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Prepublished online as a Blood First Edition Paper on June 5, 2003; DOI 10.1182/blood-2002-12-3921.
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Blood, 1 October 2003, Vol. 102, No. 7, pp. 2506-2512
IMMUNOBIOLOGY
A novel strategy using single-chain antibody to show the importance of Bcl-2 in mast cell survival
Cellina Cohen-Saidon,
Hovav Nechushtan,
Shira Kahlon,
Nadav Livni,
Ahuva Nissim, and
Ehud Razin
From the Department of Biochemistry, Hebrew University Hadassah Medical School, Jerusalem, Israel; and Felsenstein Medical Research Center, Sackler Faculty of Medicine, Tel Aviv University, Petach Tikva, Israel.
Apoptosis or programmed cell death plays an important role in a wide variety of physiologic processes and is regulated by proteins of the Bcl-2 family consisting of both antiapoptotic and proapoptotic factors. The direct involvement of the Bcl-2 protein family in the process of mast cell apoptosis has not been clarified. In the present work we have used a single-chain antibody (scFv) raised against Bcl-2 derived from a semisynthetic human phage-display antibody library. The addition of TAT sequence, which is responsible for translocation through the membrane, endows the anti-Bcl-2-scFv with the ability to penetrate living cells. Moreover, it specifically neutralizes Bcl-2 intracellularly by binding to the BH1 domain and eradicates its antiapoptotic activity in 2 types of mast cells and in a human breast cancer cell line. (Blood. 2003;102:2506-2512)
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