The N-glycolyl form of mouse sialyl Lewis X is recognized by selectins but not by HECA-452 and FH6 antibodies that were raised against human cells.
E-, P- and L-selectins critically function in lymphocyte recirculation and recruiting leukocytes to inflammatory sites. MECA-79 antibody inhibits L-selectin-mediated lymphocyte adhesion in several species and does not require sialic acid in its epitope. Many other antibodies, however, recognize human selectin ligands expressing N-acetylneuraminic acid but not mouse selectin ligands expressing N-glycolylneuraminic acid, suggesting that difference in sialic acid in sialyl Lewis X leads to differential reactivity. We found that HECA-452 and FH6 monoclonal antibodies bind Chinese hamster ovary (CHO) cells expressing N-acetylneuraminyl Lewis X oligosaccharide but not its N-glycolyl form. Moreover, synthetic N-acetylneuraminyl Lewis X oligosaccharide but not its N-glycolyl oligosaccharide inhibited HECA-452 and FH6 binding. By contrast, E-, P- and L-selectin bound to CHO cells regardless of whether they express N-acetyl or N-glycolyl form of sialyl Lewis X, showing that selectins have a broader recognition capacity than HECA-452 and FH-6 anti-sialyl Lewis x antibodies.
Mitoma
Junya
J
Glycobiology Program, Cancer Research Center, Burnham Institute for Medical Research, La Jolla, CA 92037, USA.
Miyazaki
Tatsuo
T
Sutton-Smith
Mark
M
Suzuki
Misa
M
Saito
Hideo
H
Yeh
Jiunn-Chern
JC
Kawano
Takehiro
T
Hindsgaul
Ole
O
Seeberger
Peter H
PH
Panico
Maria
M
Haslam
Stuart M
SM
Morris
Howard R
HR
Cummings
Richard D
RD
Dell
Anne
A
Fukuda
Minoru
M
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000378:metabolism*; D016466:CHO Cells; D019556:COS Cells; D002522:Chlorocebus aethiops; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D003412:Cricetulus; D019040:E-Selectin / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005647:Fucosyltransferases / Q000235:genetics; D006801:Humans; D019041:L-Selectin / Q000378:metabolism*; D013058:Mass Spectrometry; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D006899:Mixed Function Oxygenases / Q000235:genetics; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics; D019158:N-Acetylneuraminic Acid / Q000737:chemistry / Q000378:metabolism*; D009438:Neuraminic Acids / Q000378:metabolism; D009504:Neutrophils / Q000378:metabolism*; D019007:P-Selectin / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000906:Antibodies; D019040:E-Selectin; D008562:Membrane Glycoproteins; D009438:Neuraminic Acids; D019007:P-Selectin; C084882:P-selectin ligand protein; C032592:N-glycolylneuraminic acid; D019041:L-Selectin; D006899:Mixed Function Oxygenases; C074383:CMPacetylneuraminate monooxygenase; D005647:Fucosyltransferases; D017351:N-Acetylglucosaminyltransferases; C053024:beta-1,3-galactosyl-O-glycosyl-glycoprotein beta-1,6-acetylglucosaminyl transferase; D019158:N-Acetylneuraminic Acid