Clustering of adenosine A2B receptors with ectonucleotidases in caveolin-rich lipid rafts underlies immunomodulation by Leishmania amazonensis

Extracellular adenosine plays important roles in modulating the immune responses. We have previously demonstrated that infection of dendritic cells (DC) by Leishmania amazonensis leads to increased expression of CD39 and CD73 and to the selective activation of the low affinity A(2B) receptors (A(2B)R), which contributes to DC inhibition, without involvement of the high affinity A(2A)R. To understand this apparent paradox, we now characterized the alterations of both adenosine receptors in infected cells. With this aim, bone marrow-derived DC from C57BL/6J mice were infected with metacyclic promastigotes of L. amazonensis. Fluorescence microscopy revealed that L. amazonensis infection stimulates the recruitment of A(2B)R, but not of A(2A)R, to the surface of infected DC, without altering the amount of mRNA or the total A(2B)R density, an effect dependent on lipophosphoglycan (LPG). Log-phase promastigotes or axenic amastigotes of L. amazonensis do not stimulate A(2B)R recruitment. A2BR clusters are localized in caveolin-rich lipid rafts and the disruption of these membrane domains impairs A(2B)R recruitment and activation. More importantly, our results show that A(2B)R co-localize with CD39 and CD73 forming a purinergic cluster that allows for the production of extracellular adenosine in close proximity with these receptors. We conclude that A(2B)R activation by locally produced adenosine constitutes an elegant and powerful evasion mechanism used by L. amazonensis to down-modulate the DC activation.

Automated summary

The study demonstrates that infection of dendritic cells by Leishmania amazonensis leads to the recruitment of A(2B) receptors but not A(2A) receptors. This effect depends on lipophosphoglycan (LPG), and A2BR clusters are localized in caveolin-rich lipid rafts, co-localizing with CD39 and CD73 to produce extracellular adenosine.