Search

Apoptotic cells have long been considered as intrinsically tolerogenic or unable to elicit immune responses specific for dead cell-associated antigens. However, multiple stimuli can trigger a functionally peculiar type of apoptotic demise that does not go unnoticed by the adaptive arm of the immune system, which we named "immunogenic cell death" (ICD). ICD is preceded or accompanied by the emission of a series of immunostimulatory damage-associated molecular patterns (DAMPs) in a precise spatiotemporal configuration. Several anticancer agents that have been successfully employed in the clinic for decades, including various chemotherapeutics and radiotherapy, can elicit ICD. Moreover, defects in the components that underlie the capacity of the immune system to perceive cell death as immunogenic negatively influence disease outcome among cancer patients treated with ICD inducers. Thus, ICD has profound clinical and therapeutic implications. Unfortunately, the gold-standard approach to detect ICD relies on vaccination experiments involving immunocompetent murine models and syngeneic cancer cells, an approach that is incompatible with large screening campaigns. Here, we outline strategies conceived to detect surrogate markers of ICD in vitro and to screen large chemical libraries for putative ICD inducers, based on a high-content, high-throughput platform that we recently developed. Such a platform allows for the detection of multiple DAMPs, like cell surface-exposed calreticulin, extracellular ATP and high mobility group box 1 (HMGB1), and/or the processes that underlie their emission, such as endoplasmic reticulum stress, autophagy and necrotic plasma membrane permeabilization. We surmise that this technology will facilitate the development of next-generation anticancer regimens, which kill malignant cells and simultaneously convert them into a cancer-specific therapeutic vaccine. peerreview_statement: The publishing and review policy for this title is described in its Aims & Scope. aims_and_scope_url: http://www.tandfonline.com/action/journalInformation?show=aimsScope&journalCode=koni20 ispartof: OncoImmunology vol:3 issue:9 pages:12472-124508 ispartof: location:United States status: published

International audience; Human Vδ2 cells are innate-like γδ T effectors performing potent immune surveillance against tumors. The constitutive expression of NKG2A identifies a subset of Vδ2 T cells licensed with an intrinsic hyper-responsiveness against cancer. Indeed, the transcriptomic profiles of NKG2A+ and NKG2A- cells characterize two distinct "intralineages" of Vδ2 T lymphocytes that appear early during development, keep their phenotypes, and show self-renewal capabilities in adult life. The hyper-responsiveness of NKG2A+ Vδ2 T cells is counterbalanced by the inhibitory signaling delivered by human leukocyte antigen E (HLA-E) expressed on malignant cells as a tumor-escape mechanism. However, either masking or knocking out NKG2A restores the capacity of Vδ2 T cells to exert the highest effector functions even against HLA-E+ tumors. This is highly relevant in the clinic, as the different degrees of engagement of the NKG2A-HLA-E checkpoint in hepatocellular carcinoma, glioblastoma, and non-small cell lung cancer directly impact patients' overall survival. These findings open avenues for developing combined cellular and immunologic anticancer therapies.

CD8+ T cells infiltrating tumors are largely dysfunctional. Brummelman and Mazza et al. identify partially exhausted CXCR5+ TIM-3– CD8+ T cells with enhanced stem-like properties and cytotoxicity infiltrating human solid tumors. These cells express candidate immunotherapeutic targets (PD-1, TIGIT and CD27) for their reinvigoration., CD8+ T cells infiltrating tumors are largely dysfunctional, but whether a subset maintains superior functionality remains ill defined. By high-dimensional single cell analysis of millions of CD8+ T cells from 53 individuals with lung cancer, we defined those subsets that are enriched in tumors compared with cancer-free tissues and blood. Besides exhausted and activated cells, we identified CXCR5+ TIM-3– CD8+ T cells with a partial exhausted phenotype, while retaining gene networks responsible for stem-like plasticity and cytotoxicity, as revealed by single cell sequencing of the whole transcriptome. Ex vivo, CXCR5+ TIM-3– CD8+ T cells displayed enhanced self-renewal and multipotency compared with more differentiated subsets and were largely polyfunctional. Analysis of inhibitory and costimulatory receptors revealed PD-1, TIGIT, and CD27 as possible targets of immunotherapy. We thus demonstrate a hierarchy of differentiation in the context of T cell exhaustion in human cancer similar to that of chronically infected mice, which is further shown to disappear with disease progression., Graphical Abstract

