title: Purification of recombinant human interferon-epsilon and oligonucleotide microarray analysis of interferon-epsilon-regulated genes.
abstract: 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.
authors.affiliation: Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, 100 Ying-Xin Street, Xuan Wu District, Beijing 100052, China.
authors.forename: Fu-Wang
authors.initials: FW
authors.lastname: Peng
authors.forename: Zhao-Jun
authors.initials: ZJ
authors.lastname: Duan
authors.forename: Li-Shu
authors.initials: LS
authors.lastname: Zheng
authors.forename: Zhi-Ping
authors.initials: ZP
authors.lastname: Xie
authors.forename: Han-Chun
authors.initials: HC
authors.lastname: Gao
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.forename: Wu-Ping
authors.initials: WP
authors.lastname: Li
authors.forename: Yun-De
authors.initials: YD
authors.lastname: Hou
mesh_terms: 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
chemical_list: D000998:Antiviral Agents; D018076:DNA, Complementary; D007370:Interferon Type I; D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins