We studied carbohydrate specificity and isotypes of antibodies to BSA-conjugated tetrasaccharide, a repeating unit of the capsular polysaccharide of Streptococcus pneumoniae serotype 14, in mouse polyclonal sera and hybridoma-synthesized products. Natural IgM antibodies to the tetrasaccharide containing epitopes similar to surface carbohydrate structures of mammalian and human cells in low titers were determined in native mouse serum by ELISA using biotinylated tetrasaccharide and synthetic capsular polysaccharide as the solid-phase antigens. Polyclonal sera to the conjugated tetrasaccharide contained IgM and all subclasses of IgG antibodies, which were detected in a higher titer when the biotinylated tetrasaccharide was used as a solid phase antigen compared to synthetic capsular polysaccharide. Monoclonal antibodies to S. pneumoniae serotype 14 tetrasaccharide were identified in an equivalent titer using either biotinylated tetrasaccharide or synthetic capsular polysaccharide. Monoclonal antibodies obtained in vitro belonged to IgM isotype and cross-reacted with secondary full-size IgG antibodies. In the serum of mice inoculated with hybridoma, IgM and IgG2a antibodies recognizing the tetrasaccharide epitope in the structure of synthetic capsular polysaccharide were simultaneously determined.
Similar content being viewed by others
References
Akhmatova NK, Kurbatova EA, Akhmatov EA, Egorova NB, Logunov DY, Gening ML, Sukhova EV, Yashunsky DV, Tsvetkov YE, Nifantiev NE. The effect of a BSA conjugate of a synthetic hexasaccharide related to the fragment of capsular polysaccharide of Streptococcus pneumoniae type 14 on the activation of innate and adaptive immune responses. Front. Immunol. 2016;7:248. doi: https://doi.org/10.3389/fimmu.2016.00248.
Ananikov VP, Eremin DB, Yakukhnov SA, Dilman AD, Levin VV, Egorov MP, Karlov SS, Kustov LM, Tarasov AL, Greish AA, Shesterkina AA, Sakharov AM, Nysenko ZN, Sheremetev AB, Stakheev AY, Mashkovsky IS, Sukhorukov AY, Ioffe SL, Terent’ev AO, Vil VA, Tomilov YV, Novikov RA, Zlotin SG, Kucherenko AS, Ustyuzhanina NE, Krylov VB, Tsvetkov YE, Gening ML, Nifantiev NE. Organic and hybrid systems: from science to practice. Mendeleev Commun. 2017;27(5):425-438.
Broecker F, Anish C, Seeberger PH. Generation of monoclonal antibodies against defined oligosaccharide antigens. Methods Mol. Biol. 2015;1331:57-80.
Cantelli CP, da Glória Martins Teixeira M, Santos EA, da Silva HC, da Silva E Mouta S, Pimenta MM, Vianna CO, de Souza NP, Batoreu NM, Galler R, de Moraes MT. Generation of monoclonal antibodies against human recombinant interferon beta using genetic immunization with simultaneous expression of IgM and IgG isotypes. Hybridoma (Larchmt). 2009;28(3):211-214.
Gening ML, Tsvetkov YE, Nifantiev NE, Kurbatova EA. Development of approaches to a third-generation carbohydrate-conjugate vaccine against Streptococcus pneumoniae: the search for optimal oligosaccharide ligands. Russ. Chem. Rev. 2015;84(11):1100-1113.
Huflejt ME, Vuskovic M, Vasiliu D, Xu H, Obukhova P, Shilova N, Tuzikov A, Galanina O, Arun B, Lu K, Bovin N. Anti-carbohydrate antibodies of normal sera: findings, surprises and challenges. Mol. Immunol. 2009;46(15):3037-3049.
Jansen WT, Snippe H. Short-chain oligosaccharide protein conjugates as experimental pneumococcal vaccines. Indian J. Med. Res. 2004;119(Suppl):7-12.
Kurbatova EA, Akhmatova NK, Akhmatova EA, Egorova NB, Yastrebova NE, Sukhova EV, Yashunsky DV, Tsvetkov YE, Gening ML, Nifantiev NE. Neoglycoconjugate of tetrasaccharide representing one repeating unit of the Streptococcus pneumoniae type 14 capsular polysaccharide induces the production of opsonizing IgG1 antibodies and possesses the highest protective activity as compared to hexa- and octasaccharide conjugates. Front. Immunol. 2017;8:659. doi: https://doi.org/10.3389/fimmu.2017.00659.
Payelle-Brogard B, Ragimbeau J, Avrameas S, Christodoulou C. Immunoglobulin double isotype-producing hybridomas isolated from an autoimmune (NZB*NZW)F1 mouse. Hybridoma. 1998;17(3):289-297.
Safari D, Dekker HA, Joosten JA, Michalik D, de Souza AC, Adamo R, Lahmann M, Sundgren A, Oscarson S, Kamerling JP, Snippe H. Identification of the smallest structure capable of evoking opsonophagocytic antibodies against Streptococcus pneumoniae type 14. Infect. Immun. 2008;76(10):4615-4623.
Sukhova EV, Yashunsky DV, Tsvetkov YE, Nifantiev NE, Kurbatova EA. Synthesis of oligosaccharide fragments of the Streptococcus pneumoniae type 14 capsular polysaccharide and their neoglycoconjugates with bovine serum albumin. Russ. Chem. Bull. 2014;63(2):511-521.
Vidarsson G, Dekkers G, Rispens T. IgG subclasses and allotypes: from structure to effector functions. Front. Immunol. 2014;5:520. doi: https://doi.org/10.3389/fimmu.2014.00520.
Vollmers HP, Brändlein S. Natural antibodies and cancer. J. Autoimmun. 2007;29(4):295-302.
Weintraub A. Immunology of bacterial polysaccharide antigens. Carbohydr. Res. 2003;338(23):2539-2547.
Author information
Authors and Affiliations
Corresponding author
Additional information
Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 166, No. 10, pp. 470-474, October, 2018
Rights and permissions
About this article
Cite this article
Kurbatova, Е.А., Yakovleva, I.V., Akhmatova, E.А. et al. Carbohydrate Specificity and Isotypes of Monoclonal and Polyclonal Antibodies to Conjugated Tetrasaccharide, a Synthetic Analogue of Repeating Unit of Capsular Polysaccharide of Streptococcus Pneumoniae Serotype 14. Bull Exp Biol Med 166, 477–480 (2019). https://doi.org/10.1007/s10517-019-04376-5
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10517-019-04376-5