TNF-α Genotypes Research Articles

Tumor Necrosis Factor-α Gene Polymorphisms and Risk of Oral Cancer: Evidence from a Meta-analysis

Numerous studies have been conducted regarding association between TNF-α and oral cancer risk, but the results remain controversial. The present meta-analysis is performed to acquire a more precise estimation of relationships. Databases of Pubmed, the Cochrane library and the China National Knowledge Internet (CNKI) were retrieved until August 10, 2013. Pooled odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated with fixed- or random-effect models. The heterogeneity assumption was assessed by I-squared test. Among the eight included case-control studies, all were focused on TNF-α-308G>A and four also concerned the TNF-α-238G>A polymorphism. It was found that oral cancer risk were significant decreased with the TNF-α-308G>A polymorphism in the additive genetic model (GG vs. AA, OR=0.19, 95% CI: [0.04, 1.00], P=0.05, I2=68.9%) and the dominant genetic model (GG+GA vs. AA, OR=0.22, 95% CI: [0.06, 0.82], P=0.03, I2=52.4%); however, no significant association was observed in allele contrast (G vs. A, OR=0.70, 95% CI: [0.23, 2.16], P=0.54, I2=95.9%) and recessive genetic models (GG vs. GA+AA, OR=0.72, 95% CI: [0.33, 1.57], P=0.41, I2=93.1%). For the TNF-α-238G>A polymorphism, significant associations with oral cancer risk were found in the allele contrast (G vs. A, OR=2.75, 95% CI: [1.25, 6.04], P=0.01, I2=0.0%) and recessive genetic models (GG vs. GA+AA, OR=2.23, 95%CI: [1.18, 4.23], P=0.01, I2=0.0%). Conclusively, this meta-analysis indicates that TNF-α polymorphisms may contribute to the risk of oral cancer. Allele G and the GG+GA genotype of TNF-α- 308G>A may decrease the risk of oral cancer, while allele G and the GG genotype of TNF-α-238G>A may cause an increase.

P-215 Biomarkers of Neoplasm in Ulcerative-Colitis

Colorectal cancer (CRC) remains a serious complication in ulcerative colitis (UC). Biomarkers that identify patients at greatest risk for this outcome could increase the effectiveness of current surveillance methods. Our previous work compared single nucleotide polymorphisms (SNP) and methylation between UC patients who developed CRC (UC-CRC) and UC patients who did not develop CRC (UC-controls). In that work, we studied methylation in DNA extracted from tissue obtained not only in the cancer section, but also from the non-adjacent, non-neoplastic tissue. These initial studies yielded both a potential “risk” SNP in the tumor necrosis factor alpha gene (TNFα) plus 3 markers with significant alterations in methylation status, RUNX3, MINT1 and COX2. The aims of the current study were to: 1.) validate methylation markers in earlier neoplasms (low-grade dysplasia, LGD), 2.) determine if TNFα is a marker of “risk” to develop any neoplasm and 3.) determine if these markers were associated with a LGD-non-progressor phenotype. We identified/reviewed 200 UC-LGD patients for potential inclusion. After chart review and histological confirmation, 162 were selected for testing. For each sample, formalin-fixed, paraffin-embedded sections were cut both from the dysplastic area (LGD-DYS) as well as a non-adjacent, non-neoplastic area (LGD-NAN). DNA was extracted using a modified Gentra protocol and 500 ng bisulfite treated using the EZ DNA Methylation Gold Kit (Zymo Research). TNFα genotyping was completed using TaqMan assay (Applied Biosystems). Methylation testing for RUNX3, MINT1 and COX2 was done using MS-PCR as previously described by our group. Comparisons were made between LGD-NAN, UC-controls (prior data) and UC-CRC cases (prior data). LGD-NAN results were also combined with UC-CRC data and compared to UC-controls. The subset of LGD patients who did not progress to CRC in long-term follow up (LGD-NP) was compared to UC-CRC and UC-control data. Statistical analysis was completed using JMP. Table 1 summarizes the Validation (A) and the Non-progressive LGD (B) analyses. P-values and sample numbers for each comparison are provided. •Validation of methylation as a marker for concurrent neoplasm in non-adjacent mucosa: RUNX3, MINT1 and COX2 have altered methylation compared to UC-controls, even in patients with early neoplasm in the form of LGD. • TNFα-308 is associated with neoplasm in UC: Comparisons between UC-controls and any neoplasm were significant. Comparisons within neoplastic groups (UC-CRC, LGD-DYS and LGD-NP) were not significant. • Methylation in non-progressor LGD: The only variation between UC-controls and LGD-NP-NAN was methylation status of COX2, with LGD-NP-NAN more likely to be unmethylated than UC-controls. In comparisons between LGD-NP and UC-CRC (either NAN or DYS), COX2 was not significant but both RUNX3 and MINT1 were. Results in Table1A confirm our previous findings that TNFα-308 (G>A) is a risk SNP and that this methylation panel can be used to increase standard surveillance sensitivity. Although the sample set is small, the data for LGD-NP suggest that altered methylation patterns may be informative in guiding treatment of LGD. In addition, the alteration of COX2 between LGD-NP-NAN and UC-controls may suggest a possible target for chemoprevention.

