HIV Protocols pp 375-391

Part of the Methods In Molecular Biology™ book series (MIMB, volume 485)

Multiparameter Flow Cytometry Monitoring of T Cell Responses

  • Holden T. Maecker

Abstract

HIV vaccine research increasingly uses polychromatic flow cytometry as a tool to monitor T cell responses. The use of this technology allows for the analysis of highly defined subsets of cells with unique phenotypes and functions. Ultimately, such studies may identify surrogate markers of protection from disease progression. However, this powerful technology comes with a number of technical hurdles, and there is a need to standardize the assays and protocols used in clinical trial monitoring. Here an optimized protocol, with variations for specific circumstances, is presented. This protocol covers the analysis of multiple cytokines, cell surface markers, and other functional markers such as perforin, CD107, and CD154. While the protocol can be adapted to various numbers of fluorescence parameters, optimized panels of 8–10 colors are presented.

Key words

Antigen-specific intracellular staining multicolor polychromatic fixation permeabilization AIDS vaccine research 

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Holden T. Maecker
    • 1
  1. 1.Palo AltoUSA

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