title: ADAM10 inhibition of human CD30 shedding increases specificity of targeted immunotherapy in vitro.
abstract: CD30 is a transmembrane protein selectively overexpressed on many human lymphoma cells and therefore an interesting target for antibody-based immunotherapy. However, binding of therapeutic antibodies stimulates a juxtamembrane cleavage of CD30 leading to a loss of target antigen and an enhanced release of the soluble ectodomain of CD30 (sCD30). Here, we show that sCD30 binds to CD30 ligand (CD153)-expressing non-target cells. Because antibodies bind to sCD30, this results in unwanted antibody binding to these cells via sCD30 bridging. To overcome shedding-dependent damage of normal cells in CD30-specific immunotherapy, we analyzed the mechanism involved in the release. Shedding of CD30 can be enhanced by protein kinase C (PKC) activation, implicating the disintegrin metalloproteinase ADAM17 but not free cytoplasmic calcium. However, antibody-induced CD30 shedding is calcium dependent and PKC independent. This shedding involved the related metalloproteinase ADAM10 as shown by the use of the preferential ADAM10 inhibitor GI254023X and by an ADAM10-deficient cell line generated from embryonically lethal ADAM10(-/-) mouse. In coculture experiments, the antibody-induced transfer of sCD30 from the human Hodgkin's lymphoma cell line L540 to the CD30-negative but CD153-expressing human mast cell line HMC-1 was inhibited by GI254023X. These findings suggest that selective metalloproteinase inhibitors blocking antibody-induced shedding of target antigens could be of therapeutic value to increase the specificity and reduce side effects of immunotherapy with monoclonal antibodies.
authors.affiliation: Department of Internal Medicine I and Center for Molecular Medicine, University Hospital Cologne, Kerpener Strasse 62, 50924 Cologne, Germany.
authors.forename: Dennis A
authors.initials: DA
authors.lastname: Eichenauer
authors.forename: Vijaya Lakshmi
authors.initials: VL
authors.lastname: Simhadri
authors.forename: Elke Pogge
authors.initials: EP
authors.lastname: von Strandmann
authors.forename: Andreas
authors.initials: A
authors.lastname: Ludwig
authors.forename: Vance
authors.initials: V
authors.lastname: Matthews
authors.forename: Katrin S
authors.initials: KS
authors.lastname: Reiners
authors.forename: Bastian
authors.initials: B
authors.lastname: von Tresckow
authors.forename: Paul
authors.initials: P
authors.lastname: Saftig
authors.forename: Stefan
authors.initials: S
authors.lastname: Rose-John
authors.forename: Andreas
authors.initials: A
authors.lastname: Engert
authors.forename: Hinrich P
authors.initials: HP
authors.lastname: Hansen
mesh_terms: D051722:ADAM Proteins / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D000072197:ADAM10 Protein; D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D000906:Antibodies / Q000276:immunology / Q000494:pharmacology; D053285:CD30 Ligand / Q000276:immunology / Q000378:metabolism; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D045744:Cell Line, Tumor; D006801:Humans; D007167:Immunotherapy / Q000379:methods*; D017730:Ki-1 Antigen / Q000276:immunology* / Q000378:metabolism; D008223:Lymphoma / Q000201:enzymology / Q000276:immunology* / Q000628:therapy*; D008565:Membrane Proteins / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism
chemical_list: D000906:Antibodies; D053285:CD30 Ligand; D017730:Ki-1 Antigen; D008565:Membrane Proteins; D000001:Calcimycin; D011493:Protein Kinase C; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human; D002118:Calcium