The CD318/CD6 axis limits type 1 diabetes islet autoantigen-specific human T cell activation.

CD6 is a glycoprotein expressed on CD4 and CD8 T cells involved in immunoregulation. CD318 has been identified as a CD6 ligand. The role of CD318 in T cell immunity is restricted as it has only been investigated in a few mice autoimmune models but not in human diseases. CD318 expression was thought to be limited to mesenchymal-epithelial cells and, therefore, contribute to CD6-mediated T cell activation in the CD318-expressing tissue rather than through interaction with antigen-presenting cells. Here, we report CD318 expression in a subpopulation of CD318+ myeloid dendritic (mDC), whereas the other peripheral blood populations were CD318 negative. However, CD318 can be induced by activation: a subset of monocytes treated with LPS and IFNγ and in vitro monocyte derived DCs were CD318+. We also showed that recombinant CD318 inhibited T cell function. Strikingly, CD318+ DCs suppressed the proliferation of autoreactive T cells specific for GAD65, a well-known targeted self-antigen in Type 1 Diabetes (T1D). Our study provides new insight into the role of the CD318/CD6 axis in the immunopathogenesis of inflammation, suggesting a novel immunoregulatory role of CD318 in T cell-mediated autoimmune diseases and identifying a potential novel immune checkpoint inhibitor as a target for intervention in T1D which is an unmet therapeutic need.

Thrombospondin-1, CD47, and SIRPα display cell-specific molecular signatures in human islets and pancreata.

Thrombospondin-1 (TSP1) is a secreted protein minimally expressed in health but increased in disease and age. TSP1 binds to the cell membrane receptor CD47, which itself engages signal regulatory protein α (SIRPα), and the latter creates a checkpoint for immune activation. Individuals with cancer administered checkpoint-blocking molecules developed insulin-dependent diabetes. Relevant to this, CD47 blocking antibodies and SIRPα fusion proteins are in clinical trials. We characterized the molecular signature of TSP1, CD47, and SIRPα in human islets and pancreata. Fresh islets and pancreatic tissue from nondiabetic individuals were obtained. The expression of THBS1, CD47, and SIRPA was determined using single-cell mRNA sequencing, immunofluorescence microscopy, Western blot, and flow cytometry. Islets were exposed to diabetes-affiliated inflammatory cytokines and changes in protein expression were determined. CD47 mRNA was expressed in all islet cell types. THBS1 mRNA was restricted primarily to endothelial and mesenchymal cells, whereas SIRPA mRNA was found mostly in macrophages. Immunofluorescence staining showed CD47 protein expressed by ß cells and present in the exocrine pancreas. TSP1 and SIRPα proteins were not seen in islets or the exocrine pancreas. Western blot and flow cytometry confirmed immunofluorescent expression patterns. Importantly, human islets produced substantial quantities of secreted TSP1. Human pancreatic exocrine and endocrine tissue expressed CD47, whereas fresh islets displayed cell surface CD47 and secreted TSP1 at baseline and in inflammation. These findings suggest unexpected effects on islets from agents that intersect TSP1-CD47-SIRPα.NEW & NOTEWORTHY CD47 is a cell surface receptor with two primary ligands, soluble thrombospondin-1 (TSP1) and cell surface signal regulatory protein alpha (SIRPα). Both interactions provide checkpoints for immune cell activity. We determined that fresh human islets display CD47 and secrete TSP1. However, human islet endocrine cells lack SIRPα. These gene signatures are likely important given the increasing use of CD47 and SIRPα blocking molecules in individuals with cancer.

The TSP1-CD47-SIRPα interactome: an immune triangle for the checkpoint era.