No Association between Tumor Necrosis Factor-alpha Gene Polymorphisms and Lung Cancer Risk

ObjectivesThe role of genetic polymorphisms of tumor necrosis factor-alpha (TNF-α) for lung cancer development was evaluated.MethodsGenotypes of the TNF-α polymorphisms, -1210C>T, -487A>G, -417A>G, IVS1+123G>A, and IVS3+51A>G, were determined in 616 lung cancer cases and 616 lung cancer-free controls.ResultsAfter adjusting for body mass index and smoking, each TNF-α genotype or haplotype composed of five TNF-α single nucleotide polymorphisms did not show an association with lung cancer risk (p>0.05). The statistical power was found to be 88.4%, 89.3%, 93.3%, 69.7%, and 93.9% for 1210C>T, -487A>G, -417A>G, IVS1+123G>A, and IVS3+51A>G, respectively. Furthermore, the effects of each SNP or haplotype on lung cancer risk were not found to be different according to the cell type of lung cancer (p>0.05). In the repeated analysis with only subjects without other diseases related to inflammation, there was also no association between polymorphisms or haplotypes of the TNF-α gene and lung cancer risk (p>0.05).ConclusionsThis study found no association between common variants of the TNF-α gene and lung cancer risk.

Impact Of Patient/Donor Cytokine Genotypes On The Development Of Acute Graft Versus Host Disease (aGVHD) After Peripheral Blood Stem Cell Transplantation From An Identical Sibling

In spite of prophylactic measures GVHD continues to be one of the serious complications that affect transplantation-related morbidity and mortality. We have previously reported a role of cytokines in the prediction of aGVHD (Kamel et al, 54th ASH Annual meeting, December 8-11.2012.Publicaion Number 1956. Nov 2012; 120: 195 and J Leuk 2013, 1:2 http://dx.doi.org/10.4172/jlu.1000111)Apart from the triggering stimulus for cytokine production, the genotype may be a contributing factor to the actual level of the secreted cytokine. We hypothesized that the genotype of a given cytokine with its expected phenotype in patients and/or donors may impact the immune response contributing to the pathogenesis of aGVHD. We aimed to verify if the genotype of a given cytokine in patients and/or donors may help to predict the occurrence of aGVHDThe work was performed according to Helsinki declaration, the protocol was approved by the IRB of the NCI, Cairo University and an informed consent was obtained from all subjects.The study cohort included 80 patient/donor pairs who received PBSCT from an identical sibling at Nasser Institute, MOH, Egypt in the period from December 2010 –December 2012. We determined the genotype of TNFα, TGFβ, IL10, IL6 and IFNγ by SSP typing using Cytokine Genotyping Tray REF CYTGEN (one Lambda, Inc, 21001 Kittridge st., Canoga Park, CA 91303 USA).Patients were followed up for at least one year; development of GVHD was recorded.Of the 80 cases, 23 developed acute GVHD, 9 developed chronic GVHD on top.The genotypes with the corresponding phenotype are presented in table (1).TNFα was of the high producer genotype (G/A or A/A) in all patients and donors. IL6 was also of the high producer genotype (G/G or G/C) in all except one in each category which was low producer genotype (C/C). These two genes will not be discussed any further. TGFβ was of the low producer genotype (C/C G/C, C/C C/C, T/t C/C, or T/c C/C) in 22/23 patients who developed acute GVHD; the remaining patient had an intermediate producing genotype (T/C G/C). The corresponding figures for those who did not develop acute GVHD was 71/80 (88.75%) with low producing genotype, one with high producing genotype (T/C G/G) and seven with intermediate producing genotype (T/C G/C, C/C G/G or T/T G/C). All donors of patients who developed acute GVHD had the low producing genotype except one that showed an intermediate producing genotype (T/C G/C). The corresponding figures for donors of patients who did not develop GVHD included 51 (89.5%) with low producing genotype, two with high producing genotype (T/T G/G) and four with intermediate producing genotype. However, the difference did not achieve statistical significance for either patients or donors. For IL10 and IFN γ, the distribution of the various genotypes among patients/donors who developed was comparable to those who did not develop acute GVHD. ConclusionThe genotype of TNFα and IL6 are homogenous among Egyptian population; they cannot be used for discrimination between any studied disease conditions. TGFβ of high or intermediate producing genotypes are uncommon but mainly confined to patient/donor pairs with no GVHD; yet statistical significance was not achieved. IL10 and IFN γ genotypes of patients/donors are not associated with or predictive of acute GVHD. Taking in consideration the previous reports on the predictive value of both or either at the cytokine level including the work of the authors, it may be speculated that the genotype is not the main or the only predictor of the level of cytokine production in the setup of PBSCT. Disclosures:No relevant conflicts of interest to declare.

