Expression Of Factor Forkhead Box P3 Research Articles

The Regulation of FOXP3 Expression and Its Therapeutic Potential in Non-Small Lung Cancer

Non-small cell cancer (NSCLC) is today one of the most threatening malignant tumors, and it is still rarely cured by current standard therapies with a high mortality rate. The transcription factor Forkhead box p3 (FOXP3) is a potential molecular target for NSCLC since it has been identified with an oncogenic effect in the tumor microenvironment (TME). It is well known that FOXP3 expression could characterize the regulatory T cells (Treg), which plays a critical role in regulating anti-tumor immune responses in TME. Through analyzing the basic mechanisms of FOXP3 and Treg in distinct tumor tissues, they have been found with contradictory expression and the frequency in different TME. In NSCLC, the suppressive function of Treg to immune systems and frequency of FOXP3 is accumulated to promote the development of tumors. Most recently, accumulating evidence reveals the therapeutic potential of targeting FOXP3 and the related signaling pathways in the treatment of NSCLC patients. Although unsolved side effects and limitations that still must be considered when targeting FOXP3 and FOXP3-related proteins, there is no doubt that they are potential therapeutic applications for NSCLC treatment. The main objective of this review is to mention the structure, function and recent progress in treatment targeting FOXP3 and FOXP3-related signaling pathways, especially to therapies with immune checkpoint inhibitors.

FOXP3 expression in esophageal squamous cell carcinoma : Implications for cetuximab sensitivity and therapeutic strategies.

Investigating the impact of FOXP3 (transcription factor forkhead box P3) expression on the biological behavior of esophageal squamous cell carcinoma (ESCC) and its influence on the sensitivity of ESCC cells towards cetuximab-targeted (an EGFR monoclonal antibody inhibitor) therapy. Aspecifically designed recombinant FOXP3 shRNA plasmid was synthesized to target the human FOXP3 gene, and the plasmid was transfected into TE12 cells using aliposome method. Multiple assays were conducted to evaluate the effect of FOXP3 expression on ESCC cells and their response to cetuximab treatment. Proliferation activity and cetuximab sensitivity of ESCC cells were measured using the CCK‑8 assay. The invasion ability of cells was assessed using an in vitro invasion assay. Furthermore, the efficacy of cetuximab in treating ESCC was analyzed using atumorigenesis assay in nude mice. Silencing the FOXP3 gene in the TE12 cell line (shFOXP3 group) resulted in asignificant reduction in FOXP3 mRNA and protein expression (p = 0.013). The shFOXP3 group exhibited slowed cell growth (p = 0.035), decreased invasion rate (p = 0.031), and increased sensitivity to cetuximab treatment (p = 0.039) compared to the control group (shNC group). In the in vivo tumorigenesis assay, the shFOXP3 group demonstrated asignificant reduction in tumor volume and lung metastasis rate following cetuximab treatment (p = 0.028 and 0.007, respectively). High FOXP3 expression promotes the proliferation and migration of ESCC cells, while negatively affecting their sensitivity to cetuximab-targeted therapy. Consequently, targeting FOXP3 shows potential therapeutic implications for enhancing the effectiveness of cetuximab treatment in ESCC patients.

Ectopic FOXP3 Expression in Combination with TGF-β1 and IL-2 Stimulation Generates Limited Suppressive Function in Human Primary Activated Thymocytes Ex Vivo.

Regulatory T cells (Tregs), which are characterized by the expression of the transcription factor forkhead box P3 (FOXP3), are the main immune cells that induce tolerance and are regulators of immune homeostasis. Natural Treg cells (nTregs), described as CD4+CD25+FOXP3+, are generated in the thymus via activation and cytokine signaling. Transforming growth factor beta type 1 (TGF-β1) is pivotal to the generation of the nTreg lineage, its maintenance in the thymus, and to generating induced Treg cells (iTregs) in the periphery or in vitro arising from conventional T cells (Tconvs). Here, we tested whether TGF-β1 treatment, associated with interleukin-2 (IL-2) and CD3/CD28 stimulation, could generate functional Treg-like cells from human thymocytes in vitro, as it does from Tconvs. Additionally, we genetically manipulated the cells for ectopic FOXP3 expression, along with the TGF-β1 treatment. We demonstrated that TGF-β1 and ectopic FOXP3, combined with IL-2 and through CD3/CD28 activation, transformed human thymocytes into cells that expressed high levels of Treg-associated markers. However, these cells also presented a lack of homogeneous suppressive function and an unstable proinflammatory cytokine profile. Therefore, thymocyte-derived cells, activated with the same stimuli as Tconvs, were not an appropriate alternative for inducing cells with a Treg-like phenotype and function.

