Abstract |
To target CD30 on Hodgkin's disease and anaplastic large-cell lymphoma, anti-CD30 single-chain antibodies were obtained by DNA immunization of mice with the complete human CD30 cDNA. Spleens were isolated from mice with high anti-CD30 titer, and the RNA was used for the production of an scFv-displaying phage library. Specific phages were enriched by 3 rounds of panning on soluble CD30 or CD30+ K562 cells. Recombinant immunotoxins (rITs) were made from 3 ELISA-positive scFv phages by fusion to a 38 kDa truncated mutant of Pseudomonas exotoxin (PE38) with or without a KDEL mutant sequence at the C terminus. In vitro cytotoxicity of purified anti-CD30 rITs was measured on CD30-transfected A431 cells. IC50 values ranged from 3 to 7 ng/ml (50-110 pM) for PE38 rITs and 0.1 ng/ml (2 pM) for the PE38-KDEL IT on A431-CD30 cells. The parental A431 cells were resistant, indicating that the cytotoxicity was specific and CD30-mediated. rITs were tested for anti- tumor activity in a nude mouse model. A431-CD30 cells were injected s.c. on day 0; then, mice bearing measurable tumors were treated beginning on day 4 with 3 alternate daily doses i.v. Anti- tumor activity was dose-dependent and not found when irrelevant ITs were administered or when CD30- tumors were treated. Our data show that DNA immunization and antibody phage display may be useful in producing new rITs against hematologic malignancies. Published 2001 Wiley-Liss, Inc.
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Authors | H Rozemuller, P S Chowdhury, I Pastan, R J Kreitman |
Journal | International journal of cancer
(Int J Cancer)
Vol. 92
Issue 6
Pg. 861-70
(Jun 15 2001)
ISSN: 0020-7136 [Print] United States |
PMID | 11351308
(Publication Type: Journal Article)
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Chemical References |
- Bacterial Toxins
- Cancer Vaccines
- Codon
- DNA, Complementary
- Exotoxins
- Immunoglobulin Fragments
- Ki-1 Antigen
- Peptide Library
- RNA, Messenger
- Recombinant Fusion Proteins
- Recombinant Proteins
- Virulence Factors
- RNA
- DNA
- ADP Ribose Transferases
- Pseudomonas aeruginosa exotoxin A
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Topics |
- ADP Ribose Transferases
- Amino Acid Sequence
- Animals
- Bacterial Toxins
- Cancer Vaccines
- Cell Line
- Codon
- DNA
(metabolism)
- DNA, Complementary
(metabolism)
- Dose-Response Relationship, Drug
- Enzyme-Linked Immunosorbent Assay
- Exotoxins
(metabolism)
- Humans
- Immunoglobulin Fragments
(metabolism)
- Inhibitory Concentration 50
- Ki-1 Antigen
(chemistry, immunology)
- Kinetics
- Mice
- Mice, Inbred BALB C
- Mice, Inbred C57BL
- Mice, Nude
- Models, Biological
- Molecular Sequence Data
- Peptide Library
- Plasmids
(metabolism)
- RNA
(metabolism)
- RNA, Messenger
(metabolism)
- Recombinant Fusion Proteins
(metabolism)
- Recombinant Proteins
(metabolism)
- Sequence Homology, Amino Acid
- Surface Plasmon Resonance
- Time Factors
- Transfection
- Tumor Cells, Cultured
- Virulence Factors
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