Post-infusion PD-1+CD8+CAR-T cells identify patients responsive to CD19-CAR-T therapy in non-Hodgkin's lymphoma.

Chimeric antigen receptor T cell therapy (CAR-T) has revolutionized treatment for relapsed/refractory (r/r) B-cell non-Hodgkin's lymphoma (NHL). Robust biomarkers and a complete understanding of CAR-T cell function in the post-infusion phase remain limited. Here we used a 37-color spectral flow cytometry panel to perform high dimensional single cell analysis of post-infusion samples in 26 patients treated with CD28 co-stimulatory domain containing commercial CAR-T (CD28-CAR-T) for NHL and focused on computationally gated CD8+ CAR-T cells. We found that the presence of post-infusion PD-1+ CD8+ CAR-T cells at the Day 14 timepoint highly correlated with the ability to achieve complete response (CR) by 6 months. Further analysis identified multiple subtypes of CD8+ PD-1+ CAR-T cells including PD-1+ TCF1+ stem-like CAR-T cells and PD-1+ TIM3+ effector-like CAR-T cells that correlated with improved clinical outcomes such as response and progression free survival. Additionally, we identified a subset of PD-1+ CD8+ CAR+ T cells with effector-like function that was increased in patients who achieved a CR and was associated with Grade 3 or higher immune effector cell-associated neurotoxicity syndrome. Here we identified robust biomarkers of response to CD28-CAR-T and highlight the importance of PD-1 positivity in CD8+ CAR-T cells post-infusion in achieving CR.

Development and consensus of entrustable professional activities for final-year medical students in anaesthesiology.

BACKGROUND: The transfer of classic concepts of competency-based medical education into clinical practice has been proven to be difficult in the past, being described as partially fragmented, misleading and inadequate. At the beginning of training, novice doctors commonly feel overwhelmed, overloaded and exposed to extreme time pressure. The discrepancy between expected and actual clinical competence of doctors at the start of their speciality training jeopardizes patient safety. The framework of Entrustable Professional Activities (EPAs) is a promising instrument to effectively integrate competency-based training into clinical practice and may help to close this gap and consequently to improve patient safety. METHODS: For anaesthesiology, we developed 5 EPAs for final-year medical students. The EPAs comprised the following seven categories: 1. Title, 2. Specifications, 3. Limitations, 4. Competency domains, 5. Knowledge, abilities and skills, professional attitudes, 6. Assessment and 7. Entrustment. Based on a modified, online-based Delphi study, we further developed and refined these EPAs. Education experts were recruited from the alumni network of the Master of Medical Education (MME) degree course from the University of Heidelberg, Germany. RESULTS: 28 data sets were evaluated in three Delphi rounds. 82% of study participants had previous experience with EPAs. Qualitative and quantitative data formed the basis during the iterative process and resulted in complete descriptions of 5 EPAs for final-year medical students in anaesthesiology. CONCLUSIONS: Our study including the associated description of 5 EPAs represent a further step and starting point for EPA-based curricula in medical training in Germany linking undergraduate training, to residency training and continuous medical education.

Quorum Sensing and Cyclic di-GMP Exert Control Over Motility of Vibrio fischeri KB2B1.

Bacterial motility is critical for symbiotic colonization by Vibrio fischeri of its host, the squid Euprymna scolopes, facilitating movement from surface biofilms to spaces deep inside the symbiotic organ. While colonization has been studied traditionally using strain ES114, others, including KB2B1, can outcompete ES114 for colonization for a variety of reasons, including superior biofilm formation. We report here that KB2B1 also exhibits an unusual pattern of migration through a soft agar medium: whereas ES114 migrates rapidly and steadily, KB2B1 migrates slowly and then ceases migration. To better understand this phenomenon, we isolated and sequenced five motile KB2B1 suppressor mutants. One harbored a mutation in the gene for the cAMP receptor protein (crp); because this strain also exhibited a growth defect, it was not characterized further. Two other suppressors contained mutations in the quorum sensing pathway that controls bacterial bioluminescence in response to cell density, and two had mutations in the diguanylate cyclase (DGC) gene VF_1200. Subsequent analysis indicated that (1) the quorum sensing mutations shifted KB2B1 to a perceived low cell density state and (2) the high cell density state inhibited migration via the downstream regulator LitR. Similar to the initial point mutations, deletion of the VF_1200 DGC gene increased migration. Consistent with the possibility that production of the second messenger c-di-GMP inhibited the motility of KB2B1, reporter-based measurements of c-di-GMP revealed that KB2B1 produced higher levels of c-di-GMP than ES114, and overproduction of a c-di-GMP phosphodiesterase promoted migration of KB2B1. Finally, we assessed the role of viscosity in controlling the quorum sensing pathway using polyvinylpyrrolidone and found that viscosity increased light production of KB2B1 but not ES114. Together, our data indicate that while the two strains share regulators in common, they differ in the specifics of the regulatory control over downstream phenotypes such as motility.