A triple positive staining to capture B cells expressing anti-tetanus toxoid IgG in human blood

Abstract

Event Abstract Back to Event A triple positive staining to capture B cells expressing anti-tetanus toxoid IgG in human blood Eduardo Aliprandini1, Wagner Quintilio2, Jorge Kalil3, 4, Silvia B. Boscardin5 and Ana M. Moro1, 4* 1 Laboratório de Biofármacos em Células Animais, Instituto Butantan, Brazil 2 Controle de Qualidade, Instituto Butantan, Brazil 3 Instituto Butantan, Brazil 4 INCT/iii - Instituto de Investigação em Imunologia, Brazil 5 Departamento de Parasitologia, ICB/USP, Brazil Aiming to sort B cells expressing anti-tetanus toxoid IgG from human blood that could be used for heavy and light chain amplification and cloning, this work started by the preliminary detection of such cells in 10 mL of human peripheral blood. After informed consent, blood was drawn from six volunteers and the mononuclear cells (PBMC) were stained using anti CD19-FITC, anti IgG-APC, and streptavidin-PerCP-Cy5.5, following analysis by flow cytometry. Tetanus toxoid (TT) conjugated to biotin (TT-bio) was included or not in the mixture. Five donors were selected based on their high anti-TT antibody titers: 12.0, 16.1, 13.3, 13.0 and 12.2 UI/mL; a sixth donor did not receive tetanus booster immunization after infant vaccination (titer 1.2 UI/mL). Despite the fact that all donors presented protective anti-TT antibody titers (>0,1 UI/mL), neither donor (1) or (6) showed detectable anti-TT producing B-lymphocytes in the blood. The other 4 donors presented variable responses, expressed as anti-TT B lymphocytes/10^6 PBMC as 16.13, 36.18, 11.15 and 2.82. The frequency of anti-TT lymphocytes did not correlate with frequency of total lymphocytes in PBMC, which varied between 35 to 62%. The individual variability of the immunological response denotes the importance of testing several donors, even with a small volume of blood. The possibility of detection of the desired cells by the above mentioned triple staining protocol prompted us to draw a large volume of blood from the donors in order to start the sorting protocols. Acknowledgements Financial support: CAPES, FAPESP, CNPq. Keywords: antibody, Tetanus Toxoid, B lymphocytes, Flow Cytometry, immune repertoire Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Adaptive Immunity Citation: Aliprandini E, Quintilio W, Kalil J, Boscardin SB and Moro AM (2013). A triple positive staining to capture B cells expressing anti-tetanus toxoid IgG in human blood. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00567 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 08 May 2013; Published Online: 22 Aug 2013. * Correspondence: Dr. Ana M Moro, Laboratório de Biofármacos em Células Animais, Instituto Butantan, São Paulo, Brazil, anamoro@butantan.gov.br Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Eduardo Aliprandini Wagner Quintilio Jorge Kalil Silvia B Boscardin Ana M Moro Google Eduardo Aliprandini Wagner Quintilio Jorge Kalil Silvia B Boscardin Ana M Moro Google Scholar Eduardo Aliprandini Wagner Quintilio Jorge Kalil Silvia B Boscardin Ana M Moro PubMed Eduardo Aliprandini Wagner Quintilio Jorge Kalil Silvia B Boscardin Ana M Moro Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.