Abstract
We examined the sera of six patients before and after i.v. infusions of autologous chronic lymphocytic leukemia (CLL) cells transduced ex vivo with an adenovirus encoding CD154 (Ad-CD154). Five patients made high-titer antibodies against adenovirus and three made IgG reactive with a leukemia-associated surface antigen, which we identified as ROR1. Anti-ROR1 antibodies were not detected in the sera of untreated patients. We generated anti-ROR1 mAbs and found they reacted specifically with the CLL cells of all patients, but not with nonleukemic leukocytes, a wide variety of normal adult tissues, or blood mononuclear cells, including CD5(+) B cells of healthy adults. ROR1 could bind Wnt5a, which induced activation of NF-kappaB when coexpressed with ROR1 in HEK293 cells and enhanced the survival of CLL cells in vitro, an effect that could be neutralized by posttreatment anti-ROR1 antisera. We conclude that patients with CLL can break immune tolerance to ROR1, which is an oncofetal surface antigen and survival-signaling receptor in this neoplastic disease.
Publication types
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Comparative Study
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenoviridae / immunology
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Animals
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Antibodies, Monoclonal
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Antigens, Neoplasm / immunology
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Antigens, Neoplasm / metabolism
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Base Sequence
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CD40 Ligand / immunology*
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CHO Cells
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Cricetinae
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Cricetulus
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DNA Primers
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DNA, Complementary / genetics
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Enzyme-Linked Immunosorbent Assay
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Flow Cytometry
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Humans
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Immune Sera / immunology*
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Immunoblotting
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Leukemia, Lymphocytic, Chronic, B-Cell / immunology*
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Molecular Sequence Data
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NF-kappa B / metabolism
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Proto-Oncogene Proteins / metabolism*
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Receptor Protein-Tyrosine Kinases / genetics
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Receptor Protein-Tyrosine Kinases / immunology
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Receptor Protein-Tyrosine Kinases / metabolism*
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Receptor Tyrosine Kinase-like Orphan Receptors
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Sequence Analysis, DNA
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Tumor Cells, Cultured
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Wnt Proteins / metabolism*
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Wnt-5a Protein
Substances
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Antibodies, Monoclonal
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Antigens, Neoplasm
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DNA Primers
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DNA, Complementary
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Immune Sera
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NF-kappa B
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Proto-Oncogene Proteins
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WNT5A protein, human
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Wnt Proteins
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Wnt-5a Protein
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oncofetal antigens
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CD40 Ligand
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ROR1 protein, human
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Receptor Protein-Tyrosine Kinases
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Receptor Tyrosine Kinase-like Orphan Receptors