Blood -- FoxP3+CD4+ regulatory T cells play an important role in acute HIV-1 infection in humanized Rag2-/-{gamma}C-/- mice in vivo

Blood, Vol. 112, Issue 7, 2858-2868, October 1, 2008

FoxP3⁺CD4⁺ regulatory T cells play an important role in acute HIV-1 infection in humanized Rag2^–/– ${gamma}$ C^–/– mice in vivo
Blood Jiang et al. 112: 2858

Supplemental materials for: Jiang et al

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Figure S1. Antibodies, isotype controls and gating strategy for multi-color FACS analysis (JPG, 120 KB) -
(A) Live human (VLD⁻, murine CD45⁻, human CD45⁺) cells in PBMC, mesenteric lymph node (mLN), spleen, peripheral lymph node (PLN) and thymus were analyzed by multi-color FACS for expression of CD25 and FoxP3 on CD3⁺CD4⁺CD8⁻human T cells. Isotype control antibodies with appropriate labels were used for background staining. (B) Expressions of FoxP3 and CD25 on CD3⁺CD4⁺CD8⁻ T cells were analyzed with human cells from various lymphoid organs of DKO-hu mice. Numbers indicate percentages of each subset. Shown are representative data from at least three independent experiments.
Figure S2. The expression profile of human leukocyte markers on Treg cells (JPG, 70.2 KB) -
FoxP3⁺ or FoxP3⁻ T cells from spleen (Spl, A) and mesenteric lymph node (mLN, B) of DKO-hu HSC mice were analyzed. Cells shown were gated on human CD45⁺ mouse CD45⁻VLD⁻human CD3⁺CD4⁺CD8⁻ cells. Shown are representative data from at least three independent experiments.
Figure S3. Depletion of Treg by ONTAK in DKO-hu HSC mice (JPG, 97.4 KB) -
As performed in human patients, ONTAK (Denileukin diftitox) in saline was injected iv at 5 mg/kg/day over 2–3 days (10 DKO-hu HSC mice treated with ONTAK or PBS). (A) Human CD45⁺, CD3⁺CD4⁺CD8⁻ T cells in PBMC were analyzed by multi-color FACS before injection (day 0) or 3 and 6 days after injection (post-ONTAK). Relative % CD4⁺CD25^+/hi cells are shown as % relative to pretreatment. ONTAK treatment significantly reduced % CD4⁺CD25⁺ T cells (p+CD3⁺ T cells from spleen were analyzed for CD4/CD8 and CD25/FoxP3 expression. Spleen CD45⁺CD3⁺ T cells were also stained for T cell activation marker HLA-DR, relative activation of CD4 and CD8 T cells (% HLA-DR⁺) is shown.