CCL17 and CCL22 chemokines within tumor microenvironment are related to accumulation of Foxp3+ regulatory T cells in gastric cancer

It has been reported that an increased population of regulatory T cells (Tregs) is one of the reasons for impaired anti-tumor immunity. Recently, Foxp3 has been reported as a reliable marker of Tregs. The authors investigated the frequency of Foxp3(+) Tregs within CD4(+) cells in TILs, regional lymph nodes and PBLs of gastric cancer patients (n = 45). Furthermore, to elucidate the mechanisms behind Treg accumulation within tumors, they evaluated the relationship between CCL17 or CCL22 expression and the frequency of Foxp3(+) Tregs in gastric cancer. CD4(+)CD25(+)Foxp3(+) Tregs as a percentage of CD4(+) cells were counted by flow cytometry and evaluated by immunohistochemistry. Moreover, an in vitro migration assay using Tregs derived from gastric cancers was performed in the presence of CCL17 or CCL22. As a result, the frequency of Foxp3(+) Tregs in TILs was significantly higher than that in normal gastric mucosa (12.4% +/- 7.5% vs. 4.1% +/- 5.3%, p < 0.01). Importantly, the increase in Tregs in TILs occurred to the same extent in early and advanced disease. Furthermore, the frequency of CCL17(+) or CCL22(+) cells among CD14(+) cells within tumors was significantly higher than that of normal gastric mucosa, and there was a significant correlation between the frequency of CCL17(+) or CCL22(+) cells and Foxp3(+) Tregs in TILs. In addition, the in vitro migration assay indicated that Tregs were significantly induced to migrate by CCL17 or CCL22. In conclusion, CCL17 and CCL22 within the tumor are related to the increased population of Foxp3(+) Tregs, with such an observation occurring in early gastric cancer.     

食管癌患者化疗前后ＣＤ４＋ＣＤ２５ｈｉｇｈ调节性Ｔ细胞及Ｆｏｘｐ３ｍＲＮＡ的表达及其临床意义

目的：探讨晚期食管癌患者化疗前后外周血中ＣＤ４＋ＣＤ２５ｈｉｇｈ调节性Ｔ细胞及Ｆｏｘｐ３ｍＲＮＡ的表达变化及其临床意义。方法：采用流式细胞术检测６８例晚期食管癌患者化疗前后外周血ＣＤ４＋ＣＤ２５ｈｉｇｈ调节性Ｔ细胞的水平,并与４０例健康成人进行比较。同时运用ＲＴ-ＰＣＲ方法检测其中４０例患者化疗前后外周血调节性Ｔ细胞Ｆｏｘｐ３基因ｍＲＮＡ的表达情况。结果：（１）食管癌患者外周血中ＣＤ４＋ＣＤ２５ｈｉｇｈ调节性Ｔ细胞占ＣＤ４＋Ｔ细胞总数的（１.８２±０.５４）％,显著高于健康对照组的（１.５２±０.７０）％（Ｐ＜０.０１）;（２）化疗前患者外周血中ＣＤ４＋ＣＤ２５ｈｉｇｈ调节性Ｔ细胞为（１.８２±０.５４）％,明显高于化疗后的（１.６６±０.５８）％（Ｐ＜０.０5）;（３）化疗前患者调节性Ｔ细胞Ｆｏｘｐ３基因ｍＲＮＡ的相对表达量为０.３１８±０.０２７,明显高于化疗后的０.２６６±０.０２８（Ｐ＜０.０５）。结论：食管癌患者接受化疗后外周血中Ｆｏｘｐ３ｍＲＮＡ水平与ＣＤ４＋ＣＤ２５ｈｉｇｈ调节性Ｔ细胞数量表达均显著下降,推测Ｆｏｘｐ３基因可能对ＣＤ４＋ＣＤ２５ｈｉｇｈＴ细胞有重要的调节功能,从而影响食管癌患者化疗后ＣＤ４＋ＣＤ２５ｈｉｇｈ调节性Ｔ细胞的水平。     