Role of FOXP3 Expression and Serum Vitamin D and C Concentrations When Predicting Acquisition of Tolerance in Infants With Cow's Milk Allergy.

Treg cells and dietetic factors may play a significant role in the natural acquisition of tolerance in children with cow's milk allergy (CMA). The best marker for Treg lymphocytes is the transcription factor forkhead boxP3 (FOXP3). Objective: We examine the relationship between FOXP3 mRNA expression and serum concentrations of vitamins D and C and the development of different phenotypes of tolerance in children with CMA. The study group comprised 138 infants with CMA and 101 healthy infants. All children underwent oral food challenge, first with an extensively heated milk product and then with unheated products. FOXP3 mRNA expression and serum vitamin C and D concentrations were evaluated. At 2 years of life, 54 children (39.1%) still had CMA, 43 (31.2%) were unheated milk-reactive and heated milk-tolerant, while 41 (29.7%) had outgrown their allergy. The mean (SD) level of FOXP3 expression in the study group was 2.07 (1.23), which was lower than the control group value of 2.98 (1.52) (P<.001). A value below 1.45 indicated allergy. The mean serum level of vitamin D in the study group was lower than in the control group (29.67 [7.09] vs 33.35 [4.13] ng/mL; P<.001). No significant differences were found in mean serum vitamin C content. Increased FOXP3 mRNA expression can predict faster acquisition of tolerance in infants with CMA. These children have lower serum vitamin D levels than healthy children. No relationship was found between the natural history of CMA and serum vitamin C concentration.

Downregulation of FOXP3 in neutrophils by IL-8 promotes the progression of oral squamous cell carcinoma.

The aim of the present study was to investigate the effects of the transcription factor forkhead box P3 (FOXP3) in neutrophils on the progression of oral squamous cell carcinoma (OSCC). Cancer tissue samples and paracarcinoma tissues were collected from 23 patients with OSCC for the current study. In addition, SCC-9, a human tongue carcinoma cell line, was co-cultured with primary human neutrophils and treated with recombinant interleukin 8 (IL-8). The effect of FOXP3 on the proliferation of SCC-9 cells was analyzed using a Cell Counting Kit 8 assay. FOXP3 expression in neutrophils was analyzed by quantitative PCR following IL-8 treatment. FOXP3 protein expression in neutrophils and the amount of IL-8 protein in the OSCC tumor microenvironment were determined by immunofluorescence analysis. The present study demonstrated that IL-8 downregulated FOXP3 mRNA expression in neutrophils. Neutrophils and peptide P60, a specific inhibitor of FOXP3, increased proliferation of SCC-9 cells. In patients with OSCC, FOXP3 protein expression in neutrophils of the stage IV group was significantly lower compared with that of the stage II and stage III groups, while IL-8 protein expression was higher in cancer tissues compared with that in paracarcinoma tissues. In summary, IL-8 in the tumor microenvironment may recruit neutrophils, and downregulation of FOXP3 in neutrophils by IL-8 may promote the progression of OSCC.

A Phenotypic Screening Approach Using Human Treg Cells Identified Regulators of Forkhead Box p3 Expression.

Regulatory T (Treg) cells, expressing the transcription factor forkhead box p3 (FOXP3), are the key cells regulating peripheral autoreactive T lymphocytes by suppressing effector T cells. FOXP3+ Treg cells play essential roles controlling immune responses in autoimmune diseases and cancer. Several clinical approaches (e.g., polyclonal expansion of Treg cells with anti-CD3 and anti-CD28 coated beads in the presence of drugs) are under evaluation. However, expression of FOXP3, recognized as the master regulator of Treg cells, in induced Treg cells have been shown to be instable, and molecular targets involved in regulating FOXP3 expression and Treg cell function have not been well-defined. Thus, new targets directly regulating FOXP3 expression and the expression of its downstream genes (e.g., cytotoxic T-lymphocyte-associated protein 4 (CTLA4)) have the potential to stabilize the Treg cell phenotype and function. This report describes the development of an automated medium-throughput 384-well plate flow cytometry phenotypic assay meauring the protein expression of FOXP3 and CTLA4 in human Treg cells. Screening a library of 4213 structurally diverse compounds allowed us to identify a variety of compounds regulating FOXP3 and CTLA4 expression. Further evaluation of these and related small molecules, followed by confirmation using siRNA-mediated gene knockdown, revealed three targets: euchromatic histone-lysine N-methyltransferase (EHMT2) and glycogen synthase kinase 3 alpha/beta (GSK3α/β) as potent positive regulators of FOXP3 expression, and bromodomain and extra-terminal domain (BET) inhibitors as negative regulators of FOXP3 and CTLA4 expression. These targets have potential implications for establishing novel therapies for autoimmune diseases and cancer.

SIRT1-Regulated Abnormal Acetylation of FOXP3 Induces Regulatory T-Cell Function Defect in Hashimoto's Thyroiditis.

Hashimoto's thyroiditis (HT) is an autoimmune thyroid disease characterized by low expression of transcription factor Forkhead Box P3 (FOXP3) and functional deficiency of a cluster of differentiation regulatory T cells (Tregs). This study aimed to investigate the mechanism of Treg dysfunction in HT. The number of CD4+CD25+FOXP3+ T cells was determined by flow cytometry. Expression of FOXP3 and Sirtuin type 1 (SIRT1) was evaluated by Western blot analysis. Acetylation of FOXP3 was analyzed by immunoprecipitation and Western blot analysis. The suppressive function of Treg was analyzed by the 5,6-carboxyfluorescein diacetate succinimidyl ester (CFSE) assay. The percentage of CD4+CD25+FOXP3+ T cells, expression of FOXP3, and FOXP3 acetylation level in the HT group were significantly lower than in the control groups. Conversely, SIRT1 expression was significantly higher in the HT group than in the other two groups. After Ex-527 treatment, the CD4+CD25+FOXP3+ T cells percentage, FOXP3 expression, and FOXP3 acetylation level in the HT group were significantly increased. HT Tregs exhibited less suppressive activity, but Ex-527 treatment significantly increased their suppressive activity. The findings demonstrate that the reduced FOXP3 expression level and Treg function defect in HT patients are regulated by SIRT1-mediated abnormal FOXP3 acetylation. Ex-527 may upregulate the FOXP3 acetylation level and subsequently increase the number and suppressive function of Treg cells.

Preliminary study on decreasing the expression of FOXP3 with miR-155 to inhibit diffuse large B-cell lymphoma.

The aim of the present study was to analyze the association between the transcription factor forkhead box P3 (FOXP3) and diffuse large B-cell lymphoma (DLBCL), and investigate the effect of microRNA-155 (miR-155) on the generation and development of FOXP3 in DLBCL. The reverse transcription-quantitative polymerase chain reaction (RT-qPCR) technique was used to determine the expression of FOXP3 in the human DLBCL cell lines Ly1, Ly8 and Ly10, and in normal B cells. An immunohistochemical method was used to determine FOXP3 expression in 60 DLBCL tumor and adjacent tissues, and a retrospective analysis of FOXP3 expression in tumor tissues and clinical data was performed. The lentiviral transfection technique was used to silence the miR-155 gene in mouse A20 cells to analyze the influence of miR-155 on FOXP3 in DLBCL. The A20 cell line with a silenced miR-155 gene was used to perform a tumorigenicity assay in BALB/c mice, and to compare the tumorigenicity rate and the tumor growth rate. The results identified that the expression of the transcription factor FOXP3 in the human DLBCL cell lines was increased compared with normal B cells; FOXP3 in human DLBCL tumor issues was increased compared with the tumor-adjacent tissue, and the increased expression of FOXP3 was identified as an indicator of poor prognosis of patients with DLBCL in the middle and late period; FOXP3 level decreased subsequent to silencing miR-155 in A20 cells; A20 cells with the low-expression miR-155 gene were used to determine the tumorigenicity in BALB/c mice and it was identified that the tumorigenicity of the low-expression miR-155 gene group was decreased compared with the untransfected group. Therefore, miR-155 may be a regulatory factor of FOXP3, and miR-155 may be associated with the metastasis and prognosis of patients with DLBCL.

FOXP3 and CEACAM6 expression and T cell infiltration in the occurrence and development of colon cancer.

