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Imaging Mass Cytometry and Single-Cell Genomics Reveal Differential Depletion and Repletion of B-Cell Populations Following Ofatumumab Treatment in Cynomolgus Monkeys

Ofatumumab is the first, fully human, anti-CD20 monoclonal antibody in Phase 3 development for multiple sclerosis (MS). The study focused on changes in lymphocyte subsets in blood and lymphoid tissues and on potential novel biomarkers as a result of anti-CD20 antibody action in Cynomolgus monkeys tr...

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Autores principales: Theil, Diethilde, Smith, Paul, Huck, Catherine, Gilbart, Yoann, Kakarieka, Algirdas, Leppert, David, Rauld, Celine, Schmid, Cindy, Baumgartner, Reto, Stuber, Nathalie, Cordoba, Francisco, Dubost, Valerie, Darribat, Katy, Jivkov, Magali, Frieauff, Wilfried, Kneuer, Rainer, Stoeckli, Markus, Reinker, Stefan, Mansfield, Keith, Carballido, José M., Couttet, Philippe, Weckbecker, Gisbert
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6596277/
https://www.ncbi.nlm.nih.gov/pubmed/31281311
http://dx.doi.org/10.3389/fimmu.2019.01340
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author Theil, Diethilde
Smith, Paul
Huck, Catherine
Gilbart, Yoann
Kakarieka, Algirdas
Leppert, David
Rauld, Celine
Schmid, Cindy
Baumgartner, Reto
Stuber, Nathalie
Cordoba, Francisco
Dubost, Valerie
Darribat, Katy
Jivkov, Magali
Frieauff, Wilfried
Kneuer, Rainer
Stoeckli, Markus
Reinker, Stefan
Mansfield, Keith
Carballido, José M.
Couttet, Philippe
Weckbecker, Gisbert
author_facet Theil, Diethilde
Smith, Paul
Huck, Catherine
Gilbart, Yoann
Kakarieka, Algirdas
Leppert, David
Rauld, Celine
Schmid, Cindy
Baumgartner, Reto
Stuber, Nathalie
Cordoba, Francisco
Dubost, Valerie
Darribat, Katy
Jivkov, Magali
Frieauff, Wilfried
Kneuer, Rainer
Stoeckli, Markus
Reinker, Stefan
Mansfield, Keith
Carballido, José M.
Couttet, Philippe
Weckbecker, Gisbert
author_sort Theil, Diethilde
collection PubMed
description Ofatumumab is the first, fully human, anti-CD20 monoclonal antibody in Phase 3 development for multiple sclerosis (MS). The study focused on changes in lymphocyte subsets in blood and lymphoid tissues and on potential novel biomarkers as a result of anti-CD20 antibody action in Cynomolgus monkeys treated with human equivalent doses of subcutaneous (s.c.) ofatumumab on Days 0, 7, and 14. Axillary lymph nodes (LNs) and blood samples were collected at various time points until Day 90. Lymphocyte subsets were quantified by flow cytometry, while morphological and immune cell changes were assessed by imaging mass cytometry (IMC), immunohistochemistry (IHC), in situ hybridization (ISH), and transcriptome analyses using single-cell methodology. Ofatumumab treatment resulted in a potent and rapid reduction of B cells along with a simultaneous drop in CD20(+) T cell counts. At Day 21, IHC revealed B-cell depletion in the perifollicular and interfollicular area of axillary LNs, while only the core of the germinal center was depleted of CD20(+)CD21(+) cells. By Day 62, the perifollicular and interfollicular areas were abundantly infiltrated by CD21(+) B cells and this distribution returned to the baseline cytoarchitecture by Day 90. By IMC CD20(+)CD3(+)CD8(+) cells could be identified at the margin of the follicles, with a similar pattern of distribution at Day 21 and 90. Single-cell transcriptomics analysis showed that ofatumumab induced reversible changes in t-distributed stochastic neighbor embedding (t-SNE) defined B-cell subsets that may serve as biomarkers for drug action. In summary, low dose s.c. ofatumumab potently depletes both B cells and CD20(+) T cells but apparently spares marginal zone (MZ) B cells in the spleen and LN. These findings add to our molecular and tissue-architectural understanding of ofatumumab treatment effects on B-cell subsets.
