we illustrate the presence of AKAP149, AKAP79, Ezrin and the regulatory subunits of PKA in DC lipid rafts. Incubation of DCs with the type II anchoring inhibitor, AKAP-in silico (AKAP-IS), removes Ezrin and RII from the lipid raft without disrupting raft formation. Addition of a lipid raft disruptor, methyl-Β-cyclodextrin, blocks the efficacy of AKAP-IS, suggesting that the lipid raft must be intact for AKAP-IS to inhibit antigen presentation. Ezrin and AKAP79 are present in the lipid raft of stimulated KG1 cells, but Ezrin is not present in the lipid raft of unstimulated KG1 cells and AKAP79 levels are greatly diminished, suggesting that Ezrin and AKAP79 may be the key AKAPs responsible for regulating antigen presentation.

Original languageEnglish (US)
Pages (from-to)650-658
Number of pages9
JournalImmunology and Cell Biology
Volume89
Issue number5
DOIs
StatePublished - Jul 2011

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Antigen Presentation
Dendritic Cells
Protein Kinases
Lipids
Cyclic AMP-Dependent Protein Kinases
Histocytochemistry
Cyclodextrins
Computer Simulation
Western Blotting
ezrin
Peptides

Keywords

  • AKAP
  • antigen presentation
  • human dendritic cell
  • lipid raft
  • signal transduction

ASJC Scopus subject areas

  • Immunology
  • Cell Biology

Cite this

AKAPs in lipid rafts are required for optimal antigen presentation by dendritic cells. / Schillace, Robynn V.; Miller, Casey L.; Carr, Daniel.

In: Immunology and Cell Biology, Vol. 89, No. 5, 07.2011, p. 650-658.

Research output: Contribution to journalArticle

Schillace, Robynn V. ; Miller, Casey L. ; Carr, Daniel. / AKAPs in lipid rafts are required for optimal antigen presentation by dendritic cells. In: Immunology and Cell Biology. 2011 ; Vol. 89, No. 5. pp. 650-658.
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