Clinical Implication of Toll-Like Receptors (TLR2 and TLR4) in Acute Myeloid Leukemia Patients

Backgrounds: Toll-like receptors 2; 4 (TLR2;4) are an essential component of the innate immunity and play an important role in immune-surveillance and immune response to various microorganisms. This study aimed to investigate the association between TLR2 and TLR4 polymorphism and the risk of acquiring severe infections, and impact on AML patient’s outcome. Subjects and methods: Using polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP); we analyzed three SNPs in the TLR2 (Arg753Gln) and TLR4 (Asp299Gly and Thr399Ile) in 120 AML patients and 100 healthy control subjects. Results: No significant differences in genotype or alleles frequency between healthy controls and AML patients regarding TLR2 Arg753Gln, TLR4 Asp299Gly and TLR4 Thr399Ile polymorphisms (P>0.05 for all). Neutropenic fever was detected in 110 out of 120 (91.7%) of the studied AML patients. The sepsis and pneumonia were identified in 20 out of 120 patients (16.7%). The incidence of sepsis was associated with TLR2 Arg753Gln: AG genotypes, A allele and TLR4 Asp299Gly: CT genotype and C allele as compared to other genotypes and alleles. Moreover; TLR2 (Arg753Gln) GG polymorphisms significantly associated with shortest overall survival (OS) and shortest disease-free survival (DFS); while TLR4 polymorphisms affect the DSF only but not OS. In AML patients TLR2 Arg753Gln gene polymorphism is associated with high susceptibility to sepsis and TLR4 (Asp299Gly and Thr399Ile) gene polymorphism is associated with high susceptibility for both pneumonia; and sepsis. Conclusion: TLR2 Arg753Gln (AG; GG genotype) polymorphisms are associated with shortest OS and DFS. Moreover; significant association between TLR2 polymorphisms, TLR4 Arg753Gln polymorphisms and risk of severe infections in AML patients was documented.     

Clinical Implication of Toll-Like Receptors (TLR2 and TLR4) Polymorphisms in Adult Patients with Acute B-cell Lymphoblastic Leukemia

Backgrounds: Neutropenia after intensive chemotherapy of acute lymphoblastic leukemia (ALL) could lead to infectious complications that affect outcome of acute leukemia patients. Many single-nucleotide polymorphisms (SNPs) of Toll-like receptors (TLRs) can affect the genetic susceptibility to infections. We investigated the impact of different SNPs on the incidence of developing sepsis and pneumonia in patients with newly diagnosed B-ALL following induction chemotherapy. Subjects and methods: We analyzed three SNPs in the TLR2 (Arg753Gln) and TLR4 (Asp299Gly& Thr399Ile) genes using polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) in a case control study of 40 precursor B-ALL patients and 50 control subjects. The risk of developing sepsis and pneumonia were assessed by multiple logistic regression analyses. Results: The presence of the TLR-2 AG polymorphism was significantly associated with pneumonia in B-ALL patients. Furthermore, TLR4 Thr399Ile AG was a risk factor for sepsis in B-ALL patients. Moreover; Significant association between TLR-2 AA, TLR-4 CC and TL-4 AA genotypes and longer OS were detected in studied B-ALL patients. Conclusion: We concluded that TLR-4 (AG and CT) genotypes are associated with high susceptibility to sepsis and pneumonia respectively; while, TLR-2, TLR-4 AA and TLR-4 CC genotypes  could predict good B-ALL patients outcome.     

Association between Toll-like receptor gene cluster (TLR6, TLR1, and TLR10) and prostate cancer.

