RESUMEN
Tuberculosis remains one of the most important infectious diseases worldwide. Several studies have suggested that genetic factors may affect susceptibility to tuberculosis, but the specific genes involved have not yet been fully characterized. NRAMP1/SLC11 A1 and P2X(7) genes have been linked to increased risk for tuberculosis in some African and Asiatic populations. To explore the potential role of these genes in the susceptibility to pulmonary tuberculosis in a Mexican mestizo population, we evaluated the association of D543N and 3'-UTR polymorphisms in NRAMP1/SLC11 A1 and - 762 and A1513C polymorphisms in P2X(7) genes with the risk for tuberculosis. Polymerase chain reaction (PCR) amplification of genomic DNA followed by restriction fragment length polymorphism analysis, and allelic-specific PCR was employed. We found no significant differences in allelic frequency in NRAMP1/SLC11 A1 gene polymorphisms in 94 patients with tuberculosis compared to 100 healthy contacts. Similarly, no significant association of the P2X(7)-762 gene polymorphism with tuberculosis was detected. In contrast, the P2X(7) A1513C polymorphism was associated significantly with tuberculosis (P = 0.02, odds ratio = 5.28, 95% CI, 0.99-37.69), an association that had not been reported previously. However, when the function of P2X(7) was assessed by an L-selectin loss assay, we did not find significant differences in patients compared to healthy contacts or between PPD(+) and PPD(-) control individuals. This study further supports the complex role of P2X(7) gene in host regulation of Mycobacterium tuberculosis infection, and demonstrates that different associations of gene polymorphisms and tuberculosis are found in distinct racial populations.
Asunto(s)
Proteínas de Transporte de Catión/genética , Polimorfismo de Longitud del Fragmento de Restricción , Receptores Purinérgicos P2/genética , Tuberculosis Pulmonar/genética , Estudios de Casos y Controles , Proteínas de Transporte de Catión/sangre , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Genotipo , Humanos , Selectina L/sangre , Leucocitos Mononucleares/metabolismo , Masculino , Reacción en Cadena de la Polimerasa/métodos , ARN Mensajero/genética , Receptores Purinérgicos P2/sangre , Receptores Purinérgicos P2/fisiología , Receptores Purinérgicos P2X7 , Tuberculosis Pulmonar/sangreRESUMEN
P2X(7) is a channel receptor gated by adenosine triphosphate (ATP) that is involved in the killing of intracellular mycobacteria. To explore further the role of P2X(7) in immunity against Mycobacterium tuberculosis, we studied its expression and function in 19 patients with pulmonary tuberculosis (TB) and 19 healthy contacts. Flow cytometry analysis showed a similar and variable expression of P2X(7) in TB patients and healthy subjects. In contrast, P2X(7) mARN levels were significantly higher in TB patients. When the function of the P2X(7) receptor in peripheral blood mononuclear cells (PBMC) was assessed by the effect of exogenous ATP on apoptosis, the uptake of the fluorescent marker Lucifer yellow or extracellular signal regulated kinase (ERK) phosphorylation, no significant differences were detected in patients and controls. However, mRNA macroarray analysis showed that upon stimulation with ATP, the PBMC from TB patients showed a significant induction of a higher number of cytokine genes (27 of 96), and a lower number of apoptosis genes (20 of 96) compared to healthy controls (17 and 76 genes, respectively). These results suggest that although the PBMC from TB patients do not show apparent abnormalities in the expression of P2X(7), and the intracellular signals generated through it, the pattern of gene expression induced by ATP in these cells is different from that found in healthy contacts. This phenomenon suggests a defective function of P2X(7) in the immune cells from TB patients, a condition that may contribute to the inability of these patients to eliminate the mycobacteria.