HDAC5 controls the functions of Foxp3+ T‐regulatory and CD8+ T cells

Histone/protein deacetylases (HDACs) are frequently upregulated in human malignancies and have therefore become therapeutic targets in cancer therapy. However, inhibiting certain HDAC isoforms can have protolerogenic effects on the immune system, which could make it easier for tumor cells to evade the host immune system. Therefore, a better understanding of how each HDAC isoform affects immune biology is needed to develop targeted cancer therapy. Here, we studied the immune phenotype of HDAC5^–/– mice on a C57BL/6 background. While HDAC5^–/– mice replicate at expected Mendelian ratios and do not develop overt autoimmune disease, their T‐regulatory (Treg) cells show reduced suppressive function in vitro and in vivo. Likewise, CD4⁺ T‐cells lacking HDAC5 convert poorly to Tregs under appropriately polarizing conditions. To test if this attenuated Treg formation and suppressive function translated into improved anticancer immunity, we inoculated HDAC5^–/– mice and littermate controls with a lung adenocarcinoma cell line. Cumulatively, lack of HDAC5 did not lead to better anticancer immunity. We found that CD8⁺ T cells missing HDAC5 had a reduced ability to produce the cytokine, IFN‐γ, in vitro and in vivo, which may offset the benefit of weakened Treg function and formation. Taken together, targeting HDAC5 weakens suppressive function and de‐novo induction of Tregs, but also reduces the ability of CD8⁺ T cells to produce IFN‐γ.     

CD83+ dendritic cells and Foxp3+ regulatory T cells in primary lesions and regional lymph nodes are inversely correlated with prognosis of gastric cancer

Background  

Dendritic cells (DCs) are potent antigen-presenting cells that are central to the regulation, maturation, and maintenance of the cellular immune response against cancer. In contrast, CD4⁺CD25⁺ regulatory T cells (Tregs) play a central role in self-tolerance and suppress antitumor immunity. In this study, we investigated the clinical significance of mature CD83⁺ DCs and Foxp3⁺ Tregs in the primary tumor and regional lymph nodes from the viewpoint of the two opposing players in the immune responses.     

Fatty acid-binding protein 5 limits the generation of Foxp3+ regulatory T cells through regulating plasmacytoid dendritic cell function in the tumor microenvironment

Plasmacytoid dendritic cells (pDCs) promote viral elimination by producing large amounts of Type I interferon. Recent studies have shown that pDCs regulate the pathogenesis of diverse inflammatory diseases, such as cancer. Fatty acid-binding protein 5 (FABP5) is a cellular chaperone of long-chain fatty acids that induce biological responses. Although the effects of FABP-mediated lipid metabolism are well studied in various immune cells, its role in pDCs remains unclear. This study, which compares wild-type and Fabp5^?/? mice, provides the first evidence that FABP5-mediated lipid metabolism regulates the commitment of pDCs to inflammatory vs tolerogenic gene expression patterns in the tumor microenvironment and in response to toll-like receptor stimulation. Additionally, we demonstrated that FABP5 deficiency in pDCs affects the surrounding cellular environment, and that FABP5 expression in pDCs supports the appropriate generation of regulatory T cells (Tregs). Collectively, our findings reveal that pDC FABP5 acts as an important regulator of tumor immunity by controlling lipid metabolism.     

CD8+ Foxp3+ regulatory T cells mediate immunosuppression in prostate cancer.

