Diagnostic accuracy of NI-RADS for prediction of head and neck squamous cell carcinoma: a systematic review and meta-analysis.

OBJECTIVES: This study aimed to assess the diagnostic performance of NI-RADS for the prediction of recurrence in patients treated for Head and Neck Squamous Cell Carcinoma (HNSCC). METHODS: A literature search was conducted using various databases to identify relevant articles published from June 2016 onwards. We included studies reporting the diagnostic accuracy of NI-RADS in distinguishing recurrence in patients undergoing imaging surveillance, with pathologic results and/or follow-up as the reference standard. Summary estimates of diagnostic accuracy in terms of sensitivity, specificity, positive likelihood ratio (LR +), negative likelihood ratio (LR -), and diagnostic odds ratio (DOR) were calculated with the hierarchical summary receiver operating characteristic (HSROC) model. Meta-regression and subgroup analyses were conducted to investigate different clinical settings. Study quality was evaluated using the Quality Assessment of Diagnostic Accuracy Studies-2 tool. RESULTS: A total of 12 studies were included in the current meta-analysis. The pooled sensitivity and specificity were 0.69 (95% CI 0.59-0.79) and 0.94 (95% CI 0.89-0.97), respectively. For the primary site, the pooled summary estimates were 0.67 (95% CI 0.53-0.78) and 0.95 (95% CI 0.90-0.97), for the nodal sites were 0.64 (95% CI 0.44-0.80) and 0.99 (95% CI 0.98-0.99), respectively. The recurrence rate for NI-RADS categories 1-3 was 0.03 (95% CI 0.02-0.05), 0.13 (95% CI 0.10-0.15), and 0.77 (95% CI 0.73-0.81). Meta-regression revealed that the type of analysis (per person vs. per site) and number of sites (≤ 200 vs. > 200) were significant factors associated with heterogeneity. CONCLUSIONS: NI-RADS demonstrated high specificity but moderate sensitivity in patients after treatment for HNSCC. Summary estimates showed a significantly higher malignancy rate for NI-RADS category 3 compared to category 2.

CD4+CCR8+ Tregs in ovarian cancer: a potential effector Tregs for immune regulation.

BACKGROUND: Tregs are key drivers of immunosuppression in solid tumors. As an important chemokine receptor on Tregs, the regulatory effect of CCR8 on tumor immunity has received more and more attention. However, the current research on CCR8 in the immune microenvironment of ovarian cancer has not been clear. METHODS: Bioinformatics analysis was used to compare the transcriptome differences between CD4+ T cells in the peripheral circulation and infiltrated in ovarian tumor tissues. RT-PCR was used to detect the expression levels of chemokine receptor-related differential genes on CD4+ T cells in peripheral blood and ovarian tumor tissues. Multiparameter flow cytometry was used to detect the proportion and phenotypic characteristics of CD4+CCR8+ Tregs and CD4+CCR8- Tregs in different sample types. The expression level of CCR8 ligands was detected at multiple levels. To explore the important role of CCR8-CCL1 and CCR8-CCL18 axis in the migration and invasion of CD4+CCR8+ Tregs into ovarian tumor tissues by establishing a chemotaxis system in vitro. RESULTS: In this study, significantly different gene expression profiles were found between peripheral circulating CD4+ T cells and infiltrating CD4+ T cells in ovarian tumor tissues, in which chemokine-chemokine receptor signaling pathway was significantly enriched in all three groups of differential genes. The expression level of CCR8 in infiltrating CD4+ T cells of ovarian cancer tissue was significantly higher than that in peripheral blood of healthy controls and ovarian cancer patients, and high expression of CCR8 was significantly correlated with advanced tumor stage and poor differentiation. CD4+CCR8+ Tregs are the main type of infiltrating CD4+ Tregs in ovarian tumor tissues, which have stronger immunosuppressive phenotypes, secrete more inhibitory cytokines and have stronger proliferation ability. The ligands CCL1 and CCL18 corresponding to CCR8 were significantly overexpressed in ovarian tumor tissues, and the CCR8-CCL1 and CCR8-CCL18 axis played a key role in the migration and infiltration of CD4+CCR8+ Tregs into ovarian tumor tissues. CONCLUSIONS: The results of this study may help to understand the phenotypic characteristics and recruitment process of Tregs in the tumor, and provide new ideas for improving the immunosuppressive status of the ovarian cancer microenvironment.

