a-Syn and Innate Immunity Figure 5. Chemokine release profile of a-Syn-stimulated primary mixed glial cultures and isolated microglia cultures. IP-10 (A), RANTES (B), MCP-1 (C), and MIP-1a (D) were measured in supernantants of mixed-glial cultures (left) and microglia (right) after a 20-hour treatment with monomeric Wt or mutant a-Syn variants, or lipopolysaccharide (LPS). All chemokines were assayed by ELISA as described in Materials and Methods. Values are mean 6 S.E.M. (n = 2). * P,0.05, ** P,0.01. The results shown are representative of two and three independent experiments with microglia and mixed glial cultures, respectively. doi:10.1371/journal.pone.0013481.g005 Syn variants beyond purely innate pro-inflammatory responses. In this sense, IP-10/CXCL10 is pivotal in generating antigen-specific T-cells [71]. As the possible roles of adaptive immune responses in PD is gaining increasing attention [72,73], our data should contribute to a better understanding of the differential immune responses exerted by the different a-Syn variants in terms of augmented microglia and macrophage recruitment and/or activation. The strong pro-inflammatory species (IL-1b, IL-6, IP- 10/CXCL10, and RANTES/CCL5) whose release the A30P and E46K variants seem to promote in the CNS could lead to augmented macrophage recruitment and/or activation. This view is supported by the observation of upregulation of MCP-1/CCL2 and MIP-1a/CCL3 by A30P and E46K (Figures 5C and 5D). Interestingly, the large effects observed for A30P and E46K were much more pronounced in microglia, indicating a context- dependent stimulation mechanism. Further studies to asses the PLoS ONE | www.plosone.org 7 October 2010 | Volume 5 | Issue 10 | e13481