IFN-γ (+874) and not TNF-α (−308) is associated with HBV-HCC risk in India

Tumor necrosis factor (TNF)-α and interferon (IFN)-γ, the pro-inflammatory Th1 cytokines are the indispensable coordinators of the inflammatory responses involved in hepatitis B virus (HBV) pathogenesis. This study attempted to evaluate any possible association among TNF-α (-308G>A) and IFN-γ (+874T/A) genotypes, the spontaneous blood and mRNA levels and expression of their major signal transducers, namely STAT1 and NF-кB with hepatitis B virus-induced hepatocellular carcinoma (HCC) susceptibility in India. For this, 398 subjects (146 controls, 68 inactive-HBV-carriers, 64 chronic-active HBV patients, 61 HBV-cirrhotics, and 59 HBV-HCC subjects) were enrolled. Polymerase chain reaction-restriction fragment length polymorphism, allele-specific PCR, enzyme-linked immunosorbent assay, reverse transcriptase-PCR, and Western blot analysis were done for assessing polymorphism, blood levels, mRNA expression, and protein expression of signal transducers, respectively, of TNF-α and IFN-γ. The study revealed no significant association of TNF-α (-308) GA genotype, while a significant negative association of IFN-γ (+874) TA and AA genotypes, in HBV-HCC risk. Moreover, blood levels of TNF-α were significantly elevated as disease progresses to HCC, while IFN-γ levels were raised in HCC patients only. Besides, IFN-γ mRNA levels were significantly elevated in cirrhotics, with no change observed in TNF-α transcript levels. Moreover, NF-кB expression also consistently increased during HCC progression. These observations suggest a vital negative association of IFN-γ (+874) with HBV-HCC risk, with no significant association evident in TNF-α (-308). However, the TNF-α and IFN-γ levels markedly increased in HCC development.

The relationship between the polymorphism of immunity genes and both aging and age-related diseases

Aging is acommon, progressive and irreversible state of multi-cell dysfunction. Immune aging mainly includes the declines of regenerative capacity and lymphoid lineage differentiation potential, the hyporesponsive to infection and vaccination, the hyperresponsive in the context of inflammatory pathology, and the increased risk of autoimmunity. The dysfunction of aged immune system accelerates the occurrence of aging and age-related diseases. The mutation of immunity genes that affect immune responses accelerates or slows aging process and age-related diseases. The frequencies of acquired immunity genes, such as immune protective HLA II DRB1*11 and DRB*16-associated haplotype, are increased in the longevity populations. The increased susceptibility of immune inflammatory response, morbidity and mortality in the elderly is often associated with decreased frequencies of anti-inflammatory factor IL-10 -1082G allele, TNF-β1 haplotype cnd10T/C, cnd25G/G, -988C/C, -800G/A, low proinflammatory fator TNFa level related extended TNF-A genotype -1031C/C, -863C/A, -857C/C, IL-6-174 CC and IFN-γ+874 T allele as well. The innate immunity genes, such as highly expressed anti-inflammatory +896 G KIR4 allele, CCR5Δ32 variant, -765 C Cox-2 allele, -1708 G and 21 C 5-Lox alleles are detected in centenarians. In age-related diseases, a higher CMV-specific IgG antibody level in elderly individuals is associated with a decreased frequency of KIR haplotypes KIR2DS5 and A1B10 and an increased frequency of MBL2 haplotypes LYPB, LYQC and HYPD that result in the absence of MBL2 protein. The increased frequencies of CRP ATG haplotypes and CFH 402 His allele indicate high mortality in the elderly. In the present study, we review the advances in the polymorphism and haplotype of innate and adoptive immunity genes, and their association with both aging and age-related diseases. To strengthen the analysis of extended haplotypes, epigenetic studies of immunity genes and genetic study of hematopoietic stem cell senescence will be helpful to understand the accurate basis of aging-related immune genetics better.

Assessment of 11-β hydroxysteroid dehydrogenase (11-βHSD1) 4478T>G and tumor necrosis factor-α (TNF-α)-308G>A polymorphisms with obesity and insulin resistance in Asian Indians in North India

11-β hydroxysteroid dehydrogenase (11-βHSD1), tumor necrosis factor-α (TNF-α) and their role in obesity, regional adiposity and insulin resistance has been sparsely evaluated. We determined the polymorphic status of 11-βHSD1 4478T>G and TNF-α-308G>A in Asian Indians in north India. In this cross-sectional study (n = 498; 258 males, 240 females), association of genotypes (PCR–RFLP) of 11-βHSD1 and TNF-α were analyzed with obesity [BMI ≥ 25 kg/m(2), percentage body fat (%BF by DEXA); subcutaneous and intra-abdominal fat area (L(2-3) level by single slice MRI) in a sub sample] and insulin resistance. 46 percent subjects had generalized obesity, 55 % abdominal obesity and 23.8 % were insulin resistant. Frequencies (%) of [T/T] and [T/G] genotypes of 11-βHSD1 were 89.57 and 10.43 respectively. Homozygosity for 11-βHSD1 4478G/G was absent with no association with parameters of obesity and insulin resistance. Frequencies (%) of TNF-α [G] and [A] alleles were 88 and 12 respectively. Higher frequency of variant -308[A/A] was observed in females versus males (p = 0.01). Females with at least one single A allele of TNF-α-308G>A had significantly high %BF and total skinfold, whereas higher values of waist hip ratio, total cholesterol, triglycerides and VLDL were observed in males. Subjects with even a single A allele in TNF-α genotype showed higher subscapular skinfold predisposing them to truncal subcutaneous adiposity (p = 0.02). Our findings of association of TNF-α-308G>A variant in females with obesity indices suggests a gender-specific role of this polymorphism in obesity. High truncal subcutaneous adiposity is associated with A allele of TNF-α-308G>A in this population.

