Figure 2 - uploaded by Feng Yang
Content may be subject to copyright.
FOXP3 + lymphocytes in the draining lymph nodes of pancreatic cancer patients. Immunohistochemical staining of FOXP3 + lymphocytes in a positive lymph node (A: low magnification, 640; B, high magnification, 6400) and a negative lymph node(C: low magnification, 640; D: high magnification, 6400), and a comparison of the FOXP3 + lymphocyte densities in positive and negative lymph nodes (E) (p = 0.03), among different groups in positive lymph nodes (F) (p = 0.75), and among different groups in negative lymph nodes (G) (p = 0.52). doi:10.1371/journal.pone.0106741.g002
Source publication
Objective
To determine if the density of FOXP3+ lymphocytes in primary tumors and lymph nodes in pancreatic cancer correlates with the presence of lymph node metastases.
Methods
FOXP3+ lymphocyte density in primary pancreatic cancer tissue and draining lymph nodes was measured using immunohistochemistry. We analyzed the clinical and pathological a...
Similar publications
Regulatory T cells (Tregs) restrict overexuberant lymphocyte activation. While close proximity between Tregs and their suppression targets is important for optimal inhibition, and literature indicates that draining lymph nodes (LNs) may serve as a prime location for the suppression, signaling details orchestrating this event are not fully character...
Regulatory T cells (Tregs) restrict overexuberant lymphocyte activation. While close proximity between Tregs and their suppression targets is important for optimal inhibition, and literature indicates that draining lymph nodes (LNs) may serve as a prime location for the suppression, signaling details orchestrating this event are not fully character...
Citations
... In the case of PC, the density of Tregs was significantly higher in tumor tissues than that noted in the paratumoral pancreatic tissues. The density of Tregs was significantly correlated with the histological grade and lymph node metastasis (38). In a consecutive series of 92 patients with PC resection, Liu et al (39) demonstrated that patients with higher levels of intratumoral Tregs exhibited shorter DFS times than those with lower levels of Tregs (11.2 vs. 22.2 months; P<0.001). ...
Tumor-infiltrating lymphocytes (TILs) are important components of the tumor microenvironment (TME). However, the distribution characteristics of TILs and their significance in pancreatic cancer (PC) remain largely unexplored. The levels of TILs, including the total number of T cells, cluster of differentiation (CD)4+ T cells, CD8+ cytotoxic T lymphocytes (CTLs), regulatory T-cells (Tregs), programmed cell death protein 1+ T cells and programmed cell death ligand 1 (PD-L1)+ T cells, in the TME of patients with PC were detected using multiple fluorescence immunohistochemistry. The associations between the number of TILs and the clinicopathological characteristics were investigated using χ2 tests. In addition, Kaplan-Meier survival and Cox regression analyses were used to assess the prognostic value of these TIL types. Compared with paracancerous tissues, in PC tissues, the proportions of total T cells, CD4+ T cells and CD8+ CTLs were markedly decreased, while those of Tregs and PD-L1+ T cells were significantly increased. The levels of CD4+ T cell and CD8+ CTL infiltrates were inversely associated with tumor differentiation. Higher infiltrates of Tregs and PD-L1+ T cells were closely associated with advanced N and TNM stages. It is important to note that the infiltrates of total T cells, CD4+ T cells, Tregs and PD-L1+ T cells in the TME were independent risk factors for the prognosis of PC. PC was characterized by an immunosuppressive TME with a decrease in the number of CD4+ T cells and CD8+ CTLs, and an increase in the number of Tregs and PD-L1+ T cells. Overall, the number of total T cells, CD4+ T cells, Tregs and PD-L1+ T cells in the TME was a potential predictive marker for the prognosis of PC.
... In the present study, we found significantly different Treg cell counts between the TC, TP, and NAT. Interestingly, in alignment with our findings, the number of Treg cells was found to be significantly higher within the tumour sites of BCC [3,23], breast cancers [24], pancreatic cancers [25], and colorectal cancers [26] compared to their matched NATs, regardless of their association with the status of lymph node or distant metastasis. The tumour phenotypes, chemokine expression, oxidative status, metabolism, and pH in TME were suggested to influence the infiltration of Treg cells to tumour sites [25]. ...