The transcription factor forkhead box P3 (FOXP3) is involved in immune cell regulation, and carcinoembryonic antigen-related cell adhesion molecule 6 (CEACAM6) is an adhesion molecule of the immunoglobulin superfamily. These two genes are associated with cancer progression. In the current study, colon tissue specimens from 78 cases of colon cancer (including 40 of stage I–II and 38 of stage III–IV), 30 cases of colonic adenoma and 12 healthy controls were collected from the First Affiliated Hospital of Soochow University between January 2010 and December 2011. The expression of cluster of differentiation (CD) 3, CD4, CD8, CD45RO, CEACAM6 and FOXP3 in colon tissues was examined by immunohistochemical analysis. In addition, a reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay, based on SYBR Green I, was used to detect CD3, CD4, CD8, CD45RO, CEACAM6 and FOXP3 mRNA levels in the paraffin block specimens. CD3+, CD8+ and CD45RO+ T cell infiltrations in colonic adenoma were significantly higher than in normal colonic mucosa (P<0.001, P=0.001 and P<0.001, respectively). However, CD3+, CD8+ and CD45RO+ lymphocytes in stage III–IV colon cancer tissues were lower than in normal control tissues (P=0.015, P=0.002 and P=0.041, respectively); consistently, CD3+, CD4+, CD8+ and CD45RO+ lymphocytes in stage III–IV tissues were even more markedly lower compared with adenoma (P=0.001, P<0.001, P<0.001 and P<0.001, respectively). Similarly, CD3+, CD8+ and CD45RO+ T cell infiltration was lower in stage I–II cancer tissues compared with adenoma (P=0.001, P<0.001 and P<0.001). CD3+, CD4+, CD8+ and CD45RO+ T cell infiltrations were also significantly higher in stage I–II compared with stage III–IV cancer tissues (P<0.001, P=0.045, P<0.001 and P<0.001, respectively). CEACAM6 was found to gradually increase from normal colon tissue to adenoma and cancer tissue. FOXP3 was expressed more highly in stage I–II compared with normal tissues (P=0.014), and was even higher in stage III–IV (P<0.001). These results were verified using RT-qPCR, which yielded almost identical results. In summary, the current study demonstrates that FOXP3, CEACAM6 and T cell infiltration are significantly associated with the occurrence and progression of colon cancer, and that immune reactions vary between different stages of colon cancer development.

FOXP3 can modulate TAL1 transcriptional activity through interaction with LMO2

T-cell acute lymphoblastic leukemia (T-ALL) frequently involves aberrant expression of TAL1 (T-cell acute lymphocytic leukemia 1) and LMO2, oncogenic members of the TAL1 transcriptional complex. Transcriptional activity of the TAL1-complex is thought to have a pivotal role in the transformation of thymocytes and is associated with a differentiation block and self-renewal. The transcription factor Forkhead Box P3 (FOXP3) was recently described to be expressed in a variety of malignancies including T-ALL. Here we show that increased FOXP3 levels negatively correlate with expression of genes regulated by the oncogenic TAL1-complex in human T-ALL patient samples as well as a T-ALL cell line ectopically expressing FOXP3. In these cells, FOXP3 expression results in altered regulation of cell cycle progression and reduced cell viability. Finally, we demonstrate that FOXP3 binds LMO2 in vitro, resulting in decreased interaction between LMO2 and TAL1, providing a molecular mechanism for FOXP3-mediated transcriptional modulation in T-ALL. Collectively, our findings provide initial evidence for a novel role of FOXP3 as a tumor suppressor in T-ALL through modulation of TAL1 transcriptional activity.

FOXP3(+) Treg Cells and Gender Bias in Autoimmune Diseases.

CD4+CD25+ regulatory T (Treg) cells play a pivotal role in the maintenance of immune homeostasis, where the X-linked master transcription factor forkhead box P3 (FOXP3) determines Treg cell development and function. Genetic deficiency of foxp3 induces dysfunction of Treg cells and immuno-dysregulation, polyendocrinopathy, enteropathy, and X-linked syndrome in humans. Functionally deficient Treg cells or the development of exTreg cells positively correlate with autoimmune diseases, such as systemic lupus erythematosus (SLE), multiple sclerosis (MS), and ankylosing spondylitis (AS). In general, females are more susceptible to SLE and MS but less susceptible to AS, where the expression of FOXP3 and its protein complex are perturbed by multiple factors, including hormonal fluctuations, inflammatory cytokines, and danger signals. Therefore, it is critical to explore the potential molecular mechanisms involved and these differences linked to gender. Here, we review recent findings on the regulation of FOXP3 activity in Treg cells and also discuss gender difference in the determination of Treg cell function in autoimmune diseases.

ME-19 * ANALYSIS OF TREG RECRUITMENT AND FOXP3 EXPRESSION IN GLIOMAS REGARDING THE INTRAOPERATIVE FLUORESCENCE OF 5-ALA

Not only are regulatory T cells (Tregs) an essential component of the immune system in maintaining immunologic self-tolerance but they are also involved in the suppression of anti-tumor immune responses. The transcription factor forkhead box P3 (FOXP3) is widely used as a marker for the identification of Tregs being essential for the functionality. Oral 5 aminolevulinic acid (5-ALA) is used to assist surgical resection of high-grade gliomas, being known as an indicator of malignancy as it is able to pass the blood brain barrier in high-grade gliomas. Gaining more knowledge regarding the pathogenesis of gliomas, the tumor microenvironment came to the fore. In this study, patterns of Treg recruitment and FOXP3 expression in the microenvironment of gliomas were characterized, regarding the fluorescence of 5-ALA. Glioma tissue positive and negative for intraoperative fluorescence of 5-ALA was analyzed and compared. Tregs present in the tumor microenvironment were detected by double-immunohistochemistry for FOXP3 and the glioma marker, anti- glial fibrillary acidic protein (GFAP). FOXP3 mRNA expression was examined, using quantitative real-time-PCR. Tregs were found in the tumor microenvironment as well as within tumor cell islets, and FOXP3 mRNA expression in the tumors was significantly higher than in normal brain. Intriguingly, single FOXP3+ cells exhibited morphologic characteristics of glioma cells. 5-ALA positive tumor tissue showed a higher accumulation of Tregs than 5-ALA negative tissue. Gliomas recruit Tregs into their microenvironment, presumably in order to suppress immunosurveillance, thus avoiding destruction by the immune system. Endogenous FOXP3 expression in glioma cells might present a novel mechanism of immune escape. Our study showed differences between the amount of Treg and FOPX3 expression in 5-ALA positive and negative glioma tissue indicating that Tregs are more present in high-grade malignant gliomas. These findings suggest FOXP3 and Tregs as a possible therapeutic target in the therapy of gliomas.

Clinical significance of FOXP3 expression in human gliomas

Studies have demonstrated that the transcription factor forkhead box P3 (FOXP3) is expressed not only in regulatory T cells, but also in some cancer cells. This study aims to clarify whether or not FOXP3 expression occurs in human gliomas and investigate the clinical significance of this expression in gliomas. We detected FOXP3 protein expression in 40 glioma samples, 3 normal brain tissue samples, and 4 normal tonsil tissue samples using immunohistochemical staining and western blot. The expression of FOXP3 protein was also detected in five glioma cell lines by western blot. We also evaluated the association of FOXP3 expression with clinical pathological grades, prognosis, and recurrence. Western blot analysis showed that the expression of FOXP3 protein was upregulated in high-grade glioma (HGGS) samples compared with low-grade samples. The cell line U87 showed the highest FOXP3 expression, while U373 had the lowest expression. Immunohistochemical analysis detected FOXP3 protein in 35 out of the 40 (87.5%) glioma samples and high levels of FOXP3 were observed in 26 out of the 27 (96.3%) high-grade gliomas samples. Statistical analysis suggested that the upregulation of FOXP3 is significantly correlated with the histologic grade of gliomas (P<0.05) and that patients with high expression of FOXP3 protein exhibit a poorer prognosis than those with low FOXP3 expression. Our findings suggest that FOXP3 expression in glioma cells has a crucial function in the development of HGGS and is associated with the malignant biological behavior of HGGS.

