The kinetics of CD4+Foxp3+ T cell accumulation during a human cutaneous antigen-specific memory response in vivo
Milica Vukmanovic-Stejic, Elaine Agius, Nicola Booth, Padraic J. Dunne, Katie E. Lacy, John R. Reed, Toni O. Sobande, Steven Kissane, Mike Salmon, Malcolm H. Rustin, Arne N. Akbar
Milica Vukmanovic-Stejic, Elaine Agius, Nicola Booth, Padraic J. Dunne, Katie E. Lacy, John R. Reed, Toni O. Sobande, Steven Kissane, Mike Salmon, Malcolm H. Rustin, Arne N. Akbar
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Research Article

The kinetics of CD4+Foxp3+ T cell accumulation during a human cutaneous antigen-specific memory response in vivo

Abstract

Naturally occurring CD4+CD25hiFoxp3+ Tregs (nTregs) are highly proliferative in blood. However, the kinetics of their accumulation and proliferation during a localized antigen-specific T cell response is currently unknown. To explore this, we used a human experimental system whereby tuberculin purified protein derivative (PPD) was injected into the skin and the local T cell response analyzed over time. The numbers of both CD4+Foxp3– (memory) and CD4+Foxp3+ (putative nTreg) T cells increased in parallel, with the 2 populations proliferating at the same relative rate. In contrast to CD4+Foxp3– T cell populations, skin CD4+Foxp3+ T cells expressed typical Treg markers (i.e., they were CD25hi, CD127lo, CD27+, and CD39+) and did not synthesize IL-2 or IFN-γ after restimulation in vitro, indicating that they were not recently activated effector cells. To determine whether CD4+Foxp3+ T cells in skin could be induced from memory CD4+ T cells, we expanded skin-derived memory CD4+ T cells in vitro and anergized them. These cells expressed high levels of CD25 and Foxp3 and suppressed the proliferation of skin-derived responder T cells to PPD challenge. Our data therefore demonstrate that memory and CD4+ Treg populations are regulated in tandem during a secondary antigenic response. Furthermore, it is possible to isolate effector CD4+ T cell populations from inflamed tissues and manipulate them to generate Tregs with the potential to suppress inflammatory responses.

Authors

Milica Vukmanovic-Stejic, Elaine Agius, Nicola Booth, Padraic J. Dunne, Katie E. Lacy, John R. Reed, Toni O. Sobande, Steven Kissane, Mike Salmon, Malcolm H. Rustin, Arne N. Akbar

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Figure 4

Induction of Foxp3 expression in human CD4+CD25– T cells by stimulation.

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Induction of Foxp3 expression in human CD4+CD25– T cells by stimulation....
(A) CD4+CD25– (responder) T cells were isolated from PBMCs using MACS and labeled with CFSE. Cells were stimulated with magnetic beads coated with antibodies to CD3 and CD28 for 7 days. Samples were removed at regular intervals and stained for the expression of CD25, Foxp3, and Ki67. Numbers in dot plots indicate the percentage of cells in each quadrant. Dilution of the CFSE signal indicates proliferation of the cells. Percentages refer to cells that have not divided. Data shown are representative of 3 independent experiments. (B) Cells were removed from CD3/CD28-stimulated cultures on day 4 and restimulated with PMA and ionomycin before staining for Foxp3, IL-2, and IFN-γ. Histograms show cytokine production by Foxp3– and Foxp3+ cells and are representative of 3 independent experiments. Numbers indicate the percentage of cells expressing IL-2 or IFN-γ.