Association between lncRNA IFNG-AS1 SNP and Hashimoto's thyroiditis / 中华内分泌代谢杂志

Objective To assess the association between three single nucleotide polymorphisms( SNPs) ( rs10878724、 rs7980829 and rs11177020 ) of lnc IFNG-AS1 and Hashimoto's thyroiditis ( HT) susceptibility. Methods TaqMan probe technology was used to genotype the selected SNPs in a total of 179 subjects, including 70 HT cases, and 109 controls. The expression levels of lnc IFNG-AS1 and IFNG were detected by SYBR-Green qRT-PCR. Results Compared with control, not only the A allele and AA genotype frequencies of rs10878724 were significantly different in group HT ( P=0. 01, P=0. 003), but also the T allele and TT genotype frequencies of rs7980829 were significantly high in group HT. Haplotype analysis showed that the G-G-A decreased the risk of HT (P=0.04), while the haplotype of A-T-T incresed the risk of HT( P=0.01). The relative expression of both IFNG mRNA and lnc IFNG-AS1 were higher in group HT than in control( P=0. 001,P=0. 013). In HT patients, IFNG mRNA relative expression in both rs7980829-TT and rs1087872-TT were significantly higher than those of other genotypes(P=0.017,P=0.009). Conclusion The SNPs of Inc IFNG-AS1 were correlated with the expression levels of IFNG and lncRNA IFNG-AS1. Noncoding genes should be further screened as potential biomarkers in prediction of HT susceptibility.

Concordance between IFNγ gene +874 A/T polymorphism and interferon-γ expression in a TB-endemic indigenous setting

Abstract INTRODUCTION: Interferon-γ (IFN-γ) plays a crucial role in resistance to mycobacterial diseases; accordingly, variants of the gene encoding this cytokine may be associated with elevated risk of contracting pulmonary tuberculosis (TB). METHODS: Blood samples were collected from 135 Warao indigenous individuals with newly diagnosed sputum culture-positive TB. Of these, 24 were diagnosed with active tuberculosis (ATB). The study comprised 111 participants, who were grouped as follows: 1) 14 tuberculin skin test (TST)-positive Warao indigenous individuals and 4 that were QuantiFERON-TB?Gold In-Tube (QFT-IT) test-positive, collectively comprising the latent TB infection group (LTBI), n = 18), and 2) healthy controls who were QFT-IT- and TST-negative, comprising the control group (CTRL, n = 93). Detection of the IFN γ gene (IFNG) +874A/T polymorphism was performed via PCR and quantification of IFNG expression via qPCR. RESULTS: Relative to indigenous and white Americans, ATB and CTRL groups had a higher frequency of the IFNG SNP (+874A): 23 (95.8%) and 108 (97.3%), respectively. Indigenous Warao individuals homozygous for the IFNG (+874) A allele exhibited 3.59-fold increased risk of developing TB (95% confidence interval, 2.60-4.96, p =0.0001). A decreased frequency of the AT genotype was observed in individuals with pulmonary TB (4.16%) and controls (0.90%). The frequency of the TT genotype was decreased among controls (1.80%); none of the patients with TB were found to have this genotype. The differences in IFNG expression between the groups, under unstimulated and stimulated conditions, were not statistically significant. CONCLUSIONS: Preliminary results demonstrate concordance between IFNG +874 A/A genotype and low expression of IFNG.

Association of TNFA (−308G/A), IFNG (+874 A/T) and IL-10 (−819 C/T) polymorphisms with protection and susceptibility to dengue in Brazilian population