BACKGROUND: Chronic inflammation may be a risk factor for prostate cancer. Previously, we found significant associations between single nucleotide polymorphisms (SNPs) and haplotypes in Toll-like receptor (TLR) 4 and the risk of prostate cancer. TLR6, TLR1, and TLR10 are also involved in the pathogen-mediated inflammation pathway. A Swedish study observed associations between sequence variants in the TLR6-TLR1-TLR10 gene cluster and the risk of prostate cancer. We assessed if genetic polymorphisms of this gene cluster were associated with the risk of prostate cancer in a U.S. population. METHODS: In a nested case-control design within the Health Professionals Follow-Up Study, we identified 700 participants with prostate cancer who were diagnosed after they had provided a blood specimen in 1993 and by January 31, 2000. Controls were 700 age-matched men without prostate cancer who had had a prostate-specific antigen test. We genotyped 19 common (>5%) haplotype-tagging SNPs chosen from the SNPs discovered in a resequencing study spanning TLR6, TLR1, and TLR10 to test for the association between sequence variants cluster and prostate cancer. RESULTS: Neither individual SNPs nor common haplotypes in the three gene regions were associated with altered risk of prostate cancer or subgroups of aggressive prostate cancer. No effect modification was observed for age, body mass index, or family history of prostate cancer, except that TLR6_3649 showed nominally significant interaction with family history at the P < 0.05 level. CONCLUSION: Inherited sequence variants of the innate immune gene cluster TLR6-TLR1-TLR10 were not appreciably associated with the risk of prostate cancer in this cohort.     

TLR2mRNA和TLR4mRNA在蕈样肉芽肿皮损中的表达

目的:探讨Toll样受体2(TLR2) mRNA和Toll样受体4(TLR4) mRNA在蕈样肉芽肿皮损中的表达水平.方法:采用原位杂交技术检测蕈样肉芽肿皮损和正常皮肤中TLR2 mRNA和TLR4 mRNA的表达.结果:蕈样肉芽肿皮损中TLR2 mRNA和TLR4 mRNA主要表达于表皮基底上层,正常皮肤则表达于表皮的基底层.统计学分析表明TLR2 mRNA和TLR4 mRNA在正常皮肤和蕈样肉芽肿皮损处基底层和基底上层的表达均有显著性差异(P＜0.05),且TLR2 mRNA和TLR4 mRNA在蕈样肉芽肿皮损中的表达均高于在正常皮肤中的表达(P＜0.05).结论:TLR2 mRNA和TLR4 mRNA可能在蕈样肉芽肿的发生与发展过程中发挥重要作用.     

胃癌及胃粘膜不典型增生中Fas蛋白表达及其意义

目的 探讨Ｆａｓ蛋白在胃癌形成与发展过程中作用及其意义。方法 采用免疫组化染色,对２０例正常胃粘膜,３８例胃粘膜不典型生,５２例胃癌进行Ｆａｓ蛋白检测。结果 胃粘膜不典型增生与胃癌的Ｆａｓ蛋白表达均显著高于正常胃粘膜（Ｐ均〈０．０５）。Ｆａｓ蛋白表达与胃癌分期,淋巴结转移无相关性。结论 Ｆａｓ蛋白与胃癌形成有关,与胃癌进展无关。     

膀胱癌细胞钟声蛋白样受体表达与卡介苗诱导

[目的]探讨膀胱癌细胞钟声蛋白样受体(TLRs)在卡介苗(BCG)诱导的抗膀胱癌免疫反应中的作用.[方法]以不同剂量(菌数/细胞数比值)的BCG和大肠杆菌分别刺激培养人膀胱癌T24细胞株.1小时后提取细胞总RNA,用半定量RT-PCR技术检测T24细胞中TLR-2和TLR-4的表达水平.12小时后提取各孔细胞的上清液,用ELISA试剂盒测定IL-4和IL-12的浓度.[结果](1)BCG刺激后T24细胞中TLR-2和TLR-4的表达水平及IL-12浓度随着BCG剂量增加而逐渐升高,在菌数/细胞数比值为10时达到最高,随后逐渐降低,但仍高于未刺激组.(2)BCG刺激后T24细胞中IL-4的浓度与未刺激组相比无明显差别.(3)大肠杆菌刺激后各剂量组T24细胞中TLR-2和TLR-4的表达水平以及IL-4和IL-12浓度与未刺激组相比无明显差别.[结论]膀胱癌细胞TLRs表达可能与BCG诱导的抗膀胱癌免疫反应有关.     

