| Full text | |
| Author(s): |
Koike, Gabriela
[1]
;
Katz, Iana Suly Santos
[1]
;
Fernandes, Elaine Raniero
[1]
;
Guedes, Fernanda
[1]
;
Silva, Sandriana Ramos
[1]
Total Authors: 5
|
| Affiliation: | [1] Inst Pasteur, 393 Paulista Ave, BR-01311000 Sao Paulo, SP - Brazil
Total Affiliations: 1
|
| Document type: | Journal article |
| Source: | Immunobiology; v. 226, n. 2 MAR 2021. |
| Web of Science Citations: | 0 |
| Abstract | |
Rabies lyssavirus (RABV) neutralizing IgG antibodies confer protection after rabies vaccination, although how the RABV-specific antibodies neutralize the virus is still unknown. As changes in the antibody`s carbohydrate chain can interfere with its effector functions, we compared the glycosylation patterns of both neutralizing and non-neutralizing IgG1 induced by pre-exposure prophylaxis to human rabies and analyzed their influence on in vitro antibody neutralizing activities. Specific IgG1 were purified from human serum using affinity chromatography. Purity and avidity were analyzed by SDS-PAGE and indirect ELISA using NH4SCN respectively. The N-linked oligosaccharide chain of the purified IgG antibody was evaluated using a lectin-based ELISA assay with a panel of seven lectins. The activity of purified IgG1 and neutralizing IgG1 deglycosylated by PNGase F enzyme were analyzed using the rapid fluorescent focus inhibition test. The purified IgG1 showed an electrophoretic pattern compatible with human IgG. All of the antibodies recognized RABV, although neutralizing IgG1 had a higher avidity (RAI = 80%) than non-neutralizing IgG1 (RAI = 30%). The neutralizing IgG1 also showed higher binding to WFA, ECA, WGA, and ConA lectins, indicating possible different N-acetylgalactosamine, galactose, N-acetylglucosamine, and mannose contents. Non-neutralizing IgG1, on the other hand, showed strong binding at UEA-1 and SNA, which bind to fucose and sialic acid residues respectively. Different glycosylation profiles were also observed in Fab and Fc fragments from neutralizing and non-neutralizing IgG1, although the deglycosylated IgG1 lost its neutralizing activity. Our results suggest that antibody glycosylation is important for neutralizing RABV in vitro, since neutralizing IgG1 has a different glycosylation profile than non-neutralizing IgG1. Further research will be needed to better evaluate the differential glycosylation patterns between IgG1 antibodies following vaccination. (AU) | |
| FAPESP's process: | 18/17580-3 - Comparison of the neutralizing activity and the glycosylation pattern of the IgG antibodies induced by pre-exposure schedule of human rabies vaccination |
| Grantee: | Gabriela Koike |
| Support Opportunities: | Scholarships in Brazil - Scientific Initiation |