title: Monoclonal antibodies against ROR1 induce apoptosis of chronic lymphocytic leukemia (CLL) cells.
abstract: ROR1 is a receptor tyrosine kinase (RTK) recently identified to be overexpressed at the gene and protein levels in chronic lymphocytic leukemia (CLL). Monoclonal antibodies (MAbs) against RTKs have been successfully applied for therapy of solid tumors. We generated five MAbs against the Ig (n = 1), cysteine-rich (CRD) (n = 2) and kringle (KNG) (n = 2) domains, respectively, of the extracellular part of ROR1. All CLL patients (n = 20) expressed ROR1 on the surface of the leukemic cells. A significantly higher frequency of ROR1 expression was found in patients with progressive versus non-progressive disease, and in those with unmutated versus mutated IgVH genes. All five MAbs alone induced apoptosis in the absence of complement or added effector cells (Annexin-V and MTT, as well as cleavage of poly-(ADP ribose)-polymerase, caspase-8 and caspase-9) of CLL cells but not of normal B cells. Most effective were MAbs against CRD and KNG, significantly superior to rituximab (P < 0.005). Cross-linking of anti-ROR1 MAbs using the F(ab')(2) fragments of anti-Fc antibodies significantly augmented apoptosis. Two of the MAbs induced complement-dependent cytotoxicity (CDC) similar to that of rituximab and one anti-ROR1 MAb (KNG) (IgG1) showed killing activity by antibody-dependent cellular cytotoxicity. The identified ROR1 epitopes may provide a basis for generating human ROR1 MAbs for therapy.
authors.affiliation: Immune and Gene Therapy Lab, CCK, Karolinska University Hospital Solna, Karolinska Institutet, Stockholm, Sweden.
authors.forename: A H
authors.initials: AH
authors.lastname: Daneshmanesh
authors.forename: M
authors.initials: M
authors.lastname: Hojjat-Farsangi
authors.forename: A S
authors.initials: AS
authors.lastname: Khan
authors.forename: M
authors.initials: M
authors.lastname: Jeddi-Tehrani
authors.forename: M M
authors.initials: MM
authors.lastname: Akhondi
authors.forename: A A
authors.initials: AA
authors.lastname: Bayat
authors.forename: R
authors.initials: R
authors.lastname: Ghods
authors.forename: A-R
authors.initials: AR
authors.lastname: Mahmoudi
authors.forename: R
authors.initials: R
authors.lastname: Hadavi
authors.forename: A
authors.initials: A
authors.lastname: Österborg
authors.forename: F
authors.initials: F
authors.lastname: Shokri
authors.forename: H
authors.initials: H
authors.lastname: Rabbani
authors.forename: H
authors.initials: H
authors.lastname: Mellstedt
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D000917:Antibody Formation; D000920:Antibody-Dependent Cell Cytotoxicity; D017209:Apoptosis / Q000276:immunology*; D006801:Humans; D007114:Immunization; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000276:immunology* / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010446:Peptide Fragments / Q000276:immunology; D057050:Receptor Tyrosine Kinase-like Orphan Receptors / Q000276:immunology* / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D000911:Antibodies, Monoclonal; D010446:Peptide Fragments; C078716:ROR1 protein, human; D057050:Receptor Tyrosine Kinase-like Orphan Receptors