Expression of biologically active recombinant porcine interleukin-12 from Escherichia coli.
The control of viral infections is of critical importance to livestock industries worldwide and is highlighted by costly infection outbreaks, such as that seen with foot and mouth disease virus. To ameliorate the impact of increasing problems with viral infections, new vaccine and anti-viral strategies are required and a greater understanding of the anti-viral response is essential. Furthermore, in pigs, evidence is still being gathered on the components of a defined anti-viral immune response. However, this has been greatly improved by the recent cloning and expression of critical cytokines involved in the anti-viral response. To assess the use of recombinant porcine interleukin-12 (rPoIL-12) as an immunotherapeutic and immunomodulator of swine, we have cloned and expressed rPoIL-12 as a single-chain fusion protein from Esherichia coli (E. coli). The fusion encodes the p40 and p35 subunits, linked by a glycine-serine linker and expressed as a C-terminal 6xHis tagged protein. rPoIL-12 stimulated the proliferation of human lymphoblasts and its activity on porcine cells was demonstrated by the ability of rPoIL-12 to increase the mRNA expression of porcine interleukin-18 receptor-alpha (poIL-18Ralpha) from porcine peripheral blood mononuclear cells (PoPMBCs). This data supports the inclusion of E. coli produced rPoIL-12 in immunomodulation strategies in the pig.
Morris
K R
KR
CSIRO Livestock Industries, Private Bag 24, Geelong, Victoria 3220, Australia. Kirsten.Morris@csiro.au
Bruce
M P
MP
Janardhana
V
V
Thomas
J D
JD
Bean
A G D
AG
Strom
D G
DG
D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli; D005786:Gene Expression Regulation / Q000276:immunology*; D007167:Immunotherapy / Q000379:methods / Q000662:veterinary*; D018664:Interleukin-12 / Q000378:metabolism*; D053727:Interleukin-18 Receptor alpha Subunit / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism*; D034421:Sus scrofa / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
D053727:Interleukin-18 Receptor alpha Subunit; D011994:Recombinant Proteins; D014765:Viral Vaccines; D018664:Interleukin-12