Expression of NAT2 in immune system cells and the relation of NAT2 gene polymorphisms in the anti-tuberculosis therapy in Mexican mestizo population
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Arylamine N-acetyltransferase 2 (NAT2) metabolizes isoniazid (INH) and Single Nucleotide Polymorphisms (SNP) responsible for its activity has been reported. The aim of this study in the Mexican mestizo population was to evaluate NAT2 expression at the protein level in immune cells, as well as the distribution and frequency of six NAT2 SNPs and their association with anti-TB therapy, by measuring the plasma levels of INH and Acetyl-INH (AcINH). We performed genotyping assays of NAT2 SNPs in 40 TB patients and 121 healthy volunteers by real-time PCR. A method for detecting NAT2 in immune cells using flow cytometry was developed. Plasma concentrations of INH and AcINH were obtained by HPLC in TB patients and the Metabolic Ratio (MR) was calculated. The phenotypes obtained in the healthy volunteers were as follows; 18.87 %25 of subjects had the rapid acetylator phenotype, 45.45 %25 had the intermediate phenotype and 39.66 %25 exhibited the slow acetylator phenotype. In the TB patient group, 35 %25 of patients had the rapid acetylator phenotype, 32.5 %25 were intermediate and 32.5 %25 showed the slow acetylator phenotype. A higher expression level of NAT2 in innate immune cells from TB patients compared to those from healthy volunteers was detected (P < 0.013). In TB patients the MR showed a bimodal distribution with an antimode of 0.7, which was used as a threshold value for acetylator classification. A high correspondence between the rapid and slow acetylator phenotype with MR was demonstrated. In conclusion, the 282C>T, 341T>C, 481C>T, 590G>A, 803A>G, 857G>A SNPs of NAT2 gene provides accurate for prediction of the acetylator phenotype in Mexican mestizo population. A statistical difference was found in frequency of rapid metabolizer phenotype, which was higher in TB patients. In addition, the expression of NAT2 protein in immune cells can lead to further studies related to its functional role in the innate immune response against M. tuberculosis and other xenobiotics metabolized by this enzyme. © 2014, Springer Science Business Media Dordrecht.
Arylamine N-acetyltransferase 2 (NAT2) metabolizes isoniazid (INH) and Single Nucleotide Polymorphisms (SNP) responsible for its activity has been reported. The aim of this study in the Mexican mestizo population was to evaluate NAT2 expression at the protein level in immune cells, as well as the distribution and frequency of six NAT2 SNPs and their association with anti-TB therapy, by measuring the plasma levels of INH and Acetyl-INH (AcINH). We performed genotyping assays of NAT2 SNPs in 40 TB patients and 121 healthy volunteers by real-time PCR. A method for detecting NAT2 in immune cells using flow cytometry was developed. Plasma concentrations of INH and AcINH were obtained by HPLC in TB patients and the Metabolic Ratio (MR) was calculated. The phenotypes obtained in the healthy volunteers were as follows; 18.87 %25 of subjects had the rapid acetylator phenotype, 45.45 %25 had the intermediate phenotype and 39.66 %25 exhibited the slow acetylator phenotype. In the TB patient group, 35 %25 of patients had the rapid acetylator phenotype, 32.5 %25 were intermediate and 32.5 %25 showed the slow acetylator phenotype. A higher expression level of NAT2 in innate immune cells from TB patients compared to those from healthy volunteers was detected (P < 0.013). In TB patients the MR showed a bimodal distribution with an antimode of 0.7, which was used as a threshold value for acetylator classification. A high correspondence between the rapid and slow acetylator phenotype with MR was demonstrated. In conclusion, the 282C>T, 341T>C, 481C>T, 590G>A, 803A>G, 857G>A SNPs of NAT2 gene provides accurate for prediction of the acetylator phenotype in Mexican mestizo population. A statistical difference was found in frequency of rapid metabolizer phenotype, which was higher in TB patients. In addition, the expression of NAT2 protein in immune cells can lead to further studies related to its functional role in the innate immune response against M. tuberculosis and other xenobiotics metabolized by this enzyme. © 2014, Springer Science%2bBusiness Media Dordrecht.
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Acetylator phenotype; Isoniazid; NAT2; NK cells; Tuberculosis adenine; arylamine acetyltransferase; cytosine; guanine; isoniazid; thymine; xenobiotic agent; arylamine acetyltransferase; isoniazid; NAT2 protein, human; tuberculostatic agent; acetylator phenotype; adult; aged; Article; clinical article; controlled study; drug blood level; female; flow cytometry; genotype; high performance liquid chromatography; human; human cell; immunocompetent cell; innate immunity; male; Mestizo; metabolic ratio; NAT2 gene; nucleic acid base substitution; phenotype; prediction; protein expression; real time polymerase chain reaction; single nucleotide polymorphism; tuberculosis; animal; blood; CHO cell line; Cricetulus; ethnology; genetics; HeLa cell line; lymphocyte; metabolism; Mexico; middle aged; young adult; Mycobacterium tuberculosis; Adult; Animals; Antitubercular Agents; Arylamine N-Acetyltransferase; CHO Cells; Cricetulus; Female; HeLa Cells; Humans; Isoniazid; Lymphocytes; Male; Mexico; Middle Aged; Polymorphism, Single Nucleotide; Tuberculosis; Young Adult
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