Tumour necrosis factor-α plus interleukin-10 low producer phenotype predicts acute kidney injury and death in intensive care unit patients

Genetic polymorphism studies of cytokines may provide an insight into the understanding of acute kidney injury (AKI) and death in intensive care unit (ICU) patients. The aim of this study was to investigate whether the genetic polymorphisms of -308 G < A tumour necrosis factor (TNF)-α, -174 G > C interleukin (IL)-6 and -1082 G > A IL-10 may predispose ICU patients to the development of AKI and/or death. In a prospective nested case-control study, 303 ICU patients and 244 healthy individuals were evaluated. The study group included ICU patients who developed AKI (n = 139) and 164 ICU patients without AKI. The GG genotype of TNF-α (low producer phenotype) was significantly lower in the with AKI than without AKI groups and healthy individuals (55 versus 62 versus 73%, respectively; P = 0·01). When genotypes were stratified into four categories of TNF-α/IL-10 combinations, it was observed that low TNF-α plus low IL-10 producer phenotypes were more prevalent in patients with AKI, renal replacement therapy and death (P < 0·05). In logistic regression analysis, low TNF-α producer plus low IL-10 producer phenotypes remained as independent risk factors for AKI and/or death [odds ratio (OR) = 2·37, 95% confidence interval (CI): 1·16-4·84; P = 0·02] and for renal replacement therapy (RRT) and/or death (OR = 3·82, 95% CI: 1·19-12·23; P = 0·02). In this study, the combination of low TNF-α plus low IL-10 producer phenotypes was an independent risk factor to AKI and/or death and RRT and/or death in critically ill patients. Our results should be validated in a larger prospective study with long-term follow-up to emphasize the combination of these genotypes as potential risk factors to AKI in critically ill patients.

Single Nucleotide Polymorphisms in the Tumor Necrosis Factor-Alpha Gene Promoter Region Alter the Risk of Psoriasis Vulgaris and Psoriatic Arthritis: A Meta-Analysis

BackgroundIt has been confirmed that tumor necrosis factor-alpha (TNFα), a macrophage-derived pro-inflammatory cytokine, plays an important role in the pathogenesis of psoriasis vulgaris and psoriatic arthritis (PsV&PsA). In contrast, the reported association of TNFα gene promoter region single nucleotide polymorphisms (SNPs) and PsV&PsA has remained controversial. Accordingly, we performed a meta-analysis to provide new evidence that SNPs in the TNFα gene promoter region alter not only the risk of psoriasis vulgaris (PsV) or psoriatic arthritis (PsA) but also of PsV&PsA.MethodsInterrelated literature dated to October 2012 was acquired from the PubMed, ScienceDirect, and SpringerLink databases. The number of the genotypes and/or alleles for the TNFα promoter in the PsV and PsA and control subjects was obtained. Odds ratios (ORs) and 95% confidence intervals (CIs) were used to calculate the risk of PsV and/or PsA with TNFα promoter SNPs.ResultsA total of 26 papers of 2159 for PsV (2129 normal controls) and 2360 for PsA (2997 normal controls) were included in our meta-analysis. The results showed that the variant genotype and allele of TNFα -308A/G was protective in pooled groups of patients with PsV&PsA (OR = 0.682, 0.750; 95% CI, 0.596-0.779, 0.653-0.861). However, the variant genotypes and alleles of TNFα -238A/G and -857T/C had an increased risk of PsV&PsA (OR = 2.493, 2.228, 1.536, 1.486, 95% CI, 1.777-3.498, 1.628-3.049, 1.336-1.767, 1.309-1.685). Moreover, the meta-analysis revealed a significant association between TNFα -238A/G and -857T/C polymorphism and PsA susceptibility (OR = 2.242, 2.052, 1.419, 1.465; 95% CI, 1.710-2.941, 1.614-2.610, 1.214-1.658, 1.277-1.681). In contrast, the variant genotypes and alleles of TNFα -308A/G proved to be protective against PsV (OR = 0.574, 0.650, 95% CI, 0.478-0.690, 0.556-0.759), whereas TNFα -238A/G was found to have a risk association (OR = 2.636, 2.223, 95% CI, 1.523-4.561, 1.317-3.751).ConclusionsSNPs in the TNFα gene promoter region alter the risk of PsV and/or PsA.

