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Fig. 6 | Journal of Neuroinflammation

Fig. 6

From: Glial cell activation, recruitment, and survival of B-lineage cells following MCMV brain infection

Fig. 6

B cell migration towards recombinant chemokines and supernatants from stimulated glial cell cultures. CD19+ cells were isolated from the spleens of the MCMV-primed mice (7 days), and migration was assessed using chemotaxis chambers. a Migration of B cells towards select recombinant chemokines (100 ng/ml; 300 ng/ml for CXCL13). After 4 h, the migrated cells were stained using Alamar blue and read with a fluorescent plate reader (Ex550 nm Em590 nm). **p < 0.0001 vs. media alone. b Total numbers of migrated CD19+ cells were determined using flow cytometry. Data (mean ± SD) presented are representative of three independent experiments. **p = 0.0034 for CXCL9 and **p < 0.0001 for CXCL10 and CXCL13 vs. media alone. c B cell migration to recombinant CXCL13 (dose 0 to 300 ng/ml). Pooled data are (mean ± SEM) derived from two independent experiments. **p = 0.0038 for 100 ng/ml and **p < 0.0001 for 300 ng/ml vs. 0 ng/ml. d Migration of B cells towards culture supernatants obtained from IFN-γ-stimulated (10 ng/ml) or MCMV-infected (MOI = 5) primary microglial cell and primary astrocyte cultures at 48 h post-stimulation or infection. B cell migration was assessed after 4 h by flow cytometry. Data (mean ± SD) shown are representative of two separate experiments. **p < 0.0001 for astrocytes vs. untreated and **p = 0.0044 for microglia vs. untreated. e Blockade of specific migration towards MCMV-infected (MOI = 5) microglial cell supernatants. Anti-chemokine Abs were added (10 μg/ml) to supernatants from infected microglial cell cultures for 1 h prior to being loaded onto the chemotaxis chamber for assessment of effects on migration. B cell migration was assessed after 4 h by flow cytometry. Pooled data are (mean ± SEM) derived from two independent experiments. †† p = 0.0083 vs. MCMV alone

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