Detection of differential expression of mouse interferon-alpha subtypes by polymerase chain reaction using specific primers

J Immunol Methods. 2004 Jan;284(1-2):177-86. doi: 10.1016/j.jim.2003.10.012.

Abstract

Specific primers for nine mouse interferon-alpha (IFN-alpha) subtypes, namely, IFN-alpha1, IFN-alpha1-9, IFN-alpha2, IFN-alpha4, IFN-alpha5, IFN-alpha7, IFN-alpha6/8, IFN-alpha11, and IFN-alphaB, were designed and evaluated on Poly(I).Poly(C)-induced and influenza virus-infected L929 cells. Specificity of the primers was confirmed in a cross-polymerase chain reaction (cross-PCR). IFN-alpha1, IFN-alpha1-9, IFN-alpha4, IFN-alpha6/8, IFN-alpha11, and IFN-alphaB were found to be induced in L929 cells 6-9 h after Poly(I).Poly(C) treatment. The amplification of a particular subtype was not biased in the presence of excess of other templates. Differential expression of the IFN-alpha subtypes was observed in influenza A/NWS/33- and B/Lee/40-infected L929 cells. A/NWS/33 virus was found to upregulate the gene expression of IFN-alpha1, IFN-alpha4, IFN-alpha6/8, IFN-alpha11, and IFN-alphaB in L929 cells as early as 6 h after infection. In B/Lee/40-infected L929 cells, only IFN-alpha4 was upregulated. Our results suggest that the designed primers will serve as a useful tool in analyzing the expression of IFN-alpha subtypes in various systems and hence for the evaluation of their function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • DNA Primers / chemistry*
  • DNA Primers / genetics
  • Fibroblasts
  • Influenza A virus / immunology
  • Influenza B virus / immunology
  • Interferon-alpha / biosynthesis*
  • Interferon-alpha / classification
  • Interferon-alpha / genetics
  • Mice
  • RNA, Messenger / biosynthesis*
  • RNA, Messenger / genetics
  • Reverse Transcriptase Polymerase Chain Reaction / methods*

Substances

  • DNA Primers
  • Interferon-alpha
  • RNA, Messenger