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In Vitro and Ex Vivo Methodologies for T-Cell Trafficking Through Blood–Brain Barrier After TLR Activation

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Toll-Like Receptors

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2700))

Abstract

This chapter describes ex vivo isolation of human T cells and of naïve splenocytes respectively collected from multiple sclerosis patients and healthy controls and experimental autoimmune encephalomyelitis-affected mice. After the magnetic sorting of naïve and activated T helper lymphocytes, we provide details about the cell cultures to measure the interaction with extracellular matrix proteins using standard cell invasion or hand-made in vitro assays, upon different stimuli, through Toll-like receptor(s) ligands, T-cell activators, and cell adhesion molecules modulators. Finally, we describe the methods to harvest and recover T cells to evaluate the properties associated with their trafficking ability.

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Acknowledgments

This work received nonconditional support by Fondazione Cassa di Risparmio di Perugia, Project 2021.0347 (GDS).

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Authors and Affiliations

  1. Department of Translational Medicine and Surgery, Section of General Pathology, Università Cattolica del Sacro Cuore, Rome, Italy

    Camilla Moliterni, Maria Tredicine, Renato Falcicchia & Francesco Ria

  2. Department of Biology and Biotechnology Charles Darwin, University of Rome Sapienza, Rome, Italy

    Camilla Moliterni

  3. Department of Medicine and Surgery, Section of Human Anatomy, University of Perugia, Perugia, Italy

    Alessandra Pistilli, Anna Maria Stabile, Mario Rende & Gabriele Di Sante

  4. Department of Pharmaceutical Sciences, University of Perugia, Perugia, Italy

    Desirée Bartolini

Authors
  1. Camilla Moliterni
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  2. Maria Tredicine
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  3. Alessandra Pistilli
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  4. Renato Falcicchia
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  5. Desirée Bartolini
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  6. Anna Maria Stabile
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  8. Francesco Ria
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  9. Gabriele Di Sante
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Corresponding author

Correspondence to Gabriele Di Sante .

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Editors and Affiliations

  1. Department of Medicine and Surgery, University of Perugia, Perugia, Italy

    Francesca Fallarino

  2. Department of Medicine and Surgery, University of Perugia, Perugia, Italy

    Marco Gargaro

  3. Department of Medicine and Surgery, University of Perugia, Perugia, Italy

    Giorgia Manni

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Moliterni, C. et al. (2023). In Vitro and Ex Vivo Methodologies for T-Cell Trafficking Through Blood–Brain Barrier After TLR Activation. In: Fallarino, F., Gargaro, M., Manni, G. (eds) Toll-Like Receptors. Methods in Molecular Biology, vol 2700. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3366-3_12

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  • DOI: https://doi.org/10.1007/978-1-0716-3366-3_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3365-6

  • Online ISBN: 978-1-0716-3366-3

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