[Biological activities of recombinant human interferon Epsilon].
OBJECTIVE
To construct a novel recombinant rhIFN-epsilon155ser, and study its biological activities.


METHODS
The whole sequence of rhIFN-epsilon was artificially synthesized and some codons were altered according to the preferred codon using of E.coli. The sequence was cloned into plasmid vector pBV220 to express in E.coli DH5alpha. After purification and re-folding of rhIFN-epsilon155ser inclusion body, the final product was tested for its biological activities, including anti-viral, anti-proliferative and NK cell enhancing activities. At the same time, by using DNA microarray biochips, the gene expression patterns in the rhIFN-epsilon155ser and rhIFN-alpha2b treated cells were compared and analyzed.


RESULTS
The re-built rhIFN-epsilon155ser sequence was expressed in E.coli as a form of inclusion body. After purified and re-folded, the rhIFN-epsilon155ser protein reached a purity of above 95%. The rhIFN-epsilon155ser protein had a specific anti-viral activity of about 6 x 10(5) IU/mg in WISH/VSV system. Its anti-proliferative activity and NK cell enhancing activities in vitro seemed to be lower than that of rhIFN-alpha2b. Data obtained from microarray biochips indicated that there were 283 pieces increasing 2 folds and 1489 pieces decreasing 2 folds among totally 22,278 pieces of human genes were found in the rhIFN-epsilon155ser treated cells; more changes in gene expression pattern were detected in the rhIFN-alpha treated cells.


CONCLUSIONS
A novel recombinant rhIFN-epsilon155ser was constructed, which belonged to type 1 interferon. The biological activities of rhIFN-epsilon155ser were compared with rhIFN-alpha2b. The changes of gene expression pattern in the interferon treated cells were detected, analyzed and discussed.
Peng
Fu-wang
FW
State Key Laboratory for Molecular Virology and Genetic Engineering, Institute for Viral Disease Control and Prevention, Beijing 100052, China.
Gao
Han-chun
HC
Xie
Zhi-ping
ZP
Zhang
Hui
H
Li
Qi-ming
QM
Duan
Zhao-jun
ZJ
Hou
Yun-de
YD
D000998:Antiviral Agents / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects*; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D006367:HeLa Cells; D006801:Humans; D007372:Interferons / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D020014:K562 Cells; D007694:Killer Cells, Natural / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D008826:Microbial Sensitivity Tests; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology*
D000998:Antiviral Agents; C524005:IFNE protein, human; D011994:Recombinant Proteins; D007372:Interferons