PCR-Based Simultaneous Analysis of the Interferon-Alpha Family Reveals Distinct Kinetics for Early Interferons
(2008) Journal of Interferon & Cytokine Research — Vol. 28, n° 11, p. 653-660 (2008)
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- Here we describe a PCR-based analysis system that allows the simple simultaneous assessment of murine interferons (IFN)-alpha and IFN-beta induction in a single reaction. In this analysis, the so-called early IFN-alpha 4 can be distinguished from the so-called late IFN-non alpha 4 by employing a primer mixture that amplifies a part of the IFN-alpha genes in which IFN-alpha 4 exhibits a deletion of 15 nucleotides compared to IFN-non alpha 4. By including a final denaturation and a slow cooling step at the end of the PCR procedure, hybrid formation was avoided that regularly occurred when standard protocols were used. Separation of the amplification products on 4.5% agarose gels allowed the comparative assessment of the classical type I IFNs. Using this analysis system, we could show that in immortalized adult fibroblasts, IFN-beta is induced first and the two types of IFN-alpha are induced later and simultaneously. When similar fibroblasts derived from mice that lack IFN-beta were tested, the IFN response was delayed. However, now IFN-alpha 4 appeared first and apparently induced the cascade of IFN-nona4. This confirms the role of IFN-beta as master regulator of the normal IFN response.
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Lienenklaus, S., Walisko, R., Boekhorst, A. t., May, T., Samuelsson, C., Michiels, T., & Weiss, S. (2008). PCR-Based Simultaneous Analysis of the Interferon-Alpha Family Reveals Distinct Kinetics for Early Interferons. Journal of Interferon & Cytokine Research, 28(11), 653-660. https://doi.org/10.1089/jir.2008.0082 (Original work published 2008)