CD40 activation of BCP-ALL cells generates IL-10-producing, IL-12-defective APCs that induce allogeneic T-cell anergy

The use of leukemia cells as antigenpresenting cells (APCs) in immunotherapy is critically dependent on their capacity to initiate and sustain an antitumor-specific immune response. Previous studies suggested that pediatric B-cell precursor acute lymphoblastic leukemia (BCP-ALL) cells could be manipulated in vitro through the CD40-CD40L pathway to increase their immunostimulatory capacity. We extended the APC characterization of CD40L-activated BCP-ALL for their potential use in immunotherapy in a series of 19 patients. Engaging CD40 induced the up-regulation of CCR7 in 7 of 11 patients and then the migration to CCL19 in 2 of 5 patients. As accessory cells, CD40Lactivated BCP-ALL induced a strong proliferation response of naive T lymphocytes. Leukemia cells, however, were unable to sustain proliferation over time, and T cells eventually became anergic. After CD40-activation, BCP-ALL cells released substantial amounts of interleukin-10 (IL-10) but were unable to produce bioactive IL-12 or to polarize TH1 effectors. Interestingly, adding exogenous IL-12 induced the generation of interferon- (IFN- )–secreting TH1 effectors and reverted the anergic profile in a secondary response. Therefore, engaging CD40 on BCP-ALL cells is insufficient for the acquisition of full functional properties of immunostimulatory APCs. These results suggest caution against the potential use of CD40L-activated BCP-ALL cells as agents for immunotherapy unless additional stimuli, such as IL-12, are provided.Fil: D'Amico, Giovanna. Università Milano Bicocca; ItaliaFil: Vulcano, Marisa. Universidad de Buenos Aires. Facultad de Medicina; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaFil: Bugarin, Cristina. Università Milano Bicocca; ItaliaFil: Bianchi, Giancarlo. Università Milano Bicocca; ItaliaFil: Pirovano, Gisella. Università Milano Bicocca; ItaliaFil: Bonamino, Martin. Università Milano Bicocca; ItaliaFil: Marin, Virna. Università Milano Bicocca; ItaliaFil: Allavena, Paola. Università Milano Bicocca; ItaliaFil: Biagi, Ettore. Università Milano Bicocca; ItaliaFil: Biondi, Andrea. Università Milano Bicocca; Itali

Mutational analysis of ribosomal proteins in a cohort of pediatric patients with T-cell acute lymphoblastic leukemia reveals Q123R, a novel mutation in RPL10

T-cell acute lymphoblastic leukemia (T-ALL) is a subtype of ALL involving the malignant expansion of T-cell progenitors. It is driven by a number of different possible genetic lesions, including mutations in genes encoding for ribosomal proteins (RPs). These are structural constituents of ribosomes, ubiquitous effectors of protein synthesis. Albeit the R98S mutation in RPL10, recurring with a higher frequency among RP mutations, has been extensively studied, less is known about the contribution of mutations occurring in other RPs. Alterations affecting translational machinery may not be well tolerated by cells, and there may be a selective pressure that determines the emergence of mutations with a compensatory effect. To explore this hypothesis, we sequenced the exomes of a cohort of 37 pediatric patients affected by T-ALL, and analyzed them to explore the co-occurrence of mutations in genes involved in ribosome biogenesis (including RPs) and translational control, and in known T-ALL driver genes. We found that some of the mutations in these sub-classes of genes tend to cluster together in different patients, indicating that their co-occurrence may confer some kind of advantage to leukemia cells. In addition, our sequencing highlighted the presence of a novel mutation in RPL10, namely the Q123R, which we found associated with a defect in protein synthesis. Our findings indicate that genetic alterations involving ribosome biogenesis and translational control should be carefully considered in the context of precision medicine in T-ALL

Abnormal B-Cell Maturation and Increased Transitional B Cells in CBL Syndrome

CBL syndrome is a Noonan-like RASopathy with heterogeneous clinical phenotype and predisposition to juvenile myelomonocytic leukemia (JMML). Here we describe two patients with identical germline CBL mutation and clinical and immune-hematological overlapping features with autoimmune lymphoproliferative syndrome (ALPS) and B-cell expansion with NF-κB and T-cell anergy (BENTA) syndrome. Increased immature/transitional B cells can be depicted in CBL syndrome, ALPS, and BENTA. Nonetheless, our patients here described showed peculiar B-cell phenotype due to increased immature/transitional CD34+ B cells. This feature differentiates CBL syndrome from BENTA, pointing toward an abnormal proliferation of B-cell early precursors

Absent B Cells, agammaglobulinemia, and Hypertrophic Cardiomyopathy in Folliculin-interacting Protein 1 Deficiency