Role of the transcription factor forkhead box P3 in breast cancer and metastasis

Event Abstract Back to Event Role of the transcription factor forkhead box P3 in breast cancer and metastasis Lucia Sfondrini1*, Alessandra Cataldo1, Valentina Uva1, Tiziana Triulzi2, Patrizia Casalini2, Elda Tagliabue2 and Andrea Balsari1, 2 1 Università degli Studi di Milano, Dipartimento di Scienze Biomediche per la Salute, Italy 2 IRCCS Istituto Nazionale Tumori Milano, Expertimental Oncology Department, Italy Background: The transcription factor forkhead box P3 (FOXP3), which is known for its critical role in regulating the development and function of regulatory T cells, can also be expressed in cancer cells. We have previously shown that FOXP3 expression in breast cancer represents a strong prognostic negative factor for distant metastases-free survival. Thus, we investigated the role and molecular mechanism of action of FOXP3 in the metastatic process using breast cancer models. Methods and results MDA-MB-231 human breast cancer cells, expressing low levels of FOXP3, were used to establish a tet-off inducible expression system, in which full-length (WT) FOXP3 or splice variant form (Δ2) were induced upon doxycycline removal. Real Time PCR and Western Blot analysis confirmed that under this condition FOXP3 mRNA and protein levels were significantly increased in both WT and Δ2 FOXP3-tet-off MDA-MB-231 clones. In vitro, an up-regulation of WT and Δ2 FOXP3 inhibited cell proliferation but resulted in an increased migration and invasion. In vivo experiments, initially performed using Δ2 FOXP3-tet-off clones, revealed a significant increase in the number of spontaneous and experimental lung metastases, obtained by mammary fatpad and intravenous injections, respectively, when Δ2 FOX3 expression was induced by doxycycline removal. Accordingly, a significant modulation of the expression of genes relevant for dissemination and/or control of the immune response has been observed in Δ2 FOXP3-induced clones when compared to not-induced ones by gene microarrays analysis. Conclusions: These findings indicate that in breast tumor FOXP3 expression inhibits tumor growth while promotes metastatic capability. Keywords: Foxp3, breast cancer, Prognostic factor, metastases, inducible expression Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Translational immunology and immune intervention Citation: Sfondrini L, Cataldo A, Uva V, Triulzi T, Casalini P, Tagliabue E and Balsari A (2013). Role of the transcription factor forkhead box P3 in breast cancer and metastasis. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00776 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 17 Jun 2013; Published Online: 22 Aug 2013. * Correspondence: Dr. Lucia Sfondrini, Università degli Studi di Milano, Dipartimento di Scienze Biomediche per la Salute, Milan, 20133, Italy, lucia.sfondrini@unimi.it Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract Supplemental Data The Authors in Frontiers Lucia Sfondrini Alessandra Cataldo Valentina Uva Tiziana Triulzi Patrizia Casalini Elda Tagliabue Andrea Balsari Google Lucia Sfondrini Alessandra Cataldo Valentina Uva Tiziana Triulzi Patrizia Casalini Elda Tagliabue Andrea Balsari Google Scholar Lucia Sfondrini Alessandra Cataldo Valentina Uva Tiziana Triulzi Patrizia Casalini Elda Tagliabue Andrea Balsari PubMed Lucia Sfondrini Alessandra Cataldo Valentina Uva Tiziana Triulzi Patrizia Casalini Elda Tagliabue Andrea Balsari Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Indoleamine 2,3-dioxygenase and forkhead boxP3 on immune tolerance of gallbladder carcinoma cells

Objective To investigate the expressions of indoleamine 2,3-dioxygenase (IDO) and transcription factor forkhead box P3(FOXP3)in gallbladder adenocarcinoma; and to evaluate the relationship between the expressions of IDO and FOXP3 protein,and clinicopathology and lymph node metastasis of gallbladder carcinoma.Methods The expressions of IDO and FOXP3 in 51 cases of primary gallbladder cancer,90 cases of chronic cholecystitis,and 20 cases of normal gallbladder tissues were studied by immunohistochemical staining. Results The positive expression rates of IDO and FOXP3 in gallbladder carcinoma were 72.5％ (37/51) and 60.8％ (31/51),in normal gallbladder mucosa were 5％ (1/20) and 20.0％ (4/20),in cholecystitis and gallstone were 11.1％ (10/90) and 32.2％ (29/90),respectively.There were significant differences among the three groups in IDO and FOXP3 expressions (P＜0.01).The positive expression rates of IDO and FOXP3 were 7.1％ and 14.3％ for simple hyperplasia,17.7％ and 35.3％ for atypical hyperplasia,40％ and 35％ for gallbladder carcinoma (stages Ⅰ-Ⅲ),and 77.4％ and 64.5％ for gallbladder carcinoma (stages Ⅳ-Ⅴ ).There were significant differences among the four groups in IDO and FOXP3 expressions (P＜0.01).There were significant differences among the Nevin stage groups,lymph node metastasis group and gallbladder stone in IDO and FOXP3 expressions.However,there were no significant differences among the sex groups and the age groups in IDO and FOXP3 expressions (P＞0.05).In gallbladder carcinoma,the expression of IDO had a positive correlation with FOXP3 expression in lymphocyte (γ=0.406,P=0.003).Conclusion In gallbladder cancer,a high-expression of IDO and an expression of FOXP3 in lymphocyte are closely related with immune tolerance of gallbladder carcinoma.The results provide a reference for clinical use of immunotherapy in gallbladder cancer. Key words: Primary gallbladder carcinoma; Indoleamine 2,3-dioxygenase; Forkhead box P3

