Expression of fusion IL2-B7.1(IgV+C) and effects on T lymphocytes.
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.
Department of Biological Science and Engineering, School of Life Science & Technology, Xi'an Jiaotong University, Xi'an, China.
Linghong
L
Kong
Yaochen
Y
Li
Ye
Y
Yang
Kangsheng
K
Li
D000818:Animals; D018122:B7-1 Antigen / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D049109:Cell Proliferation / Q000187:drug effects; D003001:Cloning, Molecular; D019295:Computational Biology; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel; D000939:Epitopes; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007376:Interleukin-2 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D051379:Mice; D010957:Plasmids / Q000378:metabolism; D018583:Pliability / Q000187:drug effects; D011387:Prokaryotic Cells / Q000187:drug effects / Q000378:metabolism; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D013601:T-Lymphocytes / Q000166:cytology* / Q000187:drug effects
D018122:B7-1 Antigen; D000939:Epitopes; D007376:Interleukin-2; D011993:Recombinant Fusion Proteins