Circulating CD4+ Treg, CD8+ Treg, and CD3+ γδ T Cell Subpopulations in Ovarian Cancer.

Background and Objectives: Regulatory T cells (Tregs) are usually enriched in ovarian cancer (OC), and their immunosuppressive function plays a key role in tumorigenesis and progression. We mainly explored the phenotypical characterization of Treg-related markers on αß and γδ T cell subsets in patients with OC. Materials and Methods: Thirty-six untreated patients with OC at the Women's Hospital of Nanjing Medical University from September 2019 to August 2021 were enrolled. Phenotypical characterization of Tregs-related markers were detected by flow cytometry (FCM). Enzyme-linked immunosorbent assay was used to detect the levels of carbohydrate antigen (CA125) and transforming growth factor ß (TGF-ß). The level of human epididymis protein 4 (HE4) was detected by electrochemiluminescence immunoassay. Results: Circulating CD4+ Tregs, CD8+ Tregs, and CD3+γδ T cell subpopulations from OC patients have elevated Foxp3, CD25, CD122, Vδ1, and reduced CD28 expression compared to benign ovarian tumor (BOT) patients and healthy controls (HC). The upregulation of Foxp3 and Vδ1 and the downregulation of CD28 were highly specific for maintaining the immunosuppression function of CD4+ Tregs, CD3+γδ T cells, and CD8+ Tregs in OC patients. These Treg subpopulations were able to discriminate OC from BOT and HC. The levels of CA125, HE4, and TGF-ß were increased in OC patients. A significant positive correlation between Treg subpopulations and CA125, HE4, and TGF-ß was revealed. Conclusions: Proportions of CD4+ Tregs, CD8+ Tregs, and CD3+γδ T cell subsets were significantly increased in OC patients and were positively correlated with FIGO stage/metastasis status, CA125, HE4, and TGF-ß. These indicators have the potential to be used as immunosurveillance biomarkers for OC.

Distribution and functions of γδ T cells infiltrated in the ovarian cancer microenvironment.

BACKGROUND: The role of γδ T cells, innate-like lymphocytes with unrestrained MHC, in various malignancies has recently been extensively studied. However, there is limited research regarding γδ T cells in ovarian cancer (OC) patients. METHODS: Here, we investigated the distribution patterns of γδ T cells and their main subsets in peripheral blood and tumor tissues among OC patients, benign ovarian tumor (BOT) patients, and age-matched healthy controls (HC) by flow cytometry, as well as the expression levels of IFN-γ and IL-17A secreted from γδ T cells. Immunohistochemical staining was utilized to detect the numbers of γδ T cells and their main subsets in different types of ovarian tumor tissues. Additionally, we also investigated chemotaxis effects on γδ T cells, as well as their cytotoxic activity and proliferation. RESULTS: We found that the percentages of γδ T cells and Vδ1 T cells were significantly higher in OC tissues than BOT tissues and normal (N) ovarian tissues, while there were no obvious differences in peripheral blood. Meanwhile, higher numbers of γδ T cells and Vδ1 T cells were observed in OC tissues, and were positively related to advanced clinicopathological features of OC patients. Further, the levels of IFN-γ secreted by γδ T cells were relatively lower, while IL-17A was expressed at a high level in both the peripheral blood and tissues of OC patients. Chemotaxis assay revealed that supernatants derived from OC tissues possessed a stronger capacity to attract and recruit γδ T cells. However, γδ T cells sorted from OC tissues showed weakened cytotoxic activity against ovarian cancer cells, and γδ T cells cocultured with OC tissue supernatants could effectively inhibit the proliferative activity of naïve CD4+ T cells. CONCLUSIONS: These data suggested that γδ T cells might have critical roles in OC progression and potential utilization in treatment approaches or prognosis prediction.

MRI for the detection of small malignant renal masses: a systematic review and meta-analysis.

