title: Glycomics-based analysis of chicken red blood cells provides insight into the selectivity of the viral agglutination assay.
abstract: Agglutination of red blood cells (RBCs), including chicken RBCs (cRBCs), has been used extensively to estimate viral titer, to screen glycan-receptor binding preference, and to assess the protective response of vaccines. Although this assay enjoys widespread use, some virus strains do not agglutinate RBCs. To address these underlying issues and to increase the usefulness of cRBCs as tools for studying viruses, such as influenza, we analyzed the cell surface N-glycans of cRBCs. On the basis of the results obtained from complementary analytical strategies, including MS, 1D and 2D-NMR spectroscopy, exoglycosidase digestions, and HPLC profiling, we report the major glycan structures present on cRBCs. By comparing the glycan structures of cBRCs with those of representative human upper respiratory cells, we offer a possible explanation for the fact that certain influenza strains do not agglutinate cRBCs, using specific human-adapted influenza hemagglutinins as examples. Finally, recent understanding of the role of various glycan structures in high affinity binding to influenza hemagglutinins provides context to our findings. These results illustrate that the field of glycomics can provide important information with respect to the experimental systems used to characterize, detect and study viruses.
authors.affiliation: Harvard-MIT Division of Health Sciences & Technology, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Udayanath
authors.initials: U
authors.lastname: Aich
authors.forename: Nia
authors.initials: N
authors.lastname: Beckley
authors.forename: Zachary
authors.initials: Z
authors.lastname: Shriver
authors.forename: Rahul
authors.initials: R
authors.lastname: Raman
authors.forename: Karthik
authors.initials: K
authors.lastname: Viswanathan
authors.forename: Sven
authors.initials: S
authors.lastname: Hobbie
authors.forename: Ram
authors.initials: R
authors.lastname: Sasisekharan
mesh_terms: D000372:Agglutination Tests / Q000379:methods*; D000818:Animals; D002645:Chickens / Q000097:blood*; D004912:Erythrocytes / Q000737:chemistry*; D054794:Glycomics; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology*; D009980:Influenza A virus / Q000276:immunology*; D019906:Nuclear Magnetic Resonance, Biomolecular; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism; D011134:Polysaccharides / Q000032:analysis / Q000097:blood / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D062367:ortho-Aminobenzoates / Q000737:chemistry
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; D011134:Polysaccharides; D062367:ortho-Aminobenzoates; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; C000219:anthranilamide