|HAN, JAE - SEOUL NAT UNIV
|HAN, IN - SEOUL NAT UNIV
Submitted to: Korean-Japan Joint Seminar Collaborative Researches on Biological Science
Publication Type: Abstract Only
Publication Acceptance Date: 10/30/1997
Publication Date: N/A
Technical Abstract: Chicken cDNA encoding interferon-gamma (IFN-g) was cloned from mRNA made from p34 cell line, a CD4 4 chicken T-cell hybridoma, by reverse transcription-polymerase chain reaction (RT-PCR) based on the published sequence and expressed in E. coli, COS and chicken cell line. The expression of chicken IFN-g gene was examined in concanavalin A (Con A)-activated spleen and peripheral blood lymphocyte by Northern blot and RT-PCR, IFN-g mRNA was detected was early as 30 min after Con A activation of lymphocytes, reached a peak expression of 2 hr continuing to 4 hr post T cell activation followed by sharp decline, indicating that chicken IFN-g may be an early expressed gene upon mitogenic stimulation. The polyclonal antibody made against immunogenic peptide portion of IFN-y immunoprecipitated 62 kDa molecule of E. coli expressed MBP-fusion protein of IFN-g and 26 kDa secreted molecules from CD4 4 T cell hybridoma. The recombinant chicken IFN-g (rchIFN-g) inhibited vesicular stomatitis virus (VSV)-mediated cytotoxicity of chicken embryonic fibroblast (CEF) cells and unregulated the class I and II major histocompatibility complex antigen expressions on HDII macrophage cell line. These results show that rchIFN-g which was expressed in chicken cell line is bioreactive and possesses macrophage activating activity which is associated with chicken IFN-g.