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 Blood, 15 December 2008, Vol. 112, No. 13, pp. 4953-4960.
Prepublished online as a Blood First Edition Paper on September 26, 2008; DOI 10.1182/blood-2008-06-163048.

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IMMUNOBIOLOGY

FOXP3 expression accurately defines the population of intratumoral regulatory T cells that selectively accumulate in metastatic melanoma lesions

Mojgan Ahmadzadeh1, Aloisio Felipe-Silva2, Bianca Heemskerk1, Daniel J. Powell, Jr1, John R. Wunderlich1, Maria J. Merino2, and Steven A. Rosenberg1

1 Surgery Branch and 2 Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, MD

Regulatory T (Treg) cells are often found in human tumors; however, their functional characteristics have been difficult to evaluate due to low cell numbers and the inability to adequately distinguish between activated and Treg cell populations. Using a novel approach, we examined the intracellular cytokine production capacity of tumor-infiltrating T cells in the single-cell suspensions of enzymatically digested tumors to differentiate Treg cells from effector T cells. Similar to Treg cells in the peripheral blood of healthy individuals, tumor-infiltrating FOXP3+CD4 T cells, unlike FOXP3– T cells, were unable to produce IL-2 and IFN-{gamma} upon ex vivo stimulation, indicating that FOXP3 expression is a valid biological marker for human Treg cells even in the tumor microenvironment. Accordingly, we enumerated FOXP3+CD4 Treg cells in intratumoral and peritumoral sections of metastatic melanoma tumors and found a significant increase in proportion of FOXP3+CD4 Treg cells in the intratumoral compared with peritumoral areas. Moreover, their frequencies were 3- to 5-fold higher in tumors than in peripheral blood from the same patients or healthy donors, respectively. These findings demonstrate that the tumor-infiltrating CD4 Treg cell population is accurately depicted by FOXP3 expression, they selectively accumulate in tumors, and their frequency in peripheral blood does not properly reflect tumor microenvironment.


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