The use of treatments, such as programmed death protein 1 (PD1) or cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) antibodies, that loosen the natural checks upon immune cell activity to enhance cancer killing have shifted clinical practice and outcomes for the better. Accordingly, the number of antibodies and engineered proteins that interact with the ligand-receptor components of immune checkpoints continue to increase along with their use. It is tempting to view these molecular pathways simply from an immune inhibitory perspective. But this should be resisted. Checkpoint molecules can have other cardinal functions relevant to the development and use of blocking moieties. Cell receptor CD47 is an example of this. CD47 is found on the surface of all human cells. Within the checkpoint paradigm, non-immune cell CD47 signals through immune cell surface signal regulatory protein alpha (SIRPα) to limit the activity of the latter, the so-called trans signal. Even so, CD47 interacts with other cell surface and soluble molecules to regulate biogas and redox signaling, mitochondria and metabolism, self-renewal factors and multipotency, and blood flow. Further, the pedigree of checkpoint CD47 is more intricate than supposed. High-affinity interaction with soluble thrombospondin-1 (TSP1) and low-affinity interaction with same-cell SIRPα, the so-called cis signal, and non-SIRPα ectodomains on the cell membrane suggests that multiple immune checkpoints converge at and through CD47. Appreciation of this may provide latitude for pathway-specific targeting and intelligent therapeutic effect.

Analysis of Circulating Tumor DNA in Synchronous Metastatic Colorectal Cancer at Diagnosis Predicts Overall Patient Survival.

Sporadic colorectal cancer (sCRC) initially presents as metastatic tumors in 25-30% of patients. The 5-year overall survival (OS) in patients with metastatic sCRC is 50%, falling to 10% in patients presenting with synchronous metastatic disease (stage IV). In this study, we systematically analyzed the mutations of RAS, PIK3CA and BRAF genes in circulating tumor DNA (ctDNA) and tumoral tissue DNA (ttDNA) from 51 synchronous metastatic colorectal carcinoma (SMCC) patients by real-time PCR, and their relationship with the clinical, biological and histological features of disease at diagnosis. The highest frequency of mutations detected was in the KRAS gene, in tumor biopsies and plasma samples, followed by mutations of the PIK3CA, NRAS and BRAF genes. Overall, plasma systematically contained those genetic abnormalities observed in the tumor biopsy sample from the same subject, the largest discrepancies detected between the tumor biopsy and plasma from the same patient being for mutations in the KRAS and PIK3CA genes, with concordances of genotyping results between ttDNA and ctDNA at diagnosis of 75% and 84%, respectively. Of the 51 SMCC patients in the study, 25 (49%) showed mutations in at least 1 of the 4 genes analyzed in patient plasma. From the prognostic point of view, the presence and number of the most common mutations in the RAS, PIK3CA and BRAF genes in plasma from SMCC patients are independent prognostic factors for OS. Determination of the mutational status of ctDNA in SMCC could be a key tool for the clinical management of patients.

Efficacy and safety of itolizumab, a novel anti-CD6 monoclonal antibody, in patients with moderate to severe chronic plaque psoriasis: results of a double-blind, randomized, placebo-controlled, phase-III study.

BACKGROUND: Itolizumab, a humanized monoclonal antibody to CD6, is a novel therapeutic agent evaluated in chronic plaque psoriasis. OBJECTIVE: We sought to assess the safety and efficacy of itolizumab in moderate to severe chronic plaque psoriasis. METHODS: A total of 225 patients were randomized (2:2:1) to 2 different itolizumab arms (A or B; A = 4-week loading dose of 0.4 mg/kg/wk followed by 1.6 mg/kg every 2 weeks; B = 1.6/mg every 2 weeks) or placebo. At week 12, the placebo arm was switched to 1.6 mg/kg itolizumab every 2 weeks. The primary end point was the proportion of patients with at least 75% improvement in Psoriasis Area and Severity Index score at week 12. RESULTS: At week 12, 27.0% in arm A (P = .0172 vs placebo), 36.4% in B (P = .0043 vs placebo), and 2.3% in the placebo arm had at least 75% improvement in Psoriasis Area and Severity Index score. At week 28, the proportion with at least 75% improvement in Psoriasis Area and Severity Index score was comparable: 46.1%, 45.5%, and 41.9% for A, B, and placebo, respectively. In weeks 1 to 12, the incidence of all adverse events was comparable across arms (A, 43%; B, 38%; placebo, 47%) and the incidence of infections was not greater than placebo (11.1%, 8.9%, and 18.6% for A, B, and placebo). LIMITATIONS: No active comparator is a limitation. CONCLUSIONS: Itolizumab is an effective and well-tolerated novel biological therapy in moderate to severe psoriasis.

