title: Efficacy and toxicity of plasma-cell-reactive monoclonal antibodies B-B2 and B-B4 and their immunotoxins.
abstract: Immunotherapy based on the delivery of toxic agents to the tumor site using monoclonal antibodies (mAb) may be a promising modality in the treatment of hematological malignancies. In the selection of mAb, both for ex vivo but even more for in vivo therapy, not only their reactivity to the neoplastic cells should be considered, but also reactivity to other body constituents. Here we describe the screening of two human plasma-cell-reactive mAb B-B2 and B-B4, which may be used for immunotherapy of multiple myeloma. Cross-reactivity of B-B2 and B-B4 was determined by immunohistochemistry on a series of tissues. This revealed for both B-B2 and B-B4 a strong staining of epithelial cells in various organs, e.g. lung, liver, skin, kidney and gut, while only a weak and diffuse staining was seen with endothelial cells. In bone marrow reactivity was only found with plasma cells and not with hemopoietic precursors (CD34+ cells). Immunotoxins from B-B2 and B-B4 were constructed by coupling them to the plant-derived ribosome-inactivating protein saporin. Both B-B2 and B-B4 immunotoxins appeared to be efficient in specific inhibition of protein synthesis in plasma cell lines (IC50 respectively 1 nM and 0.1 nm). The immunotoxins were also tested on epithelial cell line A431, on liver cell line HepG2 and on human umbilical vein endothelial cells. The epithelial cell line A431 was reactive with both B-B2 and B-B4, but was only inhibited by B-B4 immunotoxin. Cell line HepG2 was reactive with both mAb, but was not inhibited by either immunotoxin. The endothelial cells showed no reactivity with B-B2 and B-B4 and were not inhibited by either immunotoxin. Bone marrow treated with B-B2 and B-B4 immunotoxin did not show a decrease in colonies of hemopoietic precursor cells. Incubation of multiple-myeloma-derived bone marrow with these immunotoxin resulted in a clear decrease of the number of plasma cells. From these data we conclude that B-B2 and B-B4 immunotoxin can be used for ex vivo bone marrow purging. Discrepancies were found between immunohistochemistry, binding assays and cytotoxicity assays with the mAb and the immunotoxin, which underlines the necessity for these various assays as a preclinical screening.
authors.affiliation: Department of Hematology, University Hospital, Utrecht, The Netherlands.
authors.forename: W C
authors.initials: WC
authors.lastname: Vooijs
authors.forename: J
authors.initials: J
authors.lastname: Post
authors.forename: J
authors.initials: J
authors.lastname: Wijdenes
authors.forename: H J
authors.initials: HJ
authors.lastname: Schuurman
authors.forename: A
authors.initials: A
authors.lastname: Bolognesi
authors.forename: L
authors.initials: L
authors.lastname: Polito
authors.forename: F
authors.initials: F
authors.lastname: Stirpe
authors.forename: E J
authors.initials: EJ
authors.lastname: Bast
authors.forename: G C
authors.initials: GC
authors.lastname: de Gast
mesh_terms: D000911:Antibodies, Monoclonal / Q000378:metabolism / Q000494:pharmacology* / Q000633:toxicity; D001853:Bone Marrow / Q000187:drug effects; D001854:Bone Marrow Cells; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D004727:Endothelium / Q000166:cytology / Q000187:drug effects; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects; D006801:Humans; D007150:Immunohistochemistry; D000922:Immunotoxins / Q000378:metabolism / Q000494:pharmacology* / Q000633:toxicity; D008099:Liver / Q000166:cytology / Q000187:drug effects; D009101:Multiple Myeloma / Q000188:drug therapy; D010950:Plasma Cells / Q000187:drug effects* / Q000378:metabolism; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D000922:Immunotoxins