title: Coupling of Immunostimulants to Live Cells through Metabolic Glycoengineering and Bioorthogonal Click Chemistry.
abstract: The present study investigated the potential of metabolic glycoengineering followed by bioorthogonal click chemistry for introducing into cell-surface glycans different immunomodulating molecules. Mouse tumor models EG7 and MC38-OVA were treated with Ac4GalNAz and Ac4ManNAz followed by ligation of immunostimulants to modified cell-surface glycans of the living cells through bioorthogonal click chemistry. The presence of covalently bound oligosaccharide and oligonucleotide immunostimulants could be clearly established. The activation of a reporter macrophage cell line was determined. Depending on the tumor cell line, covalently and noncovalently bound CpG activated the macrophages by between 67 and 100% over controls. EG7 cells with covalently attached immunostimulants and controls were injected subcutaneously into C57BL/6 mice. All tumor cells subjected to the complete treatment with control molecules formed tumors like nontreated cells confirming cell viability. However, when CpG oligonucleotide was linked to cell-surface glycans, tumor growth was slowed significantly (60% reduction, n = 10, by covalently bound CpG compared to noncovalently bound CpG, n = 10). When mice that had not developed large tumors were challenged with unmodified EG7 cells, no new tumors developed, suggesting protection through the immune system.
authors.affiliation: Centre de Biophysique Moléculaire, CNRS UPR4301 , Rue Charles Sadron, 45071 Orléans, France.
authors.forename: Aline
authors.initials: A
authors.lastname: Mongis
authors.affiliation: Centre de Biophysique Moléculaire, CNRS UPR4301 , Rue Charles Sadron, 45071 Orléans, France.
authors.forename: Friedrich
authors.initials: F
authors.lastname: Piller
authors.affiliation: Centre de Biophysique Moléculaire, CNRS UPR4301 , Rue Charles Sadron, 45071 Orléans, France.
authors.forename: Véronique
authors.identifier: 0000-0002-0484-8661
authors.initials: V
authors.lastname: Piller
mesh_terms: D000276:Adjuvants, Immunologic / Q000737:chemistry*; D000818:Animals; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D057930:Click Chemistry / Q000379:methods; D008262:Macrophage Activation / Q000187:drug effects; D008264:Macrophages / Q000166:cytology; D051379:Mice; D008810:Mice, Inbred C57BL; D009369:Neoplasms / Q000473:pathology / Q000628:therapy*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry* / Q000494:pharmacology / Q000627:therapeutic use; D011134:Polysaccharides / Q000737:chemistry*
chemical_list: D000276:Adjuvants, Immunologic; C408982:CPG-oligonucleotide; D009838:Oligodeoxyribonucleotides; D011134:Polysaccharides