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spelling pubmed-65962772019-07-05 Imaging Mass Cytometry and Single-Cell Genomics Reveal Differential Depletion and Repletion of B-Cell Populations Following Ofatumumab Treatment in Cynomolgus Monkeys Theil, Diethilde Smith, Paul Huck, Catherine Gilbart, Yoann Kakarieka, Algirdas Leppert, David Rauld, Celine Schmid, Cindy Baumgartner, Reto Stuber, Nathalie Cordoba, Francisco Dubost, Valerie Darribat, Katy Jivkov, Magali Frieauff, Wilfried Kneuer, Rainer Stoeckli, Markus Reinker, Stefan Mansfield, Keith Carballido, José M. Couttet, Philippe Weckbecker, Gisbert Front Immunol Immunology Ofatumumab is the first, fully human, anti-CD20 monoclonal antibody in Phase 3 development for multiple sclerosis (MS). The study focused on changes in lymphocyte subsets in blood and lymphoid tissues and on potential novel biomarkers as a result of anti-CD20 antibody action in Cynomolgus monkeys treated with human equivalent doses of subcutaneous (s.c.) ofatumumab on Days 0, 7, and 14. Axillary lymph nodes (LNs) and blood samples were collected at various time points until Day 90. Lymphocyte subsets were quantified by flow cytometry, while morphological and immune cell changes were assessed by imaging mass cytometry (IMC), immunohistochemistry (IHC), in situ hybridization (ISH), and transcriptome analyses using single-cell methodology. Ofatumumab treatment resulted in a potent and rapid reduction of B cells along with a simultaneous drop in CD20(+) T cell counts. At Day 21, IHC revealed B-cell depletion in the perifollicular and interfollicular area of axillary LNs, while only the core of the germinal center was depleted of CD20(+)CD21(+) cells. By Day 62, the perifollicular and interfollicular areas were abundantly infiltrated by CD21(+) B cells and this distribution returned to the baseline cytoarchitecture by Day 90. By IMC CD20(+)CD3(+)CD8(+) cells could be identified at the margin of the follicles, with a similar pattern of distribution at Day 21 and 90. Single-cell transcriptomics analysis showed that ofatumumab induced reversible changes in t-distributed stochastic neighbor embedding (t-SNE) defined B-cell subsets that may serve as biomarkers for drug action. In summary, low dose s.c. ofatumumab potently depletes both B cells and CD20(+) T cells but apparently spares marginal zone (MZ) B cells in the spleen and LN. These findings add to our molecular and tissue-architectural understanding of ofatumumab treatment effects on B-cell subsets. Frontiers Media S.A. 2019-06-20 /pmc/articles/PMC6596277/ /pubmed/31281311 http://dx.doi.org/10.3389/fimmu.2019.01340 Text en Copyright © 2019 Theil, Smith, Huck, Gilbart, Kakarieka, Leppert, Rauld, Schmid, Baumgartner, Stuber, Cordoba, Dubost, Darribat, Jivkov, Frieauff, Kneuer, Stoeckli, Reinker, Mansfield, Carballido, Couttet and Weckbecker. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Immunology
Theil, Diethilde
Smith, Paul
Huck, Catherine
Gilbart, Yoann
Kakarieka, Algirdas
Leppert, David
Rauld, Celine
Schmid, Cindy
Baumgartner, Reto
Stuber, Nathalie
Cordoba, Francisco
Dubost, Valerie
Darribat, Katy
Jivkov, Magali
Frieauff, Wilfried
Kneuer, Rainer
Stoeckli, Markus
Reinker, Stefan
Mansfield, Keith
Carballido, José M.
Couttet, Philippe
Weckbecker, Gisbert
Imaging Mass Cytometry and Single-Cell Genomics Reveal Differential Depletion and Repletion of B-Cell Populations Following Ofatumumab Treatment in Cynomolgus Monkeys
title Imaging Mass Cytometry and Single-Cell Genomics Reveal Differential Depletion and Repletion of B-Cell Populations Following Ofatumumab Treatment in Cynomolgus Monkeys
title_full Imaging Mass Cytometry and Single-Cell Genomics Reveal Differential Depletion and Repletion of B-Cell Populations Following Ofatumumab Treatment in Cynomolgus Monkeys
title_fullStr Imaging Mass Cytometry and Single-Cell Genomics Reveal Differential Depletion and Repletion of B-Cell Populations Following Ofatumumab Treatment in Cynomolgus Monkeys
title_full_unstemmed Imaging Mass Cytometry and Single-Cell Genomics Reveal Differential Depletion and Repletion of B-Cell Populations Following Ofatumumab Treatment in Cynomolgus Monkeys
title_short Imaging Mass Cytometry and Single-Cell Genomics Reveal Differential Depletion and Repletion of B-Cell Populations Following Ofatumumab Treatment in Cynomolgus Monkeys
title_sort imaging mass cytometry and single-cell genomics reveal differential depletion and repletion of b-cell populations following ofatumumab treatment in cynomolgus monkeys
topic Immunology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6596277/
https://www.ncbi.nlm.nih.gov/pubmed/31281311
http://dx.doi.org/10.3389/fimmu.2019.01340
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