Objective: We aimed to review the available evidence on the diagnostic performance of magnetic resonance imaging in differentiating malignant from benign small renal masses. Methods: An electronic literature search of Web of Science, MEDLINE (Ovid and PubMed), Cochrane Library, EMBASE, and Google Scholar was performed to identify relevant articles up to 31 January 2023. We included studies that reported the diagnostic accuracy of using magnetic resonance imaging to differentiate small (≤4 cm) malignant from benign renal masses. The pooled sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, and diagnostic odds ratio were calculated using the bivariate model and the hierarchical summary receiver operating characteristic model. The study quality evaluation was performed with the Quality Assessment of Diagnostic Accuracy Studies-2 tool. Results: A total of 10 studies with 860 small renal masses (815 patients) were included in the current meta-analysis. The pooled sensitivity and specificity of the studies for the detection of malignant masses were 0.85 (95% CI 0.79-0.90) and 0.83 (95% CI 0.67-0.92), respectively. Conclusions: MRI had a moderate diagnostic performance in differentiating small malignant renal masses from benign ones. Substantial heterogeneity was observed between studies for both sensitivity and specificity.

Glucose metabolism and function of CD4+ Tregs are regulated by the TLR8/mTOR signal in an environment of SKOV3 cell growth.

PURPOSE: To investigate the role of mammalian target of rapamycin (mTOR) signal in Toll-like receptor (TLR) 8-mediated regulation of glucose metabolism and its effect on reversing immunosuppression in CD4+ regulatory T-cells (Tregs) in ovarian cancer (OC). METHODS: Fluorescence-activated cell sorting was used to detect the expression levels of mTOR+ and 4E-BP1+ cells in CD4+ Tregs. The prognosis and immune infiltration analysis of mTOR mRNA in OC were performed using the TIMER and Kaplan-Meier plotter database. Furthermore, real-time polymerase chain reaction (RT-PCR) and western blot (WB) were used to detect expression levels of glucose metabolism-related genes and proteins in CD4+ Tregs. Glucose uptake and glycolysis levels were detected by colorimetry, while the effects of CD4+ Tregs on the proliferation of CD4+ T-effector cells (Teffs) were evaluated by carboxyfluorescein diacetate succinimidyl ester (CFSE). RESULTS: mTOR expression in CD4+ Tregs was significantly higher in patients with OC compared with controls and in CD4+ Tregs than in CD4+ Teffs in OC. Additionally, the expression level of mTOR mRNA was related to prognosis and immune infiltration levels in patients with OC. Blocking the mTOR signal resulted in downregulation of glucose metabolism in CD4+ Tregs. Simultaneous inhibition of the mTOR signal while activation of the TLR8 signal had a coordinated inhibitory effect on glucose metabolism and the immunosuppressive function of CD4+ Tregs. Furthermore, the mTOR signal played an essential role in TLR8-mediated reversal of immunosuppressive function in CD4+ Tregs. CONCLUSION: These findings imply that activation of the TLR8 signal inhibits glucose metabolism in CD4+ Tregs by downregulating mTOR signaling, thereby reversing the immunosuppressive function of these cells in an OC cell growth environment.

Evaluation of collateral circulation and short-term prognosis of patients with acute cerebral infarction by perfusion-weighted MRI.

