RESUMO
INTRODUCTION: The development of quantitative methods for the detection of viral nucleic acids have significantly improved our ability to manage disease progression and to assess the efficacy of antiviral treatment. Moreover, major advances in molecular technologies during the last decade have allowed the identification of new host genetic markers associated with antiviral drug response but have also strongly revolutionized the way we see and perform virus diagnostics in the coming years. Areas covered: In this review, we describe the history and development of virology diagnostic methods, dedicating particular emphasis on the gradual evolution and recent advances toward the introduction of multiparametric platforms for the syndromic diagnosis. In parallel, we outline the consolidation of viral genome quantification practice in different clinical settings. Expert commentary: More rapid, accurate and affordable molecular technology can be predictable with particular emphasis on emerging techniques (next generation sequencing, digital PCR, point of care testing and syndromic diagnosis) to simplify viral diagnosis in the next future.
Assuntos
Antígenos Virais/análise , Genoma Viral , Técnicas de Diagnóstico Molecular , Virologia/métodos , Viroses/diagnóstico , Vírus/isolamento & purificação , Ensaio de Imunoadsorção Enzimática , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Kit de Reagentes para Diagnóstico , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e Especificidade , Virologia/instrumentação , Viroses/virologia , Vírus/classificação , Vírus/genética , Vírus/imunologiaRESUMO
This study aimed to evaluate the association between the IFNL4 rs368234815 (ΔG/TT) dinucleotide polymorphism and the IFN response during chronic HIV-1 infection. We carried out genotyping analysis and measured the expression of IFN-stimulated genes (ISGs) (myxovirus resistance protein A [MxA], ISG15, ISG56, apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like [APOBEC] 3F and APOBEC3G) on peripheral blood mononuclear cells collected from naïve and HAART-treated HIV-1-infected patients. There were no statistically significant differences in endogenous ISGs mRNA levels among HIV-1-positive patients bearing different IFNL4 genotypes, suggesting that ISG expression is independent of the IFNL4 genotype in HIV-1 infection.
Assuntos
Perfilação da Expressão Gênica , Genótipo , Infecções por HIV/imunologia , Fatores Imunológicos/biossíntese , Interleucinas/genética , Polimorfismo Genético , Adulto , Feminino , Técnicas de Genotipagem , Humanos , Leucócitos Mononucleares/imunologia , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/análiseRESUMO
Given the multifactorial nature of action of type I interferon (IFN) in HIV-1 infection and the need to firmly establish the action of key components of IFN pathways, we compared the IFN stimulated gene (ISG)15 expression with that of other well-characterized ISGs, evaluating its relationship with immunosuppressive factors regulating T-cell response in HIV-1 patients. PBMC from 225 subjects were included: healthy donors (n=30), naïve (n=93) and HAART treated HIV-1 subjects (n=102). Levels of ISG15-mRNA, ISG56-mRNA, APOBEC3G/3F-mRNA, TRAIL-mRNA, IDO-mRNA, proviral load andISG15 (rs15842 and rs1921) SNPs were evaluated by using TaqMan assays. We found that ISG15, ISG56, APOBEC3G/3F levels were increased in untreated HIV-1 patients compared to healthy donors, being ISG15 the highest ISG expressed. The amount of ISG15 correlated with viral load and with CD4+ T cell counts whereas no relationship was found between all ISGs analyzed and proviral load or HIV-1 tropism. ISG15 expression was reduced following long-term antiretroviral therapy. In addition, ISG15 levels were correlated with those of TRAIL and IDO in HIV-1 viremic patients. Lastly, ISG15 SNPs had no influence on ISG15 levels. We demonstrates that ISG15 is elevated in viremic HIV-1 patients and is associated with high TRAIL and IDO levels.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Citocinas/genética , Infecções por HIV/genética , Indolamina-Pirrol 2,3,-Dioxigenase/genética , RNA Mensageiro/genética , Ligante Indutor de Apoptose Relacionado a TNF/genética , Ubiquitinas/genética , Viremia/genética , Desaminase APOBEC-3G , Proteínas Adaptadoras de Transdução de Sinal , Adulto , Terapia Antirretroviral de Alta Atividade , Contagem de Linfócito CD4 , Linfócitos T CD4-Positivos/virologia , Estudos de Casos e Controles , Citidina Desaminase/genética , Citidina Desaminase/imunologia , Citocinas/imunologia , Citosina Desaminase/genética , Citosina Desaminase/imunologia , Feminino , Regulação da Expressão Gênica , Infecções por HIV/tratamento farmacológico , Infecções por HIV/imunologia , Infecções por HIV/virologia , HIV-1/efeitos dos fármacos , HIV-1/imunologia , Humanos , Indolamina-Pirrol 2,3,-Dioxigenase/imunologia , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , RNA Mensageiro/imunologia , Proteínas de Ligação a RNA , Transdução de Sinais , Ligante Indutor de Apoptose Relacionado a TNF/imunologia , Fatores de Transcrição/genética , Fatores de Transcrição/imunologia , Ubiquitinas/imunologia , Carga Viral/efeitos dos fármacos , Tropismo Viral , Viremia/tratamento farmacológico , Viremia/imunologia , Viremia/virologiaRESUMO
Interferons (IFNs) are critically important in the control of influenza A virus infections. To better understand the pathogenic characteristics of the pandemic 2009 H1N1 influenza virus (pH1N1) from an innate immunity viewpoint, we investigated whether in vitro pH1N1 is sensitive to the antiviral activity of IFN beta, leukocyte IFN alpha, and several IFN alpha subtypes in a human lung adenocarcinoma epithelial cell line under single-growth cycle conditions. The results showed that 50% inhibitory concentration values against pH1N1 for various type I IFN preparations were higher than those against the IFN-sensitive encephalomyocarditis virus. Leukocyte IFN alpha and IFN alpha 5, 7, 8, 10, 14, 17, and 21 subtypes also appeared to be less active against pH1N1 than the Puerto Rico/8/34 H1N1 human influenza strain A. Taken together, the results provide new insights into the contributions of the various IFN alpha subtypes toward the regulation of innate immunity against pH1N1.