Fig 3.

T. b. gambiense secretomes modulate the phenotype of immature BMDCs. (A) After stimulation with LPS in the presence or absence of the T. b. gambiense secretome, BMDCs from BALB/c mice were collected in PBS containing 0.1% bovine serum albumin and 0.1% NaN₃ and incubated with MAbs for 30 min at 4°C. BMDCs gated for CD11c positivity (R2; Fig. 2) were analyzed for changes in CD40, CD80, CD86, and MHC class II molecule expression levels (mean ± SD of the percentage of cells gated in M1; Fig. 2; Table 1). (B) TNF-α, IL-10, and IL-6 in culture supernatants of BMDCs exposed to the following stimuli for 48 h were quantified using a BD CBA mouse inflammation kit: LPS with or without T. b. gambiense secretomes. Data were acquired by flow cytometry (FACSCalibur), were analyzed by BD Bioscience cytometric bead array software according to the manufacturer's instructions, and are presented as the mean ± SD cytokine concentration (pg/ml; Table 2). For both panels A and B, cells in culture medium alone (BMDCs) were used as a negative control, and those supplemented with LPS were used as a positive control. Data are presented as the mean ± SD of triplicates from three independent experiments. Statistical significance was analyzed using the Student t test; P values are indicated for each histogram.