Acute graft-versus-host disease (GVHD) remains a leading cause of morbidity and non-relapse mortality following allogeneic hematopoietic cell transplantation. The clinical staging of GVHD varies greatly between transplant centers and is frequently not agreed upon by independent reviewers. The lack of standardized approaches to handle common sources of discrepancy in GVHD grading likely contributes to why promising GVHD treatments reported from single centers have failed to show benefit in randomized multi-center clinical trials. We developed guidelines through international expert consensus opinion to standardize the diagnosis and clinical staging of GVHD for use in a large international GVHD research consortium. During the first year of use, the guidance was following discussion of complex clinical phenotypes by experienced transplant physicians and data managers. These guidelines increase the uniformity of GVHD symptom capture which may improve the reproducibility of GVHD clinical trials after further prospective validation.

CD137 is a member of the TNFR-family

IntroductionThe application of molecular technologies to identify proteins differentially expressed by transformed cells is providing large numbers of candidate antigens that can be potentially targeted to selectively eliminate tumor cells by cancer immunotherapy. 1,2 Efforts to vaccinate patients to such antigens have yielded some provocative results, but only a small subset of patients have demonstrated therapeutic responses, likely reflecting the many in vivo obstacles to generating potent responses to these proteins, particularly in patients with an established malignancy. 3 An alternative approach of isolating and expanding reactive T cells ex vivo followed by adoptive transfer into the patient circumvents many of these in vivo obstacles. Although this adoptive therapy approach has demonstrated significant clinical promise, 4 generating the large numbers of T cells required for adoptive therapy of cancer patients, particularly within the time constraints posed by progressive tumors, is often not feasible. Molecular technologies have now provided a means to more broadly capture the therapeutic potential of this treatment strategy. Genes encoding the ␣ and ␤ chains of a T-cell receptor (TCR) can be isolated from a T cell reactive with the antigen of interest and restricted to a defined HLA allele, inserted into a shuttle expression vector, and then introduced into large numbers of T cells of individual patients sharing the restricting allele and the targeted protein. 5 This approach is already being pursued clinically, 6 and the goal is to establish a library of such defined TCR genes that could provide reagents for treating a diverse set of patients and diseases. Multiple virus-and tumor-reactive TCR genes have already been successfully isolated and re-expressed in T cells, including TCR genes with specificity for HLA*0201 (HLA-A2)-restricted epitopes from melanoma antigens 7-9 and HLA-A2-and HLA*2402-restricted WT1-derived epitopes. 10,11 The avidity of a T cell for its target reflects many factors, including the affinity of the TCR for its cognate antigen 12 and the level of TCR expression. [13][14][15] One difficulty with the TCR-transfer approach is that the TCR-transduced T cells are often of lower avidity than the parental T cell from which the TCR was derived due to failure to achieve wild-type levels of TCR expression, which likely contributed to the limited efficacy observed in the recently reported clinical trial pursuing this strategy. 6 Thus, the TCR chains introduced into T cells need to be initially selected for appropriate affinity 10,16 and inserted into vectors that can achieve and maintain high-level expression. 17 However, even if these criteria are met, the introduced exogenous ␣ and ␤ chains can potentially assemble as pairs not only with each other but also with the endogenous TCR ␣ and ␤ chains, thereby reducing the number of appropriately matched exogenous ␣␤TCR pairs at the cell surface and decreasing the achievable T-cell avidity. Such mismatched pairing poses a second substantive p...

Relapse remains a leading cause of death after allogeneic hematopoietic cell transplantation (HCT) for patients with high-risk leukemias. The potentially beneficial donor T-cell-mediated graft-versus-leukemia (GVL) effect is often mitigated by concurrent graft versus host disease (GVHD). Providing T-cells that can selectively target Wilms’ Tumor Antigen 1 (WT1), a transcription factor over-expressed in leukemias that contributes to the malignant phenotype, represents a potential opportunity to promote anti-leukemic activity without inducing GVHD. HLA A*0201-restricted WT1-specific donor-derived CD8+ cytotoxic T-cell (CTL) clones were administered post-HCT to 11 relapsed or high-risk leukemia patients without any evidence of on-target toxicity. The last four treated patients received CTL clones generated with exposure to IL-21 as a means to prolong in vivo CTL survival, as IL-21 can limit terminal differentiation of antigen-specific T-cells generated in vitro. Transferred cells exhibited direct evidence of anti-leukemic activity in 2 patients: a transient response in one patient with advanced progressive disease and the induction of a prolonged remission in a patient with minimal residual disease (MRD). Additionally, three treated patients at high risk for relapse post-HCT survive without leukemia relapse, GVHD or additional anti-leukemic treatment. CTL generated in the presence of IL-21, which were transferred in these latter three patients and the patient with MRD, all remained detectable long-term and maintained/acquired in vivo phenotypic and functional characteristics associated with long-lived memory CD8+ T-cells. This study supports expanding efforts to immunologically target WT1, and provides insights into the requirements necessary to establish potent persistent T-cell responses in patients.

No abstract

In the statistics section, the equation to generate a final prediction model from the training set was incorrect. The correct sentence is below. We then created a training set at random and repeated the entire process to generate a final model: log[-log(1-p̂)] =-11.263 + 1.844(log10ST2) + 0.577(log10REG3α), where p̂ = predicted probability of 6-month NRM.

Acute graft-versus-host disease (GVHD) is treated with systemic corticosteroid immunosuppression. Clinical response after 1 week of therapy often guides further treatment decisions, but long-term outcomes vary widely among centers, and more accurate predictive tests are urgently needed. We analyzed clinical data and blood samples taken 1 week after systemic treatment of GVHD from 507 patients from 17 centers of the Mount Sinai Acute GVHD International Consortium (MAGIC), dividing them into a test cohort (n = 236) and 2 validation cohorts separated in time (n = 142 and n = 129). Initial response to systemic steroids correlated with response at 4 weeks, 1-year nonrelapse mortality (NRM), and overall survival (OS). A previously validated algorithm of 2 MAGIC biomarkers (ST2 and REG3α) consistently separated steroid-resistant patients into 2 groups with dramatically different NRM and OS ( < .001 for all 3 cohorts). High biomarker probability, resistance to steroids, and GVHD severity (Minnesota risk) were all significant predictors of NRM in multivariate analysis. A direct comparison of receiver operating characteristic curves showed that the area under the curve for biomarker probability (0.82) was significantly greater than that for steroid response (0.68, = .004) and for Minnesota risk (0.72, = .005). In conclusion, MAGIC biomarker probabilities generated after 1 week of systemic treatment of GVHD predict long-term outcomes in steroid-resistant GVHD better than clinical criteria and should prove useful in developing better treatment strategies.

Antigen-specific priming of human, naïve T-cells has been difficult to assess. Due to the low initial frequency in the naïve cell pool of specific T-cell precursors, such an analysis has been obscured by the requirements for repeated stimulations and prolonged culture time. In this protocol, we describe how to rapidly evaluate antigen-specific priming of CD8+ -cells following a single stimulation. The assay provides reference conditions, which result in the expansion of a significant population of antigen-specific T-cells from the naïve repertoire. Various conditions and modifications during the priming process (e.g. testing new cytokines, costimulators, etc.) can now be directly compared to the reference conditions. Factors relevant to achieving effective priming include the dendritic cell preparation, the T-cell preparation, the cell ratio at the time of priming, the serum source used for the experiment, and the timing of addition and concentration of the cytokines used for expansion. This protocol is relevant for human immunology, vaccine biology and drug development.