Evaluation ofFOXP3expression in canine mammary gland tumours

The expression of the transcription factor forkhead boxP3 (FOXP3) was examined in 62 canine mammary gland tumours via immunohistochemical analysis and association with the known expression of the oestrogen receptor-alpha (ERα), progesterone receptor (PR), c-erbB-2 receptor (HER2/neu), and survival. Positive staining for FOXP3 was present in 22.6% of the tumours and was associated with the histological type. Negative staining for FOXP3 was associated with positive ERα and PR expression (P < 0.001). The expression of FOXP3 in canine mammary gland tumours was significantly associated with the disease-free survival time (P = 0.029). The FOXP3 expression was not an independent prognostic factor in the multivariate analysis, though. The negative expression of FOXP3 in the canine mammary gland tumours was found to be related to histopathologic benignity and a longer survival time in canine mammary gland tumours.

Abstract 8783: Methylation of FOXP3 In Regulatory T Cells is Related to the Severity of Coronary Atherosclerosis

Objective: Regulatory T (Treg) cells play a protective role in experimental atherosclerosis. However, the role of Treg in the development of human coronary artery disease (CAD) is inconsistent. Demethylation in the DNA that encodes transcription factor forkhead box P3 ( FOXP3 ) is a prerequisite for the stable maintenance of suppressive properties in Tregs, which is also identified as the most specific marker for human Tregs. We aim to evaluate Treg levels in CAD patients by a novel method based on Treg-specific DNA demethylation within FOXP3 gene. Methods and Results: We included patients with acute myocardial infarction (AMI) and unstable angina (UA), and control subjects with normal coronary artery on angiography (NCA) in our study. Peripheral blood mononuclear cells were collected to determine Treg levels by both flow cytometry and PCR based DNA methylation analysis. We found that Treg levels that were quantified by the demethylation rate of Treg-specific demethylated region in FOXP3 gene, but not by flow cytometry, were univocally decreased in UA and AMI patients (NCA 4.22% ± 1.92%; UA 3.29% ± 1.45%; AMI 2.86% ± 1.26%). This decline was inversely related to the severity of CAD ( r =−0.308, P &lt;0.05). After adjustment for traditional risk factors, the association was still significant ( β =−0.336, P &lt;0.05). Furthermore, we detected the expression of DNA methyltransferases in CD4 + CD25 + Tregs sorted from included subjects. A significant increase of DNA methyltransferase3b was observed in CAD patients compared with normal controls. Treatment of CD4 + CD25 + Tregs with ox-LDL induced a 41% increase in the methylation of FOXP3 , accompanied with a more than 3-fold reduction of FOXP3 mRNA expression and increases in the expressions of DNA methyltransferase 3a and 3b. Moreover, the effect of ox-LDL on FOXP3 can be reversed by methyltransferase inhibitor (-)-epigallocatechin-3-gallate. Conclusion: Our data demonstrate that Treg cells are decreased in CAD patients and its reduction are associated with the severity of atherosclerosis. Ox-LDL decreases the expression of FOXP3 by DNA methylation. Thus, we confirm the important role of Treg cells in human CAD and suggest potential epigenetic regulation in the progression of atherosclerosis.

