Early and late extensive chronic graft-versus-host disease in children is characterized by different Th1/Th2 cytokine profiles: findings of the Children's Oncology Group Study ASCT0031

Abstract

Numerous mechanisms underlie chronic graft-versus-host disease (cGVHD), including skewing of Th1/Th2 cytokine expression. There are biological differences between early-onset and late-onset cGVHD. To test whether different Th1/Th2 cytokines are associated with early- or late-onset cGVHD, peripheral blood was collected from 63 children enrolled on the Children's Oncology Group Phase III trial ASCT0031 evaluating hydroxychloroquine therapy for newly diagnosed extensive cGVHD. mRNA expression of interferon (IFN)-γ and interleukin (IL)-2, -4, and -10 from stimulated peripheral blood mononuclear cells was evaluated by quantitative polymerase chain reaction. Early-onset cGVHD (n = 33) was characterized by decreased expression of IFN-γ and IL-2 mRNA after nonspecific phorbol 12-myristate 13-acetate-ionomycin stimulation. In contrast, late-onset cGVHD (n = 11) was characterized by decreased expression of IL-4 and IL-2 mRNA after anti-CD3 stimulation of T cells. Receiver-operating characteristic curve analysis revealed that IFN-γ expression was correlated with the absence of early cGVHD (area under the curve [AUC] = 0.77) and that IL-4 (AUC = 0.89) and IL-2 (AUC = 0.84) expression was correlated with the absence of late cGVHD. There was no correlation between cytokine expression and a specific immune cell subset. Increased expression of Foxp3 mRNA was seen in early-onset cGVHD and late controls. The different time-dependent cytokine profiles in patients with newly diagnosed cGVHD suggests that the mechanisms underlying cGVHD are temporally regulated. Although larger validation studies are needed, our data suggest that cytokine profiles have a potential use as biomarkers for the diagnosis of cGVHD.

Conflict of interest statement

Financial Disclosure Statement: All authors wish to disclose they do not have any existing or potential conflicts of interest.

Copyright © 2011 American Society for Blood and Marrow Transplantation. Published by Elsevier Inc. All rights reserved.

Figures

Figure 1

A) IFN-γ mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; B) IFN-γ mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; C) IL-2 mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; D) IL-2 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; E) IL-4 mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; F) IL-4 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; G) IL-10 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours.

Figure 1

A) IFN-γ mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; B) IFN-γ mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; C) IL-2 mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; D) IL-2 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; E) IL-4 mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; F) IL-4 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; G) IL-10 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours.

Figure 1

A) IFN-γ mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; B) IFN-γ mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; C) IL-2 mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; D) IL-2 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; E) IL-4 mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; F) IL-4 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; G) IL-10 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours.

Figure 1

A) IFN-γ mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; B) IFN-γ mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; C) IL-2 mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; D) IL-2 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; E) IL-4 mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; F) IL-4 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; G) IL-10 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours.

Figure 1

A) IFN-γ mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; B) IFN-γ mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; C) IL-2 mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; D) IL-2 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; E) IL-4 mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; F) IL-4 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; G) IL-10 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours.

Figure 1

A) IFN-γ mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; B) IFN-γ mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; C) IL-2 mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; D) IL-2 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; E) IL-4 mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; F) IL-4 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; G) IL-10 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours.

Figure 1

A) IFN-γ mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; B) IFN-γ mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; C) IL-2 mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; D) IL-2 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; E) IL-4 mRNA production at early and late timepoints in controls and patients with cGVHD after PI stimulation for 18 hours; F) IL-4 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours; G) IL-10 mRNA production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours.

Figure 2

A) ROC curve analysis of IFN-γ mRNA production after PI stimulation for predicting the absence of early cGVHD; B) ROC curve analysis of IL-2 mRNA production after anti-CD3 stimulation for predicting the absence of late cGVHD; C) ROC curve analysis of IL-4 mRNA production after anti-CD3 stimulation for predicting the absence of late cGVHD.

Figure 2

A) ROC curve analysis of IFN-γ mRNA production after PI stimulation for predicting the absence of early cGVHD; B) ROC curve analysis of IL-2 mRNA production after anti-CD3 stimulation for predicting the absence of late cGVHD; C) ROC curve analysis of IL-4 mRNA production after anti-CD3 stimulation for predicting the absence of late cGVHD.

Figure 2

A) ROC curve analysis of IFN-γ mRNA production after PI stimulation for predicting the absence of early cGVHD; B) ROC curve analysis of IL-2 mRNA production after anti-CD3 stimulation for predicting the absence of late cGVHD; C) ROC curve analysis of IL-4 mRNA production after anti-CD3 stimulation for predicting the absence of late cGVHD.

Figure 3

A) Foxp3 production at early and late timepoints in controls and patients with cGVHD in un-stimulated samples; B) Foxp3 production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours.

Figure 3

A) Foxp3 production at early and late timepoints in controls and patients with cGVHD in un-stimulated samples; B) Foxp3 production at early and late timepoints in controls and patients with cGVHD after anti-CD3 mAb stimulation for 18 hours.