Multiplexed immunophenotyping of human antigen-presenting cells in whole blood by polychromatic flow cytometry View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2010-02

AUTHORS

Erik Fung, Laura Esposito, John A Todd, Linda S Wicker

ABSTRACT

We describe two modular protocols for immunostaining and multiparameter flow cytometric analysis of major human antigen-presenting cells (APCs; e.g., dendritic cells, monocytes and B lymphocytes) in minimally manipulated whole blood samples. Simultaneous detection of up to eight colors is enabled by careful selection and testing of cell-subset-defining monoclonal antibodies (anchor markers) in the appropriate fluorochrome combinations, in order to show the quantification of surface expression levels of molecules involved in chemotaxis (e.g., CX(3)CR1 and CCR2), adhesion (e.g., CD11b and CD62L), antigen presentation (e.g., CD83, CD86 and CD209) and immune regulation (e.g., CD101) on circulating APCs. Each immunostaining reaction requires as little as 50-100 microl of peripheral whole blood and no density-gradient separation, and the entire procedure from preparation of reagents to flow cytometry can be completed in <5 h. More... »

PAGES

357

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nprot.2009.246

DOI

http://dx.doi.org/10.1038/nprot.2009.246

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1007656352

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/20134434


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