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Examination of Thymic Positive and Negative Selection by Flow Cytometry
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CD6 modulates thymocyte selection and peripheral T cell homeostasis.

Marc Orta-Mascaró1, Marta Consuegra-Fernández1, Esther Carreras1

  • 1Institut d'Investigacions Biomèdiques August Pi i Sunyer, 08036 Barcelona, Spain.

The Journal of Experimental Medicine
|July 6, 2016
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Summary

CD6 glycoprotein deficiency impairs T cell development and alters peripheral T cell populations. Absence of CD6 impacts thymocyte selection and regulatory T cell function, leading to exacerbated autoimmune responses.

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Area of Science:

  • Immunology
  • Cell Biology
  • Molecular Biology

Background:

  • CD6 glycoprotein is a lymphocyte surface receptor involved in T cell development and activation.
  • CD6 mediates T cell adhesion via CD166/ALCAM interaction and associates with the T cell receptor (TCR).
  • The precise role of CD6 in thymocyte development and peripheral T cell responses is not fully understood.

Purpose of the Study:

  • To investigate the in vivo consequences of CD6 deficiency on T cell development and immune responses.
  • To define the role of CD6 in thymocyte selection and peripheral T cell subset generation and function.

Main Methods:

  • Analysis of CD6-deficient (CD6(-/-)) mice.
  • Flow cytometry to assess thymocyte subsets (CD4+, CD8+).
  • In vitro calcium mobilization assays upon TCR cross-linking.
  • Bone marrow chimera experiments.
  • TCR-transgenic mouse models (OT-I and Marilyn).
  • Analysis of peripheral T cell populations (effector/memory, regulatory T cells).

Main Results:

  • CD6(-/-) thymi showed reduced CD4+ and CD8+ single-positive subsets.
  • CD6(-/-) thymocytes exhibited increased Ca(2+) mobilization to TCR cross-linking.
  • CD6(-/-) T cells displayed a selective disadvantage during development in bone marrow chimeras.
  • Abnormal T cell selection events were observed in CD6(-/-) TCR-transgenic mice.
  • Increased frequencies of antigen-experienced peripheral T cells (effector/memory and regulatory T cells) were found in CD6(-/-) mice.
  • CD6(-/-) regulatory T cells showed diminished suppressive activity.
  • CD6(-/-) mice exhibited exacerbated autoimmune responses to collagen.

Conclusions:

  • CD6 plays a critical role in modulating the threshold for thymocyte selection.
  • CD6 influences the generation and/or function of peripheral T cell subpopulations, including regulatory T cells.
  • CD6 deficiency leads to altered T cell homeostasis and increased susceptibility to autoimmune diseases.