Immune and Genetic Signatures of Breast Carcinomas Triggering Anti-Yo–Associated Paraneoplastic Cerebellar Degeneration

International audienceBackground and Objectives Paraneoplastic cerebellar degeneration (PCD) with anti-Yo antibodies is a cancer-related autoimmune disease directed against neural antigens expressed by tumor cells. A putative trigger of the immune tolerance breakdown is genetic alteration of Yo antigens. We aimed to identify the tumors' genetic and immune specificities involved in Yo-PCD pathogenesis. Methods Using clinicopathologic data, immunofluorescence (IF) imaging, and whole-transcriptome analysis, 22 breast cancers (BCs) associated with Yo-PCD were characterized in terms of oncologic characteristics, genetic alteration of Yo antigens, differential gene expression profiles, and morphofunctional specificities of their in situ antitumor immunity by comparing them with matched control BCs. Results Yo-PCD BCs were invasive carcinoma of no special type, which early metastasized to lymph nodes. They overexpressed human epidermal growth factor receptor 2 (HER2) but were hormone receptor negative. All Yo-PCD BCs carried at least 1 genetic alteration (variation or gain in copy number) on CDR2L, encoding the main Yo antigen that was found aberrantly overexpressed in Yo-PCD BCs. Analysis of the differentially expressed genes found 615 upregulated and 54 downregulated genes in Yo-PCD BCs compared with HER2-driven control BCs without PCD. Ontology enrichment analysis found significantly upregulated adaptive immune response pathways in Yo-PCD BCs. IF imaging confirmed an intense immune infiltration with an overwhelming predominance of immunoglobulin G-plasma cells. Discussion These data confirm the role of genetic alterations of Yo antigens in triggering the immune tolerance breakdown but also outline a specific biomolecular profile in Yo-PCD BCs, suggesting a cancer-specific pathogenesis

IL-2 Mediates CD4+ T Cell Help in the Breakdown of Memory-Like CD8+ T Cell Tolerance under Lymphopenic Conditions

Background: Lymphopenia results in the proliferation and differentiation of naïve T cells into memory-like cells in the apparent absence of antigenic stimulation. This response, at least in part due to a greater availability of cytokines, is thought to promote anti-self responses. Although potentially autoreactive memory-like CD8 + T cells generated in a lymphopenic environment are subject to the mechanisms of peripheral tolerance, they can induce autoimmunity in the presence of antigen-specific memory-like CD4 + T helper cells. Methodology/Principal Findings: Here, we studied the mechanisms underlying CD4 help under lymphopenic conditions in transgenic mice expressing a model antigen in the beta cells of the pancreas. Surprisingly, we found that the self-reactivity mediated by the cooperation of memory-like CD8 + and CD4 + T cells was not abrogated by CD40L blockade. In contrast, treatment with anti-IL-2 antibodies inhibited the onset of autoimmunity. IL-2 neutralization prevented the CD4-mediated differentiation of memory-like CD8 + T cells into pathogenic effectors in response to self-antigen cross-presentation. Furthermore, in the absence of helper cells, induction of IL-2 signaling by an IL-2 immune complex was sufficient to promote memory-like CD8 + T cell self-reactivity. Conclusions/Significance: IL-2 mediates the cooperation of memory-like CD4 + and CD8 + T cells in the breakdown of crosstolerance, resulting in effector cytotoxic T lymphocyte differentiation and the induction of autoimmune disease

A922 Sequential measurement of 1 hour creatinine clearance (1-CRCL) in critically ill patients at risk of acute kidney injury (AKI)

Meeting abstrac

The role of lymphopenia-induced proliferation in the breakdown of peripheral CD8+ T cell tolerance

