Glycobiology Advance Access originally published online on September 1, 2004
Glycobiology 2005 15(2):109-118; doi:10.1093/glycob/cwh146
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Glycobiology vol. 15 no. 2 © Oxford University Press 2005; all rights reserved.
Anti-
-galactosyl antibodies recognizing epitopes terminating with 1,4-linked galactose: human natural and mouse monoclonal anti-NOR and anti-P1 antibodies
2 Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Rudolf Weigl St. 12, 53-114 Wroclaw, Poland; 3 Institute of Hematology and Blood Transfusion, 00-957 Warsaw, Poland; 4 Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, v-437 Moscow, Russian Federation; and 5 Regional Center for Blood Transfusion, 50-345 Wroclaw, Poland
1 To whom correspondence should be addressed; e-mail: lisowska{at}iitd.pan.wroc.pl
Received on July 13, 2004; revised on August 30, 2004; accepted on August 30, 2004
The rare NOR erythrocytes, which are agglutinated by most human sera, contain unique glycosphingolipids (globoside elongation products) terminating with the sequence Gal
1-4GalNAcß1-3Gal- recognized by common natural human antibodies. Anti-NOR antibodies were isolated from several human sera by affinity procedures, and their specificity was tested by inhibition of antibody binding to NOR-tri-polyacrylamide (PAA) conjugate (ELISA) by the synthetic oligosaccharides, Gal1-4GalNAcß1-3Gal (NOR-tri), Gal1-4GalNAc (NOR-di), Gal1-4Galß1-3Galß1-4Glc ((Gal)3Glc), and Gal1-4Gal (P1-di). Two major types of subspecificity of anti-NOR antibodies were found. Type 1 antibodies were found to react strongly with (Gal)3Glc and NOR-tri and weakly with P1-di and NOR-di, which indicated specificity for the trisaccharide epitope Gal1-4Gal/GalNAcß1-3Gal. Type 2 antibodies were specific to Gal1-4GalNAc, because they were inhibited most strongly by NOR-tri and NOR-di and were not (or very weakly) inhibited by (Gal)3Glc and P1-di. Monoclonal anti-NOR antibodies were obtained by immunizing mice with NOR-tri-human serum albumin (HSA) conjugate and were found to have type 2 specificity. All anti-NOR antibodies reacted specifically with NOR glycolipids on thin-layer plates. The cross-reactivity of type 1 anti-NOR antibodies with Gal1-4Gal drew attention to a possible antigenic relationship between NOR and blood group P system glycolipids. The latter glycolipids include Pk (Gal1-4Galß1-4Glc-Cer) present in all normal erythrocytes and P1 (Gal1-4Galß1-4GlcNAcß1-3Galß1-4Glc-Cer) present only in P1 erythrocytes. Sera of some P2 (P1-negative) persons contain natural anti-P1 antibodies. This prompted us to test the specificity of anti-P1 antibodies. Natural human anti-P1 isolated from serum of P2 individual and mouse monoclonal anti-P1 were best inhibited by Gal1-4Galß1-4GlcNAc (P1-tri) and did not react with NOR-tri and NOR-di. Monoclonal anti-P1 bound to Pk and P1 glycolipids and not to NOR glycolipids. These results indicated an entirely different specificity of anti-NOR and anti-P1 antibodies. Human serum samples differed in the content of anti--galactosyl antibodies, including both types of anti-NOR. In the sera of some individuals, type 1 or type 2 anti-NOR antibodies dominated, and other samples contained mixtures of both types of anti-NOR. The biological significance of these new abundant anti--galactosyl antibodies still awaits elucidation.
Key words:
anti--galactosyl
/
anti-NOR
/
anti-P1
/
glycolipids
/
polyagglutination
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  M. Duk, S. Singh, V. N. Reinhold, H. Krotkiewski, E. Kurowska, and E. Lisowska Structures of unique globoside elongation products present in erythrocytes with a rare NOR phenotype Glycobiology, March 1, 2007; 17(3): 304 - 312. [Abstract] [Full Text] [PDF]
|
|