Tolerating CD47.

Cluster of differentiation 47 (CD47) occupies the outer membrane of human cells, where it binds to soluble and cell surface receptors on the same and other cells, sculpting their topography and resulting in a pleiotropic receptor-multiligand interaction network. It is a focus of drug development to temper and accentuate CD47-driven immune cell liaisons, although consideration of on-target CD47 effects remain neglected. And yet, a late clinical trial of a CD47-blocking antibody was discontinued, existent trials were restrained, and development of CD47-targeting agents halted by some pharmaceutical companies. At this point, if CD47 can be exploited for clinical advantage remains to be determined. Herein an airing is made of the seemingly conflicting actions of CD47 that reflect its position as a junction connecting receptors and signalling pathways that impact numerous human cell types. Prospects of CD47 boosting and blocking are considered along with potential therapeutic implications for autoimmune diseases and cancer.

Assessment of the performance of a symbiotic microalgal-bacterial granular sludge reactor for the removal of nitrogen and organic carbon from dairy wastewater.

Cheese whey (CW) is a nutrient deficient dairy effluent, which requires external nutrient supplementation for aerobic treatment. CW, supplemented with ammonia, can be treated using aerobic granular sludge (AGS) in a sequencing batch reactor (SBR). AGS are aggregates of microbial origin that do not coagulate under reduced hydrodynamic shear and settle significantly faster than activated sludge flocs. However, granular instability, slow granulation start-up, high energy consumption and CO2 emission have been reported as the main limitations in bacterial AGS-SBR. Algal-bacterial granular systems have shown be an innovative alternative to improve these limitations. Unfortunately, algal-bacterial granular systems for the treatment of wastewaters with higher organic loads such as CW have been poorly studied. In this study, an algal-bacterial granular system implemented in a SBR (SBRAB) for the aerobic treatment of ammonia-supplemented CW wastewaters was investigated and compared with a bacterial granular reactor (SBRB). Mass balances were used to estimate carbon and nitrogen (N) assimilation, nitrification and denitrification in both set-ups. SBRB exhibited COD and ammonia removal of 100% and 94% respectively, high nitrification (89%) and simultaneous nitrification-denitrification (SND) of 23% leading to an inorganic N removal of 30%. The efficient algal-bacterial symbiosis in granular systems completely removed COD and ammonia (100%) present in the dairy wastewater. SBRAB microalgae growth could reduce about 20% of the CO2 emissions produced by bacterial oxidation of organic compounds according to estimates based on synthesis reactions of bacterial and algal biomass, in which the amount of assimilated N determined by mass balance was taken into account. A lower nitrification (75%) and minor loss of N by denitrifying activity (<5% Ng, SND 2%) was also encountered in SBRAB as a result of its higher biomass production, which could be used for the generation of value-added products such as biofertilizers and biostimulants.

Activation of ductal progenitor-like cells from adult human pancreas requires extracellular matrix protein signaling.

