Transfer of chimeric receptor gene made of variable regions of tumor-specific antibody confers anticarbohydrate specificity on T cells

Delia Mezzanzanica, Silvana Canevari, Alessandra Mazzoni, Mariangela Figini, Maria I. Colnaghi, Tova Waks, Daniel G. Schindler, Zelig Eshhar

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

The antitumor specificity of T cells can be induced by gene transfer using a recently developed therapeutic approach (T body). In this work, we genetically conferred anticarbohydrate specificity onto T cells using the variable regions of monoclonal antibody MLuC1, which binds the LewisY (Ley) tumor-associated antigen that is overexpressed on several human carcinomas. The variable regions of MLuC1, which are in a single-chain Fv (ScFv) configuration, were cloned and spliced in a eukaryotic expression vector with both the gene encoding the signal-transducing γ-chain of the human Fcγ receptor and a flexible hinge domain. The chimeric ScFv-γ gene was expressed in a murine cytotoxic T-cell hybridoma. Transfectants receiving vector only served as a negative control (mock). Screening for functional transfectants was carried out using a tumor growth inhibition assay. The soluble form of MLuC1 ScFv was recovered from bacteria periplasm and tested for binding to Ley-expressing cells by the fluorescence-activated cell sorter analysis. Despite the low binding ability of the soluble MLuC1 ScFv, 7 of 13 genetically engineered cytotoxic T lymphocyte clones inhibited the growth of Ley-positive cells and did not affect growth of Ley-negative cells. None of the mock clones tested specifically inhibited tumor growth. These data indicate that, by chimeric MLuC1 ScFv-γ gene transfer, it is possible to confer anticarbohydrate specificity onto T cells and extend the applicability of the T-body approach to tumor-associated antigens that are naturally not recognized by T cells.

Original languageEnglish
Pages (from-to)401-407
Number of pages7
JournalCancer Gene Therapy
Volume5
Issue number6
Publication statusPublished - 1998

Fingerprint

T-Cell Antigen Receptor Specificity
Neoplasm Antibodies
Single-Chain Antibodies
Genes
Neoplasm Antigens
Growth
Clone Cells
T-Lymphocytes
Periplasm
Fc Receptors
Hybridomas
Cytotoxic T-Lymphocytes
Neoplasms
Fluorescence
Monoclonal Antibodies
Bacteria
Carcinoma

Keywords

  • Carbohydrate antigens
  • Chimeric genes
  • Lewis
  • Single-chain Fv
  • T cells
  • Tumor immunogene therapy

ASJC Scopus subject areas

  • Cancer Research
  • Genetics

Cite this

Transfer of chimeric receptor gene made of variable regions of tumor-specific antibody confers anticarbohydrate specificity on T cells. / Mezzanzanica, Delia; Canevari, Silvana; Mazzoni, Alessandra; Figini, Mariangela; Colnaghi, Maria I.; Waks, Tova; Schindler, Daniel G.; Eshhar, Zelig.

In: Cancer Gene Therapy, Vol. 5, No. 6, 1998, p. 401-407.

Research output: Contribution to journalArticle

Mezzanzanica, Delia ; Canevari, Silvana ; Mazzoni, Alessandra ; Figini, Mariangela ; Colnaghi, Maria I. ; Waks, Tova ; Schindler, Daniel G. ; Eshhar, Zelig. / Transfer of chimeric receptor gene made of variable regions of tumor-specific antibody confers anticarbohydrate specificity on T cells. In: Cancer Gene Therapy. 1998 ; Vol. 5, No. 6. pp. 401-407.
@article{02d43c6b42834ee0bb4a0cabfba12613,
title = "Transfer of chimeric receptor gene made of variable regions of tumor-specific antibody confers anticarbohydrate specificity on T cells",
abstract = "The antitumor specificity of T cells can be induced by gene transfer using a recently developed therapeutic approach (T body). In this work, we genetically conferred anticarbohydrate specificity onto T cells using the variable regions of monoclonal antibody MLuC1, which binds the LewisY (Ley) tumor-associated antigen that is overexpressed on several human carcinomas. The variable regions of MLuC1, which are in a single-chain Fv (ScFv) configuration, were cloned and spliced in a eukaryotic expression vector with both the gene encoding the signal-transducing γ-chain of the human Fcγ receptor and a flexible hinge domain. The chimeric ScFv-γ gene was expressed in a murine cytotoxic T-cell hybridoma. Transfectants receiving vector only served as a negative control (mock). Screening for functional transfectants was carried out using a tumor growth inhibition assay. The soluble form of MLuC1 ScFv was recovered from bacteria periplasm and tested for binding to Ley-expressing cells by the fluorescence-activated cell sorter analysis. Despite the low binding ability of the soluble MLuC1 ScFv, 7 of 13 genetically engineered cytotoxic T lymphocyte clones inhibited the growth of Ley-positive cells and did not affect growth of Ley-negative cells. None of the mock clones tested specifically inhibited tumor growth. These data indicate that, by chimeric MLuC1 ScFv-γ gene transfer, it is possible to confer anticarbohydrate specificity onto T cells and extend the applicability of the T-body approach to tumor-associated antigens that are naturally not recognized by T cells.",
keywords = "Carbohydrate antigens, Chimeric genes, Lewis, Single-chain Fv, T cells, Tumor immunogene therapy",
author = "Delia Mezzanzanica and Silvana Canevari and Alessandra Mazzoni and Mariangela Figini and Colnaghi, {Maria I.} and Tova Waks and Schindler, {Daniel G.} and Zelig Eshhar",
year = "1998",
language = "English",
volume = "5",
pages = "401--407",
journal = "Cancer Gene Therapy",
issn = "0929-1903",
publisher = "Nature Publishing Group",
number = "6",