In their recent paper, Weili Xu et al. [1] described the different behaviors of Vδ1pos and Vδ2pos T cell subsets in response to lifelong stress and claimed that Vδ2pos T cells are not affected by aging and Human Cytomegalovirus (HCMV) infection. While we agree that these two γδ T cell subsets diverge both in phenotype/function and in tissue distribution, we are somewhat surprised that authors did not take into account the several previously published and contradictory experimental evidence in regards to senescence of Vδ2pos T cells [2,3]. These latter studies reported that HCMV infection not only induces a clonal expansion of a distinct Vγ9neg/Vδ2pos T cell subset, but also determines a concomitant adaptive differentiation from CD27high naïve cells to CD27low/neg terminal-effectors. However, Weili Xu et al. argued that the expression and kinetics of both CD27 and CD45RA surface markers do not change and follow the homeostatic changes of Vδ2pos T cells. This statement goes in the opposite direction to previously reported findings as the CD27/CD45RA phenotype has been shown to mark the maturation and differentiation (TNaïve, TCentral-Memory, Teffector-Memory and TEffectory-Memory RA) of Vδ2pos T cells. Indeed, the different surface expression of both CD27 and CD45 parallel the progressive decrease of telomere length, the proliferative capacity as well as the different effector-functions and resistance to death of Vδ2+ T cells in response to antigens and homeostatic cytokines [4,5]. Hence, we believe that these controversial issues require further discussion beyond the unilateral conclusion given by the study of Weili Xu et al.

Background & Aims Alterations in signaling pathways that regulate resolution of inflammation (resolving pathways) contribute to pathogenesis of ulcerative colitis (UC). The resolution process is regulated by lipid mediators, such as those derived from the ω-3 docosahexaenoic acid (DHA), whose esterified form is transported by the major facilitator superfamily domain containing 2A (MFSD2A) through the endothelium of brain, retina, and placenta. We investigated if and how MFSD2A regulates lipid metabolism of gut endothelial cells to promote resolution of intestinal inflammation. Methods We performed lipidomic and functional analyses of MFSD2A in mucosal biopsies and primary human intestinal microvascular endothelial cells (HIMECs) isolated from surgical specimens from patients with active, resolving UC and healthy individuals without UC (controls). MFSD2A was knocked down in HIMECs with small hairpin RNAs or overexpressed from a lentiviral vector. Human circulating endothelial progenitor cells that overexpress MFSD2A were transferred to CD1 nude mice with dextran sodium sulfate–induced colitis, with or without oral administration of DHA. Results Colonic biopsies from patients with UC had reduced levels of inflammation-resolving DHA-derived epoxy metabolites compared to healthy colon tissues or tissues with resolution of inflammation. Production of these metabolites by HIMECs required MFSD2A, which is required for DHA retention and metabolism in the gut vasculature. In mice with colitis, transplanted endothelial progenitor cells that overexpressed MFSD2A not only localized to the inflamed mucosa but also restored the ability of the endothelium to resolve intestinal inflammation, compared with mice with colitis that did not receive MFSD2A-overexpressing endothelial progenitors. Conclusions Levels of DHA-derived epoxides are lower in colon tissues from patients with UC than healthy and resolving mucosa. Production of these metabolites by gut endothelium requires MFSD2A; endothelial progenitor cells that overexpress MFSD2A reduce colitis in mice. This pathway might be induced to resolve intestinal inflammation in patients with colitis.

