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Glial innate immunity generated by non-aggregated alpha-synuclein in mouse: differences between wild-type and Parkinson's disease-linked mutants.

Roodveldt C, Labrador-Garrido A, Gonzalez-Rey E, Fernandez-Montesinos R, Caro M, Lachaud CC, Waudby CA, Delgado M, Dobson CM, Pozo D - PLoS ONE (2010)

Bottom Line: Lately, accumulating evidence has shown the presence of extracellular α-Syn both in its aggregated and monomeric forms in cerebrospinal fluid and blood plasma.In this work, we have compared the activation profiles of non-aggregated, extracellular wild-type and PD-linked mutant α-Syn variants on primary glial and microglial cell cultures.Contrary to what had been observed using cell lines or for the case of aggregated α-Syn, we found strong differences in the immune response generated by wild-type α-Syn and the familial PD mutants (A30P, E46K and A53T).

View Article: PubMed Central - PubMed

Affiliation: CABIMER-Andalusian Center for Molecular Biology and Regenerative Medicine, Consejo Superior de Investigaciones Científicos, University of Seville-UPO-Junta de Andalucia, Seville, Spain.

ABSTRACT

Background: Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized pathologically by the presence in the brain of intracellular protein inclusions highly enriched in aggregated alpha-synuclein (α-Syn). Although it has been established that progression of the disease is accompanied by sustained activation of microglia, the underlying molecules and factors involved in these immune-triggered mechanisms remain largely unexplored. Lately, accumulating evidence has shown the presence of extracellular α-Syn both in its aggregated and monomeric forms in cerebrospinal fluid and blood plasma. However, the effect of extracellular α-Syn on cellular activation and immune mediators, as well as the impact of familial PD-linked α-Syn mutants on this stimulation, are still largely unknown.

Methods and findings: In this work, we have compared the activation profiles of non-aggregated, extracellular wild-type and PD-linked mutant α-Syn variants on primary glial and microglial cell cultures. After stimulation of cells with α-Syn, we measured the release of Th1- and Th2- type cytokines as well as IP-10/CXCL10, RANTES/CCL5, MCP-1/CCL2 and MIP-1α/CCL3 chemokines. Contrary to what had been observed using cell lines or for the case of aggregated α-Syn, we found strong differences in the immune response generated by wild-type α-Syn and the familial PD mutants (A30P, E46K and A53T).

Conclusions: These findings might contribute to explain the differences in the onset and progression of this highly debilitating disease, which could be of value in the development of rational approaches towards effective control of immune responses that are associated with PD.

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Immunoregulatory effect of α-Syn-stimulation in primary mixed                            glial and isolated microglial cultures.IL-10 release was measured by ELISA in supernatants of                            α-Syn-stimulated mixed-glial cultures (top) and microglia                            (bottom) after a 20-hour treatment with monomeric Wt or mutant                            α-Syn variants, or lipopolysaccharide (LPS). Values are mean                            ± S.E.M. (n = 4). *                            P<0.05, ** P<0.01,                            *** P<0.001. The results shown are                            representative of two or three independent experiments with microglia                            and mixed glial cultures, respectively.
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pone-0013481-g004: Immunoregulatory effect of α-Syn-stimulation in primary mixed glial and isolated microglial cultures.IL-10 release was measured by ELISA in supernatants of α-Syn-stimulated mixed-glial cultures (top) and microglia (bottom) after a 20-hour treatment with monomeric Wt or mutant α-Syn variants, or lipopolysaccharide (LPS). Values are mean ± S.E.M. (n = 4). * P<0.05, ** P<0.01, *** P<0.001. The results shown are representative of two or three independent experiments with microglia and mixed glial cultures, respectively.

Mentions: Although most studies in the past have focused on microglial production of pro-inflammatory cytokines, a large body of evidence has supported the notion that microglia also produce cytokines with anti-inflammatory or regulatory activities [25]. Indeed, a strong induction of IL-10 −recognized as an anti-inflammatory cytokine− had been observed for microglial cells stimulated with nitrated, aggregated Wt α-Syn [62]. We therefore investigated the effects of non-aggregated and unmodified α-Syn on glial secretion of IL-10 −a Th2 immunoregulator− which reduces cytokine production by Th1 cells (Figure 4). Our results show that only the A30P variant produced a significant increase in IL-10 levels in mixed glial cells (top panel) whilst, on the contrary, the A53T variant caused a significant reduction of IL-10 basal levels, also observed in microglia, likely suggesting a lack of microglial response, a differential uptake by microglia, or an effect of the uptaken α-Syn on the endogenous IL-10 when A53T is present. In this sense, it has been reported a link between α-Syn and the microglial activation features [64], including phagocytic ability.