BACKGROUND: Perfusion-weighted magnetic resonance imaging (MRI) can evaluate collateral circulation in patients with acute cerebral infarction by reflecting hemodynamic signals in the brain. The purpose of this study was to evaluate the collateral circulation and short-term prognosis of patients with acute cerebral infarction, using perfusion-weighted MRI. METHODS: The study enrolled 206 patients with acute cerebral infarction due to unilateral cerebral artery occlusion diagnosed by digital subtraction angiography (DSA) and computed tomography angiography (CTA). The relative cerebral blood volume (rCBV), relative cerebral blood flow map (rCBF), relative peak time (rTTP), and relative mean transit time (rMTT) were calculated based on the ratio of the perfusion-weighted MRI reference values of the infarcted side and the control side of the patient. According to the results of perfusion-weighted MRI, patients were divided into a high perfusion group (n=121) and a low perfusion group (n=85). The Thrombolysis in Cerebral Infarction scale proposed by Higashida et al. in 2003 (Higashida scale) was used to evaluate the establishment of collateral circulation on the day of admission and 15 days after admission. The National Institutes of Health Stroke Scale (NIHSS) score and the modified Rankin scale assessed the short-term prognosis of patients with cerebral infarction. The Spearman correlation analysis examined the correlation between the rCBV, rCBF, and NIHSS scores, and the modified Rankin scale (mRS). RESULTS: Compared with the patients in the low perfusion group, the rCBV and rCBF in the infarcted area of the patients in the high perfusion group were significantly increased and the rTTP and rMTT were significantly decreased. On day 15 after admission, the collateral circulation rate of the high perfusion group was significantly higher than that of the low perfusion group, and the NIHSS score and the mRS score were significantly lower than those scores of the low perfusion group. Perfusion-weighted MRI indexes, rCBV, and rCBF were negatively correlated with the NIHSS score and Rankin scale. CONCLUSIONS: Perfusion-weighted MRI can effectively evaluate the compensatory ability of collateral circulation and the prognosis of patients with acute cerebral infarction.

Systematic Review and Meta-Analysis of American College of Radiology TI-RADS Inter-Reader Reliability for Risk Stratification of Thyroid Nodules.

Purpose: To investigate the inter-reader agreement of using the American College of Radiology (ACR) Thyroid Imaging Reporting and Data System (TI-RADS) for risk stratification of thyroid nodules. Methods: A literature search of Web of Science, PubMed, Cochrane Library, EMBASE, and Google Scholar was performed to identify eligible articles published from inception until October 31, 2021. We included studies reporting inter-reader agreement of different radiologists who applied ACR TI-RADS for the classification of thyroid nodules. Quality assessment of the included studies was performed with the Quality Assessment of Diagnostic Accuracy Studies-2 tool and Guidelines for Reporting Reliability and Agreement Studies. The summary estimates of the inter-reader agreement were pooled with the random-effects model, and multiple subgroup analyses and meta-regression were performed to investigate various clinical settings. Results: A total of 13 studies comprising 5,238 nodules were included in the current meta-analysis and systematic review. The pooled inter-reader agreement for overall ACR TI-RADS classification was moderate (κ = 0.51, 95% CI 0.42-0.59). Substantial heterogeneity was presented throughout the studies, and meta-regression analyses suggested that the malignant rate was the significant factor. Regarding the ultrasound (US) features, the best inter-reader agreement was composition (κ = 0.58, 95% CI 0.53-0.63), followed by shape (κ = 0.57, 95% CI 0.41-0.72), echogenicity (κ = 0.50, 95% CI 0.40-0.60), echogenic foci (κ = 0.44, 95% CI 0.36-0.53), and margin (κ = 0.34, 95% CI 0.24-0.44). Conclusions: The ACR TI-RADS demonstrated moderate inter-reader agreement between radiologists for the overall classification. However, the US feature of margin only showed fair inter-reader reliability among different observers.

Distribution and Clinical Significance of IL-17A in Tumor-Infiltrating Lymphocytes of Non-Small Cell Lung Cancer Patients.

Objective: To investigate the distribution of IL-17A and its clinical significance in tumor infiltrating lymphocytes (TILs) of patients with non-small cell lung cancer (NSCLC). Methods: Expression level of IL-17A in TILs of 3 paired NSCLC and paracancerous specimens was measured by qRT-PCR. The distribution of IL-17A in immune cell subsets of 15 paired NSCLC and paracancerous specimens was examined by flow cytometry. The correlation between IL-17A and clinical features of NSCLC was identified. Results: IL-17A was significantly upregulated in TILs of NSCLC specimens than those of paracancerous ones (p < 0.0001). Meanwhile, T helper 17 cells (Th17 cells, p < 0.001), IL-17-secreting CD8+ T cells (Tc17 cells, p < 0.001) and IL-17-producing cells (γδT17 cells, p < 0.0001) were significantly abundant in TILs of NSCLC specimens than those of controls, and the higher abundance of the latter was much pronounced than that of the former two. Moreover, γδT17 cells in TILs were significantly correlated with lymphatic metastasis and CYFRA 21-1 level of NSCLC patients (p < 0.05). Conclusion: Tumor infiltrated γδT cells are the main source of IL-17 in early-stage NSCLC, and IL-17 may be a vital regulator involved in the development of NSCLC.

