Direct allorecognition

U0126 clinical trial is a vigorous reaction due to the high precursor frequency of alloreactive T cells; in this regard it is generally accepted that deletion of a substantial proportion of direct pathway alloreactive T cells will be required to ‘tip the balance’ from reactivity to regulation [12, 13]. In addition, in order to suppress the surviving alloreactive T cells by regulation one would need sufficient numbers of Tregs in the right place, at the right time, in an environment that favours regulation. Therefore, the specificity of the Tregs chosen for cellular therapy may play an important role (discussed in later sections). The main focus of this review is the clinical

application of Tregs in the setting of transplantation and the journey from bench to bedside. We will discuss the challenges that we still face in the laboratory from the isolation to the ex-vivo expansion of these cells for immunotherapy and outline the questions that still remain with regard to the clinical protocols. Moreover, human Tregs are currently less well-characterized AZD2014 solubility dmso and understood compared to mouse Tregs; we will, therefore, review briefly their biology before discussion of their clinical application. Aside from the expression of CD25 [14] and FoxP3 (outlined above), human Tregs also express Leukocyte receptor tyrosine kinase CD27 [15], CD45RA [16], CD39 [17], CD122, cytotoxic T lymphocyte antigen-4 (CTLA-4 or CD152) and the glucocorticoid-induced tumour necrosis factor receptor (GITR) family-related gene [18, 19]. However, most of these cell surface markers are not exclusive to Tregs, with some of these markers also expressed by non-regulatory CD4+ T cells, posing a challenge during the isolation process. As an example, data support the key role of FoxP3 in the development, maintenance and function of Tregs with supporting evidence that point mutations in the FoxP3 gene leads to a functional Treg deficit that is evident in patients with IPEX (immune dysregulation,

polyendocrinopathy, enteropathy, X-linked syndrome) [20]. Despite this, FoxP3 is not a sufficient marker for the isolation of Tregs, as many activated effector T cells also express FoxP3 without having a regulatory phenotype [21]. Moreover, being an intracellular protein, this marker cannot be used to isolate Tregs. What complicates the story even further is that human Tregs are heterogeneous. In contrast with mice, the combination of the marker CD45RA and the level of expression of FoxP3 delineates the human Treg compartment into naive or resting Tregs (CD45RA+FoxP3low), effector Tregs (CD45RA–FoxP3high), both of which are suppressive in vitro, and the non-suppressive, cytokine secreting non-Tregs (CD45RA–FoxP3low) [22, 23].