Choroid plexus-selective inactivation of adenosine A2A receptors protects against T cell infiltration and experimental autoimmune encephalomyelitis

Background Multiple sclerosis (MS) is one of the most common autoimmune disorders characterized by the infiltration of immune cells into the brain and demyelination. The unwanted immunosuppressive side effect of therapeutically successful natalizumab led us to focus on the choroid plexus (CP), a key site for the first wave of immune cell infiltration in experimental autoimmune encephalomyelitis (EAE), for the control of immune cells trafficking. Adenosine A(2A) receptor (A(2A)R) is emerging as a potential pharmacological target to control EAE pathogenesis. However, the cellular basis for the A(2A)R-mediated protection remains undetermined. Methods In the EAE model, we assessed A(2A)R expression and leukocyte trafficking determinants in the CP by immunohistochemistry and qPCR analyses. We determined the effect of the A(2A)R antagonist KW6002 treatment at days 8-12 or 8-14 post-immunization on T cell infiltration across the CP and EAE pathology. We determined the critical role of the CP-A(2A)R on T cell infiltration and EAE pathology by focal knock-down of CP-A(2A)R via intracerebroventricular injection of CRE-TAT recombinase into the A(2A)R(flox/flox) mice. In the cultured CP epithelium, we also evaluated the effect of overexpression of A(2A)Rs or the A(2A)R agonist CGS21680 treatment on the CP permeability and lymphocytes migration. Results We found the specific upregulation of A(2A)R in the CP associated with enhanced CP gateway activity peaked at day 12 post-immunization in EAE mice. Furthermore, the KW6002 treatment at days 8-12 or 8-14 post-immunization reduced T cell trafficking across the CP and attenuated EAE pathology. Importantly, focal CP-A(2A)R knock-down attenuated the pathogenic infiltration of Th17(+) cells across the CP via inhibiting the CCR6-CCL20 axis through NF kappa B/STAT3 pathway and protected against EAE pathology. Lastly, activation of A(2A)R in the cultured epithelium by A(2A)R overexpression or CGS21680 treatment increased the permeability of the CP epithelium and facilitated lymphocytes migration. Conclusion These findings define the CP niche as one of the primary sites of A(2A)R action, whereby A(2A)R antagonists confer protection against EAE pathology. Thus, pharmacological targeting of the CP-A(2A)R represents a novel therapeutic strategy for MS by controlling immune cell trafficking across CP.

Automated summary

A(2A)R is upregulated in the choroid plexus (CP), enhancing its gateway activity in experimental autoimmune encephalomyelitis (EAE) mice. Treatment with the A(2A)R antagonist KW6002 reduces T cell trafficking across the CP and attenuates EAE pathology. Focal knock-down of CP-A(2A)R inhibits the infiltration of Th17(+) cells across the CP through the CCR6-CCL20 axis, leading to protection against EAE pathology. Activation of A(2A)R in the cultured CP epithelium increases CP permeability and facilitates lymphocyte migration.