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. Author manuscript; available in PMC: 2017 Mar 1.
Published in final edited form as: Transplantation. 2017 Mar;101(3):541–547. doi: 10.1097/TP.0000000000001341

Fig. 1. Higher IL-10 production by B-cells from tolerant recipients was observed after CD40 ligation.

Fig. 1

A) Comparison of the distribution of total, memory, naïve and transitional B-cells within PBMCs between healthy control (HC=11) and tolerant (Tol=16) patients. Total B-cells were identified as CD20+CD19+, memory B-cells as CD27+, naïve B-cells as CD27-IgD+IgM+CD24+CD38+ and transitional B-cells as CD27-IgD+IgM+CD24hiCD38hi by surface staining. A Mann-Whitney test was used. B) Production of IL-10 and TNF-α by intracellular staining and protein levels were measured after 3 days of activation with 0.5x105 non-transfected L-cells (Basal) or human-CD40L-transfected L-cells (CD40L) in 1.0x106 PBMCs from HC=8 and Tol=10 patients. Detection limit for IL-10 and TNF-α were 2 and 15.6pg/ml, respectively. C) The expression of CD25 and CD86 on CD20+ B-cells was measured after 3 days of activation with 0.5x105 non-transfected L-cells (Basal) or human-CD40L-transfected L-cells (CD40L) in 1.0x106 PBMCs from HC=11 and Tol=14 patients. D) Up-regulation of CD40L in CD4+T-cells from healthy HC=6 and Tol=7 patients were measured after 8hrs of activation with CD3/CD28 beads (1:2 ratio). Two-way RM ANOVA test with a Sidak’s multiple comparisons test was used. For all statistical tests **** P<0.0001, *** P<0.001, ** P<0.01 and * P<0.05 were considered significant.