Figure 3: B-cell differentiation in acute leukemia is not crucially altered by membrane TLR ligation. BM CD34+ cells were purified from ProB-ALL, PreB-ALL, C-ALL, M-ALL, and AML and stimulated with Pam3CSK4 (TLR2 ligand), LPS (TLR4 ligand), Flagellin (TLR5 ligand), and MALP2 (TLR2/6 ligand) agonists for 48 h, followed by a 30 day-stromal cell coculture. Newly produced CD34+ and CD19+ cell fractions were further identified and enumerated by multiparametric flow cytometry (a). Cell frequencies within the culture for both CD34+ cells (b) and CD19+ B-lymphoid cells (c) are shown. ProB-ALL: B-cell precursor ALL with prevalence of CD34+CD10+CD19+ cells; PreB-ALL: B-cell precursor ALL with prevalence of CD3CD10+/−CD19+ cells; C-ALL: congenital ALL; M-ALL: mixed ALL; AML: acute myeloid leukemia.