Fig. 5

Exposure to α-synuclein (auto)antibodies activates RANTES secretion by astrocytes. (A) RANTES levels in cell culture supernatant of neuron-astrocyte co-cultures, with neurons expressing α-syn + NmC. RANTES was quantified at 24 h after treatment of cultures with antibodies or sera at DIV 14. Cultures were either left untreated, or were treated with a commercial anti α-syn Ab (ab138501), or with serum containing α-syn AAbs, or with serum that was depleted for α-syn AAbs. Quantification from pure cell culture medium (“medium”), and from cell culture medium plus serum (“medium + serum”) served as additional background controls. (B) RANTES levels in cell culture supernatant of astrocyte-enriched cultures (> 99% astrocytes). α-syn + EGFP or EGFP alone were expressed in astrocytes. RANTES was quantified at 24 h after treatment with the commercial anti α-syn antibody (ab138501), or with serum containing α-syn AAbs or with serum that was depleted for α-syn AAbs. (C) Percent NmC-positive cells (relative to untreated controls) upon treatment with serum containing α-syn AAb with or without anti-RANTES antibody. (D) Percent NmC-positive cells (relative to untreated controls) upon treatment with α-syn AAb containing serum with or without blockers of RANTES receptors CCR1, 3, and 5. (C) and (D) were obtained in neuron-astrocyte co-cultures. N = 4–6 biological replicates for all conditions. Statistics by one-way ANOVA with Tukey’s (A – C) or Dunnett’s (D) multiple comparison test; statistical powers (1-ß error probability): A) **, *** > 0.9, * 0.5; B) > 0.9 for all conditions; C) **, *** > 0.9; * 0.65; D) > 0.9 for all conditions. * = p < 0.05; ** = p < 0.01; *** = p < 0.001