Validated ITN FoxP3 Flow Cytometry Methodology Published, Graces Cover of Cytometry B

March 5, 2009

The ITN protocol for the measurement of Fox-P3-expressing T regulatory (Treg) cells has been published in the March 2009 issue of the peer-reviewed journal Cytometry B: Clinical Cytometry.

The manuscript, by lead authors Jean Grant and Katarzyna Bourcier, describes a standardized protocol for FoxP3 antibody staining for use in clinical trials. FoxP3, also known as forkhead box protein P3, appears to function as a master regulator in the development and function of Treg cells. Tregs are a specialized subpopulation of T cells that can suppress T cell activation and appear to be key controls in maintaining immunologic homeostasis and immune tolerance. Alterations in the Treg numbers and/or function have been associated with the development of autoimmune diseases and they have emerged as promising cellular therapeutics in a number of indications.

Cytometry B cover image

The significance of the work is highlighted by an editorial contained in the same issue by Associate Editor Francesco Lanza, entitled "Toward standardization of Foxp3+ regulatory T-cell measurement in clinical settings." Figures from the manuscript are also showcased on the cover of the March issue.

In developing the protocol, the team, led by senior author Paul K. Wallace, PhD, of Roswell Park Cancer Institute in Buffalo, NY, examined numerous variables including antibodies from several vendors, cell preparation methods and cell isolation procedures.

The resulting validated protocol was applied to clinical specimens from healthy control patients as well as those with type 1 diabetes or multiple sclerosis (MS). The authors found a significant increase in FoxP3 in cells from type 1 diabetes subjects compared to healthy controls, while those from MS subjects showed no such increase.

Details of the protocol are available in the manuscript, which may be viewed on the Cytometry B website, or may be downloaded from the ITN's online library of standardized laboratory protocols .

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