Genetic polymorphisms of tumour necrosis factor alpha (TNF‐α) promoter gene and response to TNF‐α inhibitors in Spanish patients with inflammatory bowel disease

Tumour necrosis factor alpha (TNF-α) has an important role in inflammatory response. Alterations in the regulation of TNF-α have been implicated in a variety of inflammatory disorders, including Inflammatory bowel disease (IBD). Indeed, a common treatment for IBD is the use of TNF-α inhibitors. Polymorphisms in the TNF-α promoter region are known to affect the level of gene expression. Our aim was to investigate the influence of these single nucleotide polymorphisms (SNPs) in TNF-α promoter gene play in the risk of IBD in a Spanish population and their individual response to anti-TNF-α treatment. DNA samples from patients with IBD and controls were screened for TNF-α -238G/A (rs361525) and -308G/A (rs1800629) SNPs by PCR-SSOP using a microbeads luminex assay and compared with response to TNF-α inhibitors. There were not statistical differences in -238G/A and -308G/A allele and genotype frequencies between patients. However, we found an increased frequency of -308A allele and -308GA genotype in these nonresponders patients to TNF-α inhibitors with respect to responders patients (Pc<0.05). This -308GA genotype has been classified as high producer of this cytokine. This fact could actually be interesting to explain the different response of patients with IBD with respect to TNF-α inhibitors. TNF-α promoter gene polymorphism does not seem to play a role in IBD susceptibility, but particular TNF-α genotypes may be involved in the different responses to TNF-α inhibitor treatment in Spanish patients with IBD.

Influence of polymorphisms in genes encoding immunoregulatory proteins and metabolizing enzymes on susceptibility and outcome in patients with diffuse large B-cell lymphoma treated with rituximab

We analyzed the allelic distribution of 12 candidate polymorphisms in a large retrospective study of 486 patients with diffuse large B-cell lymphoma (DLBCL) treated at Oslo University Hospital with 1056 blood donors serving as controls. Variants in TNFα (rs1800629) (GG vs. AG/AA, p < 0.001) and LTA (rs909253) (AA vs. AG/GG, p = 0.02) and deletions in GSTM1 and GSTT1 (undeleted vs. deleted, p = 0.01 and p = 0.01, respectively) were associated with increased susceptibility of developing DLBCL. IL-10 (rs1800896) variants (GG vs. AG/AA, p = 0.03) were associated with decreased susceptibility. In line with several previous reports, patients carrying the TNFα (rs1800629) A allele treated in the pre-rituximab era had inferior outcome compared to patients carrying the homozygous GG genotype (p = 0.004, n = 33). However, patients receiving at least one dose of rituximab had equal outcome regardless of their TNFα genotype (HR = 0.94, p = 0.79, n = 307). Deletion in GSTM1 was associated with inferior outcome for patients with low International Prognostic Index (IPI) score (p = 0.04). Our findings support the suggestions that polymorphisms in genes encoding immunoregulatory proteins and enzymes that metabolize carcinogens and chemotherapeutic drugs influence DLBCL susceptibility and possibly treatment outcome. The influence of polymorphisms in immunoregulatory genes on outcome in DLBCL should be reevaluated in the rituximab era.

Tumor Necrosis Factor-α Gene Polymorphisms in Korean Patients With Recurrent Spontaneous Abortion

The objective of this study was to investigate the contribution of the tumor necrosis factor-α (TNF-α) gene polymorphisms to recurrent spontaneous abortion (RSA). The study participants consisted of 357 Korean women with RSA and 236 fertile women controls. Four TNF-α gene variants of all participants were analyzed by polymerase chain reaction-restriction fragment length polymorphism assay. The TNF-α -1031T>C and TNF-α -238G>A variants increased the risk of RSA TNF-α -1031TC+CC; adjusted odds ratio [AOR], 2.292; 95% confidence interval [CI], 1.547-3.395; P < .001; TNF-α -238GA+AA; AOR, 2.327; 95% CI, 1.038-5.217; P = .040), and these data were not different in a stratified analysis according to the number of consecutive spontaneous abortions. Also, the mutant genotypes of TNF-α -1031 and TNF-α -238 showed synergistic effects on increased RSA risk (-1031TC+CC/-238GA+AA; AOR, 4.054; 95% CI, 1.520-10.812; P = .005). In haplotype analysis, there were similar trends of data for combination analysis. In conclusion, the TNF-α -1031T>C and TNF-α -238G>A variants are possible genetic risk factors for RSA.