La tolérance des lymphocytes TCD8+ est essentielle pour empêcher l'apparition d'auto-immunité mais représente un obstacle pour le développement de réponses cytotoxiques contre les tumeurs. La lymphodéplétion est utilisée comme adjuvant pour l'immunothérapie par transfert adoptif de cellules T cytotoxiques car elle améliore leur efficacité en favorisant la rupture de la tolérance périphérique. En condition lymphopénique aiguë, les lymphocytes T naïfs prolifèrent en absence apparente de stimulation antigénique et vont acquérir un phénotype et une fonctionnalité semblable aux cellules mémoires (LIP pour Lymphopenia Induced Proliferation). Les cellules mémoires ayant un seuil d'activation inférieur cellules naïves, il a été proposé que la différentiation des cellules TCD8+ potentiellement autoréactives en cellules de type mémoire dans des conditions lymphopéniques pouvait conduire à la rupture de la tolérance. Pendant ma thèse, j'ai étudié si la LIP est nécessaire pour la rupture de la tolérance croisée des cellules TCD8+ chez des souris transgéniques irradiées exprimant un antigène modèle dans les cellules β du pancréas. De manière surprenante, nous avons constaté que le blocage de la LIP ne permet pas d'inhiber l'apparition d'auto-réactivité : les lymphocytes TCD8+ qui ne transitent pas par un stade de différenciation de type mémoire parviennent à se différencier en cellules effectrices suite à la présentation croisée de l'antigène et à migrer vers le pancréas. Néanmoins, la LIP est requise pour induire de l'auto-réactivité lorsque la fréquence de lymphocytes T CD8+ est faible ; non pas à cause du déséquilibre de la population de cellules T régulatrices, mais dû à une nette augmentation du nombre de cellules TCD8+ autoréactives. Ainsi, bien que la LIP améliore les réponses auto- réactives des cellules TCD8+, la différentiation en cellules de type mémoire n'est pas indispensable pour la rupture de la tolérance croisée en condition lymphopénique induite par irradiation.The immune system has evolved multiple mechanisms of peripheral tolerance to control CD8+ T cell responses and to prevent autoimmunity. However, they also represent a barrier for the development of cytotoxic responses against tumors. Lymphodepleting protocols are currently used as adjuvants for adoptive cytotoxic T cell immunotherapy because they enhance their potency. These protocols are thought to promote the breakdown of peripheral CD8+ T cell tolerance. Under acute lymphopenic conditions, naive T cells proliferate, in the apparent absence of antigenic stimulation at least in part due to a greater availability of the cytokine IL-7. Proliferating CD8+ T cells acquire a phenotype and functionality that is similar to memory cells and are termed memory-like cells. Since memory cells have a lower activation threshold than naïve cells, it has been proposed that differentiation of potentially autoreactive CD8+ T cells into memory-like cells under lymphopenic conditions could drive the breakdown of peripheral tolerance. Here we studied whether lymphopenia induced proliferation and differentiation are required for the breakdown of CD8+ T cell cross-tolerance in irradiated transgenic mice expressing a model antigen in the beta cells of the pancreas. Surprisingly, we found that blocking lymphopenia-induced proliferation and differentiation into memory-like cells did not prevent self-reactivity. CD8+ T cells that did not differentiate into memory-like cells still became effectors upon antigen cross-presentation and migrated to the site of antigen expression. Nonetheless, LIP did enhance CD8+ T cell mediated self-reactivity at low T cell frequencies. This effect could not be explained by a Treg imbalance but by a net increase in autoreactive CD8+ T cell numbers. Thus, although LIP enhances CD8+ T cell anti-self responses, differentiation into memory-like cells is not essential for the breakdown of cross-tolerance under the lymphopenic conditions provided by irradiation

How does forest certification contribute to boreal biodiversity conservation? Standards and outcomes in Sweden and NW Russia