PURPOSE: Although elevated proportions of CD4(+)CD25(+) regulatory T (Treg) cells have been shown in several types of cancers, very little is known about the existence and function of CD8(+) Treg cells in prostate cancer. In this study, we investigated prostate tumor-derived CD8(+) Treg cells and their function. EXPERIMENTAL DESIGN: Tumor-infiltrating lymphocytes (TIL) from fresh tumor specimens of patients with prostate cancer were generated and subjected to phenotypic and suppressive function analyses. In particular, we investigated the role and function CD8(+) Treg cells in prostate cancer. RESULTS: We show that high percentages of CD4(+)CD25(+) T cells are probably present in the majority (70%) of prostate TILs. Remarkably, both CD4(+) and CD8(+) T-cell subpopulations possessed potent suppressive activity. T-cell cloning and fluorescence-activated cell sorting analyses showed the presence of CD8(+)CD25(+) Treg cell clones that expressed FoxP3 and suppressed na?ve T-cell proliferation, in addition to the previously known CD4(+)CD25(+) Treg cells. These CD8(+) Treg cells suppressed na?ve T-cell proliferation mainly through a cell contact-dependent mechanism. Importantly, the suppressive function of CD8(+) Treg cells could be reversed by human Toll-like receptor 8 (TLR8) signaling. CONCLUSION: Our study shows that like CD4(+)CD25(+) Treg cells, CD8(+) Foxp3(+) Treg cells present in prostate tumor-derived TILs suppress immune responses and that their suppressive function can be regulated by TLR8 ligands, raising the possibility that the manipulation of Treg cell function by TLR8 ligands could improve the efficacy of immunotherapy for prostate cancer patients.     

肿瘤患者外周血调节性T细胞的检测及临床意义

目的:研究肿瘤患者外周血调节性T细胞水平的变化并探讨其临床意义,旨在了解肿瘤患者的免疫功能改变,为肿瘤患者临床治疗及判断预后提供依据。方法:采用流式细胞仪对40例肿瘤患者及40例正常人外周血CD4+CD25+T细胞、CD4+CD25highT细胞及CD4+CD25+Foxp3+Treg细胞水平进行检测。结果:肿瘤患者外周血中CD4+CD25+T细胞和CD4+CD25highT细胞高于正常对照组(P<0.05);CD4+CD25+Foxp3+调节性T细胞亦明显高于对照组(P<0.01)。结论:肿瘤患者细胞免疫功能明显异常,CD4+CD25+Foxp3+调节性T细胞增多可能是肿瘤患者免疫功能受抑的重要原因之一;调节性T细胞水平检测在评价肿瘤患者疗效及判断预后方面具有一定的临床价值。     

Intratumoural FOXP3-positive regulatory T cells are associated with adverse prognosis in radically resected gastric cancer

We investigated the clinical significance of tumour-infiltrating FOXP3-positive regulatory T cells (Tregs) in radically resected (R0) gastric cancer. From a single-institution database, tumors of 110 patients who underwent R0 resection for stage II-III disease were studied for FOXP3-positive Tregs by immunohistochemistry. The observed median number of FOXP3-positive Tregs was used as the cut-point in analyses (<6 versus >or=6 count). Tregs were significantly higher in gastric carcinomas than in normal tissue (P = 0.0001). Tregs count >or=6 was significantly associated with vascular/lymphatic/perineural invasion (VELIPI) in the tumour (P = 0.03). Multivariate analysis showed association between adverse relapse-free survival and grading 3, stage III, VELIPI and Tregs count >or=6 (P = 0.02). Adverse overall survival was associated with grading 3, stage III, VELIPI and Tregs count >or=6 (P = 0.006). FOXP3-positive Tregs may be a novel marker for identifying high-risk gastric cancer patients. Present findings deserve additional investigation as Tregs may also represent an innovative therapeutic target.     

非小细胞肺癌患者外周血CD4＋CD25＋Foxp3＋调节性T细胞检测及临床意义

[目的]探讨非小细胞肺癌患者外周血CD4＋CD25＋Foxp3＋调节性T细胞（Treg）表达及临床意义。[方法]流式细胞术检测30例非小细胞肺癌组患者及12例健康志愿者外周血CD4＋CD25＋Foxp3＋Treg百分比。[结果]肺癌组外周血CD4＋CD25＋Foxp3＋细胞百分比（6.24%±2.01%）高于健康对照组（4.83%±0.85%）（P〈0.05）,外周血CD4＋CD25＋Foxp3＋细胞百分比在肺癌患者不同性别、年龄、病理、KPS及肿瘤标志物正常与否之间比较无差异（P〉0.05）,Ⅳ期患者表达高于Ⅰ～Ⅲ期患者（P〈0.05）,带瘤患者表达高于无瘤患者（P〈0.05）,有气虚证、血瘀证患者高于无气虚证、血瘀证患者（P〈0.05）。[结论]检测肺癌患者外周血CD4＋CD25＋Foxp3＋Treg表达在了解患者肿瘤进展情况和机体免疫功能状态方面有一定价值,其表达升高与患者出现气虚证和血瘀证有关。     

