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Methods of Purification of CTL-Derived Exosomes

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Cytotoxic T-Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1186))

Abstract

Exosomes are membrane nanovesicles (approximately <120 nm in size) released by most, if not all, living cells and in particular by leukocytes. They originate within the endocytic compartment by invagination of the endosome membrane. Therefore, they have a different biogenesis and molecular composition than microvesicles (>0.2 μm) shed from the plasma membrane. Although the functions of exosomes in vivo are beginning to be elucidated, increasing evidence suggests that exosomes constitute a mechanism of cell-to-cell communication, transferring antigens, proteins, mRNAs, and noncoding RNAs among cells. Interestingly, effector T cells including cytotoxic T lymphocytes (CTLs) release death-inducing molecules of the TNF superfamily through exosomes contained in their cytotoxic granules. The present chapter provides basic protocols for purification of exosomes secreted by CTLs.

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Acknowledgements

We thank the Research Specialists Olga A. Tkacheva and William J. Shufesky (University of Pittsburgh, Pittsburgh, PA, USA) for their comments. We also thank Dr. Lawrence P. Kane for his revisions. This work was supported by the National Institutes of Health grant AI077511 (to A.T.L.), and funds from the T.E. Starzl Transplantation Institute (to A.E.M.).

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Authors and Affiliations

  1. Department of Immunology, University of Pittsburgh Medical Center, University of Pittsburgh School of Medicine, University of Pittsburgh, E1017 Biomedical Sciences Tower, 200 Lothrop Street, Pittsburgh, PA, 15213, USA

    Angela Montecalvo Ph.D., Adriana T. Larregina M.D., Ph.D. & Adrian E. Morelli M.D., Ph.D.

  2. Department of Dermatology, University of Pittsburgh Medical Center, University of Pittsburgh School of Medicine, University of Pittsburgh, Pittsburgh, PA, 15213, USA

    Adriana T. Larregina M.D., Ph.D.

  3. Department of Surgery, University of Pittsburgh Medical Center, University of Pittsburgh School of Medicine, University of Pittsburgh, Pittsburgh, PA, 15213, USA

    Adrian E. Morelli M.D., Ph.D.

  4. Thomas E. Starzl Transplantation Institute, University of Pittsburgh Medical Center, University of Pittsburgh, Pittsburgh, PA, 15213, USA

    Adrian E. Morelli M.D., Ph.D.

Authors
  1. Angela Montecalvo Ph.D.
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  2. Adriana T. Larregina M.D., Ph.D.
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  3. Adrian E. Morelli M.D., Ph.D.
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Corresponding author

Correspondence to Angela Montecalvo Ph.D. .

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Editors and Affiliations

  1. Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy

    Elena Ranieri

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Montecalvo, A., Larregina, A.T., Morelli, A.E. (2014). Methods of Purification of CTL-Derived Exosomes. In: Ranieri, E. (eds) Cytotoxic T-Cells. Methods in Molecular Biology, vol 1186. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1158-5_7

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  • DOI: https://doi.org/10.1007/978-1-4939-1158-5_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1157-8

  • Online ISBN: 978-1-4939-1158-5

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