Clinicopathological and Prognostic Significance of MUC- 2, MUC-4 and MUC-5AC Expression in Japanese Gastric Carcinomas

Background: The mucin components of the gastric gel layer function as a protective and lubricatingfactor against luminal acid and proteolytic enzymes. Alteration of mucin expression in gastric preneoplasticand neoplastic lesions has suggested potential roles in neoplastic processes. This study aimed to assess theclinicopathological and prognostic significance of MUC-2, MUC-4 and MUC-5AC in Japanese gastric cancer.Methods: Expression of MUC-2, -4 and -5AC was evaluated on tissue microarrays of gastric carcinomas andadjacent non-cancerous mucosa specimens by immunohistochemistry and compared with clinicopathologicalparameters and survival time of the patients. Results: The three mucins were found to be expressed to a lesserextent in gastric carcinomas in comparison with non-cancerous mucosa (p<0.05). MUC-2 expression wasnegatively correlated with tumor size, depth of invasion, and TNM staging of gastric cancer (p<0.05), while thatof MUC-5AC was negatively associated with the depth of invasion, venous invasion, lymph node metastasis andTNM staging (p<0.05), but positively with MUC-4 and MUC-2 expression (p<0.05). There was higher MUC-2expression in intestinal- than diffuse-type carcinomas (p<0.05). Kaplan-Meier analysis indicated no relationshipbetween expression of the three mucins and the cumulative survival rate of patients, even stratified according tothe depth of invasion (p>0.05). Conclusion: Down-regulated expression of MUC-2, -4 and -5AC may be involvedin pathogenesis, invasion, metastasis or differentiation of gastric carcinoma. Their altered expression mighttherefore be employed as an indicator of pathobiological behavior.     

胃粘膜异型增生上皮p53蛋白表达的临床意义

采用免疫组化方法，对２４例常规胃镜活检的胃粘膜异型增生上皮的ｐ５３蛋白表达进行检测，并做１～２４个月的随访研究。结果表明，ｐ５３蛋白阳性的异型增生上皮的癌检出率为５７．１％（４／７），其中２例为早期胃癌，而ｐ５３阴性的异型增生上皮的癌检出率为１１．８％（２／１７），两者有显著性差异（Ｐ＜０．０５）。结果提示，胃粘膜上皮ｐ５３蛋白表达是一项恶性生物学指标，可作为早期诊断及鉴别诊断胃癌的标志     

胃癌组织中SFRP2和NDRG4的表达及其临床意义

目的探讨胃癌组织中SFRP2和NDRG4的表达及其临床意义。方法选取64例接受手术切除治疗的胃癌患者作为研究对象,取适量胃癌病灶组织作为观察组,距离肿瘤边缘5 cm以上的癌旁正常组织则为对照组。分别采用Western-blot和荧光定量PCR检测胃癌病灶组织和正常组织中SFRP2和NDRG4蛋白及mRNA含量。结果SFRP2和NDRG4在胃癌病灶组织和正常组织中均有表达,病灶组织中SFRP2和NDRG4灰度值分别为(0. 149±0. 044)和(0. 19±0. 038),明显低于正常组织中的(0. 557±0. 123)和(0. 740±0. 093);此外,胃癌病灶组织中SFRP2和NDRG4mRNA的相对表达量分别为(0. 331±0. 048)和(0. 276±0. 085),正常组织中的相对表达量则是(1. 056±0. 145)和(1. 043±0. 150)。胃癌病灶组织中的SFRP2和NDRG4蛋白含量及mRNA水平均显著低于正常组织,差异具有统计学意义(P <0. 05)。SFRP2和NDRG4联合检测敏感性和特异性分别为44. 2%和87. 6%,优于单独检测。结论胃癌中SFRP2和NDRG4表达同时下调并参与了其病理发展,联合测定上述肿瘤相关因子对提高胃癌诊断的特异性具有重要临床意义。     