Objective To evaluate gene polymorphisms and their association with susceptibility to dengue. Methods A retrospective case-control study was performed with 262 subjects, comprising 78 dengue fever (DF) patients, 49 dengue hemorrhagic fever (DHF) patients and 135 healthy controls. Genotypic and allelic profiles were identified using polymerase chain reaction based in real time and amplification-refractory mutation system. Results We observed a protective association of IL-10 (−819 C/T) C allele (P = 0.028, OR = 0.56, CI = 0.34–0.91) against DHF, while the C/T (P = 0.047, OR = 2.10, CI = 1.01–4.38) and T/T (P = 0.008, OR = 3.82, CI = 1.38–10.59) genotypes were associated with DHF and DF, respectively. The dominant model TNFA −308 GA + AA (P = 0.043, OR = 0.45, CI = 0.20–1.00) genotypes were found to have protective effect against dengue infection. A protective association among the IFNG (+874 A/T) A/T genotype against DF (P = 0.02, OR = 0.46, CI = 0.24–0.89) and DHF (P = 0.034, OR = 0.43, CI = 0.19–0.95) was observed. When the studied single-nucleotide polymorphism was analyzed in combination, the combination GTA (P = 0.022, OR = 2.95, CI = 1.18–7.41) was statistically significantly associated with susceptibility to DF and the combination GCT (P = 0.035, OR = 0.28, CI = 0.08–0.90) with protection against the development of DHF. Conclusions This research identifies the association of the IFNG (+874 A/T), TNFA (−308G/A), IL-10 (−819 C/T) genotypes as a factor for protection, susceptibility and severity to dengue.

Association of TNFA (-308G/A), IFNG (+874 A/T) and IL-10 (-819 C/T) polymorphisms with protection and susceptibility to dengue in Brazilian population

OBJECTIVE@#To evaluate gene polymorphisms and their association with susceptibility to dengue.@*METHODS@#A retrospective case-control study was performed with 262 subjects, comprising 78 dengue fever (DF) patients, 49 dengue hemorrhagic fever (DHF) patients and 135 healthy controls. Genotypic and allelic profiles were identified using polymerase chain reaction based in real time and amplification-refractory mutation system.@*RESULTS@#We observed a protective association of IL-10 (-819 C/T) C allele (P = 0.028, OR = 0.56, CI = 0.34-0.91) against DHF, while the C/T (P = 0.047, OR = 2.10, CI = 1.01-4.38) and T/T (P = 0.008, OR = 3.82, CI = 1.38-10.59) genotypes were associated with DHF and DF, respectively. The dominant model TNFA -308 GA + AA (P = 0.043, OR = 0.45, CI = 0.20-1.00) genotypes were found to have protective effect against dengue infection. A protective association among the IFNG (+874 A/T) A/T genotype against DF (P = 0.02, OR = 0.46, CI = 0.24-0.89) and DHF (P = 0.034, OR = 0.43, CI = 0.19-0.95) was observed. When the studied single-nucleotide polymorphism was analyzed in combination, the combination GTA (P = 0.022, OR = 2.95, CI = 1.18-7.41) was statistically significantly associated with susceptibility to DF and the combination GCT (P = 0.035, OR = 0.28, CI = 0.08-0.90) with protection against the development of DHF.@*CONCLUSIONS@#This research identifies the association of the IFNG (+874 A/T), TNFA (-308G/A), IL-10 (-819 C/T) genotypes as a factor for protection, susceptibility and severity to dengue.

Diagnostic accuracy of immunological methods in patients with tuberculous pleural effusion from Venezuela / Precisión diagnóstica de métodos inmunológicos en pacientes con tuberculosis pleural de Venezuela

