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Gupta S, Maecker H. Intracellular Cytokine Staining (ICS) on Human Lymphocytes or Peripheral Blood Mononuclear Cells (PBMCs). Bio Protoc. 2015;5(5):e1442doi: 10.21769/BioProtoc.1442.
Gupta, S., & Maecker, H. (2015). Intracellular Cytokine Staining (ICS) on Human Lymphocytes or Peripheral Blood Mononuclear Cells (PBMCs). Bio-protocol, 5(5), e1442. https://doi.org/10.21769/BioProtoc.1442
Gupta, Sheena, and Maecker, Holden. "Intracellular Cytokine Staining (ICS) on Human Lymphocytes or Peripheral Blood Mononuclear Cells (PBMCs)." Bio-protocol vol. 5,5 (2015): e1442. doi: https://doi.org/10.21769/BioProtoc.1442
Gupta S, Maecker H. Intracellular Cytokine Staining (ICS) on Human Lymphocytes or Peripheral Blood Mononuclear Cells (PBMCs). Bio Protoc. 2015 Apr 05;5(5):e1442. doi: 10.21769/BioProtoc.1442. eCollection 2015 Apr 05. PMID: 34604458; PMCID: PMC8443452.
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Bio Protoc. 2015 Apr 05;5(5):e1442. doi: 10.21769/BioProtoc.1442. eCollection 2015 Apr 05.

Intracellular Cytokine Staining (ICS) on Human Lymphocytes or Peripheral Blood Mononuclear Cells (PBMCs).

Bio-protocol

Sheena Gupta, Holden Maecker

Affiliations

  1. Human Immune Monitoring Center, Stanford University, Stanford, USA; Institute for Immunity, Transplantation, and Infection, Stanford University, Stanford, USA.

PMID: 34604458 PMCID: PMC8443452 DOI: 10.21769/BioProtoc.1442

Abstract

Production of cytokines plays an important role in the immune response. Cytokines are involved in many different pathways including the induction of many anti-viral proteins by IFN gamma, the induction of T cell proliferation by IL-2 and the inhibition of viral gene expression and replication by TNF alpha. Cytokines are not preformed factors but are rapidly produced and secreted in response to cellular activation. Intracellular cytokine detection by flow cytometry has emerged as the premier technique for studying cytokine production at the single-cell level. It detects the production and accumulation of cytokines within the endoplasmic reticulum after cell stimulation, allowing direct TH1 versus TH2 determination. It can also be used in combination with other flow cytometry protocols for immunophenotyping using cell surface markers or with MHC multimers to detect an antigen specific response, making it an extremely flexible and versatile method. This capability, combined with the high throughput nature of the instrumentation, gives intracellular cytokine staining an enormous advantage over existing single-cell techniques such as ELISPOT, limiting dilution, and T cell cloning. The principle steps of intracellular cytokine staining is as follows: Cells are activated for a few hours using either a specific peptide or a non-specific activation cocktail; An inhibitor of protein transport (

Copyright © 2015 The Authors; exclusive licensee Bio-protocol LLC.

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