... Interestingly, in alignment with our findings, the number of Treg cells was found to be significantly higher within the tumour sites of BCC [3,23], breast cancers [24], pancreatic cancers [25], and colorectal cancers [26] compared to their matched NATs, regardless of their association with the status of lymph node or distant metastasis. The tumour phenotypes, chemokine expression, oxidative status, metabolism, and pH in TME were suggested to influence the infiltration of Treg cells to tumour sites [25]. ...
Basal cell carcinoma (BCC) is the most common skin malignancy worldwide. Current evidence suggests tumour-infiltrating lymphocytes (TILs) may influence the clinical outcomes of patients with BCC. The present study aimed to profile the infiltrative characteristics of stromal TILs and regulatory T cells (Treg cells) in the tumour centre (TC), tumour periphery (TP), and normal adjacent tissue (NAT) of BCC. A total of 111 samples from 43 cutaneous BCC cases were examined for TIL (CD3+) and Treg cell (FOXP3+/CD3+) expression using immunohistochemical techniques. The correlations of Treg cells with TILs, invasion depth, and tumour morphological risk were analysed. We identified a high mean proportion of Treg cells within the tumour (TC = 46.9%, TP = 56.1%, NAT = 51.8%) despite a relatively low median of TILs (TC = 12.7%, TP = 10.3%, NAT = 3.6%), supporting the classification of BCC as a cold tumour. A significant positive correlation was observed between the proportion of Treg cells and sTILs (ρ = 0.325, p < 0.001), suggesting a predominant role of TILs in the infiltration of Treg cells. An inverse correlation discovered between Treg cells and tumour invasion depth (r = −0.36, p = 0.017) might indicate Treg cells’ anti-tumour capacity in BCC.
... CD3 + CD8 − (CD4 + ) T cells, including Tregs, were the most abundant T cell population within this PDAC cohort. Prior research has indicated that PDAC's distinctive immunosuppressive microenvironment is largely driven by Tregs [22,[48][49][50]. However, their functional role in the immune contexture of cancer has been increasingly described as ambiguous, also in PDAC [51,52]. ...
Pancreatic ductal adenocarcinoma (PDAC) is considered to be a poorly immunogenic cancer type that combines a low mutation burden with a strong immunosuppressive tumor microenvironment. Regulatory T cells (Tregs) are major drivers of immune suppression but their prognostic role, particularly in gastrointestinal malignancies, remains controversial. Lymphocytic infiltration in 122 PDAC samples was assessed by multispectral immunofluorescence with anti-Keratin, -CD3, -CD8, -FOXP3 and -CD163 antibodies. Differential infiltration by Tregs was analyzed in the context of transcriptomic profiles that were available for 65 tumors. High infiltration of CD3+CD8− (mainly CD4+) T cells and, especially, of the subset expressing FOXP3 (Tregs) was associated with improved patient survival, whilst cytotoxic CD3+CD8+ T cell infiltration did not have an impact on overall survival. Transcriptomic analysis revealed three signatures in PDAC tumors comprising of epithelial-mesenchymal transition (EMT)/stromal, metabolic, and secretory/pancreatic signature. However, none of these signatures explained differences in Treg infiltration. We show that Tregs associate with improved overall survival in PDAC patients. This effect was independent of cytotoxic T cell infiltration and the transcriptomic profiles of their respective tumors. These findings provide a new layer of complexity in the study of PDAC tumor microenvironment that must be considered when developing immunotherapeutic interventions for this disease.
... Tregs dampen anti-tumor immune responses as well as lead to the formation of an immunosuppressive TME, facilitating immune evasion and cancer progression [232,233]. In many cancers, including glioblastoma [234], non-lymphoma Hodgkin's [235], pancreatic ductal adenocarcinoma [236], ovarian cancer [237], lung cancer [238], and melanoma [239], an increase of FoxP3 + Tregs and a higher Treg: T effector cell (Teff) ratio in tumor is correlated with poor prognoses. Transient Treg depletion resulted in the regression of metastatic lesions in higher-stage melanoma patients. ...
