Toll-like Receptor 4 Expression In Patients Research Articles

Prognostic and clinicopathological significance of TLR4 expression in patients with breast cancer: a meta-analysis.

The prognostic value of Toll-like receptor 4 (TLR4) in breast cancer remains to be determined. Therefore, this paper aims to conduct a meta-analysis to assess the correlation between TLR4 and clinicopathological indicators as well as survival outcomes in breast cancer. Related literature retrieved from Embase, PubMed, Cochrane Library, Web of Science, China National Knowledge Infrastructure (CNKI) and China Wanfang. The search deadline is April 12, 2023. The outcome measures employed in the study comprised hazard ratio (HR), odds ratio (OR), and 95% confidence interval (CI) as effective indices. The data analysis was conducted using Stata 17.0 software. High TLR4 expression was associated with lymph node metastasis (OR=2.077, 95%CI=1.160-3.717, P= 0.014), tumor size (≥2 cm) (OR=2.194, 95%CI= 1.398-3.445, P= 0.001), PR expression (OR = 0.700, 95% CI = 0.505-0.971, P= 0.033), and clinical stage (OR = 3.578, 95%CI= 3.578-5.817, P<0.05), but not with histological grade (95%CI= 0.976-1.735, P= 0.072), ER expression (OR = 1.125, 95% CI = 0.492-2.571,P= 0.781), and HER-2 status (OR = 1.241, 95% CI = 0.733-2.101, P = 0.422). In addition, TLR4 overexpression was an independent prognostic indicator of DFS (HR= 1.480, 95%CI= 1.028- 2.130, p= 0.035) in breast cancer patients, but not related to OS(HR=1.730, 95%CI= 0.979-3.057, P= 0.059). From our main analysis results, high TLR4 expression is associated with lymph node metastasis, larger tumor size (≥2 cm), later clinical stage, negative PR expression and shorter DFS, suggesting poor prognosis in breast cancer patients.

Differential expression of TLR4 in patients of coronary artery disease with and without traditional cardiovascular risk factors

Abstract Funding Acknowledgements Type of funding sources: None. Background Atherosclerosis is inherently an inflammatory process, with a complex interplay of inflammatory markers. It is established that these inflammatory markers play an important role in patients of Coronary Artery Disease(CAD) with traditional cardiovascular risk factors. However, the role of inflammation in the atherosclerotic process in patients of CAD without traditional risk factors is still not clearly known. Purpose Our purpose was to determine whether in patients of CAD without traditional risk factors, TLR4(Toll like receptor 4) expression as a marker of inflammation is similar to that in patients of CAD with traditional risk factors. Materials and Methods This observational cross sectional study was done between July’20 to Dec’21. Equal number of patients of CAD with and without traditional cardiovascular risk factors undergoing Coronary Artery Bypass Grafting (CABG) were enrolled. The risk factors considered were Hypertension, Diabetes, Dyslipidemia, Obesity and Addiction to Tobacco and/or Alcohol. A minimum of two punch biopsy samples of aortic tissue was taken from each subject undergoing CABG. Immunohistochemistry for TLR4, was done in Ventana BenchMark GX System. The primary TLR4 antibody was procured from reputed source. Results The presence or absence of TLR4 expression was associated significantly with the Syntax scores (37.40±4.74 vs 29.5±8.71; p value=0.036), total Cholesterol (187±35.06mg/dL vs 130±35.69 mg/dL ;p value 0.010) and LDL cholesterol (118.86±28.12mg/dL vs 64.21±25.61 mg/dL; p value 0.003). TLR4 expression, however, was not significantly associated with the number of coronary vessels involved (p=0.298). TLR4 expression was also not significantly associated with any other individual risk factors. However, when the traditional risk factors were considered in combination, TLR4 expression was associated significantly with the number of risk factors present(p=0.029) the strongest being in those having 4 traditional risk factors. The level of TLR4 expression gradually declined with the decrease in the number of risk factors, having mostly weak or negative expression in patients without any traditional risk factors. Conclusion Patients with CAD without any traditional risk factors, had a less severe coronary artery disease as manifested by lower Syntax scores, and had lower degree of TLR4 expression. Patients with CAD with traditional risk factors had more severe coronary artery disease as evidenced by higher Syntax scores, and had higher degree of TLR4 expression proportional to the number of traditional risk factors present. Thus, differential TLR4 expression in CAD patients with and without traditional risk factors indicated a difference in inflammatory state between the two groups and warrants further investigation.

High TLR4 expression predicts a poor prognosis after esophagectomy for advanced thoracic esophageal squamous cell carcinoma.

