A Role for Regulatory T Cells in Cutaneous T-Cell Lymphoma; Induction of a CD4+CD25+Foxp3+ T-Cell Phenotype Associated with HTLV-1 Infection  Patrick T. Walsh, Bernice M. Benoit, Maria Wysocka, Nicole M. Dalton, Laurence A. Turka, Alain H. Rook  Journal of Investigative Dermatology  Volume 126, Issue 3, Pages 690-692 (March 2006) DOI: 10.1038/sj.jid.5700121 Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Expression of Foxp3 in PBMCs from normal and CTCL patients. RNA from both patient (n=15) and normal PBMC (n=3) samples were screened for Foxp3 expression by quantitative real-time RT-PCR. Level of Foxp3 was measured relative to levels of β-actin. Primers were obtained as TaqMan Gene expression assays with the assay numbers Hs00203958_m1 (FoxP3) and Hs99999903_m1 (β-actin) (PE Applied Biosystems, Foster City, CA). Foxp3 expression in normal samples was taken as baseline expression. Results represent mean±standard deviation level of Foxp3 expression in individual samples. Journal of Investigative Dermatology 2006 126, 690-692DOI: (10.1038/sj.jid.5700121) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Induction of a regulatory T-cell phenotype by HTLV-1 infection. (a) Percentage expression of CD25 on CD4+ T cells after coculture of CD4+CD25- T cells with either irradiated MJ cells or irradiated Jurkat cells for 8 weeks (n=4). Results expressed as mean±standard deviation. (b) Representative flow-cytometric analysis from (a) of induction of CD25 expression after 8-week coculture with irradiated MJ cells. (c) Quantitative real-time PCR analysis of Foxp3 expression on CD4+ T cells after coculture with the irradiated MJ cell line as in (a) above. Results depict mean±standard deviation expression level of Foxp3 relative to β-actin under each condition. Data are representative of four independent experiments with similar results. (d) TGF-β secretion by CD4+ T cells after coculture with irradiated MJ cells compared to TGF-β secreted by CD4+ T cells cocultured with the non-HTLV-1-positive jurkat cell line and cultures containing both irradiated cell lines alone. Results represent mean±standard deviation secretion from four independent coculture experiments. (e) Presence of HTLV-1 virus in CD4+ T cells after in-vitro coculture with irradiated MJ cell line detected by PCR of viral genome. Nonirradiated HTLV-1-positive (MJ) and -negative (Jurkat) cell lines used as controls for PCR. Journal of Investigative Dermatology 2006 126, 690-692DOI: (10.1038/sj.jid.5700121) Copyright © 2006 The Society for Investigative Dermatology, Inc Terms and Conditions