Calcitonin gene-related peptide decreases IL-1beta, IL-6 as well as Ym1, Arg1, CD163 expression in a brain tissue context-dependent manner while ameliorating experimental autoimmune encephalomyelitis

Activation states of immune cells (among them, the so-called pro- or anti-inflammatory states) influence thepathogenesis of multiple sclerosis (MS). The neuropeptide calcitonin gene-related peptide (CGRP) can exert apro- or anti-inflammatory role in a context-dependent manner. In mice CGRP was found to attenuate the de-velopment of experimental autoimmune encephalomyelitis (EAE, a common MS animal model). We analyzedCGRP effects on the expression of cytokines and markers of activation states, as well as its intracellular cascade,following intrathecal administration during EAE immunization. Real Time quantitative-PCR (RT-PCR) showedthat IL-1beta and IL-6 (associated to a pro-inflammatory state in EAE), but also Ym1 (also known as Chil3), Arg1and CD163 (associated to an anti-inflammatory state in EAE) were decreased in the encephalon (devoid ofcerebellum). In the cerebellum itself, IL-1beta and Ym1 were decreased. TNF-alpha (associated to a pro-in-flammatory state in EAE), but also IL-10 (associated to another type of anti-inflammatory state) and BDNF wereunchanged in these two regions. No changes were detected in the spinal cord. Additional tendencies toward achange (as revealed by RT-PCR) were again decreases: IL-10 in the encephalon and Arg1 in the spinal cord.CGRP decreased percentage of Ym1+/CD68+immunoreactive cells and cell density of infiltrates in the cervicalspinal cord pia mater. Instead, Ym1 in the underlying parenchyma and at thoracic and lumbar levels, as well asArg1, were unchanged. In cultured microglia the neuropeptide decreased Ym1, but not Arg1, immunoreactivity.Inducible NOS (iNOS) was unchanged in spinal cord microglia and astrocytes. The neuropeptide increased theactivation of ERK1/2 in the astrocytes of the spinal cord and in culture, but did not influence the activation ofERK1/2 or p38 in the spinal cord microglia. Finally, in areas adjacent to infiltration sites CGRP-treated microgliashowed a larger ramification radius.In conclusion, CGRP-induced EAE amelioration was associated to a concomitant, context-dependent decreasein the expression of markers belonging to both pro- or anti-inflammatory activation states of immune cells. It canbe hypothesized that CGRP-induced EAE attenuation is obtained through a novel mechanism that promotesdown-regulation of immune cell activation that facilitates the establishment of a beneficial environment in EAEprovided possibly also by other factors