Foxp3在Treg细胞的表达与结直肠癌预后的研究进展

影响结直肠癌(Colorectal cancer,CRC)患者预后的因素除了一些临床病理学因素外,各类宿主免疫反应也成为影响预后的关键因素。因而,结直肠癌治疗方式除了传统的治疗模式之外,抗肿瘤免疫治疗逐渐成为当前治疗热点。调节性T细胞(Regulatory T,Treg)通过免疫调节机制参与免疫反应过程,并发挥重要的作用。叉头转录因子3(Forkhead box P3,Foxp3)作为调节性T细胞表达的调控因子之一,对Treg细胞通过抑制多种免疫细胞的增殖分化来发挥重要的作用。Foxp3⁺Treg细胞在结直肠癌患者中的表达与预后关系密切。该综述旨在阐明Foxp3与Treg细胞的基本特征、Foxp3⁺Treg细胞发挥肿瘤抑制性作用的机制、Foxp3在Treg细胞中表达对结直肠癌患者预后的重要意义进行阐述。     

CD45RA-Foxp3high but not CD45RA+Foxp3low suppressive T regulatory cells increased in the peripheral circulation of patients with head and neck squamous cell carcinoma and correlated with tumor progression

Background

T regulatory cells (Tregs) contribute to the progression of head and neck squamous cell carcinoma (HNSCC) by suppressing antitumor immunity. However, little is known regarding the functional heterogeneity of Tregs in HNSCC patients.

Methods

Using multicolor flow cytometry, the frequency of three Treg subsets, separated on the basis of CD45RA and Foxp3, from the peripheral circulation of newly-presenting HNSCC patients (19 oral cavity squamous cell carcinoma, 20 hypopharyngeal squamous cell carcinoma, 18 nasopharyngeal squamous cell carcinoma, 19 oropharyngeal squamous cell carcinoma, and 36 laryngeal squamous cell carcinoma) were assessed with regard to 31 healthy donors and clinicopathological features. Moreover, the functional capacity of each Treg subsets was evaluated based on CD45RA and CD25 expression.

Results

The frequency of Tregs in the peripheral circulation of HNSCC patients as a whole cohort was higher than in healthy donors (P < 0.0001). However, the frequency of Tregs was similar between patients with oral cavity squamous cell carcinoma and healthy donors (P = 0.269). Further dividing Tregs into three subsets based on Foxp3 and CD45RA expression revealed that the frequency of CD45RA^-Foxp3^high Tregs and CD45RA^-Foxp3^lowCD4⁺ T cells in patients with HNSCC developing from different subsites was higher than in healthy donors (P < 0.0001, P < 0.0001), whereas the frequency of CD45RA⁺Foxp3^low Tregs was lower than in healthy donors (P < 0.0001). Functionally study revealed that CD45RA^-CD25⁺⁺⁺ Tregs significantly inhibit the proliferation of CD4⁺CD25^- T cells (P < 0.001) and secrete lower levels of cytokines (P < 0.01) compared with CD45RA^-CD25⁺⁺CD4⁺ T cells. Importantly, the frequency of CD45RA^-Foxp3^high Tregs positively correlate with tumor stage (P < 0.0001) and nodal status (P < 0.0001).

Conclusions

CD45RA^-Foxp3^high Tregs increase in the peripheral circulation of HNSCC patients, and correlate with tumor stage and nodal status; suggesting a role in tumor progression which may be manipulated by future immunotherapy.     

