Efficient elimination of chronic lymphocytic leukaemia B cells by autologous T cells with a bispecific anti-CD19/anti-CD3 single-chain antibody construct

Leukemia. 2003 May;17(5):900-9. doi: 10.1038/sj.leu.2402890.


Recently, we have shown that a novel recombinant bispecific single-chain antibody construct (bscCD19 x CD3), induces highly efficacious lymphoma-directed cytotoxicity mediated by unstimulated peripheral T lymphocytes. Functional analysis of bscCD19 x CD3 has so far been exclusively performed with human B lymphoma cell lines and T cells from healthy donors. Here we analysed the properties of bscCD19 x CD3 using primary B cells and autologous T cells from healthy volunteers or patients with B-cell chronic lymphocytic leukaemia (B-CLL). We show that bscCD19 x CD3 induces T-cell-mediated depletion of nonmalignant B cells in all four cases and depletion of primary lymphoma cells in 22 out of 25 cases. This effect could be observed at low effector-to-target (E:T) ratios and in the majority of cases without additional activation of autologous T cells by IL-2. Even in samples derived from patients heavily pretreated with different chemotherapy regimens, strong cytotoxic effects of bscCD19 x CD3 could be observed. The addition of bscCD19 x CD3 to patients' cells resulted in an upregulation of activation-specific cell surface antigens on autologous T cells and elevated levels of CD95 on lymphoma B cells. Although anti-CD95 antibody CH-11 failed to induce apoptosis in lymphoma cells, we provide evidence that B-CLL cell depletion by bscCD3 x CD3 is mediated at least in part by apoptosis via the caspase pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Aged, 80 and over
  • Annexin A5 / metabolism
  • Antibodies, Bispecific / therapeutic use*
  • Antibody Specificity
  • Antigens, CD19 / immunology*
  • B-Lymphocytes / immunology
  • CD3 Complex / immunology*
  • Caspase Inhibitors
  • Caspases / metabolism
  • Cell Death / drug effects
  • Cell Division / drug effects
  • Cytotoxicity, Immunologic*
  • Enzyme Activation / drug effects
  • Female
  • Flow Cytometry
  • Humans
  • Immunotherapy / methods
  • Interleukin-2 / metabolism
  • Leukemia, Lymphocytic, Chronic, B-Cell / immunology
  • Leukemia, Lymphocytic, Chronic, B-Cell / therapy*
  • Leukocytes, Mononuclear / metabolism
  • Lymphocyte Depletion*
  • Male
  • Middle Aged
  • T-Lymphocytes / immunology*
  • Tumor Cells, Cultured


  • Annexin A5
  • Antibodies, Bispecific
  • Antigens, CD19
  • CD3 Complex
  • Caspase Inhibitors
  • Interleukin-2
  • Caspases