Thermosensitive Liposomal Nanoplatform Based on Metal-Free Phthalocyanine with Copper(II)-Regulated Photoactivities.

This work reports the development of a multifunctional thermosensitive liposomal nanoplatform (PcS4 @Lip-FA) based on a metal-free phthalocyanine modified with tetra-sulfonates (PcPS4 ), which exhibited photodynamic and photothermal activities simultaneously. Upon irradiation with a near infrared laser, thermosensitive PcS4 @Lip-FA could release PcS4 as a result of the local hyperthermia of PcS4 . Interestingly, PcS4 could easily chelate with Cu2+ , leading to the enhancement of photothermal activity and decrease of photodynamic activity. In addition, in vivo fluorescence imaging revealed that PcS4 @Lip-FA could selectively accumulate in tumor tissue of H22 tumor-bearing mice after tail vein injection, and exhibited a significant anticancer phototherapeutic effect, with a tumor inhibition rate of 83.5 %. Therefore, PcPS4 @Lip-FA has realized fluorescence imaging-guided combined cancer treatment, providing a promising multifunctional nanoplatform for cancer diagnostics and therapy.

Diagnostic performance of ESUR scoring system for extraprostatic prostate cancer extension: A meta-analysis.

PURPOSE: We aimed to evaluate the diagnostic performance of the European Society of Urogenital Radiology (ESUR) scoring system for detection of extraprostatic extension (EPE) in prostate cancer (PCa) by performing a meta-analysis. MATERIALS AND METHODS: A literature search of MEDLINE, EMBASE, Cochrane Library, Web of Science, and Google Scholar was performed to identify relevant studies from January 2012 to December 2020. We included diagnostic accuracy studies using ESUR scoring system for detection of EPE, and with prostatectomy histopathological results as the reference standard. Quality assessment was performed with the Quality Assessment of Diagnostic Accuracy Studies-2 tool. The summary estimates of sensitivity and specificity were pooled using bivariate random-effects modeling. We conducted multiple subgroup analyses and meta-regression to explore varied clinical settings. RESULTS: 10 studies with a total of 1698 participants were included in this meta-analysis. Pooled sensitivity and specificity were 0.71 (95% CI 0.61-0.80) and 0.76 (95% CI 0.67-0.84), respectively, with the area under ROC of 0.80 (95% CI 0.77-0.84). The Higgins I2 statistics demonstrated substantial heterogeneity in both sensitivity (I2 = 86.5%) and specificity (I2 = 91.6%), meta-regression revealed that the cutoff values (ESUR score ≥ 3 vs. ESUR score ≥ 4, P = 0.02) and malignancy rate (<40% vs. ≥40%, P = 0.04) were significant factors responsible for heterogeneity. Using endorectal coil and higher field strength (3.0 T) showed no additional benefit for EPE detection. CONCLUSION: The evidence available for ESUR scoring system tends to show moderate diagnostic performance for detection of EPE, and the cutoff values (P = 0.02) and malignancy rate (P = 0.04) were significant factors contributed to the heterogeneity.

Clinical laboratory features of Meigs' syndrome: a retrospective study from 2009 to 2018.

