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Association of TLR5 gene polymorphisms in ulcerative colitis patients of north India and their role in cytokine homeostasis.

Meena NK, Ahuja V, Meena K, Paul J - PLoS ONE (2015)

Bottom Line: However, there was decreased level of TNFα (p = 0.014), IL-6 (p = 0.028) and IFNγ (p = 0.001) in patients carrying TLR5-R392X variant as compared to wild type patients.Patients carrying two simultaneous SNPs D299G in TLR4 gene and N592S in TLR5 gene showed significant decrease in the levels of TNFα (p = 0.011) and IFNγ (p = 0.016).Polymorphisms in TLR 5 genes were significantly associated with the UC in North Indian population.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, Jawaharlal Nehru University, New Delhi, India.

ABSTRACT

Background and aim: In health, TLR signaling protects the intestinal epithelial barrier and in disease, aberrant TLR signaling stimulates diverse inflammatory responses. Association of TLR polymorphisms is ethnicity dependent but how they impact the complex pathogenesis of IBD is not clearly defined. So we propose to study the status of polymorphisms in TLR family of genes and their effect on cytokines level in UC patients.

Methods: The genotypes of the six loci TLR1-R80T, TLR2-R753Q, TLR3-S258G, TLR5-R392X, TLR5-N592S and TLR6-S249P were determined in 350 controls and 328 UC patients by PCR-RFLP and sequencing. Cytokine levels were measured by ELISA in blood plasma samples. Data were analyzed statistically by SPSS software.

Results: TLR5 variants R392X and N592S showed significant association (p = 0.007, 0.021) with UC patients but TLR 1, 2, 3, 6 variants did not show any association. Unlike other studies carried out in different ethnic groups, TLR 6 (S249P) SNP was universally present in our population irrespective of disease. Genotype-phenotype correlation analysis revealed that the patients having combination of multiple SNPs both in TLR5 and TLR4 gene suffered from severe disease condition and diagnosed at an early age. The level of TNFα (p = 0.004), IL-6 (p = 0.0001) and IFNγ (p = 0.006) significantly increased in patients as compared to controls having wild genotypes for the studied SNPs. However, there was decreased level of TNFα (p = 0.014), IL-6 (p = 0.028) and IFNγ (p = 0.001) in patients carrying TLR5-R392X variant as compared to wild type patients. Patients carrying two simultaneous SNPs D299G in TLR4 gene and N592S in TLR5 gene showed significant decrease in the levels of TNFα (p = 0.011) and IFNγ (p = 0.016).

Conclusion: Polymorphisms in TLR 5 genes were significantly associated with the UC in North Indian population. The cytokine level was significantly modulated in patients with different genotypes of TLR4 and TLR5 SNPs.

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Related in: MedlinePlus

Analysis of cytokines level in UC patients and controls.TNFα, IL-6 and IFNγ levels were measured in human blood plasma samples (n = 4–8) by ELISA. Cytokines level were measured in wild type controls, and in wild type patients as well as with different genotype category of patient samples. The data is represented as mean value of 4–8 samples in each group. The significance level of α ≤0.05 was chosen for all sets. pg = picograms, ml = milliliter.
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pone.0120697.g002: Analysis of cytokines level in UC patients and controls.TNFα, IL-6 and IFNγ levels were measured in human blood plasma samples (n = 4–8) by ELISA. Cytokines level were measured in wild type controls, and in wild type patients as well as with different genotype category of patient samples. The data is represented as mean value of 4–8 samples in each group. The significance level of α ≤0.05 was chosen for all sets. pg = picograms, ml = milliliter.

Mentions: To check the role of polymorphism in cytokine homeostasis, we measured cytokine levels (TNFα, IL-6 and IFNγ) in control and UC patient samples (wild type and in individuals possessing single SNP or co-existing SNPs). All controls included for this analysis were non IBD and wild type (for all SNPs studied here). Level of TNFα (p = 0.004), IL-6 (p = 0.0001) and IFNγ (0.006) was significantly increased in wild type UC patients (wild type for all SNPs) as compared to controls (Fig. 2) suggesting that the change is due to the disease condition only. However, UC patients carrying D299G SNP in TLR4 gene revealed decreased level of TNFα (p = 0.026) as compared to wild type UC patients. Patients with R392X SNP in TLR5 gene also exhibited significantly decreased level of TNFα (p = 0.014), IL-6 (p = 0.028) and IFNγ (p = 0.001) as compared to wild type UC patients. In case of patients with N592S SNP (in TLR5 gene), TNFα (p = 0.0007) and IFNγ (p = 0.02) levels decreased significantly (Fig. 2). We were not able to measure cytokine level for T399I SNP due to insufficient number of samples carrying only this SNP.