T cells constitute an unconventional lymphocyte population with distinct functions complementary to those of CD4 and CD8 T cells. As such, they have both adaptive features, such as expression of the TCR, and innate-like functions reminiscent of NK cells, with whom they share extensive repertoires of activating and inhibitory receptors [1, 2]. Although most antigens recognized by murine T cells remain obscure, advances have been made in identifying ligands for human T cells. The majority of circulating human T lymphocytes expresses a TCR formed by the preferentially-paired V 9 and V 2 chains (here and thereafter, called V 9V 2 T cells). Instead of binding peptides associated with molecules belonging to the MHC, V 9V 2 T cells recognize pyrophosphate intermediates of the microbial or eukaryotic isoprenoid synthesis pathways, called PAgs [3, 4]. Isoprenoids are produced by one of two major pathways: the classical mevalonate pathway and the alternative deoxyxylulose (nonmevalonate) pathway of isoprenoids synthesis that is used by numerous bacterial species and by some highly significant eukaryotic pathogens but not by vertebrate cells. IPP is an intermediate metabolite that is present in both pathways, whereas HMBPP is only produced in the nonmevalonate pathway by certain microbes, including Mycobacterium tuberculosis and Listeria monocytogenes. Given this cross-reactivity between human V 9V 2 T cells and microbial and self-PAgs, there is a great interest in the scientific community of elucidating how TCR signaling can be induced by such small molecules. PAgs can directly activate V 9V 2 T cells, but such activation is greatly enhanced by monocytes and/or DCs. Hence, either PAgs are presented as cargo to the reactive TCR or their cellular processing somehow sensitizes cell recognition through the engagement of V 9V 2 TCR (i.e., by stabilizing surface expression of TCR-binding molecules) [3, 4]. A candidate molecule involved in intracellular PAg processing is the F1-ATPase, which was reported to bind directly the V 9V 2 TCR and to also interact with IPP conjugated to AMP, an adenosine derivative of IPP [5]. Thus, V 9V 2 T cells may collectively monitor multiple components of pathways regulating cholesterol biosynthesis that are altered by infection or other forms of stress. In this regard, it was found recently that PAg-induced V 9V 2 T cell activation also involves the participation of BTN3A1, a widely expressed member of the Ig superfamily [6]. Therefore, production of exogenous HMBPP or up-regulation of endogenous IPP in human cells in response to infections or noninfective immune dysregulation provokes V 9V 2 T cell reactivity, albeit at substantially different sensitivity. Although IPP was isolated from Mycobacterium smegmatis as the first natural ligand for human T cells, it soon became clear that the amounts of IPP present in bacterial extracts do not reach the minimum threshold required for inducing V 9V 2 T cell activation in several experimental systems in vitro. HMBPP is more potent than IPP in the context of in vitro activation of V 9V 2 T cells, active at picoto nanomolar concentrations, whereas IPP requires 3-log higher concentrations within micromolar ranges. Such high concentrations are not produced in physiological conditions but are reached during the course of pathologic metabolisms of stressed or infected cells. V 9V 2 T cells evolved to detect stressed cells and microbial pathogens, although only if the local amounts of IPP released from damaged tissues are at least 10,000-fold higher than the HMBPP concentrations reached during bacterial infections. This explains the different bioactivities of these two compounds [7]. The finding that V 9V 2 T cells respond differentially in vitro to increasing concentrations of structurally related PAgs indicates that such ligands constitute agonists for the TCR at different strengths. Indeed, experimental approaches aiming to define the underlying molecular basis of T lymphocyte activation demonstrate that V 9V 2 TCR can discriminate between subtle differences of substrates. By analyzing early cellular events and late effector responses of V 9V 2 T cells, it has also been reported that IPP induces a timeand dose-dependent down-modulation of the reactive TCR, whereas HMBPP induces little or no TCR