Associations between TGF-b1 G/A and TNF-a 308 G/A gene polymorphisms with airway resistance in chronic obstructive pulmonary disease

Chronic obstructive pulmonary disease (COPD) is an important morbidity and mortality cause all over the world. Although specific gene region has not been defined in the pathogenesis of COPD, cytokine gene polymorphisms like tumor necrosis factor-alfa (TNF-α) and transforming growth factor-beta1 (TGF-β1) may contribute to the development of COPD. The aim of the present study was to evaluate the associations between airway resistance with TGF-β1 G/A and TNF-α 308 G/A gene polymorphisms in COPD patients. 264 subjects were included to the study (Group 1; 75 COPD patients, Group 2; 139 subjects with at least 10 packet year smoking history without airflow obstruction, Group 3; 50 healthy subjects). Pulmonary function tests and body plethysmography to measure airway resistance were performed to the subjects. TGF-β1 800 G/A and TNF-α 308 G/A gene polymorphisms were evaluated. Chi-square, Anova and correlation analysis were used for statistical analysis. There were significant difference among COPD stages in terms of TNF-α 308 G/A polymorphism (p< 0.05). Thirteen (23.6%) stage 1 COPD patients had TNF-α 308 G/A polymorphism and the other did not have. We did not find statistically significant difference among COPD stages in terms of TGF-β1 800 G/A polymorphism (p> 0.05). TNF-α and TGF-β1 genotypes and TNF-a 308 G/A and TGF-β1 800 G/A polymorphisms were not different among study groups. Moreover, no significant differences betweeen subjects with and without increased airway resistance in terms of TNF-α 308 G/A and TGF-β1 800 G/A polymorphisms were present. These results can suggest the lack of association between TNF-α 308 G/A and TGF-β1 800 G/A gene polymorphisms with COPD development and airway resistance in Turkish population.

Polymorphisms in toll-like receptors 2, 4 and 5 are associated with Legionella pneumophila infection

To investigate whether single-nucleotide polymorphisms (SNPs) in toll-like receptors (TLR) 2, 4 and 5 affect the susceptibility of Legionella pneumophila infection in a Han Chinese population by in vitro assay. Fifty-four (n=54) healthy subjects were genotyped for SNPs (TLR2 (C597T) [rs3804099], TLR4 (G2244A), TLR4 (A2299G) [AF177765] and TLR5 (C1174T) [rs5744168]). Peripheral blood mononuclear cells (PBMCs) were obtained from these subjects and stimulated with live L. pneumophila for 24h. The mRNA expression levels of adapter protein myeloid differentiation factor 88 (MyD88) were determined using real-time reverse transcription polymerase chain reaction (RT-PCR) and the expression levels of TNF-α, IL-1β and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA). After 24h of L. pneumophila stimulation, the mRNA expression level of MyD88 was significantly lower with TLR2 (C597T) CT/TT (p=0.0482) or TLR5 (C1174T) CC homozygotes (p=0.0223) in comparison to PBMCs with other genotypes. The mRNA expression level of MyD88 was significantly higher with TLR4 (G2244A) GG/GA than AA homozygotes (p=0.0352). No significant difference was found in PBMCs with genotypes TLR4 (A2299G) AA, AG or GG (p>0.05). Supernatant from cultures of PBMCs with genotype TLR2 (C597T) CT/TT or TLR5 (C1174T) CC were found to have higher levels of TNF-α, IL-1β and IL-6 after L. pneumophila stimulation. TLR4 (G2244A) GG/GA alleles were found to have lower levels of TNF-α (p=0.0367) and higher levels of IL-6 (p=0.0317) in comparison to cells with AA alleles. No significant association was observed between the TNF-α, IL-1β and IL-6 levels and genotype TLR4 (A2299G) after L. pneumophila stimulation (p>0.05). Our findings suggested that healthy subjects who were positive for the TLR2 (C597T) CT/TT and TLR5 (C1174T) CC alleles had a superior innate immune response to L. pneumophila than other genotypes in the evaluated Han Chinese population, whereas no association was found for the TLR4 (A2299G) [AF177765] polymorphism with L. pneumophila susceptibility. It is not clear from our study if TLR4 (G2244A) [AF177765] is associated with susceptibility to L. pneumophila infection.

Association betweenTNF-αandIL-1βgenotypesvs Helicobacter pyloriinfection in Indonesia

To investigate the correlation between the Helicobacter pylori (H. pylori) infection and host genetic background of healthy populations in Indonesia. In March 2007, epidemiological studies were undertaken on the general population of a city in Indonesia (Mataram, Lombok). The participants included 107 men and 187 women, whose ages ranged from 6 to 74 years old, with an average age of 34.0 (± 14.4) (± SD). The H. pylori of subject by UBT method determination, and through the polymerase chain reaction with confronting two-pair primers (PCR-CTPP) method parsing the single nucleotide polymorphism of interleukin (IL)-8, IL-4, IL-1β, CD14, tumor necrosis factor (TNF-α) and tyrosine-protein phosphates non-receptor type 11 (PTPN11) genotypes. The experimental data were analyzed by the statistical software SAS. The H. pylori infection rates in the healthy Indonesian population studied were 8.4% for men and 12.8% for women; no obvious differences were noted for H. pylori infection rates by sex or age. TC genotypes of IL-4, TC and CC genotypes of TNF-α, and GA genotypes of PTPN11, were higher in frequency. Both CC and TC genotype of TNF-α T-1031C loci featured higher expressions in the healthy Indonesian population Indonesia studied of (OR = 1.99; 95%CI: 0.67-5.89) and (OR = 1.66; 95%CI: 0.73-3.76), respectively. C allele of IL-1β T-31C gene locus was at a higher risk (OR = 1.11; 95%CI: 0.70-1.73) of H. pylori infection, but no statistical significance was found in our study. We reveal that the association between the TNF-α and IL-1β genotypes may be the susceptibility of H. pylori in the studied population.

