RICIN-A-CHAIN CYTOTOXICITY DEPENDS ON ITS PRESENTATION TO THE CELL-MEMBRANE

被引:6
作者
CARAYON, P
BORD, A
GAILLARD, JP
VIDAL, H
GROS, P
JANSEN, FK
机构
[1] Sanofi Recherche, Centre de Montpellier, Montpellier
[2] Laboratoire Immunociblage, ORSTOM, Montpellier
关键词
D O I
10.1021/bc00020a007
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Anti-ricin-A-chain monoclonal antibodies (mAbs) were raised in order to study epitopes of the A-chain involved in the initiation of its transmembrane passage. Five nonoverlapping epitopes were selected by cluster formation using a cross-inhibition assay of mAbs from 172 specific hybridomas. For each cluster, representative high affinity mAbs were selected. These mAbs were disulfide-linked to F(ab')2 fragments of a mAb directed against the CD5 antigen. The cytotoxicity of ricin A-chain bound by affinity through different epitopes to these hybrid mAbs was explored on the CD5-positive CEM cells. Without any enhancer, the hybrids displayed cytotoxicity varying in IC50 within a range of 1-10; in the presence of the enhancer NH4Cl, this variation of activity was maintained ranging from 1 to 20; with the enhancer monensin, the variation of cytotoxicity of hybrids extended over a 1-80-fold range. These differences of cytotoxicity were not correlated with mAb properties such as the inhibition of A-chain enzymatic activity, the A-chain binding affinity, the capacity for releasing A-chain at low pH, or the capacity of hybrids for delivering A-chain on the cell surface. This lack of correlation strongly suggests that the different presentations of A-chain epitopes to the cell membrane may be the essential reason for cytotoxicity variations.
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页码:146 / 152
页数:7
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