Toll-like receptor 9 (TLR9) agonists in the treatment of cancer

Although still early in clinical development, agonists of Toll-like receptor 9 (TLR9) have demonstrated potential for the treatment of cancer. TLR9 agonists directly induce activation and maturation of plasmacytoid dendritic cells and enhance differentiation of B cells into antibody-secreting plasma cells. Preclinical and early clinical data support the use of TLR9 agonists in patients with solid tumors and hematologic malignancies. In preclinical studies, TLR9 agonists have shown activity not only as monotherapy, but also in combination with multiple other therapies, including vaccines, antibodies, cellular therapies, other immunotherapies, antiangiogenic agents, radiotherapy, cryotherapy and some chemotherapies. Phase I and II clinical trials have indicated that these agents have antitumor activity as single agents and enhance the development of antitumor T-cell responses when used as therapeutic vaccine adjuvants. The activity and safety of these novel anticancer agents are being explored in a wide range of tumor types as part of a variety of therapeutic strategies with the goal of harnessing the immune response to fight cancer.     

Increased Expression of P2RY2, CD248 and EphB1 in Gastric Cancers from Chilean Patients

Background: Gastric cancer (GC) ranks as one of the major causes of mortality due to cancer worldwide.In Chile, it is currently the leading cause of cancer death. Identification of novel molecular markers that mayhelp to improve disease diagnosis at early stages is imperative. Materials and Methods: Using whole-genomeDNA microarrays we determined differential mRNA levels in fresh human GC samples compared to adjacenthealthy mucosa from the same patients. Genes significantly overexpressed in GC were validated by RT-PCR ina group of 14 GC cases. Results: The genes CD248, NSD1, RAB17, ABCG8, Ephb1 and P2RY2 were detected asthe top overexpressed in GC biopsies. P2RY2, Ephb1 and CD248 showed the best sensitivity for GC detectionwith values of 92.9%, 85.7% and 64.3% (p<0.05), respectively. Specificity was 85.7%, 71.4% and 71.4% (p<0.05),for each respectively.     

Toll-like receptor 5 (TLR5) expression is a novel predictive marker for recurrence and survival in squamous cell carcinoma of the tongue

Background:

Toll-like receptor 5 (TLR5) is an immune receptor recognising bacterial flagellin. Activation of TLR5 results in cancer invasion and cytokine release. As certain bacteria have been linked to oral cancer, we wanted to study TLR5 expression in oral tongue squamous cell carcinoma (OTSCC).

Methods:

Samples from 119 patients with OTSCC were obtained, including 101 samples of adjacent normal lingual mucosa. The TLR5 histoscore (0–300) was assessed semiquantitatively by immunohistochemistry in a blinded manner.

Results:

Toll-like receptor 5 was expressed in 84 normal epithelia and 118 cancer samples. Expression of TLR5 was increased in cancer when compared with normal lingual epithelium (median histoscore 15 vs 135). In cancer, higher TLR5 was associated with age of >70 years at the time of diagnosis, female gender and disease recurrence. No association between TLR5 expression and tumour grade, stage or treatment was found. In multivariate analysis, TLR5 was an independent predictor of cancer mortality (hazard ratio (HR) 3.587, 95% confidence interval (CI) (1.632–7.882)) and disease recurrence (HR 4.455, 95% CI (2.168–9.158)).

Conclusion:

Toll-like receptor 5 has a previously undescribed role in the pathophysiology of OTSCC and might represent a link between bacteria and cancer. It could be a useful marker for predicting recurrence or survival of OTSCC patients.     

