Interferon b (IFNb) represents an essential element of host defense against viruses and other pathogens, exciting immediate innate antiviral and anti-proliferative activities in cells and alerts the adaptive immune system to mount an adequate T helper-1 (Th-1) cells biased immune response. Substances containing conserved molecular patterns that identify them as non-self or “strange” induce the production of IFNb. In most body cells the transcription of IFNb results in only small amount of mRNA and for this reason preferentially mRNA IFNb constitutive producing human fibroblast (FS-4) cells are useful for the rescue and isolation of full IFNb gene.

We have established an easy Reverse transcription polymerase chain reaction (RT-PCR) for isolation of entire IFNb gene from healthy peripheral blood mononuclear cells (PBMC). PBMC were isolated by Ficoll-Hypaque centrifugation from normal healthy donors and simultaneously infected with 1 multiplicity of infection (MOI) of measles virus CAM-70 vaccine strain. Non-infected PBMC were run in parallel. The total RNA was extracted with Total RNA isolation kit (Promega, USA) and employed to perform the RT-PCR using SuperScriptIII One-Step RT-PCR kit with Platinum Taq High Fidelity (Invitrogen, USA). Manufacturer’s recommendations were followed in order to obtain the complete IFN gene. The PCR product was cloned in a TOPO vector (Invitrogen, USA) and sequenced using primers flanking the entire IFNb cloned gene.