By their localization, microglia represent privileged candidates for this activity. However, as cross-presentation Rucaparib could also lead to tolerance rather than to the priming of CD8+ T cells, it

is necessary to elucidate whether microglia could participate in the cross-presentation activity and maintain CD8+ T-cell activation. We recently demonstrated that microglia cross-present exogenous soluble Ags in vitro [10]. However, the in vivo cross-presentation capacity of resident microglia, within the brain microenvironment, remains undetermined. In response to injuries, peripheral and CNS-associated APCs infiltrate the brain parenchyma and are indistinguishable from activated microglia [5, 9, 36-38], making it difficult to address the question of the cross-presentation capacity of resident microglia. We have thus set up an original protocol based on body irradiation, wherein the head is protected from irradiation. This protocol serves to remove functional peripheral and CNS-associated CD45high APCs without affecting microglial

function and resting status. In this aplasic mice model, we demonstrate for the first time that, despite the brain inhibitory constraints, resident microglia, under appropriate stimulation, cross-prime exogenous Ag to naive CD8+ T cells injected in the brain. In order to determine the capacity of STI571 cost microglia to cross-present exogenous Ag in situ, we set up a model excluding the involvement not only of peripheral APCs that infiltrate the brain in response to injuries but also

of CNS-associated APCs, without affecting microglial function and activation status. In order to eliminate peripheral APCs, mice had their entire bodies, except for their heads, exposed to 4–16 Gy irradiation. Three days later, we determined the presence of leukocytes (that express CD45), including myeloid and lymphoid APCs such as DCs, MΦs and B cells in BM, spleen and cervical LN cells by flow cytometry. The Bortezomib results showed that only 16 Gy irradiation eliminated all CD45+ cells in BM and more than 80% of CD45+ cells in spleen and cervical LN (Fig. 1A). We then observed that residual CD45+ cells in irradiated mice were unable to cross-present Ags. Mice either irradiated or not were injected 3 days later in the flank with BSA and OVA in the absence or presence of APC adjuvant (CpG-ODN, GM-CSF and sCD40L). Spleen cells were isolated one day after OVA or BSA injection and cocultured with OVA-specific OT-1 CD8+ T cells. Spleen cells from irradiated mice injected with OVA, in the absence or presence of adjuvant, failed to induce detectable amounts of IL-2 or IFN-γ by OT-1 T cells (Fig. 1B). As expected [10], spleen cells from non-irradiated OVA-injected mice induced IL-2 (27.95 ± 0.96 pg/mL; mean ± SD, n = 5) and IFN-γ (332.20 ± 64.21 pg/mL) secretion by OT1 cells (Fig. 1B) and the presence of adjuvant enhanced IFN-γ but not IL-2 secretion (1268.11 ± 69.