Cell penetrating recombinant Foxp3 protein enhances Treg function and ameliorates arthritis - PubMed (original) (raw)

Cell penetrating recombinant Foxp3 protein enhances Treg function and ameliorates arthritis

Kentaro Yomogida et al. Biochem Biophys Res Commun. 2013.

Abstract

Foxp3 is the master transcription factor for T regulatory (Treg) cell differentiation and function. This study aimed to test the therapeutic potential of cell penetrating recombinant Foxp3 protein in arthritis. Recombinant Foxp3 protein was fused to a cell penetrating polyarginine (Foxp3-11R) tag to facilitate intracellular transduction. In vitro Foxp3-11R treated CD4(+) T cells showed a 50% increase in suppressive function compared with control protein treated cells. Severity of arthritis in Foxp3-11R treated mice was significantly reduced compared with those treated with a control protein. CD4(+) T cells of lymph nodes and spleen from Foxp3-11R treated mice showed increased levels of Foxp3 expression compared with those of a control protein treated. These results demonstrated that Foxp3-11R can enhance T cell suppressive function and ameliorate experimental arthritis and suggest that cell penetrating recombinant Foxp3 is a potentially useful agent in therapy of arthritis.

Published by Elsevier Inc.

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Conflict of interest statement

Competing Interests

SW, BB, CA, TC, BM and YZ are employees of VivoScript, Inc.

Figures

Figure 1

Figure 1. Generation of cell-permeable Foxp3-11R

(A) A schematic presentation of the Foxp3 fusion protein. (B) SKG mouse splenocytes were incubated with 10 μg/ml of Foxp3-11R, ASCL1-11R or culture medium in anti-CD3/CD28 coated plates for 120 hours, and Foxp3 was stained intracellularly. Mean fluorescence of Foxp3 was compared in the three treatment group. Data are representative of three independent experiments and mean ± SD (n=3). *p<0.05. (C) Splenocytes were treated as in (B) and stained with Annexin V and propidium iodide (PI). Figure shows CD4+ T cells stained by Annexin V (pooled data of three experiments, n=6 in each group).

Figure 2

Figure 2. Foxp3-11R treatment did not upregulate CD25, but induced Treg-like cells

(A) SKG mouse splenocytes were incubated with 10 μg/ml of Foxp3-11R, ASCL1-11R or culture medium in anti-CD3/CD28 coated plates for 120 hours, and CD25 on cell surface was stained. (B) Suppressor cells (sup) were prepared by incubating SKG mouse splenocytes with 40 μg/ml of Foxp3-11R, ASCL-11R, or culture medium for 1 hour and washed twice. As responder cells (res), SKG mouse splenocytes were stained with CFSE. Suppressor cells and responder cells were mixed at various ratios. Data are representative of three independent experiments.

Figure 3

Figure 3. Foxp3-11R prevented progression of arthritis in SKG mice

(A) Seven days after arthritis induction, SKG mice were treated with Foxp3-11R or ASCL1-11R for 7 days. Fourteen days after final treatment, splenocytes were harvested and Foxp3 was stained intracellularly. Data are CD4 T cell gated and mean ± SD (n=4). *p<0.05 (B) Severity of arthritis was monitored daily after the above treatment (n=10 in each group). (p=0.78 at week 4, p=0.02 at week 6 and p=0.033 at week 8, pooled data of two experiments).

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