Ludwik Hirszfeld Institute of Immunology and Experimental Therapy

Alpha-Gal syndrome unveiled: decoding the glycan-driven red meat allergy.

α-Gal syndrome (AGS) is a type of allergy to red meat and other mammalian products that develops after a tick bite. Symptoms may include urticaria, angioedema, gastrointestinal and respiratory symptoms, and, most dangerously, anaphylactic shock. Among food allergies, it is considered unusual because: (i) the major allergen is not a protein, but a carbohydrate moiety Galα-1,3-Gal, which humans do not synthesize; (ii) it is caused by α-Gal-containing glycoconjugates injected during a tick bite; (iii) the hypersensitivity is effected by IgE antibodies which arise after a class switch from IgG and IgM-class antibodies; (iv) it seems to be triggered by α-Gal-containing glycolipids present in red meat, with delayed onset. Here, we evaluate glycans that may cause AGS, review the data about glycosyltransferases that synthesize α-Gal and related antigens and describe the immune mechanism behind AGS: sensitization and host response to ticks' saliva. In addition, we consider future research avenues that may lead to a better understanding and management of AGS as a modern dietary affliction rooted in an old evolutionary trade-off.

Correction to: Alpha-Gal syndrome unveiled: decoding the glycan-driven red meat allergy.

Specificity of human anti-NOR antibodies, a distinct species of "natural" anti-alpha-galactosyl antibodies.

Natural anti-NOR antibodies are common in human sera and agglutinate human erythrocytes of a rare NOR phenotype. The NOR phenotype-related antigens are unique neutral glycosphingolipids recognized by these antibodies and Griffonia simplicifolia IB4 isolectin (GSL-IB4). The oligosaccharide chains of NOR glycolipids are terminated by Galalpha1-4GalNAcbeta1-3Galalpha units. To characterize the specificity of anti-NOR antibodies and compare it with specificities of GSL-IB4 and known anti-Galalpha1,3Gal antibodies, alpha-galactosylated saccharides and saccharide-polyacrylamide conjugates were used. New synthetic oligosaccharides, corresponding to the terminal di- and trisaccharide sequence of NOR glycolipids and the conjugate of the NOR-tri with HSA were included. These compounds were tested by microtiter plate ELISA and hemagglutination inhibition. Anti-NOR antibodies reacted most strongly with Galalpha1-4GalNAcbeta1-3Gal (NOR-tri), and over 100 times less strongly with Galalpha1-4GalNAc (NOR-di). The antibodies reacted also with Galalpha1-4Gal and Galalpha1-4Galbeta1-4GlcNAc, similarly as with NOR-di but not with other tested compounds. In turn, anti-Galalpha1,3Gal antibodies reacted most strongly with Galalpha1-3Gal and were very weakly inhibited by the NOR-related oligosaccharides (weaker than by galactose), and NOR-tri was less active than NOR-di. GSL-IB4 reacted with all tested alpha-galactosylated saccharides and conjugates, including the similarly active NOR-tri and NOR-di. These results showed that anti-NOR represent a new species of anti-alpha-galactosyl antibodies with high affinity for the Galalpha1-4GalNAcbeta1-3Gal sequence present in rare NOR erythrocytes.

Structure of a neutral glycosphingolipid recognized by human antibodies in polyagglutinable erythrocytes from the rare NOR phenotype.

NOR is a rare inheritable polyagglutination phenomenon that has been described in two families. Our recent studies on these erythrocytes showed they contained at least two unique neutral glycosphingolipids, and based on their reactivity with Griffonia simplicifolia IB4 (GSL-IB4) isolectin (Kusnierz-Alejska, G., Duk, M., Storry, J. R., Reid, M. E., Wiecek, B., Seyfried, H., and Lisowska, E. (1999) Transfusion 39, 32-38), both oligosaccharide chains terminated with an alpha-galactose residue. The reactivity with GSL-IB4 suggested that these oligosaccharide chains terminated with a Galalpha1-->3Gal- sequence and that anti-NOR agglutinins were common human anti-Galalpha1-->3Gal xenoantibodies. In this report we describe the structure of one NOR component (NOR1) that migrated on thin-layer chromatographic plates in the region of pentaglycosylceramides. Treatment of this sample with alpha-galactosidase and beta-N-acetylhexosaminidase was followed by high-performance thin-layer chromatography with product detection by lectins and the anti-Gb4 monoclonal antibody. The results suggested that NOR1 was an alpha-galactosylated Gb4Cer with a beta-N-acetylhexosaminidase-resistant GalNAc residue. Gas phase disassembly by ion trap mass spectrometry analysis showed the sequence to be Hex1-->4HexN1-->3Hex1-->4Hex1-->4Hex linked to a ceramide composed of C18 sphingosine and a C24 monounsaturated fatty acid. Together these data indicate NOR1 to be a novel Galalpha1-->4GalNAcbeta1-->3Galalpha1-->4Galbeta1-->4 Glc-Cer structure. Additionally it has been shown that NOR glycolipids are recognized by human antibodies that were distinct from the known anti-Galalpha1-->3Gal xenoantibodies.

Red blood cell antigens responsible for inherited types of polyagglutination.

The three described types on inheritable polyagglutination are related to altered carbohydrate structures in glycoproteins or/and glycolipds on the erythrocyte surface. HEMPAS, a condition causing anemia and other pathological symptoms, is characterized by impaired biosynthesis of N-glycans, mostly those carried by band 3 and band 4.5 erythrocyte membrane proteins. Cad erythrocytes have abnormal glycophorin O-glycans, structurally related to the more common human Sd(a) and murine CT determinants, and accumulate an Sd(a)-like ganglioside. NOR erythrocytes express recently detected abnormal alpha-galactose-terminated glycosphingolipids, which strongly react with G. simplicifolia IB4 isolectin, but do not react with human anti-Galalpha1-3Gal antibodies.