Expression of indoleamine 2,3-dioxygenase predicts shorter survival in patients with vulvar squamous cell carcinoma (vSCC) not influencing on the recruitment of FOXP3-expressing regulatory T cells in cancer nests

Objective Regulatory T cells (Tregs), and the enzyme indoleamine 2,3-dioxygenase (IDO), have potential regulatory properties for immune escape in cancer. Inhibitors of IDO are available and could potentially be used in vulvar cancer if IDO was proved to drive progression of the disease. The aim of this study was to evaluate the expression of factor forkhead boxP3 (FOXP3), a marker of Tregs, and IDO in vulvar squamous cell carcinoma (vSCC), and to verify their prognostic significance. Methods 76 primary tumors and 35 lymph node metastases derived from 76 patients with full clinical history were analyzed. The intratumoral infiltration of Tregs and IDO expression within cancer were evaluated by immunohistochemistry. Results The number of Tregs in primary tumor and in corresponding lymph node metastasis was significantly correlated. Intensity of Treg infiltrates in the primary and metastatic sites was not correlated to IDO expression and had no influence on the overall patient survival. High IDO expression was associated with significantly worse overall survival among vSCC patients and was found to be an independent prognostic factor similarly to the tumor grade and patient's age. Conclusions The degree of intratumoral Treg infiltrates is an individual feature and remains stable throughout the course of the disease without impact on the patient's survival. IDO expression predicts shorter survival of vSCC patients. If immunologic tolerance of the tumor is promoted by the overexpression of IDO it will not influence the number of intratumoral Tregs. IDO expression seems to be an independent prognostic factor in patients with vSCC.

FOXP3 Expression and Overall Survival in Breast Cancer

The transcription factor forkhead box P3 (FOXP3) up- or downregulates a large number of genes and has been recently reported to be expressed in tumor cells. We investigated the prognostic importance of FOXP3 expression in patients with breast cancer. The expression patterns of FOXP3 were characterized by immunohistochemistry in primary breast carcinoma specimens from patients of the Milan 3 and 1 trials. Kaplan-Meier analysis and Cox proportional hazards regression modeling were used to assess the overall survival, distant metastasis-free survival, and local relapse cumulative incidence, according to the presence or absence of FOXP3 expression. FOXP3 expression in tumors was associated with worse overall survival probability and the risk increased with increasing FOXP3 immunostaining intensity. FOXP3 was also a strong prognostic factor for distant metastases-free survival but not for local recurrence risk. In multivariate analysis FOXP3 resulted an independent prognostic factor and the hazard ratio of FOXP3 expression and of lymph node positivity were similar. In the Milan 3 trial, the probability of 10-year survival in node-negative subgroup was 100% for FOXP3-negative and 82% for FOXP3-positive patients; in node-positive subgroup 82% for FOXP3-negative and 41% for FOXP3-positive patients. Even in the Milan 1 trial the lack of FOXP3 expression in node-positive subgroup was related to a significantly better prognosis than in FOXP3-positive patients (10-year survival probability, 89% v 59%). The data identify FOXP3 expression as a new independent prognostic factor in breast carcinoma, which might help to improve the selection of patients for appropriate therapy.

FOXP3 Expression in Human Kidney Transplant Biopsies Is Associated with Rejection and Time Post Transplant but Not with Favorable Outcomes

Expression of the transcription factor forkhead box P3 (FOXP3) in transplant biopsies is of interest due to its role in a population of regulatory T cells. We analyzed FOXP3 mRNA expression using RT-PCR in 83 renal transplant biopsies for cause in relationship to histopathology, clinical findings and expression of pathogenesis-based transcript sets assessed by microarrays. FOXP3 mRNA was higher in rejection (T-cell and antibody-mediated) than nonrejection. Surprisingly, some native kidney controls also expressed FOXP3 mRNA. Immunostaining for FOXP3 was consistent with RT-PCR, showing interstitial FOXP3+ lymphocytes, even in some native kidney controls. FOXP3 expression correlated with interstitial inflammation, tubulitis, interstitial fibrosis, tubular atrophy, C4d positivity, longer time posttransplant, younger donors, class II panel reactive antibody >20% and transcript sets reflecting inflammation and injury, but unlike these features was time dependent. In multivariate analysis, higher FOXP3 mRNA was independently associated with rejection, T-cell-associated transcripts, younger donor age and longer time posttransplant. FOXP3 expression did not correlate with favorable graft outcomes, even when the analysis was restricted to biopsies with rejection. Thus FOXP3 mRNA expression is a time-dependent feature of inflammatory infiltrates in renal tissue. We hypothesize that time-dependent entry of FOXP3-positive cells represents a mechanism for stabilizing inflammatory sites.