Meigs' syndrome (MS), a rare complication of benign ovarian tumors, is easily misdiagnosed as ovarian cancer (OC). We retrospectively reviewed the clinical laboratory data of patients diagnosed with MS from 2009 to 2018. Serum carbohydrate antigen 125 and HE4 levels were higher in the MS group than in the ovarian thecoma-fibroma (OTF) and healthy control groups (all P < 0.05). However, the serum HE4 levels were lower in the MS group than in the OC group (P < 0.001). A routine blood test showed that the absolute counts and percentages of lymphocytes were significantly lower in the MS group than in the OTF and control groups (all P < 0.05). However, these variables were higher in the MS group than in the OC group (both P < 0.05). The neutrophil-to-lymphocyte ratio (NLR) was also significantly lower, whereas the lymphocyte-to-monocyte ratio was higher in the MS group than in the OC group (both P < 0.05). The NLR, platelet-to-lymphocyte ratio, and systemic immune index were significantly higher in the MS group than in the OTF and control groups (all P < 0.05). The hypoxia-inducible factor-1 mRNA levels were also significantly higher, whereas the glucose transporter 1, lactate dehydrogenase, and enolase 1 mRNA levels were lower in peripheral CD4+ T cells obtained preoperatively in a patient with MS than those in patients with OTF, patients with OC, and controls (all P < 0.05). The expression of these four glucose metabolism genes was preferentially restored to normal levels after the tumor resection of MS (P < 0.001). These clinical laboratory features can be useful in improving the preoperative diagnostic accuracy of MS.

The oncogenic and prognostic role of PDK1 in the progression and metastasis of ovarian cancer.

Ovarian cancer (OC) is the most lethal of gynecological tumors in women. Tumor metabolism has become a new opportunity in the treatment of tumors. Pyruvate dehydrogenase kinase 1 (PDK1), as a key regulatory enzyme implicated in metabolic reprogramming of tumors, abnormally high expressed in various tumors and involved in the regulation of tumor cell biological behavior. However, the role of PDK1 in the occurrence and development of ovarian cancer remains unclear. Our team identified the expression of PDK1 in ovarian cancer cell lines and tissues through RT-PCR and immunohistochemical staining and evaluated the correlation of PDK1 expression with clinicopathologic features of patients and survival analyses. We used a variety of in vitro experiments to explore the influence of PDK1 on proliferation, invasion, migration, colony formation, apoptosis and the cell cycle of ovarian cancer cell lines CAOV3 and SKOV3. PDK1 was highly expressed in ovarian cancer cell lines and OC tissues. High expression of PDK1 was closely correlated to tumor size, FIGO stage, extraovarian metastases status and distribution. Univariate and multivariate Cox regression analysis identified that PDK1 was an independent prognostic factor for overall survival. Moreover, PDK1 was a superior predictor in prognosis of ovarian cancer and the incorporation of CA125 into PDK1 generated a predictive combination that displayed better predictive accuracy for overall survival. Downregulation of PDK1 suppressed the biological behavior of ovarian cancer cells due to S phase arrest and cellular apoptosis. PDK1 may serve as a novel prognostic biomarker, even a promising antineoplastic target of ovarian cancer.

Diagnostic Performance of Extraprostatic Extension Grading System for Detection of Extraprostatic Extension in Prostate Cancer: A Diagnostic Systematic Review and Meta-Analysis.

PURPOSE: To evaluate the diagnostic performance of the extraprostatic extension (EPE) grading system for detection of EPE in patients with prostate cancer (PCa). MATERIALS AND METHODS: We performed a literature search of Web of Science, MEDLINE (Ovid and PubMed), Cochrane Library, EMBASE, and Google Scholar to identify eligible articles published before August 31, 2021, with no language restrictions applied. We included studies using the EPE grading system for the prediction of EPE, with histopathological results as the reference standard. The pooled sensitivity, specificity, positive likelihood ratio (LR+), negative likelihood ratio (LR-), and diagnostic odds ratio (DOR) were calculated with the bivariate model. Quality assessment of included studies was performed using the Quality Assessment of Diagnostic Accuracy Studies-2 tool. RESULTS: A total of 4 studies with 1,294 patients were included in the current systematic review. The pooled sensitivity and specificity were 0.82 (95% CI 0.76-0.87) and 0.63 (95% CI 0.51-0.73), with the area under the hierarchical summary receiver operating characteristic (HSROC) curve of 0.82 (95% CI 0.79-0.85). The pooled LR+, LR-, and DOR were 2.20 (95% CI 1.70-2.86), 0.28 (95% CI 0.22-0.36), and 7.77 (95% CI 5.27-11.44), respectively. Quality assessment for included studies was high, and Deeks's funnel plot indicated that the possibility of publication bias was low (p = 0.64). CONCLUSION: The EPE grading system demonstrated high sensitivity and moderate specificity, with a good inter-reader agreement. However, this scoring system needs more studies to be validated in clinical practice.

