Abstract
Background:
Adenosine A2A receptor (A2AR) antagonists have been consistently demonstrated to protect against multiple sclerosis (MS) pathology, but A2AR knockout (A2AR-/-) mice exhibit exacerbated immune injury, raising concerns regarding the use of A2AR antagonists for MS treatment. Here, we revealed the critical involvement of A2AR-mediated interactions between Th1+ T cells and the choroid plexus (ChP) epithelium in the pathology of experimental autoimmune encephalomyelitis (EAE).
Methods:
We assessed the effects of A2AR knockout on ChP gateway activity and the interferon gamma (IFN-γ)-secreting capacity of Th1+ T cells in an EAE model by immunofluorescence, qPCR and flow cytometry (FCM). We also investigated the effects of A2AR-mediated interactions between Th1+ T cells and the ChP epithelium on ChP gateway activity in vivo via intracerebroventricular (ICV) injection of Th1+ T cells and in vitro via coculture of ChP epithelial cells and splenic Th1+ T cells. We further knocked down IFN-γ receptor 1 (IFNGR1) specifically in the ChP of A2AR-/- mice via ICV injection of AAV2/5-shRNA (IFNGR1) to disrupt the interactions between Th1+ T cells and the ChP epithelium and thus assess the roles of these interactions in the development of EAE pathology.
Results:
A2AR knockout disrupted the ChP barrier and increased T-cell infiltration across the ChP in EAE model mice. Coculture of splenic Th1+ T cells and ChP epithelial cells revealed that A2AR knockout in ChP epithelial cells strengthened the ChP barrier and attenuated T-cell migration, whereas A2AR knockout in Th1+ T cells increased the accumulation of Th1+ T cells in the ChP via the secretion of IFN-γ. Consistent with the coculture results, ICV injection of activated splenic Th1+ T cells from A2AR-/- mice increased the accumulation of T cells in the ChP to a greater extent than did injection of Th1+ T cells from A2AR+/+ mice. This effect was due to the increased secretion of IFN-γ in A2AR-/- mice compared with A2AR+/+ mice. Finally, ChP-specific knockdown of IFNGR1 attenuated A2AR knockout-induced T-cell infiltration, brain inflammation and EAE pathology.
Conclusion:
A2AR-mediated interactions between Th1+ T cells and the ChP epithelium via the secretion of IFN-γ from CD4+ T cells and the binding IFN-γ to IFNGR1 in the ChP epithelium control immune cell invasion and the development of EAE pathology in A2AR-/- mice.