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 journalArticlepeer-review

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

Keywords

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

ASJC Scopus subject areas

  • Cancer Research
  • Genetics

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