A Cytotoxic Antibody Recognizing Lacto-N-fucopentaose I (LNFP I) on Human Induced Pluripotent Stem (hiPS) Cells.
We have generated a mouse monoclonal antibody (R-17F, IgG1 subtype) specific to human induced pluripotent stem (hiPS)/embryonic stem (ES) cells by using a hiPS cell line as an antigen. Triple-color confocal immunostaining images of hiPS cells with R-17F indicated that the R-17F epitope was expressed exclusively and intensively on the cell membranes of hiPS cells and co-localized partially with those of SSEA-4 and SSEA-3. Lines of evidence suggested that the predominant part of the R-17F epitope was a glycolipid. Upon TLC blot of total lipid extracts from hiPS cells with R-17F, one major R-17F-positive band was observed at a slow migration position close to that of anti-blood group H1(O) antigen. MALDI-TOF-MS and MS(n) analyses of the purified antigen indicated that the presumptive structure of the R-17F antigen was Fuc-Hex-HexNAc-Hex-Hex-Cer. Glycan microarray analysis involving 13 different synthetic oligosaccharides indicated that R-17F bound selectively to LNFP I (Fucα1-2Galβ1-3GlcNAcβ1-3Galβ1-4Glc). A critical role of the terminal Fucα1-2 residue was confirmed by the selective disappearance of R-17F binding to the purified antigen upon α1-2 fucosidase digestion. Most interestingly, R-17F, when added to hiPS/ES cell suspensions, exhibited potent dose-dependent cytotoxicity. The cytotoxic effect was augmented markedly upon the addition of the secondary antibody (goat anti-mouse IgG1 antibody). R-17F may be beneficial for safer regenerative medicine by eliminating residual undifferentiated hiPS cells in hiPS-derived regenerative tissues, which are considered to be a strong risk factor for carcinogenesis.
['Matsumoto|Shogo|S|', 'Nakao|Hiromi|H|', 'Kawabe|Keiko|K|', 'Nonaka|Motohiro|M|', 'Toyoda|Hidenao|H|', 'Takishima|Yuto|Y|', 'Kawabata|Kenji|K|', 'Yamaguchi|Tomoko|T|', 'Furue|Miho K|MK|', 'Taki|Takao|T|', 'Okumura|Takeshi|T|', 'Yamazaki|Yuzo|Y|', 'Nakaya|Shuuichi|S|', 'Kawasaki|Nobuko|N|', 'Kawasaki|Toshisuke|T|']
D000906:Antibodies D002240:Carbohydrate Sequence D002460:Cell Line D003602:Cytotoxicity, Immunologic D006801:Humans D057026:Induced Pluripotent Stem Cells D008969:Molecular Sequence Data D009844:Oligosaccharides
cytotoxic antibody lacto-N-fucopentaose I glycolipid MALDI-TOF-MS glycan microarray carbohydrate-recognizing antibody confocal microscopy embryonic stem cell flow cytometry induced pluripotent stem cell (iPS cell) (iPSC)