Poor oral health is an independent risk factor for upper aerodigestive tract cancers, including esophageal squamous cell carcinoma (ESCC). The pattern recognition receptor Toll-like receptor 4 (TLR4) recognizes lipopolysaccharide in the cell walls of Gram-negative periodontal pathogens associated with the development and progression of ESCC. It is, therefore, plausible that TLR4 plays a crucial role in the pathogenesis of ESCC. We used an ESCC tissue microarray to confirm expression of TLR4 in patients with ESCC and to determine whether TLR4 expression status correlates with the clinicopathological features of these patients or their prognosis after esophagectomy. We also tested whether the combined expression statuses of TLR4 and TLR3 better correlate with prognosis in these patients than either parameter alone. Clinical ESCC samples from all 177 patients tested showed expression of TLR4. Moreover, high TLR4 expression (3 + and 2 +) correlated with poorer 5-year overall survival after esophagectomy than lower TLR4 expression (1 +) (p = 0.0491). Patients showing high TLR4 expression tended to have a poorer prognosis whether treated with surgery alone or with surgery and adjuvant chemotherapy. Univariate and multivariate analyses showed TLR4 expression status to be an independent prognostic factor affecting 5-year overall survival. Patients exhibiting high TLR4 expression with low TLR3 expression had a much poorer prognosis than other patients (p = < 0.0001). High TLR4 expression predicts a poor prognosis in advanced thoracic ESCC patients after esophagectomy.

Association between small intestinal bacterial overgrowth and toll-like receptor 4 in patients with pancreatic carcinoma and cholangiocarcinoma

Multiple factors have been linked to pathogenesis of pancreatic cancer and cholangiocarcinoma. Until now, few studies have investigated the role of small intestinal bacterial overgrowth (SIBO) and toll-like receptor 4 (TLR-4) signaling in these diseases. This study aimed to examine the relationship between the prevalence of SIBO and the TLR-4 expression in patients with pancreatic carcinoma and cholangiocarcinoma. A total of 90 human subjects suffering from pancreatic carcinoma (n=30), cholangiocarcinoma (n=30), and healthy controls (n=30) were enrolled in the study. A glucose hydrogen breath test (GHBT) was used to evaluate SIBO. The TLR4 protein expression was measured by immunohistochemistry (IHC). The positive rate of SIBO was 63.3% in the pancreatic cancer group and 46.7% in patients with cholangiocarcinoma, which was significantly greater than 13.3% in the healthy control group (p<0.05). An IHC analysis revealed that the TLR-4 protein expression in the SIBO-positive pancreatic carcinoma patients was significantly higher than that in the SIBO-negative patients (p<0.05), and the same result was in the cholangiocarcinoma subjects. In addition, a correlation analysis identified the positive relationship between the prevalence of SIBO and the TLR-4 protein expression in pancreatic carcinoma (r=0.489), and the same result was in the cholangiocarcinoma subjects. Our findings indicate a high prevalence of SIBO in pancreatic carcinoma and cholangiocarcinoma, and SIBO displays a positive correlation with the TLR-4 expression, suggesting that SIBO could be a risk factor for the pathogenesis of pancreatic carcinoma and cholangiocarcinoma, in which the TLR4 signaling may be involved.

Influence of simvastatin treatment on Toll-like receptor 4 in monocytes of peripheral blood in patients with sepsis and severe sepsis