}

TY - JOUR

T1 - Transfer of chimeric receptor gene made of variable regions of tumor-specific antibody confers anticarbohydrate specificity on T cells

AU - Mezzanzanica, Delia

AU - Canevari, Silvana

AU - Mazzoni, Alessandra

AU - Figini, Mariangela

AU - Colnaghi, Maria I.

AU - Waks, Tova

AU - Schindler, Daniel G.

AU - Eshhar, Zelig

PY - 1998

Y1 - 1998

N2 - The antitumor specificity of T cells can be induced by gene transfer using a recently developed therapeutic approach (T body). In this work, we genetically conferred anticarbohydrate specificity onto T cells using the variable regions of monoclonal antibody MLuC1, which binds the LewisY (Ley) tumor-associated antigen that is overexpressed on several human carcinomas. The variable regions of MLuC1, which are in a single-chain Fv (ScFv) configuration, were cloned and spliced in a eukaryotic expression vector with both the gene encoding the signal-transducing γ-chain of the human Fcγ receptor and a flexible hinge domain. The chimeric ScFv-γ gene was expressed in a murine cytotoxic T-cell hybridoma. Transfectants receiving vector only served as a negative control (mock). Screening for functional transfectants was carried out using a tumor growth inhibition assay. The soluble form of MLuC1 ScFv was recovered from bacteria periplasm and tested for binding to Ley-expressing cells by the fluorescence-activated cell sorter analysis. Despite the low binding ability of the soluble MLuC1 ScFv, 7 of 13 genetically engineered cytotoxic T lymphocyte clones inhibited the growth of Ley-positive cells and did not affect growth of Ley-negative cells. None of the mock clones tested specifically inhibited tumor growth. These data indicate that, by chimeric MLuC1 ScFv-γ gene transfer, it is possible to confer anticarbohydrate specificity onto T cells and extend the applicability of the T-body approach to tumor-associated antigens that are naturally not recognized by T cells.

AB - The antitumor specificity of T cells can be induced by gene transfer using a recently developed therapeutic approach (T body). In this work, we genetically conferred anticarbohydrate specificity onto T cells using the variable regions of monoclonal antibody MLuC1, which binds the LewisY (Ley) tumor-associated antigen that is overexpressed on several human carcinomas. The variable regions of MLuC1, which are in a single-chain Fv (ScFv) configuration, were cloned and spliced in a eukaryotic expression vector with both the gene encoding the signal-transducing γ-chain of the human Fcγ receptor and a flexible hinge domain. The chimeric ScFv-γ gene was expressed in a murine cytotoxic T-cell hybridoma. Transfectants receiving vector only served as a negative control (mock). Screening for functional transfectants was carried out using a tumor growth inhibition assay. The soluble form of MLuC1 ScFv was recovered from bacteria periplasm and tested for binding to Ley-expressing cells by the fluorescence-activated cell sorter analysis. Despite the low binding ability of the soluble MLuC1 ScFv, 7 of 13 genetically engineered cytotoxic T lymphocyte clones inhibited the growth of Ley-positive cells and did not affect growth of Ley-negative cells. None of the mock clones tested specifically inhibited tumor growth. These data indicate that, by chimeric MLuC1 ScFv-γ gene transfer, it is possible to confer anticarbohydrate specificity onto T cells and extend the applicability of the T-body approach to tumor-associated antigens that are naturally not recognized by T cells.

KW - Carbohydrate antigens

KW - Chimeric genes

KW - Lewis

KW - Single-chain Fv

KW - T cells

KW - Tumor immunogene therapy

UR - http://www.scopus.com/inward/record.url?scp=0032201554&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0032201554&partnerID=8YFLogxK

M3 - Article

VL - 5

SP - 401

EP - 407

JO - Cancer Gene Therapy

JF - Cancer Gene Therapy

SN - 0929-1903

IS - 6

ER -