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 Blood, 15 December 2008, Vol. 112, No. 13, pp. 5254-5258.
Prepublished online as a Blood First Edition Paper on October 6, 2008; DOI 10.1182/blood-2008-03-147322.

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Submitted March 25, 2008
Accepted August 25, 2008

STAT-3 and ERK 1/2 phosphorylation are critical for T cell alloactivation and graft-versus-host-disease

Sydney X Lu, Onder Alpdogan, Janine Lin, Robert Balderas, Roberto Campos-Gonzalez, Xiao Wang, Guo-Jian Gao, David Suh, Christopher King, Melanie Chow, Odette M. Smith, Vanessa M. Hubbard, Johanne L. Bautista, Javier Cabrera-Perez, Johannes L Zakrzewski, Adam A Kochman, Andrew Chow, Gregoire Altan-Bonnet, and Marcel R.M. van den Brink*

Department of Medicine and Immunology, Memorial Sloan-Kettering Cancer Center, New York, NY, United States
BD Biosciences, San Diego, CA, United States
Department of Pathology, Albert Einstein College of Medicine, New York, NY, United States
Department of Computational Biology, Memorial Sloan-Kettering Cancer Center, New York, NY, United States

* Corresponding author; email: vandenbm{at}mskcc.org.

Graft-versus-host-disease (GVHD) is a serious complication of allogeneic bone marrow transplantation, and donor T cells are indispensable for GVHD. Current therapies have limited efficacy, selectivity, and high toxicities. We used a novel flow cytometric technique for the analysis of intracellular phosphorylation events in single cells in murine BMT models to identify and validate novel GVHD drug targets1-7. This method circumvents the requirement for large numbers of purified cells, unlike western blots. We defined a signaling profile for alloactivated T cells in vivo and identified the phosphorylation of ERK1/2 and STAT-3 as important events during T cell (allo)activation in GVHD. We establish that interference with STAT-3 phosphorylation can inhibit T cell activation and proliferation in vitro and GVHD in vivo. This suggests that phospho-specific flow cytometry is useful for the identification of promising drug targets, and ERK1/2 and STAT-3 phosphorylation in alloactivated T cells may be important for GVHD.


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