Glucose metabolism characteristics and TLR8-mediated metabolic control of CD4+ Treg cells in ovarian cancer cells microenvironment.

Immunotherapy is expected to become the most promising new treatment for ovarian cancer owing to its immunogenicity. However, immunosuppression in the tumor microenvironment is a major obstacle to the efficacy of tumor therapy. Studies have found different metabolism ways of regulatory T cells (Tregs) in the cancer environment may be related to the immunosuppression and Toll-like receptor 8 (TLR8) can reverse the suppression function of Tregs. But it is still unclear that if the TLR8-mediated function reversal is associated with the change of glucose metabolism of Tregs. It was found that the positive expression rates of Glut1, HIF-1α, and Ki67 in CD4+ Treg cells of OC were significantly higher than that in benign ovarian tumor and HC, and also significantly higher than that in CD4+ Teffs of OC. What's more, compared with CD4+ Teff group, CD4+ Tregs highly expressed seven genes and three proteins related to glucose metabolism and had higher levels of glucose uptake and glycolysis. After activating TLR8 signal of CD4+ Tregs, the proliferation level of naive CD4+ T cells was higher than that of the control group. At the same time, the expression levels of eight genes and five proteins related to glucose metabolism in CD4+ Treg cells with TLR8 activated were decreased and levels of glucose uptake and glycolysis were also lower. Furthermore, TLR8 signaling also downregulated the mTOR pathway in CD4+ Tregs. CD4+ Tregs pretreated with 2-deoxy-d-Glucose (2-DG) and galloflavin also attenuated the inhibition of Teffs proliferation. Although CD4+ Tregs pretreated with 2-DG and galloflavin before activating TLR8 signal had no significant difference compared with the group only treated with inhibitors, which suggested TLR8-mediated reversal of CD4+ Treg cells inhibitory function in ovarian cancer cells co-cultured microenvironment had a causal relationship with glucose metabolism.

Implication of IL-17 producing ɑßT and γδT cells in patients with ovarian cancer.

OBJECTIVE: To investigate the expression and cellular source of IL-17A in human ovarian cancer (OC), benign ovarian tumor (BOT) and borderline ovarian tumor (OBT). METHODS: RT-PCR and immunohistochemistry were used to measure the expression level of IL-17A in human OC tissues. Find concrete source of the elevated IL-17A levels in OC tissues by flow cytometry. RESULTS: We found that IL-17A is expressed at higher levels in OC tissues than in BOT or OBT tissues at both the mRNA and protein levels. Moreover, high tumor IL-17A expression was significantly associated with poor tumor differentiation and positive lymph node status. Flow cytometric analysis demonstrated that significantly higher proportions of tumor-infiltrating IL-17A-producing CD4+ T cells (Th17), CD8+ T cells (Tc17), and γδT cells (IL-17+ γδT) were present in OC tissue compared with BOT tissue. Of these, tumor-infiltrating γδT cells were the predominant source of IL-17A in OC and BOT patients. Finally, we found that the abundance of tumor-infiltrating IL-17+ γδT cells, but not Th17 or Tc17 cells, was positively correlated with larger tumor size and lymph node metastasis in patients with advanced OC. These data suggest that increased tumor-infiltrating IL-17+ γδ T cells may be associated with cancer progression in OC patients.

Ovarian Cancer Cells Promote Glycolysis Metabolism and TLR8-Mediated Metabolic Control of Human CD4+ T Cells.

