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Aim:To construct the recombinant vectors carrying interleukin (IL) -28A,IL-28B and IL-29 cDNAs and express them in human A549 cells,and analyze their antiviral activity in vesicular stomatitis virus (VSV)-infected human immortalized amnion epithelial cell line (WISH cells).Methods:Total cell RNA was extracted from human peripheral blood mononuclear cells (PBMC) activated with poly I:C.The cDNAs encoding human IL-28A.IL-28B.and IL-29 were amplified by reversetranscription polymerase chain reaction (RT-PCR) and inserted into pcDNA3.1/V5-His-TOPO vectors.These recombinant vectors were transfected into human A549 cells by a liposome-mediated gene transfer method.Semiquantitative RT-PCR and Westem blotting were used to detect the mRNA and protein expression of IL-28A,IL-28B,and IL-29.The antiviral activity of IL-28A,IL.28B,and IL-29 was determined by a cytopathic eflfect reduction assay on WISH cells using VSV as a challenge virus.Results:The DNA sequences of the inserts were identical to the published sequences encoding IL-28A,IL-28B,and IL-29 in GenBank.It was transfected cells.Expression of all 3 interleukins in A549 cells was confirmed by Wlestem blot analysis.IL-28 and IL-29 expressed by A549 cells.1ike interferon (IFN)α-2b,were able to protect WISH cells against VSV infection.Conclusion:IL-28 and IL-29 cDNAs were successfully cloned and expressed in eukaryotic cells via transfection with pcDNA3.1/V5-His-TOPO-IL-28/IL-29.Transfection with this vector produced a specific antiviral activity similar to that of IFN-α.which will provide a new tool for the functional study of these cytokines in humans.