Effective lysis of lymphoma cells with a stabilised bispecific single-chain Fv antibody against CD19 and FcgammaRIII (CD16).
A recombinant bispecific single-chain fragment variable antibody (bsscFv), directed against the B-cell antigen CD19 and the low affinity Fc-receptor FcgammaRIII (CD16), was designed for use in the treatment of patients with leukaemias and lymphomas. The Fc-portions of whole antibodies were deliberately eliminated in this construct to avoid undesired effector functions. A stabilised bsscFv, ds[CD19 x CD16], was generated, in which disulphide bonds bridging the respective variable light (VL) and variable heavy (VH) chains were introduced into both component single-chain (sc)Fvs. After production in 293T cells and chromatographic purification, ds[CD19 x CD16] specifically and simultaneously bound both antigens. The serum stability of ds[CD19 x CD16] was increased more than threefold when compared with the unstabilised counterpart, while other biological properties were not affected by these mutations. In antibody-dependent cellular cytotoxicity experiments, ds[CD19 x CD16] mediated specific lysis of both CD19-positive malignant human B-lymphoid cell lines and primary tumour cells from patients with B-cell chronic lymphocytic leukaemia or B-cell acute lymphoblastic leukaemia. Natural killer cells, mononuclear cells (MNCs) from healthy donors and, in some instances, MNCs isolated from patients after allogeneic stem cell transplantation, were used as effectors. Thus, ds[CD19 x CD16] holds promise for the treatment of CD19(+) B-lineage malignancies.
Bruenke
Joerg
J
Chair of Genetics, University of Erlangen-Nuremberg, Erlangen, Germany.
Barbin
Karin
K
Kunert
Susanne
S
Lang
Peter
P
Pfeiffer
Matthias
M
Stieglmaier
Kristin
K
Niethammer
Dietrich
D
Stockmeyer
Bernhard
B
Peipp
Matthias
M
Repp
Roland
R
Valerius
Thomas
T
Fey
Georg H
GH
D018033:Antibodies, Bispecific / Q000276:immunology*; D000918:Antibody Specificity; D000920:Antibody-Dependent Cell Cytotoxicity / Q000276:immunology*; D018941:Antigens, CD19 / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000276:immunology; D002051:Burkitt Lymphoma / Q000276:immunology / Q000473:pathology; D002648:Child; D004306:Dose-Response Relationship, Immunologic; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000276:immunology; D007694:Killer Cells, Natural / Q000276:immunology; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000276:immunology / Q000473:pathology; D016393:Lymphoma, B-Cell / Q000276:immunology* / Q000473:pathology; D017452:Receptors, IgG / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014407:Tumor Cells, Cultured
D018033:Antibodies, Bispecific; D018941:Antigens, CD19; D000951:Antigens, Neoplasm; D007135:Immunoglobulin Variable Region; D017452:Receptors, IgG; D011993:Recombinant Fusion Proteins