Glial innate immunity generated by non-aggregated alpha-synuclein in mouse: differences between wild-type and Parkinson's disease-linked mutants.

Roodveldt C, Labrador-Garrido A, Gonzalez-Rey E, Fernandez-Montesinos R, Caro M, Lachaud CC, Waudby CA, Delgado M, Dobson CM, Pozo D - PLoS ONE (2010)

Immunoregulatory effect of α-Syn-stimulation in primary mixed                            glial and isolated microglial cultures.IL-10 release was measured by ELISA in supernatants of                            α-Syn-stimulated mixed-glial cultures (top) and microglia                            (bottom) after a 20-hour treatment with monomeric Wt or mutant                            α-Syn variants, or lipopolysaccharide (LPS). Values are mean                            ± S.E.M. (n = 4). *                            P<0.05, ** P<0.01,                            *** P<0.001. The results shown are                            representative of two or three independent experiments with microglia                            and mixed glial cultures, respectively.
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Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2964342&req=5

pone-0013481-g004: Immunoregulatory effect of α-Syn-stimulation in primary mixed glial and isolated microglial cultures.IL-10 release was measured by ELISA in supernatants of α-Syn-stimulated mixed-glial cultures (top) and microglia (bottom) after a 20-hour treatment with monomeric Wt or mutant α-Syn variants, or lipopolysaccharide (LPS). Values are mean ± S.E.M. (n = 4). * P<0.05, ** P<0.01, *** P<0.001. The results shown are representative of two or three independent experiments with microglia and mixed glial cultures, respectively.
Mentions: Although most studies in the past have focused on microglial production of pro-inflammatory cytokines, a large body of evidence has supported the notion that microglia also produce cytokines with anti-inflammatory or regulatory activities [25]. Indeed, a strong induction of IL-10 −recognized as an anti-inflammatory cytokine− had been observed for microglial cells stimulated with nitrated, aggregated Wt α-Syn [62]. We therefore investigated the effects of non-aggregated and unmodified α-Syn on glial secretion of IL-10 −a Th2 immunoregulator− which reduces cytokine production by Th1 cells (Figure 4). Our results show that only the A30P variant produced a significant increase in IL-10 levels in mixed glial cells (top panel) whilst, on the contrary, the A53T variant caused a significant reduction of IL-10 basal levels, also observed in microglia, likely suggesting a lack of microglial response, a differential uptake by microglia, or an effect of the uptaken α-Syn on the endogenous IL-10 when A53T is present. In this sense, it has been reported a link between α-Syn and the microglial activation features [64], including phagocytic ability.

Bottom Line: Lately, accumulating evidence has shown the presence of extracellular α-Syn both in its aggregated and monomeric forms in cerebrospinal fluid and blood plasma.In this work, we have compared the activation profiles of non-aggregated, extracellular wild-type and PD-linked mutant α-Syn variants on primary glial and microglial cell cultures.Contrary to what had been observed using cell lines or for the case of aggregated α-Syn, we found strong differences in the immune response generated by wild-type α-Syn and the familial PD mutants (A30P, E46K and A53T).

View Article: PubMed Central - PubMed

Affiliation: CABIMER-Andalusian Center for Molecular Biology and Regenerative Medicine, Consejo Superior de Investigaciones Científicos, University of Seville-UPO-Junta de Andalucia, Seville, Spain.

ABSTRACT

Background: Parkinson's disease (PD) is a progressive neurodegenerative disorder characterized pathologically by the presence in the brain of intracellular protein inclusions highly enriched in aggregated alpha-synuclein (α-Syn). Although it has been established that progression of the disease is accompanied by sustained activation of microglia, the underlying molecules and factors involved in these immune-triggered mechanisms remain largely unexplored. Lately, accumulating evidence has shown the presence of extracellular α-Syn both in its aggregated and monomeric forms in cerebrospinal fluid and blood plasma. However, the effect of extracellular α-Syn on cellular activation and immune mediators, as well as the impact of familial PD-linked α-Syn mutants on this stimulation, are still largely unknown.

Methods and findings: In this work, we have compared the activation profiles of non-aggregated, extracellular wild-type and PD-linked mutant α-Syn variants on primary glial and microglial cell cultures. After stimulation of cells with α-Syn, we measured the release of Th1- and Th2- type cytokines as well as IP-10/CXCL10, RANTES/CCL5, MCP-1/CCL2 and MIP-1α/CCL3 chemokines. Contrary to what had been observed using cell lines or for the case of aggregated α-Syn, we found strong differences in the immune response generated by wild-type α-Syn and the familial PD mutants (A30P, E46K and A53T).

Conclusions: These findings might contribute to explain the differences in the onset and progression of this highly debilitating disease, which could be of value in the development of rational approaches towards effective control of immune responses that are associated with PD.

Show MeSH
Related in: MedlinePlus