Role of tumor necrosis factor-α -308 G/A promoter polymorphism in gastric cancer

Background/Aim:Gastric cancer (GC) is the fourth most common cancer and the second most common cause of cancer death world-wide after lung cancer. It is a multifactorial disease with the involvement of both genetic and environmental risk factors. Genetic variation in genes encoding cytokines and their receptors, determine the intensity of the inflammatory response, which may contribute to individual differences in severity of outcome of the disease. Tumor necrosis factor alpha (TNF-α) is a potent pro-inflammatory cytokine and acid inhibitor. A bi allelic G to A polymorphism at -308 upstream from the transcription initiation site of the promoter is associated with elevated TNF levels. The present study is aimed at evaluating the role of TNF-α-308 (G → A) gene polymorphism and susceptibility to GC.Subjects and Methods:A case-control study was carried out in 114 GC patients and 229 healthy control subjects. TNF-α genotyping at position-308 (G → A) was carried out by amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) method followed by agarose gel electrophoresis.Results:The distribution of TNF-α genotypes at -308 (G → A) were GG 28.07%, GA 66.67% and AA 5.26% in GC patients and GG 33.19%, GA 55.89% and AA 10.92% in control subjects. The frequencies of alleles G and A were 0.614 and 0.386 in GC patients and 0.611 and 0.389 in control subjects respectively.Conclusion:The study showed no significant difference in the distribution of genotype and allelic frequencies between GC patients and control subject.

Parity-dependent association between TNF-α and LTF gene polymorphisms and clinical mastitis in dairy cattle.

BackgroundOne major problem in dairy cattle husbandry is the prevalence of udder infections. In today’s breeding programmes, top priority is being given to making animal evaluation more cost-effective and reliable and less time-consuming. We proposed tumor necrosis factor α (TNF-α), lactoferrin (LTF) and macrophage-expressed lysozyme (mLYZ) genes as potential DNA markers in the improvement of immunity to mastitis.This study included 588 Polish Holstein-Friesian cows kept on one farm located in the north-western region of Poland. All clinical cases of mastitis in the herd under study were recorded by a qualified veterinarian employed by the farm. The following indicators were applied to determine udder immunity to mastitis in the cows under study: morbidity rate (MR), duration of mastitis (DM) and extent of mastitis (EM). TNF-α, mLYZ and LTF genotypes were identified by real-time PCR method, using SimpleProbe technology. Due to the very low frequency of mLYZ allele T, the gene was excluded from further analysis.A statistical analysis of associations between TNF-α and LTF genes and immunity to mastitis were performed using three models: 1) a parity-averaged model including only additive effects of the genes; 2) a parity-averaged model including both additive and epistatic effects of the genes; and 3) a parity-specific model including only additive effects of the genes.ResultsWith the first and second models it was revealed that the genes effects on the applied indicators of immunity to mastitis were non-significant whereas with the third one the effects were found to be statistically significant. Particularly noteworthy was the finding that the effects of TNF-α and LTF varied depending on age (parity). The alleles which were linked to high immunity to mastitis in lower parities appeared to be less favourable in higher parities.ConclusionsThese interactions might be related to inflamm-ageing, that is an increased susceptibility to infection due to immune system deregulation that progresses with age. Such pattern of interactions makes it impossible to use the genes in question in marker-assisted selection aimed at reducing heritable susceptibility to mastitis. This is because the immune mechanisms behind resistance to infections proved to be too complex.

TNF-α -857C&gt;T Genotype is Predictive of Clinical Response after Treatment with Definitive 5-Fluorouracil/cisplatin-based Chemoradiotherapy in Japanese Patients with Esophageal Squamous Cell Carcinoma