Forest Stewardship Council (FSC) is one of the leading forest certification schemes. While many studies concern political aspects and social outcomes of FSC, little is known about the contribution of certification to biodiversity conservation. In Europe, the Russian Federation and Sweden have the largest areas of FSC certified forest. We assessed the potential of FSC certification for boreal biodiversity conservation in terms of standard content, and outcomes as habitat area set aside and habitat network functionality. First, we compared the biodiversity conservation indicators at different spatial scales in Swedish and Russian FSC standards. Second, focusing on one large state forest management unit in each country, we compared the areas of formally and voluntarily set aside forests for biodiversity conservation. Third, we evaluated the structural habitat connectivity by applying morphological spatial pattern analysis, and potential functional connectivity by using habitat suitability index modelling for virtual species. The Russian standard included indicators for all spatial scales of biodiversity conservation, from tree and stand to land-scape and ecoregions. The Swedish standard focused mainly on stand and tree scales. The area of voluntary set-asides for FSC was similar in Sweden and Russia, while formal protection in the Russian case study was three times higher than in the Swedish one. Swedish set-aside core areas were two orders of magnitude smaller, had much lower structural and potential functional connectivity and were located in a fragmented forestland holding. We conclude that to understand the potential of FSC certification for biodiversity conservation both the standard content, and its implementation on the ground, need to be assessed. We discuss the potential of FSC certification for biodiversity conservation with different levels of ambition. We stress the need for developing rapid assessment tools to evaluate outcomes of FSC for biodiversity conservation on the ground, which could be used by forest managers and FSC-auditors toward adaptive governance and management

Transient microfluidic compartmentalization using actionable microfilaments for biochemical assays, cell culture and organs-on-chip

International audienceWe report here a simple yet robust transient compartmentalization system for microfluidic platforms. Cylindrical microfilaments made of commercially available fishing lines are embedded in a microfluidic chamber and employed as removable walls, dividing the chamber into several compartments. These partitions allow tight sealing for hours, and can be removed at any time by longitudinal sliding with minimal hydrodynamic perturbation. This allows the easy implementation of various functions, previously impossible or requiring more complex instrumentation. In this study, we demonstrate the applications of our strategy, firstly to trigger chemical diffusion, then to make surface co-coating or cell co-culture on a two-dimensional substrate, and finally to form multiple cell-laden hydrogel compartments for three-dimensional cell co-culture in a microfluidic device. This technology provides easy and low-cost solutions, without the use of pneumatic valves or external equipment, for constructing well-controlled microenvironments for biochemical and cellular assays

Antibiotics do not prevent antigen-driven proliferation and differentiation of Clone 4 CD8⁺ T cells into effectors in irradiated InsHA mice.

<p><b>(A)</b> Irradiated and antibiotic-treated irradiated groups of InsHA mice adoptively transferred with 5x10⁶ CFSE-labeled naïve Clone 4 Thy1.1⁺ CD8⁺ and 5x10⁶ HNT CD4⁺ T cells were sacrificed on day 10 after transfer. CFSE fluorescence intensity on gated CD8⁺ Thy1.1⁺ donor lymphocytes in LN and pLN of single representative mice are shown in left and central panels. Percentages of highly proliferating cells, means ± SD (n = 4), of one experiment out three are presented. The presence of CD8⁺ Thy1.1⁺ donor T cells in the pancreas is shown in right panels. Numbers indicate FACS event counts in the depicted gates as means ± SD (n = 4) of one representative experiment out three. <b>(B)</b> Phenotype of CD8⁺ Thy1.1⁺ donor lymphocytes in the pLN of individual InsHA mice from groups described in panel A. Percentages of CD25⁺ Granzyme B⁺ of the donor T cells from pooled pLN of one experiment out of two.</p

Irradiation-induced systemic LPS translocation is prevented by antibiotics in BALB/c mice.

<p>Sera from BALB/c mice were collected 24h (IRR 24h) or 48h (IRR 48h) after irradiation. Antibiotic-treated BALB/c mice were irradiated 8 days later and sera collected 24h after irradiation (Antibx + IRR 24h). Sera collected 24h after Ultrapure LPS i.p. injection from non-irradiated BALB/c mice served as positive control (LPS 24h). Sera from non-irradiated mice served as negative control (Non IRR). Concentration of LBP in serum is presented as means ± SD (n = 4–7) from 3 independent experiments.</p