Ductal progenitor-like cells are a sub-population of ductal cells in the adult human pancreas that have the potential to contribute to regenerative medicine. However, the microenvironmental cues that regulate their activation are poorly understood. Here, we establish a 3-dimensional suspension culture system containing six defined soluble factors in which primary human ductal progenitor-like and ductal non-progenitor cells survive but do not proliferate. Expansion and polarization occur when suspension cells are provided with a low concentration (5% v/v) of Matrigel, a sarcoma cell product enriched in many extracellular matrix (ECM) proteins. Screening of ECM proteins identified that collagen IV can partially recapitulate the effects of Matrigel. Inhibition of integrin α1ß1, a major collagen IV receptor, negates collagen IV- and Matrigel-stimulated effects. These results demonstrate that collagen IV is a key ECM protein that stimulates the expansion and polarization of human ductal progenitor-like and ductal non-progenitor cells via integrin α1ß1 receptor signaling.

Artisanal trawl fisheries as a sentinel of marine litter pollution.

Systematic seafloor surveys are a highly desirable method of marine litter monitoring, but the high costs involved in seafloor sampling are not a trivial handicap. In the present work, we explore the opportunity provided by the artisanal trawling fisheries to obtain systematic data on marine litter in the Gulf of Cadiz between 2019 and 2021. We find that plastic was the most frequent material, with a prevalence of single-use and fishing-related items. Litter densities decreased with increasing distance to shore with a seasonal migration of the main litter hotspots. During pre-lockdown and post-lockdown stages derived from COVID-19, marine litter density decreased by 65 %, likely related to the decline in tourism and outdoor recreational activities. A continuous collaboration of 33 % of the local fleet would imply a removal of hundreds of thousands of items each year. The artisanal trawl fishing sector can play a unique role of monitoring marine litter on the seabed.

Murine Mammary Carcinoma Induces Chronic Systemic Inflammation and Immunosuppression in BALB/c Mice.

PURPOSE: The F3II cell line is a highly invasive variant of mammary carcinoma. Although it is frequently used as a model to evaluate the efficacy of immunotherapy, its impact on the immune system remains poorly understood. The main objectives of this study were to evaluate the effects of F3II tumors on the development of chronic inflammation and to characterize tumor-associated immunosuppression. METHODS: Following the experimental implantation of F3II tumors in BALB/c mice, alterations in the liver and spleen anatomy and the numbers of circulating leukocytes, myeloid-derived suppressor cells (MDSCs), and regulatory T cells were measured using hematological techniques, histopathological analysis, and flow cytometry. The capacity of the F3II tumor-bearing mice to reject MB16F10 allogeneic tumor transplantation was also evaluated. In addition, the restoration of immune parameters in tumor-bearing mice was evaluated after standard breast cancer chemotherapy and surgical tumor excision. RESULTS: F3II tumor implantation increased the levels of chronic inflammatory markers, such as the neutrophil-to-lymphocyte and platelet-to-lymphocyte ratios, and caused myeloid alterations, including extramedullary granulopoiesis and megakaryopoiesis, along with the recruitment of MDSCs to the spleen. Chemotherapy or surgical F3II tumor removal completely rescued the tumor-associated extramedullary granulopoiesis and megakaryopoiesis. Notably, the presence of F3II tumors reduced the capacity of BALB/c mice to reject MB16F10 allogeneic tumor transplantation. CONCLUSION: These results support the occurrence of F3II tumor-mediated immune cell dysfunction, which mimics the immune alterations characterized by chronic systemic inflammation and immunosuppression observed in breast cancer in clinical settings. Thus, the F3II tumor model is relevant for evaluating novel breast cancer immunotherapies and combinations in preclinical studies. This model could also be useful for identifying appropriate therapeutic targets and developing proof-of-concept experiments in the future.

Safety, immunogenicity and preliminary efficacy of multiple-site vaccination with an Epidermal Growth Factor (EGF) based cancer vaccine in advanced non small cell lung cancer (NSCLC) patients.

