Document Detail


Expression of fusion IL2-B7.1(IgV+C) and effects on T lymphocytes.
MedLine Citation:
PMID:  18059527     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The search for an effective immunotherapeutic treatment for tumors is an important area of cancer research. To prepare a more effective form of the bifunctional fusion protein IL2-B7.1(IgV+C) and analyze its effect on the stimulation of T lymphocyte proliferation, we used DNAStar 5.03 software to predict the structural diversity and biochemical character of IL2-B7.1(IgV+C). We then prepared fusion protein IL2-B7.1(IgV+C) by establishing its prokaryotic expression system, and tested its effect on the stimulation of T lymphocytes in vitro. The results indicated that IL2-B7.1(IgV+C) correctly formed a secondary structure in which both IL2 and B7.1(IgV+C) maintained their original hydrophilicity and epitopes. Western blot analysis revealed that IL2-B7.1(IgV+C) was efficiently expressed. Our analysis of CTLL-2 and T-cell proliferation showed that recombinant human (rh) IL2-B7.1(IgV+C) exerted the combined stimulating effects of both rhIL2 and rh B7.1(IgV+C) on cell proliferation, and that these effects could be blocked by adding either anti-IL2 or anti-B7.1 monoclonal antibodies. A >2-fold increase in [3H]TdR incorporation compared with that of cells treated with recombinant protein IL2, or B7.1(IgV+C) alone, revealed that rhIL2-B7.1(IgV+C) had dose-dependent synergetic effects on T-cell activation in the presence of anti-CD3 monoclonal antibody. We concluded that the augmented potency of rhIL2-B7.1(IgV+C) resulted in a stronger stimulation of T-cell proliferation than either rhB7.1(IgV+C) or rhIL2 alone.
Authors:
Linghong Kong; Yaochen Li; Ye Yang; Kangsheng Li
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biochemistry and cell biology = Biochimie et biologie cellulaire     Volume:  85     ISSN:  0829-8211     ISO Abbreviation:  Biochem. Cell Biol.     Publication Date:  2007 Dec 
Date Detail:
Created Date:  2007-12-06     Completed Date:  2008-03-07     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8606068     Medline TA:  Biochem Cell Biol     Country:  Canada    
Other Details:
Languages:  eng     Pagination:  685-95     Citation Subset:  IM    
Affiliation:
Department of Biological Science and Engineering, School of Life Science & Technology, Xi'an Jiaotong University, Xi'an, China.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antigens, CD80 / chemistry,  genetics,  metabolism*,  pharmacology
Cell Proliferation / drug effects
Cloning, Molecular
Computational Biology
Dose-Response Relationship, Drug
Electrophoresis, Agar Gel
Epitopes
Humans
Hydrophobicity
Interleukin-2 / chemistry,  genetics,  metabolism*,  pharmacology
Mice
Plasmids / metabolism
Pliability / drug effects
Prokaryotic Cells / drug effects,  metabolism
Protein Structure, Secondary
Recombinant Fusion Proteins / chemistry,  genetics,  metabolism*,  pharmacology
T-Lymphocytes / cytology*,  drug effects
Chemical
Reg. No./Substance:
0/Antigens, CD80; 0/Epitopes; 0/Interleukin-2; 0/Recombinant Fusion Proteins

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