Abstract
We determined the efficacy of HB21(Fv)PE40, a single-chain immunotoxin made by fusing the variable regions of a monoclonal antibody directed at the human transferrin receptor (TfR) with a truncated mutant of Pseudomonas exotoxin (PE), against metastatic human colon carcinoma KM12L4 cells growing in the liver or subcutis of nude mice. Organ-specific modulation of TfR expression was examined by immunohistochemistry and flow cytometry using anti-human CD71 antibody. KM12L4 cells expressed human TfR and were lysed in vitro by HB21(Fv)PE40 but not LMB-7 (a control immunotoxin specific for a Lewis Y-related carbohydrate antigen). KM12L4 cells growing in the liver expressed higher levels of TfR than cells growing s.c. Systemic administration of HB21(Fv)PE40 eliminated KM12L4 liver metastasis, whereas administration of LMB-7 did not. Treatment of mice with HB21(Fv)PE40 only delayed the growth of s.c. tumors. KM12L4 cells recovered from liver metastases, expressed higher levels of TfR, and were more sensitive to lysis by HB21(Fv)PE40 than KM12L4 cells recovered from s.c. tumors. Indeed, collectively, the data show that the expression level of the TfR by human colon cancer cells is modulated by the organ microenvironment which can be advantageous for the use of therapeutic immunotoxins.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Antibodies, Monoclonal / genetics
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Antibodies, Monoclonal / immunology
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Antigens, CD / analysis
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Antigens, Differentiation, B-Lymphocyte / analysis
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Cell Survival / drug effects
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Colonic Neoplasms / metabolism*
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Colonic Neoplasms / pathology
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Dose-Response Relationship, Drug
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Humans
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Immunoglobulin Fragments / genetics
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Immunoglobulin Fragments / immunology
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Immunohistochemistry
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Immunotoxins / genetics
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Immunotoxins / therapeutic use*
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Liver Neoplasms / metabolism
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Liver Neoplasms / prevention & control
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Liver Neoplasms / secondary
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Male
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Mice
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Mice, Nude
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Neoplasm Transplantation
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Neoplasms, Experimental / metabolism
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Neoplasms, Experimental / pathology
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Neoplasms, Experimental / prevention & control*
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Receptors, Transferrin / biosynthesis*
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Receptors, Transferrin / genetics
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Receptors, Transferrin / immunology
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / immunology
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Recombinant Fusion Proteins / therapeutic use
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Transplantation, Heterologous
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Tumor Cells, Cultured / cytology
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Tumor Cells, Cultured / drug effects
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Tumor Cells, Cultured / metabolism
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Xenograft Model Antitumor Assays
Substances
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Antibodies, Monoclonal
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Antigens, CD
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Antigens, Differentiation, B-Lymphocyte
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CD71 antigen
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Immunoglobulin Fragments
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Immunotoxins
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Receptors, Transferrin
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Recombinant Fusion Proteins
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immunoglobulin Fv