[Cell Cycle 8:16, 2578-2585; 15 August 2009]; ©2009 Landes Bioscience
Report
CD13+CD4+CD25hi regulatory T cells exhibit higher suppressive function and increase with tumor stage in non-small cell lung cancer patients Songwen Ju,1,† Hongxia Qiu,3,† Xiaoyue Zhou,1 Biqin Zhu,1 Xin Lv,1 Xinen Huang,4 Juan Li,5 Yu Zhang,1 Linxiang Liu,1 Yan Ge,2 Daniel E. Johnson,6 Songguang Ju2,* and Yongqian Shu1,* 1Cancer Center; and 3Department of Hematology; The First Affiliated Hospital of Nanjing Medical University; Nanjing, China; 2Department of Immunology; School of Medicine of Soochow University; Suzhou, China; 4Department of Internal Medicine; Tumor Hospital of Jiangsu Province; Nanjing, China; 5Department of Oncology; The Second Affiliated Hospital of Nanjing Medical University; Nanjing, China;6Division of Hematology/Oncology; Department of Medicine; University of Pittsburgh; Pittsburgh, PA USA †These
authors contributed equally to this work.
Abbreviations: Treg, regulatory T cell; TILs, tumor infiltrating lymphocytes; NSCLC, non-small cell lung cancer; PBMCs, peripheral blood mononuclear cells; mTGFβ1, membrane-bound transforming growth factor β1; CFSE, carboxylfluorescein succinimidyl ester; MFI, mean fluorescent intensity; UICC-TNM, union internationale centre le cancer-tumor node metastasis; APN, aminopeptidase N Key words: treg, non-small cell lung cancer, CD13, staging, aminopeptidase N
Regulatory T cells (Tregs) in peripheral blood and tumor infiltrating lymphocytes (TILs) play crucial roles in suppressing anti-tumor immune responses in cancer patients, and correlate with clinical outcomes. We identified an important subpopulation, CD13+CD4+CD25hiTreg cells, among CD4+CD25hiTreg cells in the peripheral blood of non-small cell lung cancer (NSCLC) patients. Peripheral blood mononuclear cells (PBMCs) were isolated from patients with NSCLC (n = 72) or from healthy donors (n = 30). Flow cytometric analyses were performed to study the expression of cell-surface or intracellular markers on the CD4+CD25hiTreg cells. The immune suppressive function of CD13+CD4+CD25hiTreg cells was evaluated by co-culturing with CD4+CD25-T cells that were activated by PHA. Our data showed that, compared with CD4+CD25Low/-T cells, CD13 expression was enriched on CD4+CD25hiTreg cells. The CD13+CD4+CD25hiTreg cells also expressed higher levels of Foxp3, CTLA-4, membrane-bound transforming growth factor β1 (mTGFβ1) and B7-H1, and are more suppressive to CD25 expression and proliferation of CD4+CD25-T cells. Additionally, we showed that the expression of Foxp3, CTLA-4, B7-H1, mTGFβ1 and the secretion of TGFβ1 and IL-10 on *Correspondence to: Songguang Ju; Immunology Department; School of Medicine; Soochow University; No. 199 Renai Road; Suzhou Industrial Park; Suzhou, China; Email:
[email protected]/ Yongqian Shu; Cancer Center; The First Affiliated Hospital of Nanjing Medical University; No. 300 Guangzhou Road; Nanjing, China; Email:
[email protected] Submitted: 04/30/09; Revised: 06/15/09; Accepted: 06/19/09 Previously published online as a Cell Cycle E-publication: http://www.landesbioscience.com/journals/cc/article/9302
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CD13+CD4+CD25hiTreg cells was significantly suppressed by anti-CD13 mAb (WM15), and the ability of these cells to suppress CD25 expression and proliferation of CD4+CD25-T cells was inhibited by WM15 as well. Interestingly, the percentage of CD13+CD4+CD25hiTreg cells among the CD4+CD25hiTreg population increased significantly and correlated with pathological stage in NSCLC: healthy donor (9.84% ± 2.23%)
