A comparison of the anti-inflammatory and immuno-stimulatory activities of orf virus and ovine interleukin-10.

Orf virus causes pustular skin lesions (orf) in sheep, goats and humans. The virus encodes an interleukin-10 (orfvIL-10) that is identical in amino acid composition to ovine IL-10 (ovIL-10) over the C terminal two-thirds of the polypeptide, but not in the N terminal third. The immuno-suppressive and immuno-stimulatory activities of orfvIL-10 and ovIL-10 were compared. Both orfvIL-10 and ovIL-10 inhibited TNF-alpha and IL-8 cytokine production from stimulated ovine macrophages and keratinocytes and IFN-gamma and GM-CSF production from peripheral blood lymphocytes. OrfvIL-10 and ovIL-10 co-stimulated both ovine and murine mast cell proliferation in conjunction with IL-3 (ovine) or IL-4 (murine). Isoleucine at position 87 (Ile(87)) of the mature human IL-10 (huIL-10) has been reported as essential for the immuno-stimulatory activity of huIL-10. In spite of the differences in amino acids within the N-terminal third of orfvIL-10 compared with ovIL-10 and substitution of Ile(87) with Ala(87) in ovIL-10, these variants of ovIL-10 and orfvIL-10 all co-stimulated mast cell proliferation and inhibited macrophage IL-8 production. As ovIL-10 and orfvIL-10 have a similar structure to huIL-10 and conserved receptor-binding residues, it was concluded that Ile(87) is not essential for IL-10 immuno-stimulatory activity. Finally, ovine keratinocytes do not express ovIL-10. This might explain why orf virus has evolved a viral IL-10.

Infection with recombinant orf viruses demonstrates that the viral interleukin-10 is a virulence factor.

Orf virus is the prototype parapoxvirus that causes the contagious skin disease orf. It encodes an orthologue of the cytokine interleukin (IL)-10. Recombinant orf viruses were constructed in which the viral interleukin-10 (vorfIL-10) was disabled (vorfIL-10ko) and reinserted (vorfrevIL-10) at the same locus and compared to wild-type virus for their ability to induce skin lesions in sheep. After either primary infection or reinfection, smaller less severe lesions were recorded in the vorfIL-10ko-infected animals compared with either of the vorfIL-10-intact virus-infected animals. Thus, the vorfIL-10ko virus was attenuated compared with the vorfIL-10 intact viruses, demonstrating that orf virus IL-10 is a virulence factor. The virus IL-10 is one of several virulence or immuno-modulatory factors expressed by orf virus. Removal of any one of these genes would be expected to have only a partial effect on virulence, which is what was observed in this study with vorfIL-10.