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In Vitro and In Vivo Assessment of T, B and Myeloid Cells Suppressive Activity and Humoral Responses from Transplant Recipients
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
In Vitro and In Vivo Assessment of T, B and Myeloid Cells Suppressive Activity and Humoral Responses from Transplant Recipients
DOI:

18:48 min

August 12, 2017

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Chapters

  • 00:00Title
  • 00:37LEW.1W to LEW.1A Allograft Procedure
  • 02:18Isolation of Allograft APCs (pDCs)
  • 06:12Isolation of Responder Effector T Cells and Regulatory T Cells (CD4+ and CD8+CD45RClow)
  • 10:30CFSE Labeling of Responder Cells to Measure their Proliferation
  • 12:09FACS Staining and Analysis After 6 Days of Coculture
  • 13:49In Vivo Assessment of the Cells Supressive Activity by Adoptive Cell Transfer in a Heart Grafted Recipient
  • 16:33Representative Results
  • 18:03Conclusion

Summary

Automatic Translation

Here, we present a protocol to induce tolerance in transplantation, and assess in vitro and in vivo the suppressive capacity of distinct cell subsets from the recipient and the immune status of the recipient toward donor or exogenous antigens.

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