A subpopulation of cells in many cancers has stem cell traits, mediates metastasis, and contributes to treatment resistance. These cells are considered as cancer stem cells (CSCs). CSC properties of tumor cells are immensely regulated by close interactions with tumor microenvironment components such as mesenchymal stem cells, tumor related fibroblasts, adipocytes, endothelial cells, and immune cells via the intricate network of cytokines, chemokines, and growth factors. Inflammatory cytokines including interleukin (IL)-1, IL-6, and IL-8 play a major role in these interactions via the activation of signal transduction pathways like Stat3/NF-κB etc. in stromal and tumor cells. The activation of these pathways increases the release of more cytokines, resulting in positive feedback loops which help in CSC self-renewal. The pathways controlled by these cytokine loops are similar to those that are active during chronic inflammation and wound healing, suggesting that they might have critical role in establishing relationship between inflammation and cancer. Anti-inflammatory drugs have been identified to inhibit these cytokines and their receptor mediated pathways. These agents have the potential to target CSCs by inhibiting signals from the tumor microenvironment and considered to be a potential candidate for future therapeutics. The significance of cytokines released from the tumor microenvironment in different phases of cancer, as well as their potential application in cancer therapeutics is discussed in this article.
... 25 This is consistent with other reports showing that T reg infiltration in the TIME of PDAC patients is associated with poor prognosis. [26][27][28][29] Understanding the effect of immunosuppression in PDAC and how it is altered by current therapies is critical design efficient combination therapeutic approaches to improve survival. 30 While studies in mouse models have demonstrated a favorable impact of radiation on the TIME in PDAC, the impact of neoadjuvant CRT on human PDAC and on the associated TIME is not well established. ...
... T regs are known to infiltrate pancreatic malignancies very early in oncogenesis and play a key role in establishing an immunosuppressed TIME. 57,58 T reg accumulation also correlates with shortened survival in several PDAC studies, both in tissue [26][27][28][29] and in blood. [59][60][61] Thus, the accumulation of T regs during CRT may be indicative of a more aggressive cancer. ...
Patients with pancreatic ductal adenocarcinoma (PDAC) have a grim prognosis despite complete surgical resection and intense systemic therapies. While immunotherapies have been beneficial with many different types of solid tumors, they have almost uniformly failed in the treatment of PDAC. Understanding how therapies affect the tumor immune microenvironment (TIME) can provide insights for the development of strategies to treat PDAC. We used quantitative multiplexed immunofluorescence (qmIF) quantitative spatial analysis (qSA), and immunogenomic (IG) analysis to analyze formalin-fixed paraffin embedded (FFPE) primary tumor specimens from 44 patients with PDAC including 18 treated with neoadjuvant chemoradiation (CRT) and 26 patients receiving no treatment (NT) and compared them with tissues from 40 treatment-naïve melanoma patients. We find that relative to NT tumors, CD3⁺ T cell infiltration was increased in CRT treated tumors (p = .0006), including increases in CD3⁺CD8⁺ cytotoxic T cells (CTLs, p = .0079), CD3⁺CD4⁺FOXP3⁻ T helper cells (Th, p = .0010), and CD3⁺CD4⁺FOXP3⁺ regulatory T cells (Tregs, p = .0089) with no difference in CD68⁺ macrophages. IG analysis from micro-dissected tissues indicated overexpression of genes involved in antigen presentation, T cell activation, and inflammation in CRT treated tumors. Among treated patients, a higher ratio of Tregs to total T cells was associated with shorter survival time (p = .0121). Despite comparable levels of infiltrating T cells in CRT PDACs to melanoma, PDACs displayed distinct spatial profiles with less T cell clustering as defined by nearest neighbor analysis (p < .001). These findings demonstrate that, while CRT can achieve high T cell densities in PDAC compared to melanoma, phenotype and spatial organization of T cells may limit benefit of T cell infiltration in this immunotherapy-resistant tumor.
... 35 Also, previous work suggests the role of FOXP3 in tumour immunity in cancer patients. 36 The lymphocyte density of FOXP3 + was observed to be related to LNM and further implicated with the development of PC. 37 Meanwhile, it has been reported that FOXP3 was regulated by lncRNA EGFR-AS1 and thus is involved in the regulation of NSCLC progression. 17 Moreover, LINC00261 overexpression decreased the expression of FOXO3 to inhibit the oncogenic phenotype of PC cells in regulation of miR-552-5p. ...
