Functional Fv fragment of an antibody specific for CD28: Fv‐mediated co‐stimulation of T cells

FEBS Letters

Katayose, Yu²  Matsuno, Seiki²  Kumagai, Izumi¹  Sakurai, Naoki²  Asano, Ryutaro¹  Arai, Takashi¹  Suzuki, Masanori²  Yoshida, Hiroshi³  Kudo, Toshio³  Takemura, Shin-ichi¹  Tsumoto, Kouhei¹  Kodama, Hideaki² 

[1] Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University, Aoba-yama 07, Aoba-Ku, Sendai 980-8579, Japan;Cell Resource Center for Biomedical Research, Institute of Development, Aging and Cancer, Tohoku University, 4-1 Seiryomachi, Aoba-Ku, Sendai 980-8575, Japan;First Department of Surgery, Tohoku University School of Medicine, Aoba-Ku, Sendai 980-8579, Japan

关键词: CD28-B7;    Co-stimulation;    Fv;    Bacterial expression;    Immunotherapy;    TCR;    T cell receptor;    mAb;    monoclonal antibody;    VH;    heavy chain variable domains;    VL;    light chain variable domains;    PBMC;    peripheral blood mononuclear cell;    MUC1;    mucin core protein 1;    bsAb;    bispecific antibody;    IL;    interleukin;    T-LAK;    lymphokine-activated killer with T cell phenotype;    Fv;    complex of VH and VL domains;    PCR;    polymerase chain reaction;    BS;    bacterial supernatant;    PP;    periplasmic;    SDS–PAGE;    sodium dodecyl sulfate–polyacrylamide gel electrophoresis;    PBS;    phosphate-buffered saline;    ELISA;    enzyme-linked immunosorbent assay;    BrdU;    bromodeoxyuridine;    MTS;    (3-(4;    5-dimethylthiazole-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazorium;    inner salt;   

DOI  :  10.1016/S0014-5793(00)01741-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The most predominant co-stimulation pathway, which is critical for T cell activation and proliferation, is the CD28-B7 pathway. The anti-CD28 monoclonal antibody (mAb) also provides a co-stimulatory signal to T cells. In order to construct a functional Fv fragment (complex of VH and VL domains) of anti-CD28 antibody using a bacterial expression system, cDNA encoding the variable regions of immunoglobulin from 15E8 hybridoma cells was cloned and expressed in Escherichia coli. The Fv fragment was obtained as a soluble protein from the periplasmic fraction and showed a binding pattern similar to parental IgG. The Fv fragment induced proliferation of peripheral blood mononuclear cells in the presence of anti-CD3 or anti-CD2 mAb and enhanced anti-tumor activity of anti-MUC1×anti-CD3 bispecific antibody when tested with lymphokine-activated killer cells with T cell phenotype. Thus, the anti-CD28 Fv fragment will be promising not only for the study of co-stimulation, but also for cancer immunotherapy.

【 授权许可】

Unknown