Association of interleukins genes polymorphisms with multi-drug resistant tuberculosis in Ukrainian population

INTRODUCTION: Multi-drug resistant tuberculosis (MDR TB) is a significant health problem in some parts of the world. Three major cytokines involved in TB immunopathogenesis include IL-2, IL-4 and IL-10. The susceptibility to MDR TB may be genetically determined. The aim of the study was to assess the association of IL-2, IL-4, IL-10 gene polymorphisms with multi-drug resistant tuberculosis (MDR TB) in Ukrainian population. MATERIAL AND METHODS: We observed 140 patients suffering from infiltrative pulmonary tuberculosis (PT) and 30 apparently healthy subjects. The patients were assigned to two groups whether they suffer or do not suffer from pulmonary MDR TB. Interleukin gene (IL) polymorphisms, particularly T330G polymorphism in the IL-2 gene, C589T polymorphism in the IL-4 gene and G1082A polymorphism in the IL-10 gene were studied through polymerase chain reaction. Circulating levels of IL-2, IL-4 and IL-10 in venous blood were estimated using ELISA. RESULTS: Prior to treatment, patients with PT showed significant increase of IL-2 levels and decrease of IL-4 and IL-10 levels compared to apparently healthy subjects. Circulating IL-4 and IL-10 levels were significantly decreased whilst serum IL-2 level was significantly increased in patients with MDR TB compared to non-MDR TB. Low IL-4 and IL-10 secretion and considerable IL-2 alterations were shown to be significantly associated with mutations of homozygous and heterozygous genotypes affecting C589T polymorphism in the IL-4 gene, G1082A polymorphism in the IL-10 gene and T330G polymorphism in the IL-2 gene in patients with PT. CONCLUSIONS: Heterozygous genotype and mutations homozygous genotypes gene in polymorphisms determining specified cytokines’ production is a PT risk factor and may lead to disease progression into chronic phase. Heterozygous genotype of aforementioned cytokine genetic polymorphisms was significantly the most frequent in patients with MDR TB.INTRODUCTION: Multi-drug resistant tuberculosis (MDR TB) is a significant health problem in some parts of the world. Three major cytokines involved in TB immunopathogenesis include IL-2, IL-4 and IL-10. The susceptibility to MDR TB may be genetically determined. The aim of the study was to assess the association of IL-2, IL-4, IL-10 gene polymorphisms with multi-drug resistant tuberculosis (MDR TB) in Ukrainian population. MATERIAL AND METHODS: We observed 140 patients suffering from infiltrative pulmonary tuberculosis (PT) and 30 apparently healthy subjects. The patients were assigned to two groups whether they suffer or do not suffer from pulmonary MDR TB. Interleukin gene (IL) polymorphisms, particularly T330G polymorphism in the IL-2 gene, C589T polymorphism in the IL-4 gene and G1082A polymorphism in the IL-10 gene were studied through polymerase chain reaction. Circulating levels of IL-2, IL-4 and IL-10 in venous blood were estimated using ELISA. RESULTS: Prior to treatment, patients with PT showed significant increase of IL-2 levels and decrease of IL-4 and IL-10 levels compared to apparently healthy subjects. Circulating IL-4 and IL-10 levels were significantly decreased whilst serum IL-2 level was significantly increased in patients with MDR TB compared to non-MDR TB. Low IL-4 and IL-10 secretion and considerable IL-2 alterations were shown to be significantly associated with mutations of homozygous and heterozygous genotypes affecting C589T polymorphism in the IL-4 gene, G1082A polymorphism in the IL-10 gene and T330G polymorphism in the IL-2 gene in patients with PT. CONCLUSIONS: Heterozygous genotype and mutations homozygous genotypes gene in polymorphisms determining specified cytokines’ production is a PT risk factor and may lead to disease progression into chronic phase. Heterozygous genotype of aforementioned cytokine genetic polymorphisms was significantly the most frequent in patients with MDR TB