Long non‐coding RNAs (lncRNAs) biological functions and molecular mechanisms associated with pancreatic cancer (PC) remain to be poorly elucidated. We aimed to clarify the role of lncRNA LINC00261 (LINC00261) in PC and confirm its regulatory mechanisms. Bioinformatics analysis, RNA pull‐down and RIP assays were performed to investigate relationship between LINC00261 and forkhead box P3 (FOXP3). Further, dual‐luciferase reporter gene and ChIP assays were employed to confirm the relationship among LINC00261, FOXP3 and sterol carrier protein‐2 (SCP2). PC cells were introduced with a series of vectors to verify the effects of LINC00261 and SCP2 on the viability, cell cycle progression, migration and angiogenesis of PC cells. Nude mice with the xenograft tumour were used to evaluate the effects LINC00261 on the tumourigenicity. LINC00261 was lowly expressed in PC tissues and cells. SCP2 was inhibited by LINC00261 through FOXP3. Functionally, upregulated LINC00261 or downregulated SCP2 led to reduced cell viability, migration, angiogenesis and tumourigenicity potentials. This study demonstrated the inhibitory role of LINC00261 in the angiogenesis and cell cycle progression of PC cells. It acts through the negative regulation of SCP2 via targeting FOXP3. Findings in this study highlight a potentially biomarker for PC treatment.
... It is important to note that previous studies have also reported an association between high FoxP3 + lymphocyte infiltrations and advanced CRC [33,55]. Additionally, the presence of FoxP3 + T lymphocyte infiltrates were associated with poor prognosis in different types of cancers, including breast [26], lung [27], pancreatic [56], ovarian [57], and cervical [28] cancers. These data show that the presence of FoxP3 + T lymphocytes is not always associated with a good prognosis. ...
Tumor-infiltrating lymphocytes include heterogeneous populations of T lymphocytes that play crucial roles in the tumor immune response; importantly, their presence in the tumor tissue may predict clinical outcomes. Therefore, we herein studied the prognostic significance of the presence and location of CD3+, CD8+, and FoxP3+ T lymphocytes in colorectal cancer samples. In the intratumor analysis, our data did not reveal any association between lymphocyte infiltrations with clinical or pathological data. However, in the tumor margins, we found that the presence of high infiltrations of CD3+, CD8+, or FoxP3+ T lymphocytes were associated with TNM stages I-II (p = 0.021, p = 0.022, and p = 0.012, respectively) and absence of lymph node metastases (p = 0.010, p = 0.003, and p = 0.004, respectively). Despite these associations with good prognostic indicators, we were not able to find any statistically significant alterations in the overall survival of the patients, even though high infiltrations of FoxP3+ T lymphocytes in the tumor margins resulted in an increased overall survival of 14 months. Taken together, these data show that the presence of CD3+, CD8+, or FoxP3+T lymphocyte infiltrates in the tumor margins are associated with the pathogenesis of CRC, but only high Foxp3+ T lymphocyte infiltrations in the tumor invasive margins are inclined to indicate favorable prognosis.
... The elevated abundance of FOXP3 + Tregs is generally associated with a poor prognosis in most non-mucosal-derived solid tumors (Chaudhary and Elkord 2016;Shang et al. 2015). This association between tumorinfiltrating Treg abundance and prognosis is particularly true when using the ratio between Tregs and conventional T cells, where a higher ratio is significantly correlated with a worse prognosis in breast cancer, lung cancer, melanoma, pancreatic cancer, and ovarian cancer (Curiel et al. 2004;Jiang et al. 2014;Leffers et al. 2009;Sayour et al. 2015;Tang et al. 2014;Tao et al. 2012;Yang et al. 2006). While Tregs tend to be higher in the peripheral blood in cancer patients, this is not always associated with the abundance of tumor-infiltrating Tregs (Adeegbe and Nishikawa 2013;Togashi et al. 2019). ...
