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Generation of T. cruzi-Specific Primary CD4+ T Cell Lines from Peripheral Blood Mononuclear Cells Isolated from Chagas Disease Patients

  • Gonzalo R. AcevedoEmail author
  • Paula B. Alcaráz
  • Clemencia Pinilla
  • Karina A. Gómez
Protocol
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Part of the Methods in Molecular Biology book series (MIMB, volume 1955)

Abstract

Human CD8+ and CD4+ T cell lines and clones are valuable tools to explore the role of these cells in the context of diseases, especially in cases in which the main underlying actor is the immune response, like Chagas disease. These cell lines and clones provide a good experimental system to address the phenotypic and functional features of specific T cell subpopulations and furthermore settle the framework necessary for analyzing their antigen/peptide specificity.

This chapter details a culture method for the establishment of T. cruzi-specific memory T cell lines from mononuclear cells isolated from Chagas disease patients’ peripheral blood. The presented protocol comprises (1) enrichment of memory CD4+ T cells, (2) stimulation with parasite lysate, (3) evaluation of specificity, and (4) expansion and maintenance of specific T cell lines.

Key words

Antigen-specific T cells T cell line Memory C4 T cells Trypanosoma cruzi Chagas disease 

Notes

Acknowledgments

The funds that permitted the development of the method hereby described were granted by the Consejo Nacional de Investigaciones Científicas y Técnicas (PIP 112-200801-02915), the Agencia Nacional de Promoción Científica y Tecnológica (PICT2014-1026), the AJ Roemmers Foundation (Clinical Research Grant for Young Investigators 2016–2018), and the National Institute of Allergy and Infectious Diseases (5R21AI109439-02).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Gonzalo R. Acevedo
    • 1
    Email author
  • Paula B. Alcaráz
    • 1
  • Clemencia Pinilla
    • 2
  • Karina A. Gómez
    • 1
  1. 1.Laboratorio de Inmunología de las Infecciones por TripanosomátidosInstituto de Investigaciones en Ingeniería Genética y Biología Molecular (INGEBI), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)Buenos AiresArgentina
  2. 2.Torrey Pines Institute for Molecular Studies (TPIMS)Port St. LucieUSA

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