Combined CD4+ donor lymphocyte infusion and low-dose recombinant IL-2 expand FOXP3+ regulatory T cells following allogeneic hematopoietic stem cell transplantation - PubMed (original) (raw)

Combined CD4+ donor lymphocyte infusion and low-dose recombinant IL-2 expand FOXP3+ regulatory T cells following allogeneic hematopoietic stem cell transplantation

Emmanuel Zorn et al. Biol Blood Marrow Transplant. 2009 Mar.

Abstract

CD4(+)CD25(+)FOXP3(+) regulatory T cells (Treg) successfully control graft-versus-host-disease (GVHD) in animal models. In humans, incomplete reconstitution of Treg after allogeneic hematopoietic stem cell transplantation (HSCT) has been associated with chronic GVHD (cGVHD). Recent studies have demonstrated that interleukin (IL)-2 infusions expand Treg in vivo. However, the effectiveness of this therapy depends on the number of cells capable of responding to IL-2. We examined the effect of low-dose IL-2 infusions on Treg populations after HSCT in patients who also received infusions of donor CD4(+) lymphocytes. Utilizing FOXP3 as a Treg marker, we found that patients who received CD4+DLI concomitantly with IL-2 had greater expansion of Treg compared to patients who received IL-2 (P = .03) or CD4(+)DLI alone (P = .001). FOXP3 expression correlated with absolute CD4(+)CD25(+) cell counts. Moreover, expanded CD4(+)CD25(+) T cells displayed normal suppressive function and treatment with CD4(+)DLI and IL-2 was not associated with GVHD. This study suggests that administration of low-dose IL-2 combined with adoptive CD4(+) cellular therapy may provide a mechanism to expand Treg in vivo.

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Figures

Figure 1. Increased FOXP3 gene expression following CD4+ DLI plus low dose IL-2

FOXP3 expression was assessed by real time PCR in PBMC from patients who received CD4+ DLI (black lines), low-dose IL-2 (blue lines) or combined CD4+ DLI and low-dose IL-2 (red lines). FOXP3 expression levels were normalized based on the expression of TFRC (transferrin receptor).

Figure 2. Intracellular staining for FOXP3 protein in patients who received CD4+ DLI and IL-2

Patient PBMC were tested for expression of cell membrane CD4 and intracellular FOXP3. Samples were collected before patients received CD4+DLI and IL-2 and between 13 to 77 days after starting therapy. Dot-plots are shown after gating on lymphocyte populations.

Figure 3. Correlation of FOXP3 gene expression and CD4+CD25+ Treg and total CD4 T cells in peripheral blood after CD4+ DLI and IL-2

A. FOXP3 gene expression (circles) was assessed by quantitative PCR in PBMC collected from patients who received CD4+DLI + IL-2 and is shown together with absolute blood CD4+CD25+ T cell counts (triangles; cells/μl). Spearman correlation coefficient r=0.79, p=0.0003. B. FOXP3 gene expression is depicted (circles) together with absolute blood CD4+ T cell counts (squares; cells/μl) for all patients treated with CD4+DLI + IL-2. Spearman correlation coefficient r=0.76, p=0.0006.

Figure 4. Suppressive activity of CD4+CD25+ Treg following IL-2 and CD4+ DLI combined therapy

Suppressive activity of purified CD4+CD25+ T cells was assessed for patients 1, 2, 3 and 5 using samples collected after DLI and during IL-2 treatment. Suppression by Treg purified from 4 healthy donors was also measured. Solid lines represent median values for the 2 groups.

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Combined CD4+ donor lymphocyte infusion and low-dose recombinant IL-2 expand FOXP3+ regulatory T cells following allogeneic hematopoietic stem cell transplantation - PubMed (original) (raw)