Toll样受体2/4在非哺乳期乳腺炎组织中的表达及意义

目的探讨Toll样受体2(TLR2)和Toll样受体4(TLR4)在不同病理类型及不同临床分期非哺乳期乳腺炎(NPM)中的表达及其意义。 方法本研究为回顾性研究。选取2012年1月至2017年1月在南京医科大学附属淮安第一医院行手术治疗的141例NPM患者及10例乳腺纤维瘤患者(对照组)的病理切片进行TLR2、TLR4表达水平的免疫组织化学检测。根据术后病理结果将NPM分为肉芽肿性乳腺炎(GM)59例，浆细胞性乳腺炎(PCM)50例，其他类型乳腺炎32例，并将其中GM、PCM与对照组进行对比研究。根据临床分期将141例患者分为急性期(21例)、亚急性期(72例)、慢性期(48例)，并与对照组进行对比研究。通过检测TLR2、TLR4在不同病理类型及不同临床分期NPM中的表达水平，同时结合临床资料进行统计分析。多组间TLR2、TLR4表达水平的比较采用单因素方差分析，方差整齐时两两比较采用LSD法，方差不齐时两两比较采用Tamhane’s法；GM与PCM患者临床特征的比较采用χ²检验。 结果GM组TLR2、TLR4表达水平分别为15.82±4.96和27.27±7.70，PCM组TLR2、TLR4表达水平分别为15.29±4.14和26.25±6.63，对照组TLR2、TLR4表达水平分别为6.12±0.81和6.40±1.18。3组相比，TLR2、TLR4表达水平的差异均有统计学意义(F＝21.613、39.746，P均<0.001)，其中GM、PCM组TLR2、TLR4表达水平均高于对照组(P均<0.050)。并且，急性期、亚急性期、慢性期NPM及对照组患者相比，TLR2、TLR4表达水平的差异均有统计学意义(F＝190.112、246.965，P均<0.001)，其中急性期NPM患者TLR2、TLR4表达水平分别为23.65±2.32和40.10±2.22，高于亚急性期NPM的12.35±2.44和23.14±4.56(P均<0.001)，也高于慢性期NPM的17.19±2.36和29.36±2.17(P均<0.001)。与PCM患者相比，GM患者下肢结节红斑的发生率较高[28.8%(17/59)比12.0%(6/50)，χ²＝4.596，P＝0.032]。 结论TLR2和TLR4在急性期NPM中显著高表达，TLR2、TLR4信号途径可能参与NPM的发生、发展；GM与PCM患者的下肢结节红斑发生率存在差异。     

Increased responsiveness to TLR2 and TLR4 ligands during dimethylsulfoxide-induced neutrophil-like differentiation of HL-60 myeloid leukemia cells

Neutrophils express functional toll-like receptor2 (TLR2) and 4 (TLR4) that are crucial for the production of inflammatory cytokines. Here, we show that dimethylsulfoxide-induced neutrophil-like differentiation of promyelocytic HL-60 cells (dHL-60) results in cells that respond to TLR2 and TLR4 ligands similarly to primary neutrophils. Consistent with the increased responsiveness of the cells to TLR2 ligand, the TLR2 gene was strongly up-regulated in dHL-60 cells. On the other hand, increased surface expression of LPS receptor complex, TLR4/MD2/CD14, was observed without affecting TLR4 gene expression. Thus, the data demonstrate a higher responsiveness of dHL-60 cells to TLR2 and TLR4 ligands because of increased TLR2 and MD2/CD14 gene expression.     

Clinical Significance of Ephrin (Eph)-A1, -A2, -A4, -A5 and -A7 Receptors in Pancreatic Ductal Adenocarcinoma

