Genetic construction and characterization of a fusion protein consisting of a chimeric F(ab') with specificity for carcinomas and human IL-2

J Immunol. 1991 Apr 1;146(7):2446-52.

Abstract

A genetic construct was created incorporating gene fragments encoding the H chain V region of the human carcinoma specific antibody L6, the CH1 domain of human IgG1, a linker region, and human IL-2. This construct was cotransfected with a chimeric L6 L chain construct into the murine myeloma cell line Ag8.653 for expression. First round clones produced the fusion protein at an estimated 5 to 10 micrograms/ml based on idiotypic reactivity. Dual binding activity was demonstrated through specific interaction with the L6 Ag on human tumor cells and the IL-2R on activated human T cells. The IL-2 portion of the molecule was shown to support the growth of the IL-2-dependent T cell line CTLL2, and the qualitative nature of the IL-2 signal was found to be the same as rIL-2 with respect to induction of tyrosine-phosphorylation of intracellular protein substrates. Tumor cells coated with the fusion protein were shown to cause T cell proliferation and the presence of the fusion protein was found to enhance cell-mediated destruction of human tumor cells.

MeSH terms

  • Amino Acid Sequence
  • Antibodies, Monoclonal / genetics
  • Antibodies, Neoplasm / genetics*
  • Base Sequence
  • Carcinoma / immunology*
  • Cloning, Molecular
  • Humans
  • Immunoglobulin Fab Fragments / genetics
  • Interleukin-2 / genetics*
  • Lymphocyte Activation
  • Molecular Sequence Data
  • Phosphotyrosine
  • Receptors, Interleukin-2 / metabolism
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • Restriction Mapping
  • Signal Transduction
  • T-Lymphocytes / metabolism
  • Tyrosine / analogs & derivatives
  • Tyrosine / metabolism

Substances

  • Antibodies, Monoclonal
  • Antibodies, Neoplasm
  • Immunoglobulin Fab Fragments
  • Interleukin-2
  • Receptors, Interleukin-2
  • Recombinant Fusion Proteins
  • Phosphotyrosine
  • Tyrosine

Associated data

  • GENBANK/M55920
  • GENBANK/M55921
  • GENBANK/M55922
  • GENBANK/M55923
  • GENBANK/M55924
  • GENBANK/M55925
  • GENBANK/M55926
  • GENBANK/M55927
  • GENBANK/M55928
  • GENBANK/M64694