title: Preparation and In Vitro Evaluation of Antitumor Activity of TGFαL3-SEB as a Ligand-Targeted Superantigen.
abstract: Tumor-targeted superantigens (TTSs) have been used to treat a variety of tumors in preclinical studies. The TTS utilizes the powerful T-cell activation strategy by means of staphylococcal enterotoxins (SEs) as superantigens (Sags) to target tumor cells. Monoclonal antibodies and tumor-related ligands have been used as targeting molecules of Sag. In this study, we assessed the antitumor potency of tumor-targeted superantigen (TTS) strategy to design and produce fusion protein as a new antitumor candidate. The third loop (L3) of transforming growth factor α (TGF-α) was genetically conjugated to staphylococcal enterotoxin type B (TGFαL3-SEB), and its in vitro antitumor activity against murine breast cancer cells (A431 cell line) was evaluated. We designed and prepared TGFαL3-SEB chimeric protein and evaluated superantigenic activity, binding property to cancer cells, overexpression of epidermal growth factor receptor (EGFR), and in vitro antitumor activities. Cloning of tgfαl3-seb was confirmed by colony-polymerase chain reaction, enzymatic digestion, and sequencing. The recombinant TGFαL3-SEB fusion protein with molecular weight of 31 kDa was expressed and confirmed by anti-His Western-blot analysis. The TGFαL3-SEB fusion protein attached to A431 cell line with proper affinity and induced dose-dependent cytotoxicity against EGFR-expressing cancer cells in vitro. The TGFαL3-SEB chimeric protein exhibited potent in vitro antitumor activity. Our findings indicated that TGFαL3-SEB may be a promising anticancer candidate in cancer immunotherapy, and further studies are required to explore its potential in vivo therapeutic applications.
authors.affiliation: Bacteriology Department, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Forough
authors.initials: F
authors.lastname: Yousefi
authors.affiliation: Bacteriology Department, Pasteur Institute of Iran, Tehran, Iran sadatff@yahoo.com mousavi@pasteur.ac.ir.
authors.forename: Seyed Fazlollah
authors.initials: SF
authors.lastname: Mousavi
authors.affiliation: Bacteriology Department, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Seyed Davar
authors.initials: SD
authors.lastname: Siadat
authors.affiliation: Bacteriology Department, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mohammad Mehdi
authors.initials: MM
authors.lastname: Aslani
authors.affiliation: Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
authors.forename: Jafar
authors.initials: J
authors.lastname: Amani
authors.affiliation: Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
authors.forename: Hamid Sedighian
authors.initials: HS
authors.lastname: Rad
authors.affiliation: Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran imanifouladi.a@gmail.com imanifouladi.a@bmsu.ac.ir.
authors.forename: Abbas Ali Imani
authors.initials: AA
authors.lastname: Fooladi
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004354:Drug Screening Assays, Antitumor; D004768:Enterotoxins / Q000235:genetics; D066246:ErbB Receptors / Q000378:metabolism; D007167:Immunotherapy; D020128:Inhibitory Concentration 50; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*; D013211:Staphylococcus aureus / Q000235:genetics; D018089:Superantigens / Q000096:biosynthesis / Q000494:pharmacology*; D016211:Transforming Growth Factor alpha / Q000235:genetics
chemical_list: D000970:Antineoplastic Agents; D004768:Enterotoxins; D011993:Recombinant Fusion Proteins; D018089:Superantigens; D016211:Transforming Growth Factor alpha; C031818:enterotoxin B, staphylococcal; C512478:EGFR protein, human; D066246:ErbB Receptors
keywords: breast cancer; cloning and expression; immunotherapy; staphylococcal enterotoxin type B (SEB); transforming growth factor α (TGF-α)