Association of TLR5 gene polymorphisms in ulcerative colitis patients of north India and their role in cytokine homeostasis.

Meena NK, Ahuja V, Meena K, Paul J - PLoS ONE (2015)

Analysis of cytokines level in UC patients and controls.TNFα, IL-6 and IFNγ levels were measured in human blood plasma samples (n = 4–8) by ELISA. Cytokines level were measured in wild type controls, and in wild type patients as well as with different genotype category of patient samples. The data is represented as mean value of 4–8 samples in each group. The significance level of α ≤0.05 was chosen for all sets. pg = picograms, ml = milliliter.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4366177&req=5

pone.0120697.g002: Analysis of cytokines level in UC patients and controls.TNFα, IL-6 and IFNγ levels were measured in human blood plasma samples (n = 4–8) by ELISA. Cytokines level were measured in wild type controls, and in wild type patients as well as with different genotype category of patient samples. The data is represented as mean value of 4–8 samples in each group. The significance level of α ≤0.05 was chosen for all sets. pg = picograms, ml = milliliter.
Mentions: To check the role of polymorphism in cytokine homeostasis, we measured cytokine levels (TNFα, IL-6 and IFNγ) in control and UC patient samples (wild type and in individuals possessing single SNP or co-existing SNPs). All controls included for this analysis were non IBD and wild type (for all SNPs studied here). Level of TNFα (p = 0.004), IL-6 (p = 0.0001) and IFNγ (0.006) was significantly increased in wild type UC patients (wild type for all SNPs) as compared to controls (Fig. 2) suggesting that the change is due to the disease condition only. However, UC patients carrying D299G SNP in TLR4 gene revealed decreased level of TNFα (p = 0.026) as compared to wild type UC patients. Patients with R392X SNP in TLR5 gene also exhibited significantly decreased level of TNFα (p = 0.014), IL-6 (p = 0.028) and IFNγ (p = 0.001) as compared to wild type UC patients. In case of patients with N592S SNP (in TLR5 gene), TNFα (p = 0.0007) and IFNγ (p = 0.02) levels decreased significantly (Fig. 2). We were not able to measure cytokine level for T399I SNP due to insufficient number of samples carrying only this SNP.

Bottom Line: However, there was decreased level of TNFα (p = 0.014), IL-6 (p = 0.028) and IFNγ (p = 0.001) in patients carrying TLR5-R392X variant as compared to wild type patients.Patients carrying two simultaneous SNPs D299G in TLR4 gene and N592S in TLR5 gene showed significant decrease in the levels of TNFα (p = 0.011) and IFNγ (p = 0.016).Polymorphisms in TLR 5 genes were significantly associated with the UC in North Indian population.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, Jawaharlal Nehru University, New Delhi, India.

ABSTRACT

Background and aim: In health, TLR signaling protects the intestinal epithelial barrier and in disease, aberrant TLR signaling stimulates diverse inflammatory responses. Association of TLR polymorphisms is ethnicity dependent but how they impact the complex pathogenesis of IBD is not clearly defined. So we propose to study the status of polymorphisms in TLR family of genes and their effect on cytokines level in UC patients.

Methods: The genotypes of the six loci TLR1-R80T, TLR2-R753Q, TLR3-S258G, TLR5-R392X, TLR5-N592S and TLR6-S249P were determined in 350 controls and 328 UC patients by PCR-RFLP and sequencing. Cytokine levels were measured by ELISA in blood plasma samples. Data were analyzed statistically by SPSS software.

Results: TLR5 variants R392X and N592S showed significant association (p = 0.007, 0.021) with UC patients but TLR 1, 2, 3, 6 variants did not show any association. Unlike other studies carried out in different ethnic groups, TLR 6 (S249P) SNP was universally present in our population irrespective of disease. Genotype-phenotype correlation analysis revealed that the patients having combination of multiple SNPs both in TLR5 and TLR4 gene suffered from severe disease condition and diagnosed at an early age. The level of TNFα (p = 0.004), IL-6 (p = 0.0001) and IFNγ (p = 0.006) significantly increased in patients as compared to controls having wild genotypes for the studied SNPs. However, there was decreased level of TNFα (p = 0.014), IL-6 (p = 0.028) and IFNγ (p = 0.001) in patients carrying TLR5-R392X variant as compared to wild type patients. Patients carrying two simultaneous SNPs D299G in TLR4 gene and N592S in TLR5 gene showed significant decrease in the levels of TNFα (p = 0.011) and IFNγ (p = 0.016).

Conclusion: Polymorphisms in TLR 5 genes were significantly associated with the UC in North Indian population. The cytokine level was significantly modulated in patients with different genotypes of TLR4 and TLR5 SNPs.

Show MeSH
Related in: MedlinePlus