The prognosis of patients with advanced non small cell lung (NSCLC) cancer remains dismal. Epidermal Growth Factor Receptor is over-expressed in many epithelial derived tumors and its role in the development and progression of NSCLC is widely documented. CimaVax-EGF is a therapeutic cancer vaccine composed by human recombinant Epidermal Growth Factor (EGF) conjugated to a carrier protein, P64K from Neisseria Meningitides. The vaccine is intended to induce antibodies against self EGF that would block EGF-EGFR interaction. CimaVax-EGF has been evaluated so far in more than 1000 advanced NSCLC patients, as second line therapy. Two separate studies were compared to assess the impact of high dose vaccination at multiple anatomic sites in terms of immunogenicity, safety and preliminary efficacy in stage IIIb/IV NSCLC patients. In both clinical trials, patients started vaccination 1 month after finishing first line chemotherapy. Vaccination at 4 sites with 2.4 mg of EGF (high dose) was very safe. The most frequent adverse events were grade 1 or 2 injection site reactions, fever, headache and vomiting. Patients had a trend toward higher antibody response. The percent of very good responders significantly augmented and there was a faster decrease of circulating EGF. All vaccinated patients and those classified as good responders immunized with high dose at 4 sites, had a large tendency to improved survival.

Breaking and restoring immune tolerance to pancreatic beta-cells in type 1 diabetes.

PURPOSE OF REVIEW: Type 1 diabetes (T1D) results from the loss of immune tolerance to pancreatic beta-cells leading to their destruction. Immune intervention therapies tested in T1D so far delayed progression but failed to restore tolerance, which partly explains their lack of durable clinical efficacy. RECENT FINDINGS: The role of beta-cells and islets themselves in dialogue with their micro- and macro-environment including the immune system and the intestinal microbiome is increasingly evident. Indeed, islets can both maintain and break immune tolerance. Some recent immune therapies in cancer that block immune regulation also break tolerance. Induction of immune tolerance requires activating immune activation too, whereas immune suppression precludes this process. Immunotherapy alone my not suffice without engaging islets to restore tolerance and preserve beta-cell function. SUMMARY: New insight into the role of islet tissue and its interaction with its environment in preserving or breaking tolerance has contributed to understand the development of islet autoimmunity and T1D. Knowing which factors in islets and the immune system contribute to maintaining, breaking, and restoring the balance in the immune system is critical to prevent initiation and reverse disease progression, and guides the design of novel tolerogenic strategies for durable therapeutic intervention and remission that target both the immune system and distressed islets.

Aberrant expression of CD6 on B-cell subsets from patients with Sjögren's syndrome.

CD6 is one of a pair of related genes encoding CD5-associated receptors on all T cells and a subset of B cells. The current availability of "T1h", a humanized anti-CD6 monoclonal antibody for B cell-mediated autoimmune disorders revives analysis of the B-cell subset expression of CD6, particularly in primary Sjögren's syndrome (SS). Refined phenotype of B-lymphocytes peripheral blood (PB), bone marrow and tonsils revealed that the overlap between the expression of CD6 is less close to that of CD5 than currently acknowledged. In contrast to CD5, CD6 is absent on transitional B cells, while present on mature and memory B cells. Interestingly, the PB proportion of CD6(+) B cells is decreased in patients with primary SS, as opposed to those with rheumatoid arthritis. The reduction in primary SS does not result from the shedding of CD6 from the membrane of B cells, but from the lowering of memory B lymphocytes. It may result from the ability of CD6 to make transmigration of CD27(+) memory B cells into the salivary glands (SGs) easier. Consistent with this view is our finding that CD166 (one of the ligands for CD6) is highly expressed on epithelial cells of patients' SGs. This study is relevant in that the humanized T1h anti-CD6 becomes an alternative to anti-CD20 for treatment of primary SS.

Anticoccidial activity of maslinic acid against infection with Eimeria tenella in chickens.

