An ex vivo tumor fragment platform to dissect response to PD-1 blockade in cancer

An ex vivo platform of patient-derived tumor fragments enables the assessment of intratumoral immune reactivation after PD-1 blockade that is predictive of clinical outcomes in patients with cancer.Inhibitors of the PD-1-PD-L1 axis have been approved as therapy for many human cancers. In spite of the evidence for their widespread clinical activity, little is known about the immunological alterations that occur in human cancer tissue after PD-1 blockade. We developed and employed a patient-derived tumor fragment platform to dissect the early immunological response of human tumor tissue to ex vivo PD-1 blockade. We observed that the capacity of immune cells to be reactivated ex vivo was predictive of clinical response, and perturbation analyses identified tumor-resident T cells as a key component of this immunological response. In addition, through combined analysis of baseline properties and immune response capacity, we identified a new subgroup of infiltrated tumors that lacks the capacity to respond to PD-1 blockade. Finally, the baseline presence of tertiary lymphoid structures and their components correlated with the capacity of cancers to undergo intratumoral immune cell reactivation.Immunobiology of allogeneic stem cell transplantation and immunotherapy of hematological disease

An HLA-A*11:01-binding neoantigen from mutated NPM1 as target for TCR gene therapy in AML

Simple Summary: Acute myeloid leukemia (AML) is an aggressive hematological malignancy with poor prognosis. For AML relapses after chemotherapy, new and effective therapies are needed. In 30-35% of AMLs, a frameshift mutation in the nucleophosmin 1 gene (dNPM1) creates potential neoantigens that are attractive targets for immunotherapy. We previously isolated a T-cell receptor (TCR) that targets an HLA-A*02:01-binding dNPM1 neoantigen on primary AML. Here, we investigated whether AVEEVSLRK is another dNPM1 neoantigen that can be targeted by TCR gene transfer. We isolated various T-cells, cloned the HLA-A*11:01-restricted TCR from one T-cell clone and, upon transfer to CD8 cells, demonstrated targeting of dNPM1 primary AMLs in vitro. However, the TCR failed to mediate an anti-tumor effect in immunodeficient mice engrafted with dNPM1 OCI-AML3 cells. Our results demonstrate that AVEEVSLRK is an HLA-A*11:01-binding neoantigen on dNPM1 AML. Whether the isolated TCR is of sufficient affinity to treat patients remains uncertain.Acute myeloid leukemia (AML) is a hematological malignancy caused by clonal expansion of myeloid progenitor cells. Most patients with AML respond to chemotherapy, but relapses often occur and infer a very poor prognosis. Thirty to thirty-five percent of AMLs carry a four base pair insertion in the nucleophosmin 1 gene (NPM1) with a C-terminal alternative reading frame of 11 amino acids. We previously identified various neopeptides from the alternative reading frame of mutant NPM1 (dNPM1) on primary AML and isolated an HLA-A*02:01-restricted T-cell receptor (TCR) that enables human T-cells to kill AML cells upon retroviral gene transfer. Here, we isolated T-cells recognizing the dNPM1 peptide AVEEVSLRK presented in HLA-A*11:01. The TCR cloned from a T-cell clone recognizing HLA-A*11:01+ primary AML cells conferred in vitro recognition and lysis of AML upon transfer to CD8 cells, but failed to induce an anti-tumor effect in immunodeficient NSG mice engrafted with dNPM1 OCI-AML3 cells. In conclusion, our data show that AVEEVSLRK is a dNPM1 neoantigen on HLA-A*11:01+ primary AMLs. CD8 cells transduced with an HLA-A*11:01-restricted TCR for dNPM1 were reactive against AML in vitro. The absence of reactivity in a preclinical mouse model requires further preclinical testing to predict the potential efficacy of this TCR in clinical development.Immunobiology of allogeneic stem cell transplantation and immunotherapy of hematological disease

Translation and adaptation of a questionnaire to assess the group processes of rehabilitation team conferences

Objective: To investigate the internal consistency, the domain structure and the influence of social desirability with regard to a questionnaire translated and adapted to assess the quality of rehabilitation team conferences in the Netherlands. Study design: A questionnaire to determine group decision-making processes was translated and adapted to rehabilitation and completed by 44 rehabilitation professionals. Results: The internal consistency of the domains Personal participation, Negative socio-emotional behaviour, Result satisfaction and Process satisfaction was found to be satisfactory (Cronbach's α ranges from 0.70 to 0.84). The domain structure is confirmed by item-total and item-rest correlations. From the original English questionnaire, one question concerning the domain Personal participation was omitted. The domain Informal leadership has been deleted from the questionnaire, because informal leadership is not an issue in a situation in which the Chairman is already known. Response to the questionnaire did not seem to be biased by social desirability. Conclusion: The translated and adapted questionnaire can be used to assess the group processes of rehabilitation team conferences. Results from the literature concerning the original questionnaire suggest that the translated and adapted questionnaire might be able to detect changes in the group process of rehabilitation team conferences

Development and implementation of the Rehabilitation Activities Profile for children: impact on the rehabilitation team

Objective: To describe the changes in functioning of the rehabilitation team induced by the Rehabilitation Activities Profile for children (Children's RAP), an instrument designed to improve interdisciplinary communication in paediatric rehabilitation. Design: Multiple case-study design. Subjects: Seven paediatric rehabilitation teams. Intervention: A two-year project to develop and implement the Children's RAP. Data collection: During the project, data were gathered from observations, documents and informal interviews. After the project, formal interviews were held with team members and parents, and a focus group meeting with representatives of the teams was organized. Data analysis: Data were analysed by the method of analytic induction. The analysis was checked by an independent researcher. The preliminary results and conclusions were discussed in detail with participating teams. Results: Development and implementation, as well as the changes induced by the project, varied between teams. Changes were observed for individual team members, for the team as a whole and for the children and their parents. However, changes for individual team members occurred relatively quickly, in comparison with the other changes. To achieve an optimal interdisciplinary team approach all changes are necessary. Therefore, we postulated four hierarchical steps in the development of an interdisciplinary team approach: (1) process-oriented approach, (2) result-oriented approach, (3) problem-oriented approach, and (4) interdisciplinary team approach. Conclusion: It took a considerable amount of time to achieve the interdisciplinary team approach by implementing the Children's RAP. However, the first steps are not only rewarding in themselves, but also prerequisites for further improvement. © Arnold 2002