An immunosuppressive microenvironment is a major obstacle for successful tumor immunotherapy. Elucidating the regulatory mechanisms of energy metabolism and functionality in CD4+ T cells will provide insights for the development of novel immunotherapies for ovarian cancer (OC). An Agilent microarray was used to detect differences in gene expression between peripheral CD4+ T cells from five OC patients and those from five healthy controls. Functional pathway analysis was performed for differentially expressed genes. Gene expression profiles revealed significant differences in expression levels of 5,175 genes in peripheral CD4+ T cells from five patients with OC. Functional analysis indicated that the most significantly enriched pathways were metabolic pathways. Furthermore, eight glycolysis-related genes all showed significantly increased expression in peripheral CD4+ T cells of OC patients. Moreover, we established a coculture system of human CD4+ T cells with the OC cell line SKOV3, and then treated them with toll-like receptor 8 (TLR8) ligand ssRNA 40. Coculturing with SKOV3 cells could increase the expression of the eight glycolysis-related genes, promote glucose uptake and glycolysis in CD4+ T cells, induce the differentiation of CD4+ CD25+ Foxp3+ T cells, and enhance the suppression of naïve CD4+ T cells. Additionally, activated TLR8 signaling could mediate the reprogramming of glycolysis metabolism and function in CD4+ T cells. Overall, our study indicates that the SKOV3 coculture environment could regulate the glycolysis metabolism and function of CD4+ T cells, and also that TLR8 mediated the metabolic control of glycolysis in CD4+ T cells cocultured with SKOV3 cells. This provides a new direction for immunotherapy investigations in OC.

In Silico Screening of Circulating MicroRNAs as Potential Biomarkers for the Diagnosis of Ovarian Cancer.

Current screening tests for the diagnosis of ovarian cancer (OC) face enduring challenges. However, microRNAs (miRNAs) are stable in the circulation and may be promising molecular biomarkers for OC prediction. Circulating miRNA expression profiles in OC were analyzed using sequencing data from the Gene Expression Omnibus database. Differentially expressed miRNAs were generated from GSE94533, of which some were selected as candidate miRNAs based on an electronic search of the literature and comprehensive evaluation. A meta-analysis was preformed to integrate an evaluation index for these miRNAs in diagnosing OC patients. An independent validation set (GSE106817) was also conducted to further confirm the roles of these miRNAs. We identified four MIR200 members (MIR200A, MIR200B, MIR200C, and MIR429) and MIR25 as being differentially expressed among malignant or benign ovarian tumor patients and healthy controls. In the meta-analysis, these five miRNAs yielded a pooled area under the receiver operating characteristic (ROC) curve (AUC) of 0.78 (sensitivity: 64%, specificity: 88%) in discriminating OC from healthy controls, while the four MIR200 members demonstrated a summary AUC of 0.81 (sensitivity: 92%, specificity: 69%) in differing OC cases from patients with benign disease. In the validation set, differential expression and ROC curve analyses of these miRNAs were consistent except for MIR25. The circulating MIR200 family has the potential to become reliable and noninvasive biomarkers for OC diagnosis. Studies with larger cohorts are warranted to validate the applicability of these miRNAs.

Aberrant Peripheral Immune Function in a Good Syndrome Patient.

Good's syndrome (GS) is often accompanied by recurrent respiratory infections and chronic diarrhea. The main purpose was to evaluate the peripheral immune status of a GS patient after thymoma resection. Twenty healthy volunteers were recruited as healthy controls (HCs). Flow cytometry was applied to determine the proportions of circuiting CD4+ T cells, CD8+ T cells, γδT cells, and regulatory T (Treg) cells in our GS patient. We also examined the proliferation capability of ex vivo CD4+ T cells and detected the levels of cytokines interferon- (IFN-) γ and interleukin-17A secreted by ex vivo immune cells from this GS patient. Compared with healthy control subjects, this GS patient had fewer B cells, an inverted ratio of CD4+/CD8+ cells, and more Treg cells in his peripheral blood. Additionally, the patient's Vδ2 T cell levels were significantly decreased despite having a normal percentage of γδT cells. Ex vivo peripheral CD4+ T cells from the patient showed insufficient proliferation and division potential as well as excessive expression of PD-1. Moreover, IFN-γ was predominantly derived from CD8+ T cells in this GS patient, rather than from CD4+ T cells and γδT cells. This GS patient had impaired T and B cell immunological alternations and cytokine disruptions after thymectomy. Detailed research should focus on therapies that can adjust the immune status in such patients for a better outcome.