Apoptosis and complement-mediated lysis of myeloma cells by polyclonal rabbit antithymocyte globulin.
Current monoclonal antibody therapies for multiple myeloma have had limited success, perhaps due to narrow target specificity. We have previously described the ability of polyclonal rabbit antithymocyte globulin (rATG) to induce caspase- and cathepsin-mediated apoptosis in human B and plasma cells. We now extend this observation to myeloma cells. Complement independent cell death was measured after addition of rATG (1-1000 microg/mL) to cultures of myeloma cell lines or primary CD138+ isolates from patient bone marrow aspirates. rATG induced significant levels of apoptosis in myeloma cells as assayed by caspase induction, annexin V binding, subdiploid DNA fragmentation, plasma-membrane permeability, and loss of mitochondrial-membrane potential. Addition of complement greatly augmented myeloma-cell death. Binding of rATG to individual myeloma cell-surface proteins, primarily CD38, CD52, CD126, and CD138, was demonstrated by competitive inhibition experiments with targeted monoclonal antibodies. Three pathways of cell death were identified involving caspase activation, cathepsin D, and the genistein sensitive tyrosine kinase pathway. Fab'2 fragments of rATG had reduced proapoptotic activity, which was restored by coincubation with Fc fragments, and anti-CD32 or anti-CD64 antibodies. We conclude that rATG is an effective agent for in vitro induction of apoptosis in multiple myeloma, and that exploratory clinical trials may be warranted.
Zand
Martin S
MS
Division of Nephrology, University of Rochester Medical Center, 601 Elmwood Ave, Box 675, Rochester, NY 14642, USA. martin_zand@urmc.rochester.edu
Vo
Thuong
T
Pellegrin
Tina
T
Felgar
Raymond
R
Liesveld
Jane L
JL
Ifthikharuddin
Jainulabdeen J
JJ
Abboud
Camille N
CN
Sanz
Ignacio
I
Huggins
Jennifer
J
D000818:Animals; D000912:Antibodies, Neoplasm / Q000276:immunology; D000961:Antilymphocyte Serum / Q000276:immunology / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001854:Bone Marrow Cells / Q000187:drug effects; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D006801:Humans; D009101:Multiple Myeloma / Q000473:pathology*; D011817:Rabbits
D000912:Antibodies, Neoplasm; D000961:Antilymphocyte Serum