Foxp3在卵巢癌组织中的表达及临床意义

目的 探讨叉状头-翅膀状螺旋转录因子p3（Forkhead box protein 3,Foxp3）在卵巢癌组织中的表达及其与预后的关系。方法 采用免疫组化染色法检测卵巢癌组织芯片中45例癌组织及7例正常卵巢组织的Foxp3表达。应用秩和检验比较不同病理类型卵巢癌Foxp3表达的差异;卡方检验分析Foxp3表达与临床病理参数的关系;Kaplan-Meier法分析Foxp3表达与卵巢癌预后关系;Cox比例风险回归模型评价影响患者预后的因素。结果 Foxp3阳性着色主要定位于细胞核。Foxp3表达与FIGO分期有关（P＜0.05）,与年龄、病理类型、分化程度、肿瘤长径、淋巴结转移等均无关（P＞0.05）。Kaplan-Meier法生存分析显示Foxp3低表达者（H-score≤90）中位生存时间（OS）为6183个月,显著高于高表达者（H-score＞90）的25.03个月（P＜0.05）。Cox比例风险模型显示,术后未化疗（HR=6.603,95％CI： 1.147～32.057）、FIGO分期较高（HR=3.861,95％CI： 1.427～10.417） 为预测卵巢癌不良预后的独立因素。结论 卵巢癌组织中Foxp3表达与FIGO分期相关,并可以作为卵巢癌预后判断的参考指标。     

Proliferation of CD4+CD25high+Foxp3+ regulatory T lymphocytes in ex vivo expanded ascitic fluid from primary and recurrent ovarian carcinoma

Objective

Regulatory T lymphocytes evoke the immune tolerance by suppressing and inactivating cytotoxic T lymphocytes. The objective of this study was to compare the proportion of regulatory T lymphocytes, precisely defined as CD4⁺CD25^high+Foxp3⁺ T lymphocytes, in primary and recurrent ovarian carcinoma before and after ex vivo expansion of ascites with interleukin-2 (IL-2).

Methods

Ascitic fluid samples were obtained from 26 patients with ovarian carcinoma. Lymphocytes were isolated from ascites and cell markers were analyzed by flow cytometry using anti-CD3/CD4/CD8/CD16/CD56/CD25 and anti-Foxp3 antibodies. Lymphocytes were incubated for 2 to 3 weeks and expanded ex vivo by IL-2 stimulation and their phenotypes were analyzed by flow cytometry.

Results

Following ex vivo expansion, ascitic fluid lymphocytes increased by a greater extent in the recurrent group than in the primary group. The proportion of ex vivo-expanded lymphocytes changed as follows; CD4⁺ T lymphocytes increased, CD8⁺ T lymphocytes decreased, and the proportion of CD3^-CD16⁺56⁺ NK cells was unchanged. The proportion of CD4⁺CD25^high+Foxp3⁺ regulatory T lymphocytes in CD4⁺ T lymphocytes increased after ex vivo expansion in both groups, but to a greater degree in the recurrent group.

Conclusion

This study showed that regulatory T lymphocytes, neither cytotoxic T lymphocytes nor NK cells, were extensively increased after ex vivo expansion, especially in recurrent ovarian carcinoma. These results may provide information that helps to guide the future development of adoptive immunotherapy against ovarian carcinoma.     

Expression of Fas/FasL in CD8+ T and CD3+ Foxp3+ Treg Cells - Relationship with Apoptosis of Circulating CD8+ T Cells in Hepatocellular Carcinoma Patients