Agammaglobulinemia is the most profound primary antibody deficiency that can occur due to an early termination of B-cell development. We here investigated 3 novel patients, including the first known adult, from unrelated families with agammaglobulinemia, recurrent infections, and hypertrophic cardiomyopathy (HCM). Two of them also presented with intermittent or severe chronic neutropenia. We identified homozygous or compound-heterozygous variants in the gene for folliculin interacting protein 1 (FNIP1), leading to loss of the FNIP1 protein. B-cell metabolism, including mitochondrial numbers and activity and phosphatidylinositol 3-kinase/AKT pathway, was impaired. These defects recapitulated the Fnip1-/- animal model. Moreover, we identified either uniparental disomy or copy-number variants (CNVs) in 2 patients, expanding the variant spectrum of this novel inborn error of immunity. The results indicate that FNIP1 deficiency can be caused by complex genetic mechanisms and support the clinical utility of exome sequencing and CNV analysis in patients with broad phenotypes, including agammaglobulinemia and HCM. FNIP1 deficiency is a novel inborn error of immunity characterized by early and severe B-cell development defect, agammaglobulinemia, variable neutropenia, and HCM. Our findings elucidate a functional and relevant role of FNIP1 in B-cell development and metabolism and potentially neutrophil activity

Adolescentes y drogas : su relación con la delincuencia

Esta investigación describe la relación con las drogas de una muestra de 286 Adolescentes en Conflicto con la Ley de la ciudad de Valencia (España). Tiene por objeto analizar relación entre trayectoria delictiva y consumo de drogas. Los resultados informan que a mayor incidencia delictiva, mayor dependencia. En cuanto a patrones de consumo por tipo de sustancia, el cannabis se asocia a trayectorias delictivas iniciales y la cocaína a trayectorias consolidadas. El consumo se produce en escenarios habituales diurnos, se extiende a otros de ocio nocturno, y particularmente forma parte de la vida cotidiana de los adolescentes en conflicto con la ley de trayectoria consolidada

Consórcios intermunicipais de saúde: uma análise à luz da teoria dos jogos

O presente estudo analisa a formação e a sustentabilidade político-financeira dos Consórcios Intermunicipais de Saúde por meio de um jogo dinâmico de dois períodos com informação imperfeita, no qual dois prefeitos de municípios decidem quanto à adesão e à permanência no consórcio. São analisados dois modelos. No primeiro admite-se a suspensão do atendimento nas instalações do consórcio à sua população do município que abandonar a parceria. Já no segundo, considerando o princípio constitucional que estabelece o acesso universal às ações e serviços de saúde, relaxa-se essa hipótese. O estudo evidencia uma forte tendência à instabilidade da associação no segundo modelo.<br>The present article studies a new organizational structure in which neighboring health districts in Brazil form a partnership to jointly supply costly specialized health care services. Each district partially funds the organization. Depending on the partnership contract, a free rider problem arises. A district has the incentive to withdraw from the partnership if it can still benefit from its services, especially when political pressures for competing expenditures arise. The main conclusion is that the partnership sustainability is strongly influenced by the punishment mechanisms to a defaulting member, the gains from joint provision of health services and the overall economic environment

Either IL-7 activation of JAK-STAT or BEZ inhibition of PI3K-AKT-mTOR pathways dominates the single-cell phosphosignature of ex vivo treated pediatric T-cell acute lymphoblastic leukemia cells

: T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive cancer arising from lymphoblasts of T-cell origin. While T-ALL accounts for only 15% of childhood and 25% of adult ALL, 30% of patients relapse with a poor outcome. Targeted therapy of resistant and high-risk pediatric T-ALLs is therefore urgently needed, together with precision medicine tools allowing the testing of efficacy in patient samples. Furthermore, leukemic cell heterogeneity requires drug response assessment at the single-cell level. Here, we used single-cell mass cytometry to study signal transduction pathways such as the JAK-STAT, PI3K-AKT-mTOR and MEK-ERK pathways in 16 diagnostic and 5 relapsed TALL primary samples and investigated the in vitro response of cells to Interleukin-7 (IL-7) and the inhibitor BEZ-235. T-ALL cells showed upregulated activity of the PI3K-AKTmTOR and MEK-ERK pathways and increased proliferation and translation markers. We found that perturbation induced by the ex vivo administration of either IL-7 or BEZ-235 reveals a high degree of exclusivity with respect to the phospho-protein responsiveness to these agents. Notably, these response signatures were maintained from diagnosis to relapse in individual patients. In conclusion, we demonstrated the power of mass cytometry single-cell profiling of signal transduction pathways in T-ALL. Taking advantage of this advanced approach, we were able to identify distinct clusters with different responsiveness to IL-7 and BEZ-235 that can persist at relapse. Collectively our observations can contribute to a better understanding of the complex signaling network governing T-ALL behavior and its correlation with influence on the response to therapy