Journal of Biochemistry Advance Access originally published online on March 6, 2009
Journal of Biochemistry 2009 145(6):799-810; doi:10.1093/jb/mvp039
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Effective Induction of Cell Death on Adult T-Cell Leukaemia Cells by HLA-DRβ-Specific Small Antibody Fragment Isolated from Human Antibody Phage Library
1Department of Bioengineering, Faculty of Engineering, Kagoshima University, Korimoto 1-21-40, Kagoshima, Kagoshima 890-0065; 2Center for Chronic Viral Diseases, Graduate School of Medical and Dental Sciences, Kagoshima University, Sakuragaoka 8-35-1, Kagoshima, Kagoshima 890-8544; 3Nanostructured and Advanced Materials Course, Graduate School of Science and Engineering, Kagoshima University, Korimoto 1-21-40, Kagoshima, Kagoshima 890-0065; and 4The Chemo-Sero-Therapeutic Research Institute, Kyokushi, Kikuchi, Kumamoto 869-1298, Japan
*To whom correspondence should be addressed. Tel: +81-99-285-8346, Fax: +81-99-285-8346, E-mail: yito{at}be.kagoshima-u.ac.jp
Received December 19, 2008; Accepted February 23, 2009
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Abstract |
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By a biopanning method using cell sorter, we quickly isolated an antibody phage clone (S1T-A3) specific to human T-lymphotropic virus type 1-carrying T-cell line S1T from a human single chain Fv (scFv) antibody phage library. This scFv antibody bound to HTLV-1-carrying T-cell lines including MT-2, MT-4 and M8166 other than S1T, but not to non-HTLV-1-carrying T-cell lymphomas such as Jurkat and MOLT4 cells. Interestingly, this antibody induced the cell death on S1T cells very quickly (< 30 min). We tried to identify the target molecules by western blotting and mass spectrometric analysis, revealing that the target antigen was HLA class II DR. The cell death was induced only in dimmer form of scFv (diabody) and at 15-fold lower concentration than that of a fusion protein of scFv and human IgG Fc [(scFv)2-Fc] or anti HLA-DR mouse whole antibody L243. Thus, S1T-A3 diabody is a small antibody fragment with agonistic activity to induce cell death through HLA-DR. This is the first report elucidating that diabody specific to HLA-DR is effective to induce the cell death in T-cell malignancy especially adult T-cell leukaemic cell line.
Key Words: adult T-cell leukaemia, human antibody, scFv, phage library, diabody
Abbreviations: APC, antigen presenting cells; ATL, adult T-cell leukaemia; FACS, fluorescence-activated cell sorter; FBS, fetal bovine serum; FITC, fluorescent isothiocyanate; HLA, human leucocyte antigen; HTLV-1, human T-cell leukaemia virus type 1; mAb, monoclonal antibody; MHC, major histocompatibility complex; PE, phycoerythrin; PI, propidium iodide; RMF, relative mean fluorescence; scFv, single chain Fv; SPR, surface plasmon resonance