Expression and functional characterization of recombinant chicken interferon-gamma.
A cDNA encoding chicken interferon-gamma (chIFN-gamma) was cloned from a CD4+ T-cell hybridoma by reverse transcription-polymerase chain reaction (RT-PCR) and expressed in Escherichia coli, COS- and CEC-32 fibroblast cell lines. In general, recombinant chicken IFN-gamma (rchIFN-gamma) expressed in the COS- and CEC-32 cell lines showed high bioactivity in vitro. The kinetics of IFN-gamma gene expression were examined in concanavalin A (Con A)-activated spleen lymphocytes by Northern blot and RT-PCR. IFN-gamma mRNA was detected as early as 30 min after Con A activation, reached peak expression at 2 h and then decreased starting at 4 h post Con A activation. A rabbit serum made to a synthetic peptide of IFN-gamma immunoprecipitated a 60 kDa E. coli maltose-binding fusion protein of recombinant IFN-gamma (MBP-IFN) and a 26-27 kDa secreted protein from COS cells and Con A-activated spleen cells. IFN-gamma inhibited vesicular stomatitis virus (VSV) mediated cytotoxicity of chicken embryonic fibroblast (CEF) cells and upregulated the expression of many macrophage cell surface antigens, including class I and class II major histocompatibility complex (MHC) proteins. These results show that chicken IFN-gamma possesses anti-viral activity and immunoregulates macrophage activities.
Song
K D
KD
Department of Animal Science and Technology, College of Agriculture and Life Sciences, Seoul National University, Suweon, South Korea.
Lillehoj
H S
HS
Choi
K D
KD
Zarlenga
D
D
Han
J Y
JY
D000818:Animals; D002645:Chickens; D003001:Cloning, Molecular; D003208:Concanavalin A / Q000494:pharmacology; D007371:Interferon-gamma / Q000096:biosynthesis / Q000494:pharmacology*; D008213:Lymphocyte Activation; D008262:Macrophage Activation / Q000187:drug effects; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits; D011994:Recombinant Proteins
D012333:RNA, Messenger; D011994:Recombinant Proteins; D003208:Concanavalin A; D007371:Interferon-gamma