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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1-3
pubmed:dateCreated
2002-9-25
pubmed:abstractText
Interleukin-21 (IL-21) has recently been identified as a novel 4-helix-bundle type I cytokine possessing a cytokine receptor gamma chain essential for the immune response. We report the preparation and functional characterization of Escherichia coli-expressed recombinant human IL-21 (rIL-21). The rIL-21, expressed as insoluble inclusion bodies in E. coli, was solubilized and then refolded by using a modified dialysis method. The introduction of redox reagents during refolding led to a dramatic increase in the refolding efficiency. Circular dichroism spectrum analysis showed that the refolded rIL-21 had an alpha-helix as a secondary structure, which is a characteristic of type I cytokines. Flow cytometry confirmed previous reports that rIL-21 binds to CD3-activated T cells (T-LAK) and to cell lines Raji, HL60, and Jurkat. rIL-21 stimulated the proliferation of T-LAK but not peripheral blood mononuclear cells, and this effect seems to be identical to that of co-stimulation with anti-CD3 antibody. Growth inhibition assay indicated that enhancement of the cytotoxicity of T-LAK to the human bile duct carcinoma TFK-1 depended on the concentration of rIL-21. Thus, refolded rIL-21 had activity identical to that of authentic IL-21 and enhanced the anti-tumor activity of T-LAK. These conclusions suggest the potential use of the refolded cytokine in adoptive immunotherapy using T-LAK cells and in the discovery of other functions of the cytokine.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
0014-5793
pubmed:author
pubmed:issnType
Print
pubmed:day
25
pubmed:volume
528
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
70-6
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:12297282-Antineoplastic Agents, pubmed-meshheading:12297282-Base Sequence, pubmed-meshheading:12297282-Cytotoxicity, Immunologic, pubmed-meshheading:12297282-DNA, Recombinant, pubmed-meshheading:12297282-Escherichia coli, pubmed-meshheading:12297282-Humans, pubmed-meshheading:12297282-Immunotherapy, Adoptive, pubmed-meshheading:12297282-Inclusion Bodies, pubmed-meshheading:12297282-Interleukins, pubmed-meshheading:12297282-Killer Cells, Lymphokine-Activated, pubmed-meshheading:12297282-Molecular Sequence Data, pubmed-meshheading:12297282-Neoplasms, pubmed-meshheading:12297282-Protein Engineering, pubmed-meshheading:12297282-Protein Folding, pubmed-meshheading:12297282-Recombinant Proteins, pubmed-meshheading:12297282-T-Lymphocyte Subsets, pubmed-meshheading:12297282-Tumor Cells, Cultured
pubmed:year
2002
pubmed:articleTitle
Antitumor activity of interleukin-21 prepared by novel refolding procedure from inclusion bodies expressed in Escherichia coli.
pubmed:affiliation
Cell Resource Center for Biomedical Research, Institute of Development, Aging, and Cancer, Tohoku University, 4-1 Seiryomachi, Aoba-ku, Sendai 980-8575, Japan.
pubmed:publicationType
Journal Article, In Vitro, Research Support, Non-U.S. Gov't