FIG. 4.
B cells do not contribute to IFN-γ production after LVS infection. C57BL/6J mice were infected with 105 CFU of LVS i.d. Single-cell suspensions of pooled splenocytes from five naive and LVS-infected mice were prepared on day 4 and enriched for B cells by use of anti-CD19+ MACS beads. The percentage of IFN-γ+ enriched B cells was assessed by ICS and flow cytometry (B), and the percentage of IFN-γ expression in total splenocytes, obtained and analyzed prior to B-cell enrichment, is shown as a positive control (A). (C) The amount of IFN-γ in supernatants after 3 days of ex vivo culture was assessed by ELISA. Data indicate means ± standard deviations for three individual cultures established using pooled cells as described above. Results shown were derived from a single experiment of this design and are consistent with similar results obtained using unseparated cells.