PMC3896844_01
PMC3896844_02
CF patient A (the subject of this case report), but not CF patient B (another patient with CF lung disease of a similar degree of severity) had reduced T cell responses compared with control. (A) IL-17 and IFN-gamma production by T cells from mononuclear cells following overnight stimulation. T cells from patient A produced deficient IL-17 and IFN-gamma compared to control. (B) Naive T cells were isolated from patients A, B and a healthy control subject using magnetic sorting and differentiated into Th1 and Th17 lineages. Shown are representative flow cytometry plots of production from differentiated Th1 and Th17 cells, respectively. Patient A, but not patient B, produced reduced levels of IFN-gamma and IL-17 compared with control. (C) Naive T cells were differentiated into regulatory T cells. Shown are representative flow cytometry plots of Foxp3 expression. Patient A, but not patient B, produced reduced levels of Foxp3 compared with control. Representative of 2 - 3 independent experiments.
1710-1492-10-2-1.jpg
PMC3896844_06_1710-1492-10-2-1.jpg
F1
Th1 and Th17 differentiation (Figures 1B, C), but compared to control showed somewhat more Th17 and less Th1, consistent with our recent report of an innate predisposition of naive CF helper T cells to differentiate towards the Th17 phenotype. Thus we cannot suggest that the T cell exhaustion described in patient A is an event that occurs in all patients at this stage of CF lung disease.
PMC3896844_03
PMC3896844_04
Naive T cells from Patient A, but not Patient B, showed marked induction of expression of the T cell exhaustion markers LAG-3 and CTLA-4 following stimulation with anti-CD3 and anti-CD28 antibodies. Representative plots of the expression of the T cell exhaustion markers LAG-3 and CTLA-4 on unstimulated naive T cells from patients A and B, and on T cells following stimulation with anti-CD3 and anti-CD28 antibodies. T cells from patient A, but not patient B, demonstrated a marked induction of the T cell exhaustion markers LAG-3 and CTLA-4. Representative of 2 - 3 independent experiments.
1710-1492-10-2-2.jpg
PMC3896844_06_1710-1492-10-2-2.jpg
F2
Unlike the subject of our present report (patient A), whose T cells demonstrated a marked induction of the T cell exhaustion markers LAG-3 and CTLA-4, pre-transplant patient B showed only a mild induction of LAG-3 and CTLA-4 (Figure 2).
PMC3896844_05
PMC3896844_06
CF patient A (the subject of this case report), but not CF patient B (another patient with CF lung disease of a similar degree of severity) had reduced T cell responses compared with control. (A) IL-17 and IFN-gamma production by T cells from mononuclear cells following overnight stimulation. T cells from patient A produced deficient IL-17 and IFN-gamma compared to control. (B) Naive T cells were isolated from patients A, B and a healthy control subject using magnetic sorting and differentiated into Th1 and Th17 lineages. Shown are representative flow cytometry plots of production from differentiated Th1 and Th17 cells, respectively. Patient A, but not patient B, produced reduced levels of IFN-gamma and IL-17 compared with control. (C) Naive T cells were differentiated into regulatory T cells. Shown are representative flow cytometry plots of Foxp3 expression. Patient A, but not patient B, produced reduced levels of Foxp3 compared with control. Representative of 2 - 3 independent experiments.
1710-1492-10-2-1.jpg
PMC3896844_06_1710-1492-10-2-1.jpg
F1
Th1 and Th17 differentiation (Figures 1B, C), but compared to control showed somewhat more Th17 and less Th1, consistent with our recent report of an innate predisposition of naive CF helper T cells to differentiate towards the Th17 phenotype. Thus we cannot suggest that the T cell exhaustion described in patient A is an event that occurs in all patients at this stage of CF lung disease.
Naive T cells from Patient A, but not Patient B, showed marked induction of expression of the T cell exhaustion markers LAG-3 and CTLA-4 following stimulation with anti-CD3 and anti-CD28 antibodies. Representative plots of the expression of the T cell exhaustion markers LAG-3 and CTLA-4 on unstimulated naive T cells from patients A and B, and on T cells following stimulation with anti-CD3 and anti-CD28 antibodies. T cells from patient A, but not patient B, demonstrated a marked induction of the T cell exhaustion markers LAG-3 and CTLA-4. Representative of 2 - 3 independent experiments.
1710-1492-10-2-2.jpg
PMC3896844_06_1710-1492-10-2-2.jpg
F2
Unlike the subject of our present report (patient A), whose T cells demonstrated a marked induction of the T cell exhaustion markers LAG-3 and CTLA-4, pre-transplant patient B showed only a mild induction of LAG-3 and CTLA-4 (Figure 2).