T-cell-mediated lysis of B cells induced by a CD19xCD3 bispecific single-chain antibody is perforin dependent and death receptor independent

Cancer Immunol Immunother. 2004 Jul;53(7):625-32. doi: 10.1007/s00262-003-0496-2. Epub 2004 Feb 6.

Abstract

A recently developed bispecific antibody construct, directed against CD19 and CD3 (bscCD19xCD3), induces T-cell-mediated lysis of allogeneic and autologous B cells in a specific and highly efficient manner. Since knowledge of the molecular mechanisms underlying this lysis is limited, a study on bscCD19xCD3-activated T-cell-effector pathways was performed. BscCD19xCD3-induced lysis of target B-cell lines Nalm-6, Daudi, and Raji and of autologous primary B cells is caused by the perforin-dependent granule-exocytosis pathway but not by the death ligands FasL, TRAIL, or TNF-alpha. When activated by bscCD19xCD3 and Raji cells, T cells express FasL mRNA, but incubation of Raji cells with cell-free supernatants from cytotoxicity experiments caused an upregulation of c-Flipl, possibly accounting for the cells' insensitivity toward death-receptor-mediated lysis. In addition to granule exocytosis, Raji cells are lysed by at least one mechanism independent of perforin, which requires transport through the T cell's Golgi apparatus.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Bispecific / immunology*
  • Antigens, CD19 / immunology*
  • Apoptosis
  • Apoptosis Regulatory Proteins
  • B-Lymphocytes / immunology*
  • CASP8 and FADD-Like Apoptosis Regulating Protein
  • CD3 Complex / immunology*
  • Carrier Proteins / metabolism
  • Cytoplasmic Granules / metabolism*
  • Cytotoxicity, Immunologic
  • Exocytosis
  • Fas Ligand Protein
  • Humans
  • Immunotherapy
  • Intracellular Signaling Peptides and Proteins*
  • Ligands
  • Lymphocyte Activation
  • Membrane Glycoproteins / genetics
  • Membrane Glycoproteins / metabolism*
  • Perforin
  • Pore Forming Cytotoxic Proteins
  • T-Lymphocytes / immunology*
  • T-Lymphocytes, Cytotoxic / immunology
  • T-Lymphocytes, Cytotoxic / pathology
  • TNF-Related Apoptosis-Inducing Ligand
  • Tumor Cells, Cultured
  • Tumor Necrosis Factor-alpha / metabolism
  • Up-Regulation

Substances

  • Antibodies, Bispecific
  • Antigens, CD19
  • Apoptosis Regulatory Proteins
  • CASP8 and FADD-Like Apoptosis Regulating Protein
  • CD3 Complex
  • CFLAR protein, human
  • Carrier Proteins
  • FASLG protein, human
  • Fas Ligand Protein
  • Intracellular Signaling Peptides and Proteins
  • Ligands
  • Membrane Glycoproteins
  • Pore Forming Cytotoxic Proteins
  • TNF-Related Apoptosis-Inducing Ligand
  • TNFSF10 protein, human
  • Tumor Necrosis Factor-alpha
  • Perforin