Identification of their epitope reveals the structural basis for the mechanism of action of the immunosuppressive antibodies basiliximab and daclizumab
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Identification of their epitope reveals the structural basis for the mechanism of action of the immunosuppressive antibodies basiliximab and daclizumab. / Binder, Mascha; Vögtle, Friederike-Nora; Michelfelder, Stefan; Müller, Fabian; Illerhaus, Gerald; Sundararajan, Sangeeth; Mertelsmann, Roland; Trepel, Martin.
In: CANCER RES, Vol. 67, No. 8, 15.04.2007, p. 3518-23.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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T1 - Identification of their epitope reveals the structural basis for the mechanism of action of the immunosuppressive antibodies basiliximab and daclizumab
AU - Binder, Mascha
AU - Vögtle, Friederike-Nora
AU - Michelfelder, Stefan
AU - Müller, Fabian
AU - Illerhaus, Gerald
AU - Sundararajan, Sangeeth
AU - Mertelsmann, Roland
AU - Trepel, Martin
PY - 2007/4/15
Y1 - 2007/4/15
N2 - Interleukin-2 (IL-2) and its receptor (IL-2R) play a major role in cellular immunity. The monoclonal antibodies basiliximab and daclizumab directed against the IL-2R subunit CD25 are widely used to prevent graft or host rejection after allogeneic tissue transplantation. Although these antibodies have been used for this purpose for many years, their common epitope within the CD25 protein is unknown. We screened a random phage display library to isolate peptides specifically binding to basiliximab. A striking amino acid sequence motif was enriched. This motif is homologous to the peptide ERIYHFV comprising amino acid positions 116 to 122 within the extracellular domain of CD25, suggesting that this is the basiliximab epitope. Basiliximab and daclizumab binding of selected phage was specific, as no binding was observed to isotype antibody controls. Phage binding could be inhibited by the cognate peptide. In cells expressing mutant CD25, binding of basiliximab was abolished when two or more amino acids of the suspected epitope were changed. In contrast, basiliximab binding remained unaffected in cells expressing CD25 versions with mutations outside this epitope. We therefore conclude that the (116)ERIYHFV(122) string within CD25 is the epitope recognized by basiliximab and daclizumab. This epitope overlaps with the interaction site of CD25 and IL-2, thus revealing the structural basis for the inhibition of IL-2R binding by this class of immunosuppressive antibodies.
AB - Interleukin-2 (IL-2) and its receptor (IL-2R) play a major role in cellular immunity. The monoclonal antibodies basiliximab and daclizumab directed against the IL-2R subunit CD25 are widely used to prevent graft or host rejection after allogeneic tissue transplantation. Although these antibodies have been used for this purpose for many years, their common epitope within the CD25 protein is unknown. We screened a random phage display library to isolate peptides specifically binding to basiliximab. A striking amino acid sequence motif was enriched. This motif is homologous to the peptide ERIYHFV comprising amino acid positions 116 to 122 within the extracellular domain of CD25, suggesting that this is the basiliximab epitope. Basiliximab and daclizumab binding of selected phage was specific, as no binding was observed to isotype antibody controls. Phage binding could be inhibited by the cognate peptide. In cells expressing mutant CD25, binding of basiliximab was abolished when two or more amino acids of the suspected epitope were changed. In contrast, basiliximab binding remained unaffected in cells expressing CD25 versions with mutations outside this epitope. We therefore conclude that the (116)ERIYHFV(122) string within CD25 is the epitope recognized by basiliximab and daclizumab. This epitope overlaps with the interaction site of CD25 and IL-2, thus revealing the structural basis for the inhibition of IL-2R binding by this class of immunosuppressive antibodies.
KW - Amino Acid Motifs
KW - Amino Acid Sequence
KW - Antibodies, Monoclonal
KW - Antibodies, Monoclonal, Humanized
KW - Binding Sites
KW - Cell Line
KW - Epitopes
KW - Humans
KW - Immunoglobulin G
KW - Immunosuppressive Agents
KW - Interleukin-2
KW - Interleukin-2 Receptor alpha Subunit
KW - Models, Molecular
KW - Molecular Sequence Data
KW - Peptide Library
KW - Peptides
KW - Protein Conformation
KW - Receptors, Interleukin-2
KW - Recombinant Fusion Proteins
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1158/0008-5472.CAN-06-3919
DO - 10.1158/0008-5472.CAN-06-3919
M3 - SCORING: Journal article
C2 - 17440057
VL - 67
SP - 3518
EP - 3523
JO - CANCER RES
JF - CANCER RES
SN - 0008-5472
IS - 8
ER -