Considerations for Monitoring Immune Cells by Flow Cytometry, Signal Transduction Pathways, Intracellular Proteins, and Tetramers

Date/Time: 08 November 2012, 1:00 PM
Speaker: Mirja Wine, Ph.D.
Speaker Affiliation: Beckman Coulter
Venue: BSB 320N

For more info: Alexandra Gurary (gurary@hawaii.edu)
Description: Monitoring immune cells by flow cytometry can take many forms. In
this presentation we will discuss 2 applications, methods for
monitoring cell signaling and intracellular proteins, and the use of
tetramers to evaluate antigen specific T cells.

Cell signaling cytometry is a powerful technique that can be used to
study normal signaling pathways, abnormalities in signaling such as
those involved in the progression of leukemia, as well as monitoring
effects of drugs on these pathways. The availability of antibodies
specifically reactive with phospho-epitopes and intracellular proteins
provides specific probes that can be used to monitor multiple signal
transduction pathways simultaneously. However, successful use of these
probes to intracellular targets has required the development of cell
fixation and permeabilization techniques and products that preserve
phospho-epitopes, cellular light scatter properties, as well as cell
surface CD molecules.

Tetramers have been successfully used to analyze antigen-specific T
cells for many years. Despite this, standardized procedures have yet
to be established. Beckman Coulter is in collaboration with The
Cancer Immunotherapy Consortium of the Cancer Research Institute
(CIC-CRI) HLA-peptide multimer (MULTIMER) proficiency panel study,
which consists of 27 laboratories from around the world participating
in a series of coordinated harmonization efforts intended to i)
identify critical performance variables, ii) optimize these variables,
iii) evaluate the impact of harmonizing their use and iv) to provide
good estimates of assay performance characteristics typically found in
representative labs. These efforts should decrease the variability
seen between labs and improve results.

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