Quantitative Analysis of p40/p46 and p69/p71 Forms of 2′,5′-Oligoadenylate Synthetase mRNA by Competitive PCR and Its Clinical Application

Abstract

AbstractBackground: 2′,5′-Oligoadenylate synthetases (2-5AS) are type I interferon (IFN)-induced proteins with antiviral capacity. Three major forms of 2-5AS with distinct enzymatic activities have been described in IFN-treated human cells. We measured distinct forms of 2-5AS mRNA to analyze the relationship with its enzymatic activity and response to IFN therapy in chronic hepatitis C.Methods: We established a method to quantify p40/p46 and p69/p71 forms of 2-5AS mRNA by use of reverse transcription followed by competitive PCR. The 2-5AS mRNA concentrations were measured in peripheral blood mononuclear cells from 40 patients with chronic hepatitis C and 28 control individuals.Results: Reconstitution experiments and comparison with Northern blot analyses revealed that our method accurately and linearly quantified 2-5AS mRNA. 2-5AS mRNA concentrations and 2-5AS enzymatic activity were correlated (P <0.03). Our data demonstrated a correlation in 2-5AS mRNA between p40/p46 and p69/p71 (P <0.02), indicating a similar regulation of the expression of these genes. Our data also demonstrated that pretreatment concentrations of 2-5AS mRNA correlated with responses to IFN therapy in chronic hepatitis C.Conclusions: Our method for measuring 2-5AS mRNA concentrations could provide an important marker for selecting patients for IFN therapy and may be useful for the development of more effective therapeutic strategies for chronic hepatitis C.