In recent years, better diagnostics for tuberculosis (TB) has received increasing attention, especially the diagnosis of tuberculous pleural effusion, which is difficult and at present the main tool in TPE diagnostic is pleural effusion smear and culture, but unfortunately, sensitivities are low, therefore better TPE diagnostic tools are needed. The aim of this study was to find a diagnostic algorithm to assess the progress in TPE diagnostic at the Hospital Vargas de Caracas, that permits identification of the majority of patients, at a satisfactory cost-benefit ratio, evaluating the levels of IFN-g and IL-12p40 in pleural effusion and serum, as well as the antibody reactivity in order to compare it with microbiological tests. A total of 60 individuals with pleural effusion were studied; 20 patients with tuberculous pleural effusion (TPE) formed the patient group and 40 patients with non-tuberculous pleural effusion (NTPE) formed the control group. The levels of IFN-g and IL-12p40 in effusion and serum and class and subclasses of IgG reactivity to Mycobacterium tuberculosis antigens were measured by ELISA. The utility of these methods for diagnosis of TPE was evaluated using receiver operating characteristic (ROC) curve analysis. The results of the 11 immunological methods evaluated showed that the anti-PPD IgG2 method was able to reach the highest specificity of 95% (CI: 88.3-101.8), positive predictive value (PPV)=75 (at 30% sensitivity); while that the overall sensitivity of methods was between 95% and 30%, of these, two methods reached higher sensitivities; increased levels of pleural IFN-g, with a sensitivity of 95% (CI: 85.5-104.5) with the highest negative predictive value (NPV)=97, (at 82.5% specificity), followed by decreased levels of serum IL-12p40 with a sensitivity of 95% (CI: 85.5-104.5), NPV=95.2 (at 50% specificity). In contrast, microbiological methods showed that smear had a sensitivity of only 20%, while smear plus ...

Recientemente existe un gran interés hacia un mejor y más rápido diagnóstico de tuberculosis (TB), especialmente de tuberculosis pleural, el cual es difícil. Al presente las principales herramientas diagnósticas son la baciloscopia y el cultivo de líquido pleural; desafortunadamente, las sensibilidades de estos métodos son bajas, por lo que el desarrollo de nuevas herramientas diagnósticas es necesario. El objetivo del presente estudio consistió en encontrar un algoritmo que permita la rápida identificación de la mayoría de los pacientes con TB pleural que ingresan en el Hospital Vargas de Caracas a un buen costo-beneficio. Para esto se evaluaron los niveles de las citocinas Interferón-gamma (IFN-g) y la Interleucina 12p40 (IL-12p40) en líquido pleural y suero, así como la reactividad de anticuerpos contra antígenos de Mycobacterium tuberculosis. Se estudiaron 60 individuos con derrame pleural; 20 individuos con líquido pleural tuberculoso (LPT) conformaron el grupo de pacientes y 40 individuos con líquido pleural no tuberculoso (LPNT) el grupo de controles. La técnica de inmunoensayo de ELISA fue utilizada para medir los niveles de IFN-g y IL-12p40; así como las reactividades de los diversos isotipos y subclases de inmunoglobulina G (IgG) frente a antígenos del bacilo. La utilidad de los métodos fue evaluada utilizando el análisis de las curvas ROC (receiver operating characteristic). Los resultados de los 11 métodos inmunológicos evaluados mostraron que el método IgG2 anti-PPD alcanzó la mayor especificidad de 95%, (CI: 88,3-101,8) con un valor predictivo positivo (VPP) de 75. La sensibilidad de los métodos estuvo entre 30% y 95%; dos métodos alcanzaron altas sensibilidades: los altos niveles de IFN-g en líquido pleural, con sensibilidad de 95% (CI: 85,5-104,5), con un valor predictivo negativo (VPN) de 97, seguido de los bajos niveles de IL-12p40 en suero, con una sensibilidad de 95% (CI: 85,5-104,5) con un VPN de 95,2. En contraste, ...

Associação do polimorfismo de ifng +874T/A com a cardiopatia chagásica crônica / Association of ifng +874 T/A polymorphism with chronic chagas' disease