Background: Genotypes of tumor necrosis factor alpha (TNF-α) and its surface receptors, TNFRSF1A and TNFRSF1B, have been examined in terms of the progression, metastasis, clinical efficacy, and prognosis of various cancers; however, little is known about their effects on clinical outcome in patients with esophageal squamous cell carcinoma (ESCC). In this study, TNF-α and TNFRSF1A genotypes were retrospectively evaluated in terms of predicting clinical response, long-term survival, and severe acute toxicities in 46 male Japanese ESCC patients treated with definitive 5-fluorouracil (5-FU)/cisplatin (CDDP)-based chemoradiotherapy (CRT).Methods: A course consisted of the continuous infusion of 5-FU at 400 mg/m2/day for days 1-5 and 8-12, the infusion of CDDP at 40 mg/m2/day on days 1 and 8, and radiation at 2 Gy/day on days 1-5, 8-12, and 15-19, with a second course being repeated after a 2-week interval. The TNF-α -1031T>C (rs1799964), -863C>A (rs1800630), -857C>T (rs1799724), -308G>A (rs1800629), -238G>A (rs361525), TNFRSF1A -609G>T (rs4149570), and 36A>G (rs767455) genotypes were evaluated.Results: The TNF-α -857C>T genotype was found to be predictive of clinical response, i.e., complete response or not (P = 0.010, Fisher's exact test), but had no effect on long-term survival (CC-857 vs. CT-857 + TT-857, P = 0.072, Fisher's exact test, P = 0.070, Log-rank test).Conclusions: The TNF-α -857C>T genotype was found to be predictive of clinical response and was more likely to predict long-term survival in Japanese ESCC patients receiving definitive 5-FU/CDDP-based CRT. Further clinical investigations with a larger number of patients or experiments in vitro should be performed to assess the predictive value of this genotype following CRT.

The Synergy of ‐260T T CD14 and ‐308GG TNF‐α Genotypes in Survival of Critically Ill Patients

Literature suggests that the analysis of several polymorphic genetic markers is more informative than the analysis of a single polymorphism. In this study, we tested whether the shared inheritance of TLR2 and TLR4 and TNF-α allelic variants may act in synergy with -260C>T CD14 SNP on the outcome from critical conditions. We monitored 524 critically ill patients from South Brazilian, daily from the ICU admission to their discharge from hospital, or death. Our results revealed that TLR2, TLR4 or TNF-α SNPs alone did not show a significant role in the outcome from critical illness. However, when we performed a combined analysis with the CD14 inheritance, we detected a significant higher survivor rate in -260TT CD14/-308GG TNF-α individuals (P = 0.037). In the adjusted analysis including the main clinical predictors to mortality, we observed that -260TT/-308GG double-genotype was a significant protective factor towards survival (P = 0.046). An increased probability for survival of -260TT/-308GG was also observed by 'pathway genetic load' analysis (unweighted: P = 0.041; weighted: P = 0.036). When we applied a hazard function analysis with the -260TT/-308GG variable as a discriminating factor, -260TT/-308GG patients group had, in fact, a higher survivor rate (P = 0.024). Connected to the beneficial effect of -260TT CD14, the -308GG TNF-α genotype was protective against the reported over expression of TNF-α caused by -308A rare allele. Results support the hypothesis that the interaction between -260C>T CD14 and -308G>A TNF-α functional SNPs may be synergistically influencing the outcome of critically ill patients.

Haploidentical Allogeneic Hematopoietic Stem Cell Transplantation Using the Donors with the HLA-B*5801-TNFα -308A Haplotype Produced Higher Frequency of Untoward Effects in Acute Lymphocytic Leukemia Patients

Abstract 2002▪▪This icon denotes a clinically relevant abstractIn this study, we investigated the clinical characteristics of acute lymphoblastic leukemia (ALL) patients using sibling donors with HLA-B*5801-TNFα-308A haplotype (B5801-TNF2) for haploidentical or identical hematopoietic stem cell transplantation (HSCT). A total of 136 B-ALL cases and 29 T/NK-ALL cases were recruited. DNA samples from the patients and their siblings were assayed for genotyping of HLA and TNFα at -308 (rs1800629). B5801-TNF2 haplotype in donors was determined by analyzing TNFα -308 and HLA-B genotypes in patients and their family members. Outcomes within 2 years including overall mortality and non-relapse mortality, disease course and complications within 100 days were compared among patients using related donors with HLA-B*5801-TNFα-308A haplotype for haploidentical HSCT (B5801-TNF2+HID group), those using related donor without HLA-B*5801-TNFα-308A haplotype for haploidentical HSCT (B5801-TNF2-HID group), and those using related sibling either B5801-TNF2+ or B5801-TNF2- haplotype for identical HSCT (SID group). In the 165 sibling donors after HSCT, 35 were found to carry HLA-B*5801-TNFα-308A haplotype. There were 21 cases in B5801-TNF2+HID group, 100 cases in B5801-TNF2-HID group, and 44 cases in SID group. Compared patients in B5801-TNF2-HID and SID groups, patients in B5801-TNF2+HID group had higher overall mortality (adjusted P=0.039) and non-relapse mortality within 2 years (adjusted P=0.001), delayed platelet engraftment (adjusted P<0.0001), higher incidences of severe acute graft-versus-host disease (P=0.007), severe late onset hemorrhagic cystitis (P=0.002), blood stream infection (P=0.017), and invasive fungal disease (P=0.004) within 100 days. Therefore, sibling donors carrying HLA-B*5801-TNFα-308A haplotype for haploidentical HSCT caused higher mortality rate and higher frequency of severe complications in ALL recipients. Keywords:HLA-B*5801-TNFα-308A haplotype; acute lymphoblastic leukemia; allogeneic haploidentical hematopoietic stem cell transplantation; allogeneic identical hematopoietic stem cell transplantation Disclosures:No relevant conflicts of interest to declare.