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In vitro assay at evaluere virkningen af immunoregulerende Pathways på HIV-specifikke CD4 T-celle effektorfunktion
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Immunology and Infection
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JoVE Journal Immunology and Infection
In Vitro Assay to Evaluate the Impact of Immunoregulatory Pathways on HIV-specific CD4 T Cell Effector Function

In vitro assay at evaluere virkningen af immunoregulerende Pathways på HIV-specifikke CD4 T-celle effektorfunktion

DOI:
10.3791/50821-v

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09:26 min

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October 15, 2013

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, , , ,
1The Ragon Institute of MGH, MIT and Harvard, 2Centre de Recherche du Centre Hospitalier de l’Université de Montréal (CRCHUM)

Chapters

  • 00:05Title
  • 01:27Depletion and Isolation of PBMCs via Ficoll Separation
  • 02:08Antibody Blockade and Antigen-Specific Stimulation of Cells
  • 02:46Collection and Analysis of Supernatant
  • 03:15Collection and Phenotypic Analysis of Cells via Flow Cytometry
  • 04:09Analysis of Cells via qRT-PCR
  • 05:15Adaptation for Intracellular Cytokine Staining at 48 Hours
  • 05:48Adaptation for Sorting Cells
  • 06:55Results: HIV-specific CD4 T Cell Effector Function
  • 09:03Conclusion

Summary

Automatic Translation

Automatic Translation

Vi udviklede en in vitro assay til at undersøge den rolle immunoregulatoriske veje i reguleringen af cytokinudskillelse af HIV-specifikke CD4 T-celler.

Tags

In Vitro AssayHIVCD4 T CellsEffector FunctionImmunoregulatory PathwaysPD-1IL-10T Cell ExhaustionCytokine ProductionPBMCFlow CytometryQRT-PCRGag Peptide Pool

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