Naturally occurring anti-band 3 antibody binds to apoptotic human T-lymphoid cell line Jurkat through sialylated poly-N-acetyllactosaminyl saccharide chains on the cell surface.
Human T-lymphoid cell line Jurkat was treated with actinomycin D (ActD) and cycloheximide (CHX). The induction of apoptosis was confirmed by the chromatin condensation and DNA ladder fragmentation. Anti-band 3 IgG, purified from normal human plasma, bound to the ActD- or CHX-treated cells, and the binding was correlated to the degree of apoptosis. Antioxidants, N-acetylcysteine, pilloridine dithiocarbamate, and trolox, inhibited neither induction of DNA fragmentation of ActD-treated cells nor anti-band 3 IgG binding to ActD-treated cells, indicating that formation of the anti-band 3 IgG binding sites on the apoptotic cell surface is caused by nonoxidative mechanism. When Jurkat cells were treated with endo-beta-galactosidase to cleave sialylated poly-N-acetyllactosaminyl saccharide chains from the cell surface before induction of apoptosis, the binding of anti-band 3 IgG was abolished. The results indicate that sialylated poly-N-acetyllactosaminyl saccharide chains on the cell surface are requisite for the binding of anti-band 3 IgG to apoptotic cells.
Ando
K
K
School of Pharmacy, Tokyo University of Pharmacy and Life Science, 1432-1 Horinouchi, Hachioji, Tokyo, 192-0392, Japan.
Hagiwara
T
T
Beppu
M
M
Kikugawa
K
K
D000606:Amino Sugars / Q000737:chemistry / Q000276:immunology*; D001457:Anion Exchange Protein 1, Erythrocyte / Q000276:immunology*; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis; D001666:Binding Sites, Antibody; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects; D003513:Cycloheximide / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D006801:Humans; D019169:Jurkat Cells; D018384:Oxidative Stress
D000606:Amino Sugars; D001457:Anion Exchange Protein 1, Erythrocyte; D000975:Antioxidants; D003609:Dactinomycin; C000458:N-acetyllactosamine; D003513:Cycloheximide