A doença de Chagas é uma zoonose causada pelo parasita hemoflagelado Trypanosoma cruzi. Atualmente afeta cerca de 10 milhões de pessoas nas Américas, e aproximadamente 90 milhões de pessoas se encontram em áreas de risco. A cardiopatia chagásica crônica (CCC) se desenvolve em um terço dos indivíduos infectados e corresponde a uma miocardite crônica fibrosante intensa e os possíveis fatores que podem contribuir para o desenvolvimento desta podem estar relacionados com a carga parasitária, a cepa do parasita, a autoimunidade e fatores genéticos do parasita e do hospedeiro. Citocinas e quimiocinas têm um importante papel no desenvolvimento de uma resposta imune protetora contra o parasita, porém esta pode estar envolvida com o aumento da inflamação encontrada no miocárdio de pacientes com CCC. O principal objetivo deste estudo foi identificar a freqüência dos genótipos e alelos para o SNP no IFNG na posição +874T/A através da técnica de ARMS - PCR, em 102 pacientes soropositivos para o T. cruzi apresentando a CCC, 86 pacientes soropositivos sem cardiopatia aparente e 179 controles soronegativos para T. cruzi. Não há diferença estatística quando pacientes com CCC foram comparados com indivíduos saudáveis, indicando que o polimorfismo para IFNG na posição +874T/A não parece ter influencia na infecção. Contudo, observamos que o genótipo AA foi mais freqüentemente encontrado em pacientes portadores da CCC do que naqueles portadores da forma indeterminada, sugerindo que indivíduos com este genótipo estão mais susceptíveis ao aparecimento de lesões cardíacas e ao adoecimento (p=0,024)...

Chagas disease is a zoonosis caused by the parasite hemoflagellate Trypanosoma cruzi. Currently affects approximately 10 millions people in the Americas, where approximately 90 millions people are at risk areas. The Chronic chagasic cardiomyopathy (CCC) develops in one third of infected individuals and represents a severe chronic myocarditi. Possible factors that may contribute to the development of several cariopathy can be related to parasite load and difference on strains, autoimmunity and the genetics of the parasite and host. Cytokines and chemokinesplay an important role in the development of a protective immune response against the parasite, but it may be involved with increased inflammation found in the myocardium of CCC. The main objective of this study was to identify the frequency of genotypes and alleles for the SNP in the IFNG position +874 T/A by ARMS-PCR technique in 102 patients seropositive for T. cruzi presenting the CCC, 86seropositive patients without apparent heart disease and 179 negative controls for T. cruzi. There was no statistical difference when CCC patients were compared withhealthy subjects, indicating that the polymorphism at the position +874 T/A does not seem to influence the infection. However, we observed that the AA genotype was more frequently found among patients with CCC than in patients without apparent heart disease, suggesting that individuals with this genotype are more susceptible to develop cardiac illness (p=0,024)...

Morphometric Analysis of Granulomas Induced by Mycobacterium bovis Suggests an Influence of IFN-Gamma on the Generation and Modulation upon Granulomatous Inflammatory Response in the Different Tissues / El Análisis Morfométrico de Granulomas Inducidos por Mycobacterium bovis Sugiere la Influencia del IFN-Gamma en la Generación y Modulación de la Respuesta Inflamatoria Granulomatosa en Diferentes Tejidos

There is evidence in both human and experimental infection caused by Mycobacterium tuberculosis, that immunologic factors influence susceptibility to infection and the progress of the disease. The present study aims to evaluate the role of IFN- g upon inflammatory granulomatous response against M. bovis. To pursue that, C57BL/6 mice lacking the genes for synthesis of IFN- g (IFN- g-/-) and their wild-type counterparts (IFN- g+/+) were intravenously inoculated with M. bovis. The ability of M. bovis to survive and replicate in the liver and lungs was evaluated by counting colony-forming unit (CFU). The histopathological features of granulomatous inflammatory response in the liver and lungs were analyzed during the infection by M. bovis. Granuloma parameters such as, size (sectional area), granuloma volume, volume density, and numerical density were calculated in each point of infection. Bacillary load was higher in both organs of the animals that were IFN- g-/- than in IFN- g+/+ mice. Granulomas were observed in the IFN- g-/- mice after 30 days of infection and were detected earlier in controls (15 days of infection). Hepatic granulomas persisted in the IFN- g-/- mice, but in the IFN- g+/+ mice control of the inflammation. In conclusion, IFN- g influenced the multiplication of M. bovis, as well as modulated the granulomatous inflammation.