To investigate the influence of simvastatin treatment on Toll-like receptor 4 (TLR4) in monocytes of peripheral blood in patients with sepsis and severe sepsis and its significance. A prospective randomized controlled trial was conducted. 106 patients with sepsis and 92 patients with severe sepsis admitted to Department of Critical Care Medicine of Henan Provincial People's Hospital from August 2013 to June 2015 were enrolled. These two groups of patients were randomized into conventional treatment group and simvastatin group. All patients received treatment according to the 2012 International Sepsis Treatment Guidelines, including anti-infection drugs, nutritional support, and palliative treatment, and the patients with severe sepsis were given early goal-directed therapy (EGDT). The patients in simvastatin group received simvastatin 40 mg daily orally for at least 15 days. The peripheral blood was collected and the monocytes were isolated at 1, 5, 10, 15 days after intensive care unit (ICU) admission. TLR4 expression on the surface of TLR4/CD14(+) double positive monocytes was determined by flow cytometry, and adverse reaction was observed during treatment. TLR4 expression on the surface of monocytes showed a tendency of decreasing with prolongation of simvastatin treatment in the simvastatin group in patients with sepsis (n = 59) or severe sepsis (n = 54). However, in patients with sepsis, TLR4 level was significantly decreased from 10 days in simvastatin group as compared with that of conventional therapy group (n = 47), and it was decreased up to 15 days [mean fluorescence intensity (MFI): 21 (19, 28) vs. 27 (25, 33) at 10 days, Z = 2.198, P = 0.021; 16 (15, 21) vs. 26 (23, 34) at 15 days, Z = 4.611, P = 0.002]. In patients with severe sepsis, there was no significant difference in TLR4 level at different time points between simvastatin group and conventional treatment group (n = 38) [MFI: 55 (52, 63) vs. 56 (48, 65) at 1 day, Z = 0.313, P = 0.692; 47 (42, 56) vs. 49 (41, 58) at 5 days, Z = 0.827, P = 0.533; 40 (35, 42) vs. 42 (37, 45) at 10 days, Z = 1.012, P = 0.301; 33 (30, 38) vs. 38 (35, 41) at 15 days, Z = 0.539, P = 0.571]. No adverse reaction related with simvastatin was found during treatment in patients with sepsis or severe sepsis. Statins could significantly down-regulate the TLR4 expression on peripheral blood monocytes in septic patients, while it showed no significant influence on TLR4 expression in patients with severe sepsis. A different effect of statins on TLR4 expression and the downstream inflammation process in sepsis and severe sepsis patients might partially explain the discrepancy in previous reports about the therapeutic effect of statins therapy in sepsis and severe sepsis patients.

Knockdown of toll-like receptor 4 inhibits human NSCLC cancer cell growth and inflammatory cytokine secretion in vitro and in vivo.

Toll-like receptor 4 (TLR4)-mediated signaling has been implicated in tumor cell invasion, survival and metastasis in several types of cancers. However, the expression of TLR4 in patients with non-small cell lung cancer (NSCLC) and its biological function in the development and progression of NSCLC have not been elucidated to date. Here, we sought to characterize the expression of TLR4 in patients with NSCLC and to investigate the biological roles of TLR4 in lung metastasis, cell invasion and survival. In this study, we found that TLR4 expression was elevated in most patients with NSCLC, and its expression levels correlated with key pathological characteristics, including tumor differentiation, stage and metastasis. Our data also showed that downregulation of TLR4 expression using an RNA silencing approach in A549 tumor cells significantly suppressed cell proliferation, cell migration and cell invasion, and induced tumor apoptosis in vitro, and suppressed tumor growth in vivo. In addition, we also found that downregulation of TLR4 expression significantly decreased cell TNF-α and IL-6 levels. Furthermore, we found that knockdown of TLR4 was able to significantly suppress constitutive phosphorylation of Akt and PI3K, which may contribute to the inhibition of tumor growth. These data suggest that TLR4 plays an important role in tumorigenic properties of human NSCLC, and that RNA interference-directed targeting of TLR4 could be used as a potential anticancer therapeutic target in NSCLC.

Toll-like receptor-4 expression by hepatic progenitor cells and biliary epithelial cells in HCV-related chronic liver disease

Notwithstanding numerous evidences implicating toll-like receptor-4 (TLR4) in the pathogenesis of chronic hepatitis C virus (HCV) infection, the localization and level of TLR4 expression in the liver of patients with hepatitis C have never been investigated. We aimed to evaluate, by means of immunohistochemistry and real-time PCR (rt-PCR), hepatic TLR4 expression in patients with chronic HCV infection. Fifty patients who had undergone liver biopsy and 11 patients transplanted because of chronic HCV infection, and 12 controls free of liver disease, were included in the study. Each case was analyzed by immunohistochemistry for TLR4, α–smooth muscle actin and cytokeratin-7 (CK-7), and a subgroup of patients and all controls by rt-PCR for TLR4. Immunohistochemistry for α-smooth muscle actin was used to derive a score of activation of hepatic stellate cells and portal/septal myofibroblasts, while immunohistochemistry for CK-7 was used to evaluate and count hepatic progenitor cells, interlobular bile ducts and intermediate hepatocytes. In patients, the parenchymal elements responsible for the highest TLR4 level of expression were hepatic progenitor cells and biliary epithelial cells of interlobular bile ducts. Double-labeling experiments between anti-TLR4 and anti-CK7, anti-CD133, anti-CD44, anti-neural cell adhesion molecule, anti-epithelial cell adhesion molecule and anti-sex determining region Y-box 9, confirmed these findings. TLR4-positive hepatic progenitor cells and interlobular bile ducts were significantly correlated with the stage of liver disease (P<0.001), the grade of inflammation (P<0.001), and the activity of portal/septal myofibroblasts (P<0.001). rt-PCR study confirmed an increased TLR4 expression in the 26 patients analyzed with respect to controls (P<0.001). TLR4 expression positively correlated with fibrosis (P<0.05) and inflammation (P<0.05). The present results suggest that TLR4 expression by hepatic progenitor cells and biliary epithelial cells contributes to the progression of liver damage in the course of chronic HCV-related infection.