Związek wariantów polimorficznych genów kodujących interleukiny z występowaniem wielolekoopornej gruźlicy w populacji Ukrainy

WSTĘP: Gruźlica wielolekooporna (MDR-TB) stanowi poważny problem zdrowotny w pewnych regionach świata. Interleukiny 2 (IL-2), 4 (IL-4) i 10 (IL-10) odgrywają istotną rolę w immunopatogenezie gruźlicy. Podatność na gruźlicę wielolekooporną może być genetycznie uwarunkowana. Celem badania była ocena związku pomiędzy polimorfizmem genów IL-2, IL-4, IL-10 a występowaniem gruźlicy wielolekoopornej w populacji ukraińskiej. MATERIAŁ I METODY: Do badania włączono 140 chorych na gruźlicę naciekową i 30 osób zdrowych (grupa kontrolna). Wyodrębniono grupę chorych na gruźlicę wielolekooporną (MDR TB) i gruźlicę z zachowaną opornością na leki przeciwgruźlicze (non-MDR TB). Zbadano polimorfizm T330G genu IL-2, C589T genu IL-4 i G1082A genu IL-10 przy zastosowaniu łańcuchowej reakcji polimerazy. Stężenia IL-2, IL-4 and IL-10 w surowicy oznaczono za pomocą testu ELISA. WYNIKI: Przed leczeniem u chorych na gruźlicę stężenia w surowicy IL-2 były wyższe, a IL-4 i IL-10 niższe w porównaniu z grupą kontrolną. Stężenia IL-4 i IL-10 były istotnie niższe, podczas gdy stężenie IL-2 było istotnie wyższe w grupie MDR TB w porównaniu z pozostałymi chorymi (non- MDR TB). Opisane zmiany związane były z homozygotycznymi lub heterozygotycznymi mutacjami polimorficznymi C589T genu IL-4, G1082A genu IL-10 i T330G genu IL-2. WNIOSKI: Mutacje genów badanych cytokin mogą stanowić czynnik ryzyka gruźlicy i prowadzić do progresji i przewlekania się choroby. W grupie MDR TB genotypy heterozygotyczne badanych cytokin występowały najczęściej.WSTĘP: Gruźlica wielolekooporna (MDR-TB) stanowi poważny problem zdrowotny w pewnych regionach świata. Interleukiny 2 (IL-2), 4 (IL-4) i 10 (IL-10) odgrywają istotną rolę w immunopatogenezie gruźlicy. Podatność na gruźlicę wielolekooporną może być genetycznie uwarunkowana. Celem badania była ocena związku pomiędzy polimorfizmem genów IL-2, IL-4, IL-10 a występowaniem gruźlicy wielolekoopornej w populacji ukraińskiej. MATERIAŁ I METODY: Do badania włączono 140 chorych na gruźlicę naciekową i 30 osób zdrowych (grupa kontrolna). Wyodrębniono grupę chorych na gruźlicę wielolekooporną (MDR TB) i gruźlicę z zachowaną opornością na leki przeciwgruźlicze (non-MDR TB). Zbadano polimorfizm T330G genu IL-2, C589T genu IL-4 i G1082A genu IL-10 przy zastosowaniu łańcuchowej reakcji polimerazy. Stężenia IL-2, IL-4 and IL-10 w surowicy oznaczono za pomocą testu ELISA. WYNIKI: Przed leczeniem u chorych na gruźlicę stężenia w surowicy IL-2 były wyższe, a IL-4 i IL-10 niższe w porównaniu z grupą kontrolną. Stężenia IL-4 i IL-10 były istotnie niższe, podczas gdy stężenie IL-2 było istotnie wyższe w grupie MDR TB w porównaniu z pozostałymi chorymi (non- MDR TB). Opisane zmiany związane były z homozygotycznymi lub heterozygotycznymi mutacjami polimorficznymi C589T genu IL-4, G1082A genu IL-10 i T330G genu IL-2. WNIOSKI: Mutacje genów badanych cytokin mogą stanowić czynnik ryzyka gruźlicy i prowadzić do progresji i przewlekania się choroby. W grupie MDR TB genotypy heterozygotyczne badanych cytokin występowały najczęściej

