Fig 5.

Spleen cells from 5-w.p.i. LP-BM5 B6 mice inhibit B-cell proliferation as alternatively assessed by flow cytometry. CFSE-labeled B6 responder cells were cultured for 72 h for LPS stimulation and 96 h for α-CD40 and IL-4 stimulation in the presence of either nonfractionated splenic suppressor cells from uninfected mice or nonfractionated or Ly6G⁺-depleted CD11b⁺-enriched cells obtained from 5-w.p.i. LP-BM5 mice. At the termination of incubation, the cells were stained with α-CD19 fluorochrome-conjugated MAb, and the CFSE dilution was assessed by FACS. The percentages of CD19⁺ cells divided and proliferation indices were obtained with the FlowJo proliferation platform (see Materials and Methods), with the Δ values obtained by subtracting the medium-stimulated responder values for the percentages of cells divided or the division index values. The presented pattern of results is representative of one additional experiment for each type of B-cell stimulation.