Ephrin (Eph) receptors have been reported to be frequently overexpressed in a wide variety of cancer types, being associated with tumor growth, invasion, metastasis and angiogenesis. The aim of the present study was to evaluate the clinical significance of Eph-A1, -A2, -A4, -A5 and -A7 expression in pancreatic ductal adenocarcinoma. Eph-A1, -A2, -A4, -A5 and -A7 expression and staining intensity were assessed immunohistochemically in tumoral samples of 67 pancreatic adenocarcinoma patients and were statistically analyzed in relation to clinicopathological characteristics, tumor proliferative capacity and patients’ survival. Eph receptors were abundantly expressed in pancreatic ductal adenocarcinoma cases examined. Eph-A1 staining intensity was significantly associated with tumor size (pT, p = 0.008) and tumor histopathological stage (pStage, p = 0.012). Eph-A2 expression was significantly associated with patients’ age (p = 0.007), while Eph-A4 and Eph-A5 with tumor proliferative capacity (p = 0.019 and p = 0.011, respectively). Pancreatic adenocarcinoma patients with moderate/intense Eph-A5 or Eph-A7 staining presented significantly shorter survival times compared to those with negative/mild one (log-rank test, p = 0.024 and p = 0.009, respectively). Multivariate analysis identified Eph-A5 and Eph-A7 staining intensity as independent prognostic factors (p = 0.048 and p = 0.004, respectively). In conclusion, the present study revealed that Eph receptors were associated with pancreatic cancer characteristics, supporting evidence for their potential clinical application in management and prognosis of pancreatic adenocarcinoma patients.     

TLR3和TLR4基因多态性与EBV相关胃癌易感性的关系

目的 探讨TLR3c.1377和TLR4Asp299Gly基因多态性与EBV感染在EBV相关胃癌（EBVassociated gastric carcinoma, EBVaGC）发生中的相关性。方法 选用41例EBVaGC组织、62例EBV阴性胃癌（EBV-negative gastric carcinoma, EBVnGC）组织以及64例健康人群血标本作为研究对象,采用PCR结合限制性长度多态性分析（reaction-restriction fragment length polymorphism, RFLP)技术检测TLR3c.1377 和TLR4 Asp299Gly基因多态性,并对实验结果进行统计分析。结果 （1）胃癌组与对照组比较TLR3c.1377基因型频率差异有统计学意义（P=0.025）,胃癌组T等位基因频率明显高于对照组（45.6% vs. 29.7%,P=0.004),T等位基因携带者的患病风险明显高于非携带者(OR=2.435,P=0.008）;（2）EBVaGC、EBVnGC以及对照组间TLR3c.1377基因型、等位基因频率差异无统计学意义（P>0.05）。（3）EBVaGC组,EBVnGC组以及正常对照组所有个体TLR4 Asp299Gly基因型均为Asp/Asp纯合子（P>0.05）。结论 TLR3c.1377基因多态性与胃癌易感性有关,T等位基因为胃癌的危险因子,而C等位基因为一保护基因;TLR3c.1377基因多态性与EBVaGC易感性无明显相关。     

幽门螺杆菌感染与慢性胃炎、溃疡病、异型增生、胃癌的关系及PCNA表达

目的　探讨幽门螺杆菌感染的不同胃粘膜增殖性病变的演进中增殖细胞核抗原 (PCNA)的表达情况及其意义。方法　对 15 0例病理证实的不同胃粘膜病变用免疫组化的方法检测PCNA标记指数 (LI) ,Warthin -starry(W -S)法检测HP感染。结果　PCNA -LI在浅表性胃炎为 2 2 0 0± 16 95、萎缩肠化性胃炎 46 45± 19 10、溃疡病 45 75± 18 15、异型增生 61 0 6± 2 0 67。在萎缩肠化性胃炎、溃疡病、异型增生及胃癌组织中均高于浅表性胃炎 (P <0 0 5 )。萎缩肠化型胃炎、溃疡病、异型增生及胃癌组PCNA阳性表达中HP感染者多于阴性者。结论　PCNA表达与胃粘膜增殖和恶化有关。HP感染与胃粘膜增殖和恶化有关     

胃癌雌激素受体的临床生物学意义

应用免疫组织化学技术检测１１４例胃癌组织的雌激素受体（ＥＲ）。结果提示胃癌可能是雌激素依赖性肿瘤，ＥＲ可作为胃癌的生物学标记。