Changes in nitric oxide synthase and nitrite and nitrate serum levels in patients with or without multidrug-resistant tuberculosis undergoing the intensive phase of antituberculosis therapy

Objective/background: There is a paucity of published data on the effect of tuberculosis (TB) chemotherapy on nitric oxide (NO) synthesis and metabolism in newly diagnosed and relapsed patients with or without multidrug-resistant TB (MDR-TB). Methods: The pattern of NO response in 140 patients with pulmonary TB, including 74 with MDR-TB (1st group) and 66 without MDR-TB (2nd group) has been studied and compared with the NO status of 30 healthy donors (3rd group). Patients comprised those with newly diagnosed pulmonary TB (Subgroups 1B and 2B) and recurrent or relapsed TB (Subgroups 1A and 2A). The NO status was assessed by measuring inducible NO synthase (iNOS), nitrites, and nitrates levels. This was measured prior to treatment initiation and 2 months after the prescribed chemotherapy. Results: Increased levels of NO indices were found in patients with TB when compared with healthy controls—1st group: iNOS, 231.6 ± 6.65pmol/min/mgB; nitrites, 5.626 ± 0.15 μmol/L; and nitrates, 62.89 ± 1.42 μmol/L (Subgroup 1A: iNOS, 208.40 ± 8.26pmol/min/mgB; nitrites, 5.027 ± 0.17 μmol/L; and nitrates, 59.29 ± 1.79 μmol/L and Subgroup 1B: iNOS, 260.4 ± 8.56pmol/min/mgB; nitrites, 6.371 ± 0.19 μmol/L; and nitrates, 67.36 ± 2.03 μmol/L); 2nd group: iNOS, 286.3 ± 5.92pmol/min/mgB; nitrites, 6.747 ± 0.17 μmol/L; and nitrates, 72.02 ± 1.43 μmol/L (Subgroup 2A: iNOS, 260.9 ± 14.12pmol/min/mgB; nitrites, 5.686 ± 0.20 μmol/L; and nitrates, 66.26 ± 1.89 μmol/L and Subgroup 2B: iNOS, 293.7 ± 6.13pmol/min/mgB; nitrites, 7.059 ± 0.19 μmol/L; and nitrates, 73.72 ± 1.71 μmol/L) versus healthy controls (iNOS, 81.03 ± 2.36pmol/min/mgB; nitrites, 3.83 ± 0.093 μmol/L; and nitrates, 37.98 ± 1.30 μmol/L). After 2 months of chemotherapy, a significant decrease in NO indicators was observed in the patients with TB, particularly in those without MDR-TB—1st group: iNOS, 114.9 ± 3.2pmol/min/mgB; nitrites, 4.21 ± 0.13 μmol/L; and nitrates, 46.65 ± 1.04 μmol/L (Subgroup 1A: iNOS, 125.3 ± 4.5pmol/min/mgB; nitrites, 4.42 ± 0.14 μmol/L; and nitrates, 49.38 ± 1.30 μmol/L and Subgroup 1B: iNOS, 102 ± 3.53pmol/min/mgB; nitrites, 3.93 ± 0.13 μmol/L; and nitrates, 43.26 ± 1.50 μmol/L) and 2nd group: iNOS, 91.4 ± 2.53pmol/min/mgB; nitrites, 3.67 ± 0.09 μmol/L; and nitrates, 35.65 ± 1.06 μmol/L (Subgroup 2A: iNOS, 106.7 ± 5.2pmol/min/mgB; nitrites, 4.04 ± 0.19 μmol/L; and nitrates-40.53 ± 1.83 μmol/L and Subgroup 2B, iNOS, 86.7 ± 2.59pmol/min/mgB; nitrites, 3.56 ± 0.1 μmol/L; and nitrates, 34.22 ± 1.19 μmol/L). The decline in NO activity was less prominent in patients with recurrent TB and MDR-TB, which suggests lower level of immunologic and reparative processes in such patients. Conclusion: In patients with pulmonary TB, significantly higher levels of NO activity were observed as compared with the levels in healthy individuals. In patients with recurrent TB and MDR-TB, significantly lower levels of NO indicators were observed in comparison with patients with newly diagnosed pulmonary TB. After 2 months on chemotherapy, a significant decrease in iNOS activity and NO metabolites was observed in patients with pulmonary TB, but the decrease in NO indicators was manifested mostly in the newly diagnosed pulmonary TB patients and patients without MDR-TB as opposed to patients with recurrent TB and MDR-TB, which suggests lower levels of immunologic and reparative processes in such patients. Therefore, the levels of nitrites and nitrates as well as iNOS activity may serve as additional diagnostic criteria to differentiate MDR-TB from nonresistant TB in patients with relapsed and newly diagnosed TB. Easily assessed NO-related markers can also serve as predictors of treatment outcome because patients with drug-susceptible strains had lower NO output approaching levels found in controls

