RESUMEN
The hepatitis C virus (HCV) core protein possesses amino acid sequences highly conserved among HCV isolates and has proved useful for various diagnostic tests. To date, no information has been published regarding the development of an immunochromatographic test for HCV core antigen detection. Therefore, the aim of this research was to develop a rapid, easily performed, highly sensitive and specific test for detection of the HCV core antigen, based on the immunochromatographic strip. The genomic region encoding the core protein (amino acids 1-136) of the hepatitis C virus was expressed in Escherichia coli as a recombinant fusion protein with glutathione S-transferase (GST) cloned into the prokaryotic expression vector pET42a and was confirmed by immunological detection with HCV positive serum. Positive reactions were detected weakly at a 1:15 dilution of the serum and more strongly in 1:10, 1:5, 1:2 and 1:1 dilutions, by the immunochromatographic test. In addition, the test was capable of detecting 0.25-12.0 microg of the recombinant protein. This immunochromatographic technique opens new perspectives for the diagnosis of hepatitis C during the early seroconversion phase and for a rapid core antigen detection.
Asunto(s)
Hepacivirus/aislamiento & purificación , Hepatitis C/diagnóstico , Proteínas del Núcleo Viral/análisis , Animales , Cromatografía de Afinidad/métodos , Humanos , Inmunoensayo/métodos , RatonesRESUMEN
HIV patients are predisposed to the development of hypertriglyceridemia and hypercholesterolemia as a result of both viral infection and HIV infection therapy, especially the protease inhibitors. Chemokines and cytokines are present at sites of inflammation and can influence the nature of the inflammatory response in atherosclerosis. We investigated the correlation between biochemical variables and beta-chemokines (MIP-1alpha and RANTES) and the apolipoprotein E genotype in HIV-infected individuals. The apolipoproteins were measured by nephelometry. Triglycerides and total cholesterol were determined by standard enzymatic procedures. The beta-chemokines were detected by ELISA. The genetic category of CCR5 and apolipoprotein E were determined by PCR amplification and restriction enzymes. Immunological and virological profiles were assessed by TCD(4)+ and TCD(8)+ lymphocyte counts and viral load quantification. Positive correlations were found between apo E and CD(8)+ (p = 0.035), apo E and viral load (p = 0.018), MIP-1alpha and triglycerides (p = 0.039) and MIP-1a and VLDL (p = 0.040). Negative correlations were found between viral load and CD(4)+ (p = 0.05) and RANTES and CD(4)+ (p = 0.029). The beta-chemokine levels may influence lipid metabolism in HIV-infected individuals.
Asunto(s)
Apolipoproteínas E/sangre , Quimiocina CCL5/sangre , Infecciones por VIH/sangre , Lipoproteínas/sangre , Proteínas Inflamatorias de Macrófagos/sangre , Adulto , Apolipoproteínas E/genética , Biomarcadores/sangre , Relación CD4-CD8 , Quimiocina CCL3 , Quimiocina CCL4 , Femenino , Genotipo , Humanos , Masculino , Receptores CCR5/sangre , Carga ViralRESUMEN
HIV patients are predisposed to the development of hypertriglyceridemia and hypercholesterolemia as a result of both viral infection and HIV infection therapy, especially the protease inhibitors. Chemokines and cytokines are present at sites of inflammation and can influence the nature of the inflammatory response in atherosclerosis. We investigated the correlation between biochemical variables and beta-chemokines (MIP-1alpha and RANTES) and the apolipoprotein E genotype in HIV-infected individuals. The apolipoproteins were measured by nephelometry. Triglycerides and total cholesterol were determined by standard enzymatic procedures. The beta-chemokines were detected by ELISA. The genetic category of CCR5 and apolipoprotein E were determined by PCR amplification and restriction enzymes. Immunological and virological profiles were assessed by TCD4+ and TCD8+ lymphocyte counts and viral load quantification. Positive correlations were found between apo E and CD8+ (p = 0.035), apo E and viral load (p = 0.018), MIP-1alpha and triglycerides (p = 0.039) and MIP-1a and VLDL (p = 0.040). Negative correlations were found between viral load and CD4+ (p = 0.05) and RANTES and CD4+ (p = 0.029). The beta-chemokine levels may influence lipid metabolism in HIV-infected individuals.
Asunto(s)
Humanos , Masculino , Femenino , Adulto , Persona de Mediana Edad , Apolipoproteínas E/sangre , Quimiocina CCL5 , Infecciones por VIH/sangre , Lipoproteínas/sangre , Proteínas Inflamatorias de Macrófagos , Apolipoproteínas E/genética , Apolipoproteínas E/metabolismo , Biomarcadores/sangre , Quimiocina CCL5 , Ensayo de Inmunoadsorción Enzimática , Genotipo , Infecciones por VIH/metabolismo , Lipoproteínas/metabolismo , Proteínas Inflamatorias de Macrófagos , Nefelometría y Turbidimetría , Reacción en Cadena de la Polimerasa , /sangre , Carga ViralRESUMEN
Neste trabalho, investigamos concentração da vitamina B12 e folato, considerando-se a influência dos genótipos da metilenotetrahidrofolato redutase, o perfil imunológico e a terapia antiretroviral utilizada na população brasileira portadora do HIV. Um grupo de 86 indivíduos portadores do HIV-1 e 29 doadores de sangue foram recrutados para compor a casuística. Entre os infectados pelo HIV-1, observou-se menor concentração de B12 no grupo com maior número de linfócitos TCD4+. Não encontramos diferença na distribuição genotípica para as mutações MTHFR C677T e A1298C entre infectados e não infectados pelo HIV-1. Indivíduos portadores do HIV, genótipo C677C, apresentaram concentrações menores de B12 em relação ao grupo controle de mesmo genótipo. A terapia antiretroviral não mostrou qualquer influência nos valores de folato e vitamina B12. Estudos adicionais são necessários para reavaliar a prevalência de menores concentrações de B12 e folato e de hiperhomocisteinemia na população portadora do HIV sob a ótica do uso de HAART e da melhoria na sobrevida dos pacientes.
Asunto(s)
Humanos , Masculino , Femenino , Adulto , Persona de Mediana Edad , Ácido Fólico , Infecciones por VIH , Homocisteína , Metilenotetrahidrofolato Reductasa (NADPH2) , Vitamina B 12 , Alelos , Antirretrovirales , Biomarcadores , Estudios de Casos y Controles , Recuento de Linfocito CD4 , Genotipo , Factores de RiesgoRESUMEN
In this study we sought to investigate the B12 and folate levels regarding the influence of methylenetetrahydrofolate reductase genotypes, immunological profile and antiretroviral therapy in the Brazilian HIV-infected population. The study group comprised 89 HIV-infected individuals and 29 blood donors. There was a decrease in the B12 levels in the HIV-infected group with higher TCD4+ lymphocyte counts. No differences in the genotype distribution for methylenetetrahydrofolate reductase polymorphisms between the HIV-infected individuals and the controls were found. HIV-infected individuals carrying the C677C genotype presented lower B12 levels (313.91 +/- 154.05) than those with the same genotype in the control group (408.27 +/- 207.69). Also, the antiretroviral therapy was not a source of variation of the folate and B12 serum levels. Further studies are needed to reanalyze the prevalence of low levels of folate and B12 and hyperhomocysteinemia among HIV-infected patients with regard to the use of HAART and the increased life expectancy of such patients.