A cell-based in vitro assay for testing of immunological integrity of Tetanus toxoid vaccine antigen

Abstract Vaccines containing inactivated toxins confer protection by eliciting a neutralizing antibody response against bacterial toxins such as tetanus and diphtheria. At present, release of tetanus toxoid (TT) and diphtheria toxoid (DT)-containing vaccines relies on in vivo experiments showing the...

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Autores principales:	Olga Ticha, Dido Klemm, Lukas Moos, Isabelle Bekeredjian-Ding
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Immunologic diseases. Allergy RC581-607 Neoplasms. Tumors. Oncology. Including cancer and carcinogens RC254-282
Acceso en línea:	https://doaj.org/article/ba79510f5f9d4ec98012d881b5343114
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spelling	oai:doaj.org-article:ba79510f5f9d4ec98012d881b53431142021-12-02T17:45:02ZA cell-based in vitro assay for testing of immunological integrity of Tetanus toxoid vaccine antigen10.1038/s41541-021-00344-12059-0105https://doaj.org/article/ba79510f5f9d4ec98012d881b53431142021-06-01T00:00:00Zhttps://doi.org/10.1038/s41541-021-00344-1https://doaj.org/toc/2059-0105Abstract Vaccines containing inactivated toxins confer protection by eliciting a neutralizing antibody response against bacterial toxins such as tetanus and diphtheria. At present, release of tetanus toxoid (TT) and diphtheria toxoid (DT)-containing vaccines relies on in vivo experiments showing the protective vaccine response. The aim of this study was to develop a reliable in vitro assay for TT vaccine antigen characterization with the potential of replacing in vivo potency experiments. To this end, we exploited that TT elicits a recall response in vaccinated donors: human peripheral blood mononuclear cells (PBMC) were stimulated with alum-adsorbed TT bulk antigen and low concentrations of TLR9 ligand; induction of TT-specific IgG was quantified via ELISpot after 5 days. Proof-of-concept was obtained using paired samples from donors before and after vaccination; anti-TT IgG was only detected in PBMC collected after booster vaccination; specificity was demonstrated with DT stimulation as control. Notably, when using PBMC from buffy coats, the specific response to TT was reproducible in 30% of cells; responsiveness correlated with higher numbers of switched memory B cells. Consecutive results showed that TT-specific IgG was also detectable when PBMC were stimulated with DTaP final vaccine product. Thus, the assay provides a viable means to test B-cell differentiation and induction of TT-specific IgG secretion using bulk antigen and final vaccine. However, prequalification of PBMC is required for reliable performance. Along with physicochemical and immunochemical methods, the functional assay could represent a complementary tool to replace in vivo potency assays in batch release of TT-containing vaccines.Olga TichaDido KlemmLukas MoosIsabelle Bekeredjian-DingNature PortfolioarticleImmunologic diseases. AllergyRC581-607Neoplasms. Tumors. Oncology. Including cancer and carcinogensRC254-282ENnpj Vaccines, Vol 6, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Immunologic diseases. Allergy RC581-607 Neoplasms. Tumors. Oncology. Including cancer and carcinogens RC254-282
spellingShingle	Immunologic diseases. Allergy RC581-607 Neoplasms. Tumors. Oncology. Including cancer and carcinogens RC254-282 Olga Ticha Dido Klemm Lukas Moos Isabelle Bekeredjian-Ding A cell-based in vitro assay for testing of immunological integrity of Tetanus toxoid vaccine antigen
description	Abstract Vaccines containing inactivated toxins confer protection by eliciting a neutralizing antibody response against bacterial toxins such as tetanus and diphtheria. At present, release of tetanus toxoid (TT) and diphtheria toxoid (DT)-containing vaccines relies on in vivo experiments showing the protective vaccine response. The aim of this study was to develop a reliable in vitro assay for TT vaccine antigen characterization with the potential of replacing in vivo potency experiments. To this end, we exploited that TT elicits a recall response in vaccinated donors: human peripheral blood mononuclear cells (PBMC) were stimulated with alum-adsorbed TT bulk antigen and low concentrations of TLR9 ligand; induction of TT-specific IgG was quantified via ELISpot after 5 days. Proof-of-concept was obtained using paired samples from donors before and after vaccination; anti-TT IgG was only detected in PBMC collected after booster vaccination; specificity was demonstrated with DT stimulation as control. Notably, when using PBMC from buffy coats, the specific response to TT was reproducible in 30% of cells; responsiveness correlated with higher numbers of switched memory B cells. Consecutive results showed that TT-specific IgG was also detectable when PBMC were stimulated with DTaP final vaccine product. Thus, the assay provides a viable means to test B-cell differentiation and induction of TT-specific IgG secretion using bulk antigen and final vaccine. However, prequalification of PBMC is required for reliable performance. Along with physicochemical and immunochemical methods, the functional assay could represent a complementary tool to replace in vivo potency assays in batch release of TT-containing vaccines.
format	article
author	Olga Ticha Dido Klemm Lukas Moos Isabelle Bekeredjian-Ding
author_facet	Olga Ticha Dido Klemm Lukas Moos Isabelle Bekeredjian-Ding
author_sort	Olga Ticha
title	A cell-based in vitro assay for testing of immunological integrity of Tetanus toxoid vaccine antigen
title_short	A cell-based in vitro assay for testing of immunological integrity of Tetanus toxoid vaccine antigen
title_full	A cell-based in vitro assay for testing of immunological integrity of Tetanus toxoid vaccine antigen
title_fullStr	A cell-based in vitro assay for testing of immunological integrity of Tetanus toxoid vaccine antigen
title_full_unstemmed	A cell-based in vitro assay for testing of immunological integrity of Tetanus toxoid vaccine antigen
title_sort	cell-based in vitro assay for testing of immunological integrity of tetanus toxoid vaccine antigen
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/ba79510f5f9d4ec98012d881b5343114
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A cell-based in vitro assay for testing of immunological integrity of Tetanus toxoid vaccine antigen

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