Regulatory T cells (Tregs) are critical in maintaining immune homeostasis under various pathophysiological conditions. A growing body of evidence demonstrates that Tregs play an important role in cancer progression and that they do so by suppressing cancer-directed immune responses. Tregs have been targeted for destruction by exploiting antibodies against and small-molecule inhibitors of several molecules that are highly expressed in Tregs—including immune checkpoint molecules, chemokine receptors, and metabolites. To date, these strategies have had only limited antitumor efficacy, yet they have also created significant risk of autoimmunity because most of them do not differentiate Tregs in tumors from those in normal tissues. Currently, immune checkpoint inhibitor (ICI)-based cancer immunotherapies have revolutionized cancer treatment, but the resistance to ICI is common and the elevation of Tregs is one of the most important mechanisms. Therapeutic strategies that can selectively eliminate Tregs in the tumor (i.e. therapies that do not run the risk of causing autoimmunity by affecting normal tissue), are urgently needed for the development of cancer immunotherapies. This chapter discusses specific properties of human Tregs under the context of cancer and the various ways to target Treg for cancer immunotherapy.
... A high number of regulatory T cells and higher ratio of Treg cells to Teff cells (effector T cells) have been found to correlate with poor prognosis and reduced survival of patients with many different types of cancer, including ovarian cancer [24] , pancreatic cancer [17] , lung cancer T1, glioblastoma [38] , and other types of cancer. In this article, we will illustrate this phenomenon numerically for the first time. ...
Recent advances in the field of regulatory T cells reveal that these cells play a vital role during immunotherapy. For example, a higher ratio between regulatory T cells and effector T cells within tumor tissue is associated with worse prognoses in many cancers, including ovarian cancer (Leffers et al., 2009), lung cancer (Tao et al., 2012), glioblastoma (Sayour et al., 2015). On the other hand, the tug of war between regulatory T cells and effector T cells for interleukin-2 may chisel the immune response against cancer. In this work, we study a mathematical model that investigates the role of regulatory T cells during immunotherapy. We demonstrate mathematically, for the first time, that the initial ratio between regulatory T cells and effector T cells does impact the tumor recurrence time. We also demonstrate the effectiveness of utilization of IL-2 may flip the outcome of immunotherapy, providing further evidence that it may be clinically viable to modulate the consumption of IL-2 by Tregs.
... Although Tregs and Th17 cells are both generated from naïve T cells, each has an opposite function and pathogenesis. Most studies regarding the accumulation of Tregs and Th17 cells in cancer have been related to the rate, subpopulation, and relationship to cancer progression/prognosis [17][18][19][20][21][22][23], while their distribution, the role of balance, and potential cancerrelated mechanism remained unclear. Through the study of BTC, the progression of which is closely related to the IL-6/TGF-β1 axis [6], we hypothesized that differences in the cytokines secreted from cancer cells would induce hetero-differentiation of T cells in the tumor microenvironment and provoke different immune responses. ...
Background/Aim
We previously demonstrated that inflammatory cytokine interleukin-6 (IL-6) was produced during cancer progression, worked together with transforming growth factor-beta 1 (TGF-β1), and induced the epithelial–mesenchymal transition (EMT) with chemo-resistance against gemcitabine (GR) at the invasion front of biliary tract cancers (BTCs). However, the significance of cytokine-induced T cell accumulation at the tumor microenvironment in biliary tract cancer (BTC) is not well understood. Because these cytokines (IL-6 and TGF-β1) are able to differentiate naïve T cells into Foxp3-expressing T cells (Tregs) and/or IL-17–producing T helper 17 (Th17) cells, we investigated the relationship between heterogeneous, cancer-producing cytokines and T cell differentiation.
Methods
In total, 127 curative resected specimens from patients with BTCs at Osaka University Hospital between 2000 and 2012 were evaluated for IL-6, TGF-β1, Tregs, and Th17 cells by immunohistochemistry. The ability of BTC–GR cells to undergo T cell differentiation was investigated in vitro.
Results
Tregs accumulated at the tumor center and Th17 cells accumulated at the invasion front during cancer progression and/or metastasis; each signaled poor prognosis. Treg accumulation was related to TGF-β1 expression by cancer cells, and Th17 cell accumulation was related to IL-6 expression by cancer cells, in resected specimens; this was confirmed in vitro. Compared with parent cells, GR cells produced IL-6 but not TGF-β1 in a time-dependent manner, had EMT features, and induced T cell differentiation to Th17 cells but not Tregs.
Conclusion
Cytokines produced by cancer cells (IL-6 and TGF-β1) induced heterogeneity of Tregs and Th17 cells in the tumor microenvironment, supporting progression of BTC.