Existen evidencias, tanto en humanos como en modelos experimentales, que factores inmunológicos influencian. durante la infección causada por la Mycobacterium tuberculosis, tanto la infección como la progresión de la enfermedad. Este estudio se propone evaluar el papel de IFN-g en la respuesta inflamatoria granulomatosa contra M. bovis. Para ello, se inocularon ratones C57BL/6 knockout para IFN- g (IFN- g-/-) y los correspondientes salvajes (IFN- g+/+) con M. bovis. Evaluamos la capacidad de la M. bovis de sobrevivir y replicarse en el hígado y pulmones mediante la cuantificación de unidades formadoras de colonias (CPU). También analizamos los aspectos histopatológicos de la respuesta inflamatoria granulomatosa en el hígado y los pulmones durante la infección con M. bovis. Para cada punto de infección, se calcularon los parámetros del granuloma, tales como el tamaño (área de sección), el volumen del granuloma, la densidad de volumen y densidad numérica. La carga bacilar fue mayor en los dos órganos estudiados procedentes de los ratones IFN- g-/-. Los granulomas de los ratones controles se detectaron a los 15 días, mientras que los de los ratones IFN-g no se detectaron hasta los 30 días post infección. Los granulomas hepáticos persistieron en los ratones IFN- g-/-. Como conclusión es posible afirmar que el IFN- g influencia la multiplicación por M. bovis, así como también modula la inflamación granulomatosa.

Cell-type specific regulation of thrombospondin-1 expression and its promoter activity by regulatory agents

Thrombospondin-1 (TSP-1), a multifunctional protein that is able to function as a negative regulator of solid tumor progression and angiogenesis, is normally present at a very low level but rapidly elevated in pathological tissues. To understand the cellular regulation of TSP-1 expression, the mode of it's expression in Hep3B, SK-HEP-1, and porcine aortic endothelial (PAE) cells was examined in the presence of all-trans retinoic acid (ATRA), interleukin-6 (IL-6), interferon-gamma (IFN-gamma), or phorbol 12-myristate 13-acetate (PMA). ATRA or IL-6 induced a dose-dependent increase of TSP-1 protein and mRNA levels in PAE cells, while they negatively regulated TSP-1 expression in the Hep3B and SK-HEP-1 cells. In contrast, PMA showed just the opposite effects on the TSP-1 expression in the same cells. IFN-gamma had little effect on TSP-1 level in Hep3B and PAE cells. The TSP-1 expression in SK-HEP-1 cells by these agents showed a close resemblance to that of liver cells rather than that of the endothelial cell line. Possible TSP-1 promoter-mediated responses by ATRA, IL-6, IFN-gamma, or PMA in Hep3B and PAE cells examined with luciferase activity of TSP-LUC reporter plasmid showed that levels of TSP-1 promoter activity were lower than that of the expressed TSP-1 protein and mRNA levels. Transfection of c-Jun and/or RARalpha expression vectors into Hep3B and PAE cells resulted in the enhanced TSP-1 promoter activity as well as the increments of of its protein and mRNA level. These results suggest that regulatory agents-induced TSP-1 expression may be attributed to mRNA stability and/or translational activation in concert with transcriptional activation and TSP-1 expression may be independently controlled via each signal pathway stimulated by PMA or ATRA.

Partial purification and characterization of a novel murine factor that augments the expression of class I MHC antigens on tumor cells

A soluble factor which augments the expression of major histocompatibility complex class I (MHC I) antigens on a number of murine tumor cell lines, has been isolated from the culture supernatants of mixed lymphocyte reaction of spleen cells derived from C57B1/6, Balb/c and Swiss mice. The factor, termed MHC-augmenting factor (MHC-AF) has been partially purified by Sephadex G-100 column chromatography and reverse phase HPLC. MHC-AF activity is associated with an 18 kDa molecule. MHC-AF activity was resistant to pH 2.0 treatment and partially purified MHC-AF preparations did not have any activity in L929 cell/vesicular stomatitis virus (VSV) interferon bioassay system. Antibodies to IFN-gamma did not block the activity of MHC-AF. These results indicate that a MHC-AF distinct from IFN-gamma, is produced by mouse spleen cells undergoing a mixed lymphocyte reaction.