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Methods of Purification of CTL-Derived Exosomes

  • Angela MontecalvoEmail author
  • Adriana T. Larregina
  • Adrian E. Morelli
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

Exosomes T cells 

Notes

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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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Angela Montecalvo
    • 1
    Email author
  • Adriana T. Larregina
    • 1
    • 2
  • Adrian E. Morelli
    • 1
    • 3
    • 4
  1. 1.Department of Immunology, University of Pittsburgh Medical Center, University of Pittsburgh School of MedicineUniversity of PittsburghPittsburghUSA
  2. 2.Department of Dermatology, University of Pittsburgh Medical Center, University of Pittsburgh School of MedicineUniversity of PittsburghPittsburghUSA
  3. 3.Department of Surgery, University of Pittsburgh Medical Center, University of Pittsburgh School of MedicineUniversity of PittsburghPittsburghUSA
  4. 4.Thomas E. Starzl Transplantation Institute, University of Pittsburgh Medical CenterUniversity of PittsburghPittsburghUSA

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