Purification of recombinant human interferon-epsilon and oligonucleotide microarray analysis of interferon-epsilon-regulated genes.
Recently identified interferon-epsilon (IFN-epsilon) belongs to type I interferons. IFN-epsilon is highly and constitutively expressed in the brain, but its biochemical and biological characteristics are poorly understood. In this study, full-length IFN-epsilon cDNA was cloned from human peripheral blood lymphocyte by RT-PCR, and was expressed in Escherichia coli (E. coli). Reverse phase high pressure liquid chromatography was used to purify recombinant human IFN-epsilon (rhIFN-epsilon) and to facilitate refolding of the protein. About 0.8mg of highly purified rhIFN-epsilon protein was obtained from 100ml of E. coli culture. Functional study of rhIFN-epsilon demonstrated that the antiviral activity of rhIFN-epsilon was 6+/-0.5x10(5)IU/mg, which was lower than that of rhIFN-alpha-2b in the WISH-VSV (WISH cells infected with vesicular stomatitis virus) assay system. As for the activity to promote NK cytotoxicity and antiproliferation activities, rhIFN-epsilon was about 60 times less potent than rhIFN-alpha-2b. However, oligonucleotide microarray analyses revealed dramatic differences in gene expression profiles of cultured human cells treated with IFN-epsilon and IFN-alpha-2b. Particularly, differential regulation of genes related to central nervous system by rhIFN-epsilon suggests a role for IFN-epsilon in maintenance of the structure and function of brain.
Peng
Fu-Wang
FW
Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, 100 Ying-Xin Street, Xuan Wu District, Beijing 100052, China.
Duan
Zhao-Jun
ZJ
Zheng
Li-Shu
LS
Xie
Zhi-Ping
ZP
Gao
Han-Chun
HC
Zhang
Hui
H
Li
Wu-Ping
WP
Hou
Yun-De
YD
D000998:Antiviral Agents / Q000302:isolation & purification / Q000494:pharmacology; D001483:Base Sequence; D001921:Brain / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D003602:Cytotoxicity, Immunologic / Q000187:drug effects; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D066298:In Vitro Techniques; D007370:Interferon Type I / Q000235:genetics* / Q000302:isolation & purification* / Q000494:pharmacology; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000493:pharmacokinetics; D007694:Killer Cells, Natural / Q000187:drug effects / Q000276:immunology; D020411:Oligonucleotide Array Sequence Analysis; D011994:Recombinant Proteins
D000998:Antiviral Agents; D018076:DNA, Complementary; D007370:Interferon Type I; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins