ABSTRACT
<p><b>OBJECTIVE</b>To prepare human interferon-k (hIFN-kappa) and study its biological activities.</p><p><b>METHODS</b>Whole length of hIFN-kappa's cDNA was cloned, and its sequence was chemically synthesized according to the optimized codons of E.coli, then was expressed in E.coli DH5alpha. After purified, the rhIFN-kappa protein was tested for its various kinds of biological activities.</p><p><b>RESULTS</b>The purity of rhIFN-kappa was above 90%. In WHIS-VSV system, the antiviral activity of rhIFN-kappa was 2.0 x 10(6) IU/mg. Compared with rhIFN-alpha-2b, the biological activities of rhIFN-kappa were all feeble, including antiviral activity, promoting NK cell activity and anti-proliferation activity.</p><p><b>CONCLUSION</b>Antiviral activities of rhIFN-kappa on cell lines of different species are different, different viruses show different sensitivity to rhIFN-kappa.</p>
Subject(s)
Animals , Humans , Antiviral Agents , Pharmacology , Cell Line , Cell Proliferation , Chlorocebus aethiops , Cloning, Molecular , DNA, Complementary , Genetics , Electrophoresis, Polyacrylamide Gel , Escherichia coli , Genetics , Gene Expression , Interferon Type I , Genetics , Pharmacology , K562 Cells , Killer Cells, Natural , Cell Biology , Allergy and Immunology , Microbial Sensitivity Tests , Plasmids , Genetics , Recombinant Proteins , Metabolism , Pharmacology , Vero CellsABSTRACT
<p><b>OBJECTIVE</b>To construct a novel recombinant rhIFN-epsilon155ser, and study its biological activities.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>