Baumgartner Joel, Wilson Cara, Palmer Brent, Richter Don, Banerjee Anirban, McCarter Martin
Department of Surgery, University of Colorado at Denver and Health Sciences Center, Denver, Colorado 80262, USA.
J Surg Res. 2007 Jul;141(1):72-7. doi: 10.1016/j.jss.2007.03.053.
The immune response to melanoma is rarely curative, suggesting the emergence of immunosuppression. FOXP3-expressing regulatory T cells (T(reg) cells) function to suppress immune responses. The objective of this study was to determine if melanoma evades immune surveillance, in part, by inducing T(reg) cells.
Peripheral blood mononuclear cells (PBMCs) were isolated and exposed to melanoma-conditioned media (MCM) or control media for 1 week. The induction of T(reg) cells in these PBMCs was determined by measuring the proportion of CD25(+)FOXP3(+) T cells in all CD4(+) T cells by flow cytometry. FOXP3 expression was determined by mean fluorescence intensity (MFI) and Western blot. Supernatant cytokines were determined by ELISA.
Normal PBMCs exposed to MCM revealed higher proportions of T(reg) cells than those exposed to control media after 6 days (3.4% versus 1.3%, respectively, P < 0.02). The expression of FOXP3 in T(reg) cells from PBMCs exposed to MCM increased over time by MFI and Western blot but was not significantly different than those exposed to control media. The level of IL-10 and TGF-beta in supernatants after 6 days growth was higher in MCM than control media, but this did not reach statistical significance.
Exposure of PBMCs to melanoma results in induction of FOXP3(+) T(reg) cells.
对黑色素瘤的免疫反应很少能治愈，这表明免疫抑制的出现。表达叉头框蛋白3（FOXP3）的调节性T细胞（Treg细胞）具有抑制免疫反应的功能。本研究的目的是确定黑色素瘤是否部分通过诱导Treg细胞来逃避免疫监视。
分离外周血单个核细胞（PBMC），并将其暴露于黑色素瘤条件培养基（MCM）或对照培养基中1周。通过流式细胞术测量所有CD4+T细胞中CD25+FOXP3+T细胞的比例，以确定这些PBMC中Treg细胞的诱导情况。通过平均荧光强度（MFI）和蛋白质免疫印迹法测定FOXP3的表达。通过酶联免疫吸附测定法测定上清液中的细胞因子。
暴露于MCM的正常PBMC在6天后显示出比暴露于对照培养基的PBMC更高比例的Treg细胞（分别为3.4%和1.3%，P<0.02）。通过MFI和蛋白质免疫印迹法，暴露于MCM的PBMC中Treg细胞的FOXP3表达随时间增加，但与暴露于对照培养基的PBMC相比无显著差异。生长6天后，MCM中上清液的白细胞介素-10（IL-10）和转化生长因子-β（TGF-β）水平高于对照培养基，但未达到统计学显著性。
PBMC暴露于黑色素瘤会导致FOXP3+Treg细胞的诱导。