Aims: Dysfunction of the host immune system in cancer patients can be due to a number of factors, includinglymphocyte apoptosis. Several studies showed that Foxp3+T cells take part in inducing this process by expressingFasL in tumor patients. However, the relationship between apoptosis, CD8+T cells and Foxp3+T cells in HCCpatients is still unclear. The present study was designed to investigate the correlation between apoptosis levelsand Fas/FasL expression in CD8+T lymphocytes and Foxp3+T cells in patients with HCC. Methods: CD8+T cellsand CD3+Foxp3+T cells were tested from peripheral blood of HCC patients and normal controls and subjectedto multicolor flow cytometry. The expression of an apoptosis marker (annexin V) and the death receptor Fas inCD8+T cells and FasL in CD3+Foxp3+T cells were evaluated. Serum TGF-β1 levels in patients with HCC weremeasured by enzyme-linked immunosorbent assay. The relationship between apoptosis and Fas expression, aswell as FasL expression in CD3+Foxp3+T cells was then evaluated. Results: The frequency of CD8+T cells bindingannexin V and Fas expression in CD8+T cells, were all higher in HCC patients than normal controls and theproportion of apoptotic CD8+T cells correlated with their Fas expression. Serum TGF-β1 levels correlated inverselywith CD3+Foxp3+T cells. Conclusions: Fas/FasL interactions might lead to excessive turnover of CD8+T cellsand reduce anti-tumor immune responses in patients with HCC. Further investigations of apoptosis inductionin Fas+CD8+T cells in vitro are required.     

Foxp3+Treg和IDO+APC细胞在不同病变宫颈组织中的表达

目的:检测免疫抑制细胞Foxp3+Treg和免疫蛋白IDO在不同病变宫颈组织中的表达情况.方法:采用免疫细胞化学及蛋白印记(Western blot)检测Foxp3蛋白及IDO蛋白在宫颈癌细胞系HeLa、SiHa及C33A中的表达与分布;采用免疫组织化学法检测Foxp3和IDO在20例正常宫颈组织及45例不同程度宫颈病...     

ICOSL expressed in triple-negative breast cancer can induce Foxp3+ Treg cell differentiation and reverse p38 pathway activation

Inducible costimulator ligand (ICOSL) expressed on cancer cells has immunoregulatory functions in various malignancies. However, the role of ICOSL in triple-negative breast cancer (TNBC) remains unclear. In this study, the role and expression of ICOSL in TNBC were analyzed using the cBioPortal and GEPIA databases. Then the role of ICOSL in Foxp3+ Treg cell differentiation, reversal of p38 pathway activation and cell proliferation, migration and apoptosis was determined in vitro. Finally, the effect of ICOSL expression on TNBC progression was verified in a nude mouse model of TNBC. We here observed that ICOSL expression in TNBC was found to be related to relapse-free survival, and Treg abundance was positively correlated with ICOSL expression, as demonstrated by database analyses. In vitro experiments showed that ICOSL overexpression (OE) in MDA-MB-231 cells induced cocultured T cells to differentiate into Foxp3+ Treg cells and promoted secretion of the tumor-promoting factors IL-10 and IL-4. Furthermore, in vitro experiments showed that ICOSL reversed p38 phosphorylation and promoted the proliferation, invasion, and metastasis of MDA-MB-231 ICOSL-OE cells. Finally, tumor progression was found to be promoted by ICOSL expression in a TNBC nude mouse model. Together, ICOSL expression can enhance tumor cell growth by inducing Foxp3+ Treg cell differentiation and reversing p38 pathway activation in TNBC.     

核叉头转录蛋白3(Foxp3)在胃间质瘤组织中的表达及其临床病理意义

目的:利用免疫组化技术评估Foxp3在胃间质瘤组织中的表达,并进一步分析Foxp3的表达是否与胃间质瘤患者的临床病理因素和预后相关。方法:收集96例胃间质瘤患者肿瘤及瘤旁组织标本,采用免疫组化方法检测肿瘤及瘤旁组织中Foxp3的表达。进一步分析Foxp3表达水平与患者临床病理因素及预后的关系。结果:与瘤旁正常组织相比,Foxp3在胃间质瘤组织中呈高表达。Foxp3表达与核分裂象、肿瘤大小、肿瘤坏死以及肿瘤危险度分级呈正相关。单因素分析显示Foxp3高表达与总生存率呈负相关,多因素分析显示Foxp3是影响胃间质瘤患者预后的独立危险因素。结论:Foxp3高表达与胃间质瘤患者预后不良有关,Foxp3可能是胃间质瘤潜在的诊断标志物和治疗靶点。