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Four newly developed monoclonal antibodies (MAbs) are characterized using flowcytometry, enzyme-linked immunoadsorbent assay (ELISA), immunoprecipitation and Western blots, carbohydrate epitope mapping, glycosidase cleavage, and competition binding assays. Their effects on selectin binding to myeloid cells was tested. These MAbs react only with myeloid cells. MAbs CI-1, BU60, and HIM95 recognize epitopes expressed by CD11/CD18 (beta2) integrins, while HI247 and CSLEX1 do not. The epitopes require Lewis x [Galbeta1-4 (Fucalpha1-3)GlcNAc] based on reactivity with oligosaccharide-polyacrylamide-biotin or oligosaccharide-BSA conjugates. MAb HI247 recognizes a related structure, sialyl-Lewis x, NeuAcalpha2-3GaLbeta1-4(Fucalpha1-3)GlcNAc. The three MAbs against Lewis x show some minor differences in their reactivity such as recognizing their antigens on CD11/CD18 integrins after endo-beta-galactosidase treatment and recognizing free Lewis x. The hydroxyl group on C-3 of the terminal galactose is important for recognition by MAb CI-1, BU60, and HIM95 as its substitution with sulfo group of sialic acid abolishes the binding of these MAbs. The C-3 sialic acid is crucial for the binding of MAb HI247. Its replacement by sulphate or its cleavage by sialidase eliminates recognition by this MAb. MAbs HI247 and CSLEX-1 did not react in ELISA with immobilized CD11/CD18, suggesting that the majority of sialyl Lewis x on CD11/CD18 molecules may have sialic acid 6-linked rather than 3-linked to galactose. Unexpectedly, MAb BU60 inhibited binding of P-selectin mu chain chimera to HL-60 or U937 cells, while CI-1, HIM95 and three other defined anti-Lewis x MAbs (6C7, M6-1 and LeuM1) did not. MAb HI247 inhibited binding of both E- and P-selectin chimeras to these cell lines more effectively than several characterized MAbs (CSLEX-1, FH6, HECA-452) to sialyl Lewis x and related oligosaccharides. Certain combinations of these anticarbohydrate MAbs had additive inhibitory effects on selectin binding, suggesting a potential application of these new MAbs in cell adhesion/migration and tumor metastasis studies.
K Zhang, and
H E Chuluyan, and
D Hardie, and
D C Shen, and
R Larsen, and
A Issekutz
September 1986,
The Journal of biological chemistry,
K Zhang, and
H E Chuluyan, and
D Hardie, and
D C Shen, and
R Larsen, and
A Issekutz
April 1996,
The Journal of biological chemistry,
K Zhang, and
H E Chuluyan, and
D Hardie, and
D C Shen, and
R Larsen, and
A Issekutz
March 1985,
Biochemical and biophysical research communications,
K Zhang, and
H E Chuluyan, and
D Hardie, and
D C Shen, and
R Larsen, and
A Issekutz
September 1984,
Cancer research,
K Zhang, and
H E Chuluyan, and
D Hardie, and
D C Shen, and
R Larsen, and
A Issekutz
October 2005,
Journal of immunological methods,
K Zhang, and
H E Chuluyan, and
D Hardie, and
D C Shen, and
R Larsen, and
A Issekutz
February 2001,
The Journal of veterinary medical science,
K Zhang, and
H E Chuluyan, and
D Hardie, and
D C Shen, and
R Larsen, and
A Issekutz
January 1988,
Current topics in microbiology and immunology,
K Zhang, and
H E Chuluyan, and
D Hardie, and
D C Shen, and
R Larsen, and
A Issekutz
July 1985,
Biochimica et biophysica acta,
K Zhang, and
H E Chuluyan, and
D Hardie, and
D C Shen, and
R Larsen, and
A Issekutz
March 1991,
European journal of biochemistry,
K Zhang, and
H E Chuluyan, and
D Hardie, and
D C Shen, and
R Larsen, and
A Issekutz
July 1987,
Biulleten' eksperimental'noi biologii i meditsiny,
