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Title: A subtraction tolerization method of immunization allowed for Wilms' tumor protein-1 (WT1) identification in melanoma and discovery of an antitumor peptide sequence
Authors: Massaoka, Mariana H. [UNIFESP]
Matsuo, Alisson L. [UNIFESP]
Figueiredo, Carlos R. [UNIFESP]
Girola, Natalia [UNIFESP]
Farias, Camyla F. [UNIFESP]
Pasqualoto, Kerly
Travassos, Luiz R. [UNIFESP]
Universidade Federal de São Paulo (UNIFESP)
Butantan Inst
Keywords: Immunization protocol
Wilms' tumor protein 1 (WT1)
Malignant melanoma
Antitumor peptide
Issue Date: 1-Dec-2014
Publisher: Elsevier B.V.
Citation: Journal of Immunological Methods. Amsterdam: Elsevier B.V., v. 414, p. 11-19, 2014.
Abstract: On searching for melanoma transcription factors in a project focusing on internal antitumor peptide sequences from transcription factors, we found that a highly immunogenic component emerged upon using a subtraction tolerization method of immunization. While several conventional immunization procedures using whole melanoma cells induced a plethora of low affinity antibodies of various specificities, the subtraction tolerization method efficiently elicited mono-specific antibodies that recognized Wilms' tumor protein 1 (WT1), which is known as an important marker in melanoma prognosis and treatment for the tolerization step, pre-immunization of Balb/c mice with a membrane-rich preparation of glioblastoma U87 cells was used. the subsequent immunizations with SK-MEL-28 melanoma cells elicited antibodies strongly reacting with 50 and 55 kDa proteins, identified as WT1. Remarkably, this was the only component strongly reactive with these antibodies in a melanoma cell lysate. WT1 was then chosen as a target for selecting internally bioactive peptides. A hydrophilic Trojan peptide containing most of the zinc finger-2 domain of WT1 was synthesized and shown to inhibit SK-MEL-28 melanoma growth in vitro. the peptide WT1-pTj was also protective in vivo in a metastatic melanoma model and peptide-stimulated syngeneic dendritic cells reproduced the anti-melanoma effect of the unprotected peptide. Identification of antitumor peptides derived from major transcription factors represents a new tool to be explored in cancer research aiming at new therapeutic drugs. (C) 2014 Elsevier B.V. All rights reserved.
ISSN: 0022-1759
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