Skip to main content
Figure 1 | Journal of Neuroinflammation

Figure 1

From: Inhibition of store-operated calcium entry in microglia by helminth factors: implications for immune suppression in neurocysticercosis

Figure 1

Effect of helminth soluble factors on cytokine production by microglia. Microglia were pulsed with medium alone, helminth soluble factors (HSFs) at 25 μg/ml or various Toll-like receptor (TLR) ligands (lipopolysaccharide (LPS; 10 ng/ml), Pam3Cys4 (Pam3; 10 ng/ml), double-stranded RNA (dsRNA; 10 ng/ml), single-stranded RNA (ssRNA; 10 ng/ml), CpG site DNA (1 μM) or HSFs) before the addition of the respective TLR agonists in the medium. Cells were cultured for a 24-hour period. The cytokine contents of interleukin 6 (IL-6) (A) and tumor necrosis factor α (TNF-α) (B) in culture supernatants were assayed by using specific sandwich enzyme-linked immunosorbent assays (ELISAs) as recommended by the manufacturer (BD Biosciences or R&D Systems (Minneapolis, MN, USA)). The mean ± SE concentration of cytokines in five independent experiments for IL-6 and three independent experiments for TNF-α was determined. (C) Microglia were pulsed with medium alone or with HSF at various concentrations (2.5 μg/ml, 5 μg/ml, 12.5 μg/ml, 25 μg/ml or 50 μg/ml) before the addition of LPS (L; 10 ng/ml) to the medium. The cytokine contents of IL-6 in culture supernatants were assayed at 24 hours as described above. (D) Cells were pulsed with HSF with or without RNase, DNase or glycosidase (O-glycosidase; Sigma-Aldrich) to modify glycan moieties 20 minutes prior to addition of LPS at 10 ng/ml. Culture supernatants were collected after 24 hours, and IL-6 concentration was measured by ELISA. (E) Microglia were pulsed with medium alone, LPS (10 ng/ml) and HSF (25 μg/ml) with or without LPS (10 ng/ml) in the medium for 24 hours. An MTT assay was performed to detect the cell viability. Absorption values at 570 nm were normalized to untreated cells. Data are presented as the percentage of untreated cells, with viability referred to as 100%. Significant differences were measured by Student’s t-test and are denoted by asterisks (***P < 0.001). NS, Nonstimulated.

Back to article page