FOXP3 Defines Regulatory T Cells in Human Tumor and Autoimmune Disease
ABSTRACT Activated T cells may express FOXP3. It is thought that FOXP3 is not a specific marker to determine regulatory T cells (Treg) in humans. Here, we examined the functional phenotype and cytokine profile of the in vitro induced FOXP3(+) T cells, primary FOXP3(+) and FOXP3(-) T cells in patients with ulcerative colitis and tumors including colon carcinoma, melanoma, hepatic carcinoma, ovarian carcinoma, pancreatic cancer, and renal cell carcinoma. We observed similar levels of suppressive capacity of primary FOXP3(+) T cells in blood, tumors, and colitic tissues. Compared with primary FOXP3(-) T cells in the same microenvironment, these primary FOXP3(+) T cells expressed minimal levels of effector cytokines, negligible amount of cytotoxic molecule granzyme B, and levels of suppressive molecules interleukin-10 and PD-1. Although the in vitro activated T cells expressed FOXP3, these induced FOXP3(+) T cells expressed high levels of multiple effector cytokines and were not functionally suppressive. The data reinforce the fact that FOXP3 remains an accurate marker to define primary Tregs in patients with cancer and autoimmune disease. We suggest that the combination of FOXP3 and cytokine profile is useful for further functionally distinguishing primary Tregs from activated conventional T cells.
- SourceAvailable from: Shalaka Hampras
[Show abstract] [Hide abstract]
- "For this study, CD3 + CD4 + CD25 + FOXP3 + was used as the principle definition of Treg cells, and results are expressed as percentage of total CD3 + cells. Foxp3 is considered to be highly specific marker for Treg cells  . At the end of one year of follow-up time (April 2010), 82 (76%) of sample donors had returned their epidemiological questionnaires to the DBBR for data processing. "
ABSTRACT: Immunosuppressive regulatory T (Treg) cells play an important role in antitumor immunity, self-tolerance, transplantation tolerance, and attenuation of allergic response. Higher proportion of Treg cells has been observed in peripheral blood of cancer cases compared to controls. Little is known about potential epidemiological predictors of Treg cell levels in healthy individuals. We conducted a cross-sectional study including 75 healthy women, between 20 and 80 years of age, who participated in the Data Bank and BioRepository (DBBR) program at Roswell Park Cancer Institute (RPCI), Buffalo, NY, USA. Peripheral blood levels of CD4(+)CD25(+)FOXP3(+) Treg cells were measured using flow cytometric analysis. A range of risk factors was evaluated using Wilcoxon Rank-Sum test, Kruskal-Wallis test, and linear regression. Age, smoking, medications for treatment of osteoporosis, postmenopausal status, body mass index (BMI), and hormone replacement therapy (HRT) were found to be significant positive predictors of Treg cell levels in peripheral blood (P ≤ 0.05). Higher education, exercise, age at first birth, oral contraceptives, and use of Ibuprofen were found be significant (P < 0.05) negative predictors of Treg levels. Thus, various epidemiological risk factors might explain interindividual variation in immune response to pathological conditions, including cancer.Journal of Cancer Epidemiology 08/2012; 2012:191090. DOI:10.1155/2012/191090
[Show abstract] [Hide abstract]
- "0.7 [0.5–3.1] CDAI 234 [194–256] 327 [312–374] HBI 9     12            "
ABSTRACT: Anti-TNF-α antibodies has been suggested to modulate regulatory T cell (Treg) percentages in rheumatoid arthritis, but results from studies of Crohn's disease (CD) are conflicting. We investigated dynamic changes of circulating Tregs in CD during treatment with the anti-TNF-α-antibody adalimumab (Humira®, Abbott Laboratories A/S, Emdrupvej 28C, DK-2100 Copenhagen). Blood samples from 26 CD patients were analysed using flow cytometry before and 1 and 26 weeks after initiation of adalimumab treatment to determine the percentage of Tregs among CD4+ T cells. In spite of a significant decline in disease activity scores and biochemical markers of inflammation, during the first week of treatment, we did not observe early modulating effects of adalimumab on Treg percentages. However, we found a long-term increase in Treg percentages in responders who had low Treg percentages (<5%) at baseline (p = 0.04). Treg percentage was inversely associated with disease activity (CD activity index or CDAI) (Spearman's rank correlation, ρ = -0.47, p = 0.02). High Treg percentages among CD4+ T cells at baseline predicted clinical response to adalimumab. Adalimumab treatment did not induce early modulatory effects on Treg percentage, even in responders. This finding suggests that adalimumab does not have a direct or selective effect on Tregs. However, Treg percentage was associated with disease activity and high Treg percentage predicted response to adalimumab.Scandinavian Journal of Gastroenterology 07/2011; 46(10):1206-14. DOI:10.3109/00365521.2011.603157 · 2.33 Impact Factor
[Show abstract] [Hide abstract]
- "FOXP3 may be transiently expressed in activated T cells, and its presence has also been reported in tumor cells (Ebert et al., 2008). Furthermore, FOXP3 is a nuclear protein, and its intracellular location limits its usefulness in Treg isolation, although it has often been applied to confirm the identity of Treg (Kryczek et al., 2009). "
ABSTRACT: The ectonucleotidase CD39 is an enzyme involved in adenosine production. Its surface expression on human regulatory T cells (Treg) allows for their flow-cytometry-based isolation from peripheral blood. To further develop and improve this method on a scale supporting translational studies, we introduced capture of CD39(+) Treg on magnetic immunobeads. Peripheral blood mononuclear cells (PBMC) obtained from healthy donors were used for negative selection of CD4(+) T cells on AutoMACS using antibodies (Abs) specific for all lineage(+) cells. CD4(+)CD39(+) Treg were captured by biotin-conjugated anti-CD39 Abs and anti-biotin Ab-coated magnetic beads. Isolated CD4(+)CD39(+) T cells were phenotyped by flow cytometry for Treg-associated markers: CD39, CD73, FOXP3, CD25, CTLA-4, CCR4, CD45RO and CD121a or for the absence of CD127 and CD49d. CFSE-based proliferation assays and ATP hydrolysis were used to measure Treg functions. The purity, recovery and viability of the separated CD4(+)CD39(+) T cells were satisfactory. The isolated CD4(+)CD39(+) T cell population consisted of FOXP3(+)CD25(+) T cells which hydrolyzed exogenous ATP and suppressed autologous CD4(+) T cell proliferation and of FOXP3(neg)CD25(neg) T cells without suppressor function. The same two subsets were detectable by flow cytometry in normal PBMC, gating on CD4(+)CD39(+), CD4(+)CD127(neg), CD4(+)CD49d(neg) or CD4(+)CD25(high) Treg. CD4(+)CD39(+) Treg capture on immunobeads led to a discovery of two CD39(+) subsets. Similar to CD39(+) Treg in the peripheral blood, half of these cells are CD25(+)FOXP3(+) active suppressor cells, while the other half are CD25(neg)FOXP3(neg) and do not mediate suppression.Journal of immunological methods 06/2011; 369(1-2):59-68. DOI:10.1016/j.jim.2011.04.004 · 2.01 Impact Factor