Morphological changes in experimental tuberculosis resulting from treatment with quercetin and polyvinylpyrrolidone

Research objective: Morphological study of tissue necrosis stages in experimental organ-preserving tuberculosis pharmacotherapy using Quercetin and Polyvinylpyrrolidone (QP). Background and methods: 32 laboratory mice of C57BL/6JLacSto strain were used in the experiment. The animals were divided into five groups, six to seven mice in each: group 1- Mycobacterium tuberculosis (MBT) uninfected mice; group 2- MBT infected mice; group 3- MBT infected and treated with antituberculosis preparation (ATP); group 4- MBT infected and QP treated; group 5- MBT infected and treated with ATP and QP. The mice were infected through caudal vein injection with MTB H37Rv strain. The preparation QP, which belongs to the capillary-stabilizing-remedy group, was used for the research. The ATP were izoniazid and streptomycin. Results: QP produced a strict delineation of caseous necrosis from the unaffected parts of the connective tissue with fibrosis in the center and a large number of Langerhans cells, which was not observed in the control groups without QP. The combination of QP and ATP had more pronounced effects. In MBT-infected mice, where QP was not used, unlike the group where QP was used, adipose dystrophy of hepatocytes was observed. Thus, the hepatoprotective effect of QP against TB can be suggested. Conclusion: QP produces a clear delineation of caseous necrosis from an uninfected tissue by connective-tissue formation, and by forming fibrotic tissue in the center of epithelioid cells that prevents further TB dissemination by enhancing TB pharmacotherapy

Changes in nitric oxide synthase and nitrite and nitrate serum levels in patients with or without MDR-TB undergoing the intensive phase of anti-tuberculosis therapy

Background: There is a paucity of published data on the effect of TB chemotherapy on nitric oxide (NO) synthesis and metabolism in newly diagnosed and relapsed patients with or without multi-drug resistant tuberculosis (MDRTB). Methods: The pattern of NO response in 140 patients with pulmonary TB, including 74 with MDR-TB and 66 without MDR-TB has been studied and compared to the NO status of 30 healthy donors. Patients comprised those with newly diagnosed TB and recurrent or relapsed TB. The NO status was assessed by measuring inducible NO synthase (iNOS) and nitrites and nitrates levels. This was measured prior to treatment initiation and two months after the prescribed chemotherapy. Results: Increased levels of NO indices were found in patients with tuberculosis when compared to healthy controls. After two months of chemotherapy a significant decrease in NO indicators was observed in the patients with TB, particularly in those without MDR-TB and newly diagnosed TB. The decline in NO activity was less prominent in patients with recurrent TB and MDR-TB, which suggests lower level of immunologic and reparative processes in such patients. Conclusion: Changes in serum levels of nitrites and nitrates as well as iNOS activity in neutrophils may serve as diagnostic criteria to differentiate various clinical forms of TB and help as prognostic tool to predict treatment outcome