Bispecific anti-HER2 and CD16 single-chain antibody production prolongs the use of stem cell-like cell transplantation against HER2-overexpressing cancer.
Humanized antibodies vary and have certain effects, but they are expensive and require repeated administration. We developed cells which constantly express a humanized antibody, and we performed anticancer humanized antibody therapy involving cell transplantation. Genes with the same amino acid sequence as that of the variable region of trastuzumab (Herceptin) as the humanized anti-HER2 monoclonal antibody were produced by overlap-PCR and were connected to the anti-human CD16 antibody [anti-HER2+anti-CD16 single-chain antibody (anti-HER2+CD16 scAb)]. For transplantation, stem cell-like cells that are immunologically tolerant and do not transform into cancer [mouse embryo fibroblast cell line C3H10T1/2 (10T1/2)] were used. The antibody was incorporated into 10T1/2 (antibody-expressing cells) using the pMX-IRES-EGFP retroviral vector. Cell supernatants and human monocytes were exposed to the human breast cancer strain HTB131 expressing HER2, and the in vitro antibody-dependent cell-mediated cytotoxicity (ADCC) effects were evaluated. After the transplantation of antibody-expressing and HTB131 cells into SCID mice, human monocytes were intermittently administered, and the in vivo ADCC effects were evaluated. We found that the ex vivo dead cell rate was 15.4% for Herceptin, 5.6% for anti-HER2+CD16 scAb, 1.5% for anti-CD16 scAb, and 2.1% for the control, demonstrating the antitumor effects of anti-HER2+CD16 scAb. In an antibody-expressing cell transplantation model, the inhibitory effects of this antibody on HTB131 cell establishment were observed. In conclusion, the establishment of breast cancer cells in the peritoneum was inhibited by the transplantation of antibody-expressing cells. Since this method requires cell transplantation only once, the drug cost may be reduced.
Kasuya
Kazuhiko
K
Department of Digestive Surgery, Hachioji Medical Center of Tokyo Medical University, Hachioji, Tokyo 193-0998, Japan. kasuya-k@jcom.home.ne.jp
Shimazu
Motohide
M
Suzuki
Minako
M
Itoi
Takao
T
Aoki
Tatsuya
T
Tsuchida
Akihiko
A
D000818:Animals; D018033:Antibodies, Bispecific / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D061067:Antibodies, Monoclonal, Humanized; D000920:Antibody-Dependent Cell Cytotoxicity; D001943:Breast Neoplasms / Q000201:enzymology / Q000276:immunology / Q000628:therapy*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D005260:Female; D058851:GPI-Linked Proteins; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D016133:Polymerase Chain Reaction; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D017452:Receptors, IgG / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D057127:Single-Chain Antibodies / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D033581:Stem Cell Transplantation / Q000379:methods*; D013234:Stem Cells / Q000276:immunology; D016019:Survival Analysis; D000068878:Trastuzumab; D023041:Xenograft Model Antitumor Assays / Q000379:methods
D018033:Antibodies, Bispecific; D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D058851:GPI-Linked Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D017452:Receptors, IgG; D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies; D000068878:Trastuzumab; C428200:FCGR3B protein, human; C508053:ERBB2 protein, human