Toll-like receptor-4 expression by hepatic progenitor cells and biliary epithelial cells in HCV-related chronic liver desease

Background . Toll-like receptor-4 (TLR4) is a transmembrane pattern recognition receptor that plays a key role in innate immunity by triggering inflammatory responses to Gram negative bacteria lipopolysaccharide (LPS) (1). Since liver is the main clearance organ for LPS, which is excreted in large amounts in the bile (2), it is not surprising that TLR4 has been involved in the pathogenesis of most liver diseases (3). Numerous evidences suggest a role for TLR4 in the pathogenesis of chronic hepatitis C virus (HCV) infection (4) and hepatic fibrosis (5), but the localization and level of TLR4 expression in the liver of patients with hepatitis C have never been investigated. Aim and methods. We aimed to evaluate, by means of immunohistochemistry (IHC) and real-time polymerase chain reaction (rt-PCR), hepatic TLR4 expression in patients with chronic HCV infection. Sixty-one patients with chronic HCV infection, and 12 controls free of liver disease, were included in the study. Each case was analyzed by IHC for TLR4, α–smooth muscle actin (αSMA) and cytokeratin-7 (CK-7), and a subgroup of patients and all controls by rt-PCR for TLR4. A score of activation of portal/septal myofibroblasts and lobular hepatic stellate cells (HSCs) was evaluated by IHC for α-SMA, whereas IHC for CK-7 was analysed in order to count hepatic progenitor cells (HPCs), interlobular bile ducts and intermediate hepatocytes. Results . The parenchimal elements responsible for the highest TLR4 level of expression were HPCs and biliary epithelial cells (BECs) of interlobular bile ducts in the infected group. Double-labeling experiments with anti-TLR4, anti-CK7 and anti-CD133 confirmed this finding. TLR4-positive HPCs and interlobular bile ducts were significantly correlated with the stage of liver disease (p<0.001), the grade of inflammation (p<0.001), and with the activity of portal/septal myofibroblasts (p<0.001). Rt-PCR study confirmed an increased TLR4 expression in the 26 patients analyzed with respect to controls (p<0.001). TLR4 expression positively correlated with fibrosis (p<0.05) and inflammation (p<0.05). Conclusions . The expression of TLR4 in HPCs and BECs in HCV-related liver damage significantly correlates with inflammation, activation of portal/septal myofibroblasts and fibrosis. 1) Beutler. Nature 2004;430:257-63; 2) Van Bossuyt et al. J Hepatol 1988;7:325-37; 3) Seki et al. Hepatology 2008;48:322-35; 4) Machida et al. J Virol 2006;80:866-74. 5) Seki et al. Nat Med 2007;13:1324-32.

Elevated Toll-like receptor 4 (TLR4) in hepatocytes in patients with chronic hepatitis B and its clinical significance

Objective To investigate the role of TLR4 in the pathogenesis of chronic hepatitis B(CHB) by study the expression of TLR4 in liver tissues in patients with CHB, and the relationship among TLR4 and serum HBV DNA level, clinical severity degrees and histo-logical grades and stages. Methods Expression of TLR4 in liver tissues was semi-quantitatively determined by immunohistochemistry and e-valuated by a scoring system in 75 patients with CHB and 10 health controls. Results The positive staining of TLR4 mainly located in the cytoplasm and some on cell membrane of bepatocytes. Expression of TLR4 in the liver tissues of patients with CHB was stronger than that of health controls. The scores of TLR4 expression in patients with mild, moderate and severe CHB were 1.0±0.5,2.3±0.5 and 2.9±0.4. The scores increased gradually and significantly along with the increase of clinical severity degrees( F = 104.8, P<0.01). The scores of TLR4 expression in the liver tissues of patients with CHB were positively correlated with the clinical severity degrees (r=0.838, P<0.01) and histological grades (r=0.579, P<0.05), but not correlated with Lg (serum HBV DNA) or histological stages. Conclusion TLR4 was up-regulated in the hepatocytes of patients with CHB. There may be a role of TLR4 in the pathogenesis of CHB. Key words: Hepatits B, chronic; Receptors, cell surface