Increased plasma level of soluble urokinase-type plasminogen activator receptor (suPAR) was associated recently with focal segmental glomerulosclerosis (FSGS). In addition, different clinical studies observed increased concentration of suPAR in various glomerular diseases and in other human pathologies with nephrotic syndromes such as HIV and Hantavirus infection, diabetes and cardiovascular disorders. Here, we show that suPAR induces nephrin down-modulation in human podocytes. This phenomenon is mediated only by full-length suPAR, is time-and dose-dependent and is associated with the suppression of Wilms’ tumor 1 (WT-1) transcription factor expression. Moreover, an antagonist of αvβ3 integrin RGDfv blocked suPAR-induced suppression of nephrin. These in vitro data were confirmed in an in vivo uPAR knock out Plaur−/− mice model by demonstrating that the infusion of suPAR inhibits expression of nephrin and WT-1 in podocytes and induces proteinuria. This study unveiled that interaction of full-length suPAR with αvβ3 integrin expressed on podocytes results in down-modulation of nephrin that may affect kidney functionality in different human pathologies characterized by increased concentration of suPAR.

Rationale The liver-specific natural killer (NK) cell population is critical for local innate immune responses, but the mechanisms that lead to their selective homing and the definition of their functionally relevance remain enigmatic. Objectives We took advantage of the availability of healthy human liver to rigorously define the mechanisms regulating the homing of NK cells to liver and the repertoire of receptors that distinguish liver-resident NK (lr-NK) cells from circulating counterparts. Findings Nearly 50% of the entire liver NK cell population is composed of functionally relevant CD56 bright lr-NK cells that localize within hepatic sinusoids. CD56 bright lr-NK cells express CD69, CCR5 and CXCR6 and this unique repertoire of chemokine receptors is functionally critical as it determines selective migration in response to the chemotactic stimuli exerted by CCL3, CCL5 and CXCL16. Here, we also show that hepatic sinusoids express CCL3 pos Kupffer cells, CXCL16 pos endothelial cells and CCL5 pos T and NK lymphocytes. The selective presence of these chemokines in sinusoidal spaces creates a unique tissue niche for lr-CD56 bright NK cells that constitutively express CCR5 and CXCR6. CD56 bright lr-NK cells co-exist with CD56 dim conventional NK (c-NK) cells that are, interestingly, transcriptionally and phenotypically similar to their peripheral circulating counterparts. Indeed, CD56 dim c-NK cells lack expression of CD69, CCR5, and CXCR6 but express selectins, integrins and CX 3 CR1. Conclusion Our findings disclosing the phenotypic and functional differences between lr-Nk cells and c-NK cells are critical to distinguish liver-specific innate immune responses. Hence, any therapeutic attempts at modifying the large population of CD56 bright lr-NK cells will require modification of hepatic CCR5 and CXCR6.

Background HIV-1 Protease Inhibitors, namely PIs, originally designed to inhibit HIV-1 aspartic protease, can modulate the immune response by mechanisms largely unknown, and independent from their activity on viral replication. Here, we analyzed the ability of PIs to interfere with differentiation program of monocytes toward dendritic cell (DCs) lineage, a key process in the inflammatory response. Methodology/Principal Findings Monocytes from healthy donors were isolated and induced to differentiate in vitro in the presence or absence of saquinavir, ritonavir, nelfinavir, indinavir or amprenavir (sqv, rtv, nlfv, idv, apv, respectively). These drugs demonstrated a differential ability to sustain the generation of immature DCs (iDCs) with an altered phenotype, including low levels of CD1a, CD86, CD36 and CD209. DCs generated in the presence of rtv also failed to acquire the typical phenotype of mature DCs (mDCs), and secreted lower amounts of IL-12 and IL-15. Accordingly, these aberrant mDCs failed to support activation of autologous Natural Killer (NK) cells, and resulted highly susceptible to NK cell-mediated cytotoxicity. Conclusions/Significance Our findings uncover novel functional properties of PIs within the DC-NK cell cross-talk, unveiling the heterogeneous ability of members of this class drugs to drive the generation of atypical monocyte-derived DCs (MDDCs) showing an aberrant phenotype, a failure to respond appropriately to bacterial endotoxin, a weak ability to prime autologous NK cells, and a high susceptibility to NK cell killing. These unexpected properties might contribute to limit inflammation and viral spreading in HIV-1 infected patients under PIs treatment, and open novel therapeutical perspectives for this class drugs as immunomodulators in autoimmunity and cancer.