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In Vitro Assay of Plasmodium-Infected Red Blood Cell Killing by Cytotoxic Lymphocytes
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Imunologia e Infecção
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JoVE Journal Imunologia e Infecção
In Vitro Assay of Plasmodium-Infected Red Blood Cell Killing by Cytotoxic Lymphocytes

In Vitro Assay of Plasmodium-Infected Red Blood Cell Killing by Cytotoxic Lymphocytes

DOI:
10.3791/63987-v

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08:20 min

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August 17, 2022

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, , ,
1Instituto René Rachou,Fundação Oswaldo Cruz, 2Departamento de Bioquímica e Imunologia,Universidade Federal de Minas Gerais

Capítulos

  • 00:04Introduction
  • 00:58Experimental Malaria Infection in Mouse
  • 01:49Obtaining Fresh Whole Mouse Splenocytes
  • 03:30Cytotoxic Effector Cell Purification: CD8a+ T Cell Negative Selection
  • 04:22P. yoelii Infected RBC Isolation
  • 05:21CFSE Labeling of RBCs, Preparation for Flow Cytometry and Cytotoxic Lymphocyte Coculture
  • 06:22Results: P. yoelii – iRBC Enrichment and Assessment of cell lysis Percentage of P. yoelii – iRBCs by CD8+ lymphocytes
  • 07:39Conclusion

Summary

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Here we describe a new method to help elucidate the mechanisms of cellular immunity to Plasmodium during the blood stage of infection. This is an in vitro assay that measures infected red blood cell killing by cytotoxic lymphocytes. 

Tags

Keywords: In Vitro AssayPlasmodium-infected Red Blood CellsCytotoxic LymphocytesCell-mediated ImmunityMalaria PathogenesisProtective Malaria ImmunitySpleen IsolationRBC LysisMACS Column Purification

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