Establishing long-term allograft acceptance without the requirement for continuous immunosuppression, a condition known as allograft tolerance, is a highly desirable therapeutic goal in solid organ transplantation. Determining which recipients would benefit from withdrawal or minimization of immunosuppression would be greatly facilitated by biomarkers predictive of tolerance. In this study, we identified the largest reported cohort to our knowledge of tolerant renal transplant recipients, as defined by stable graft function and receiving no immunosuppression for more than 1 year, and compared their gene expression profiles and peripheral blood lymphocyte subsets with those of subjects with stable graft function who are receiving immunosuppressive drugs as well as healthy controls. In addition to being associated with clinical and phenotypic parameters, renal allograft tolerance was strongly associated with a B cell signature using several assays. Tolerant subjects showed increased expression of multiple B cell differentiation genes, and a set of just 3 of these genes distinguished tolerant from nontolerant recipients in a unique test set of samples. This B cell signature was associated with upregulation of CD20 mRNA in urine sediment cells and elevated numbers of peripheral blood naive and transitional B cells in tolerant participants compared with those receiving immunosuppression. These results point to a critical role for B cells in regulating alloimmunity and provide a candidate set of genes for wider-scale screening of renal transplant recipients.

Type 1 diabetes is a chronic autoimmune disease mediated by autoreactive T-cells. Several experimental therapies targeting T-cells are in clinical trials. To understand how these therapies affect T-cell responses in vivo, assays that directly measure human T-cell function are needed. In a blinded, multicenter, case-controlled study conducted by the Immune Tolerance Network, we tested responses in an immunoblot and T-cell proliferative assay to distinguish type 1 diabetic patients from healthy control subjects. Peripheral blood cells from 39 healthy control subjects selected for DR4 and 23 subjects with recently diagnosed type 1 diabetes were studied. Autoantibody responses were measured in serum samples. Positive responses in both assays were more common in peripheral blood mononuclear cells from new-onset type 1 diabetic patients compared with control subjects. The proliferative, immunoblot, and autoantibody assays had sensitivities of 58, 91, and 78% with specificities of 94, 83, and 85%, respectively. When cellular assays were combined with autoantibody measurements, the sensitivity of the measurements was 75% with 100% specificity. We conclude that cellular assays performed on peripheral blood have a high degree of accuracy in discriminating responses in subjects with type 1 diabetes from healthy control subjects. They may be useful for assessment of cellular autoimmune responses involved in type 1 diabetes.

3MONDAY during liver injury. Addition of activated (a) HpSC, but not quiescent HpSC, signifi cantly inhibited allo-DC induced-T cell proliferative responses (MLR) in a dose dependent manner, which was associated with enhanced T cell apoptosis (TUNEL). Neutralization of B7-H1 by anti-B7-H1 mAb signifi cantly reduced the HpSC-induced T cell apoptosis and reversed the inhibition of T cell proliferation, suggesting a key role of B7-H1. To evaluate this in vivo, BALB/c islets (300) were co-transplanted with 3 x 105 activated HpSC (B6) into STZ-induced diabetic B6 recipients. Co-transplant with HpSC effectively protects islet allografts from rejection. This was associated with reduction of graft infi ltrating T cells and enhancement of apoptotic activity. Co-transplant with HpSC from B7-H1-/-livers markedly lost their islet graft protective capacity, associated with less apoptosis of infi ltrating cells. To determine the subsets of apoptotic T cells, T cells were isolated from spleen, draining lymph nodes, and grafts for phenotype and function analyses. On POD 8, a marked reduction of graft infi ltrating CD4+ (30.7%) and CD8+ T cells (31.3%) was seen in HpSC co-transplant group, as compared to islets alone, which was further progressed thereafter. CD8 T cells dropped ~7 folds on POD 140. CD4+/CD8+ ratio was increased from 0.5 at the early POD to 2.0 in the long-term survival grafts. Adoptive transfer of CFSE-labeled DES transgenic T cells was used to track the response and fate of antigen-specifi c CD8+ T cells. The results showed active division of DES+ cells within the allograft in both the islet only and HpSC cotransplantation groups. However, accumulation of these DES+ cells was signifi cantly lesser in the HpSC co-transplantation group as compared to that in the islet only group (1.9¡Á103 vs. 10¡Á103 cells per graft). These fi ndings suggest that HpSC may not inhibit activation of antigen-specifi c CD8+ T cells, but induce their robust apoptotic death, which is partially mediated by inducibly expressed B7-H1 on HpSC. This could be one of the important mechanisms by which tolerance in the peripheral organs is maintained.

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Background Current clinical trial assessments of cognitive function and activities of daily living are based on periodic assessments conducted under controlled clinical conditions using standardized measures. This approach is highly correlated with diagnosis of Alzheimer’s disease (AD) but has varied relevance to the real‐life challenges of those living with the disease and their caregivers. The use of digital technologies in clinical trials is actively encouraged. The opportunity to embed these tools in clinical trials in parallel with traditional measures is needed to move the field forward. This pilot study provides a framework to build on this approach and expand its use in research and patient care management. PFA‐AD will enable: 1) identification of key behavioral challenges that are relevant to the person with AD and their caregivers; 2) assessment of therapeutic effect in real time across multiple domains of behavioral function; and 3) innovate clinical trial measures of daily activities that are personalized and relevant to the patient and caregiver. Method PFA‐AD proof‐of‐concept development was embedded as part of a clinical trial (Clinicaltrials.gov: NCT03748303) to further develop allopregnanolone, a first‐in‐class regenerative therapeutic for AD. This study is enrolling 12 persons with AD to assess the intramuscular formulation of allopregnanolone. Participants are being asked to download the “Allo IM app” and interact with it daily during 12‐weeks. The platform/app created by MyOwnMed,Inc supports simple digital measures of lifestyle and daily function defined by the team interacting with patients. Content is personalized for each patient/caregiver based on their input. Wearable technologies have been integrated to capture physical activity and sleep, using available interfaces that will be seamlessly integrate into the app. Result Analysis of PFA‐AD data will include: 1) App usability; 2) Fidelity of personalized measures; 3) Correlation of personalized measures with standardized assessments; 4) Feasibility for inclusion in clinical trials. Conclusion AD is personal and assessment of functional capacity can be too. The PFA‐AD application brings the power of precision analytics to those living with AD and their caregivers. Outcomes of this research will advance the development of a technology‐based tool to enable assessments of real‐life function that really matters, in real‐time.