In addition, whether polyclonal Tregs or antigen-specific Tregs are used will influence the dose. Of note, studies using antigen-specific Tregs showed that lower numbers were able to achieve the LEE011 same functional efficacy as larger numbers of polyclonal Tregs [86, 87]. Finally, whether a single injection or multiple injections are required

is a matter of debate and may be determined in a Phase II efficacy study, where patient outcomes should also be measured and an in-depth patient monitoring planned. The use of molecular diagnostic tools can help to assess the increased expression of biomarkers of operational tolerance in patients receiving cellular therapy and whether these can be used as surrogate end-points of efficacy [101-103]. The same approach can be used selleck to define whether or not the patients have decreased expression of biomarkers of acute rejection [104, 105].

Furthermore, phenotypic analysis of patient PBMCs, using flow cytometric analysis, can determine whether or not the number of Tregs has increased or the composition of the T cell compartment has changed as a result of the intervention [106]. Using the same analysis, the cytokine profile of the cells that have been phenotyped can be analysed to establish their plasticity. Finally, lymphocyte compartments can be monitored after specific interventions, as has been shown useful when associating expansion of lymphocyte

subsets, in this case naive B cells, in peripheral blood of patients in whom better outcomes were noted [107]. In spite of the potential concerns and controversies outlined with regard to Treg isolation and expansion protocols and the optimal clinical protocol, clinical Oxymatrine trials are under way to test the therapeutic potential of Tregs. Beneficial effects of Treg infusions on allograft survival were first reported in bone marrow transplantation models in which donor Tregs reduced the incidence of GVHD. The first human trial using Treg cell therapy by Trzonkowski et al. [108] involved two patients. The first patient had chronic GVHD 2 years post-bone marrow transplantation. After receiving 0·1 × 106/kg FACS purified ex-vivo-expanded Tregs from the donor, the symptoms subsided and the patient was withdrawn successfully from immunosuppression without evidence of recurrence. The second patient had acute GVHD at 1 month post-transplantation, which was treated with several infusions of expanded donor Tregs. Despite initial and transitory improvement, the disease progressed and resulted ultimately in the patient’s death. This was the first report to show that adoptive transfer of Tregs is well tolerated and thus was a major breakthrough.