We propose maslinic acid (2-alpha, 3-beta-dihydroxiolean-12-en-28-oic acid), found in the leaves and fruit of the olive tree (Olea europaea L.), as a new natural coccidiostatic product against Eimeria tenella. Its action in infected animals has been compared with animals treated with sodium salinomycin. The lesion index (LS), the oocyst index (OI) and the anticoccidial index (ACI) were studied with regard to the weight of the chicks. The ACI for maslinic acid was 210.27 and for sodium salinomycin 173.09. Similarly, both LS and OI decreased in the groups treated with maslinic acid. A considerable increase in weight was found in the chicks treated with maslinic acid compared with those in the control group. Histopathological studies of the caecum at 120 h post-infection showed that the infection rate decreased significantly in chicks treated with maslinic acid.

Salivary Alpha-Amylase Mediates the Increase in Hunger Levels in Adolescents with Excess Weight after Viewing Food Images.

Background: Salivary alpha-amylase (sAA) initiates the digestion process in the mouth and its levels might influence feelings of hunger and the propensity toward obesity. This study aims to evaluate basal differences in sAA between adolescents with excess weight (EW) and normal weight (NW), and the associations between sAA levels and feelings of hunger after viewing food images. Methods: Adolescents (13-18 years old) classified as EW (n = 30) or NW (n = 30) participated in the study. Saliva samples were collected before the administration of a food-choice task. Hunger was evaluated before and after the food-choice task. Results: EW adolescents showed lower basal sAA levels than NW adolescents and a greater increase in hunger levels after viewing food images. In addition, sAA levels had a significant inverse relationship with the increase in hunger in EW adolescents, but not in NW adolescents. Finally, significant inverse associations between sAA, BMI, and body fat percentage were found. Conclusions: Levels of hunger and changes therein, after viewing food are dependent on sAA levels in EW adolescents. This finding indicates that sAA levels may be a mediator of feelings of hunger in individuals with overweight in the context of viewing food cues, suggesting the utility of the sAA enzyme as a marker of hunger and propensity toward obesity.

[Assessment of stress in childhood: Children's Daily Stress Inventory (Inventario Infantil de Estresores Cotidiano, IIEC)]. / Evaluación del estrés infantil: Inventario Infantil de Estresores Cotidianos (IIEC).

The present study introduces the Children's Daily Stress Inventory (Inventario Infantil de Estresores Cotidianos, IIEC) as a measure that assesses daily stress in primary school children. The inventory was applied to a sample of 1094 primary school students. The final version includes 25 dichotomic items covering the areas of health, school/peers, and family. The score is obtained by adding the total of positive answers. Analyses of items, reliability and several external pieces of evidence of validity based on relations with other variables are presented. The results show adequate psychometric properties for the assessment of daily stress in children.

Assessment of silt from sand and gravel processing as a suitable sub-soil material in land restoration: A glasshouse study.

Annually, sand and gravel processing generates approximately 20 million tonnes of non-commercial by-product as fine silt particles (<63 µm) which constitutes approximately 20% of quarry production in the UK. This study is significant as it investigated the use of quarry silt as a sub-soil medium to partially substitute soil-forming materials whilst facilitating successful post-restoration crop establishment. In a glasshouse pot experiment, top-soil and sub-soil layering was simulated, generating an artificial sub-soil medium by mixing two quarry non-commercial by-products, i.e. silt and overburden. These were blended in three ratios (100:0, 70:30, 50:50). Pots were packed to two bulk densities (1.3 and 1.5 g cm-3) and sown with three cover crops used in the early restoration process namely winter rye (Secale cereale), white mustard (Sinapis alba) and a grassland seed mixture (Lolium perenne, Phleum pratense, Poa pratensis, Festuca rubra). Three weeks into growth, the first signs of nitrogen (N) deficiency were observed in mustard plants, with phosphorus (P) and potassium (K) deficiencies observed at 35 days. Rye exhibited minor N deficiency symptoms four weeks into growth, whilst the grassland mixture showed no deficiency symptoms. The 70:30 silt:overburden sub-soil blend resulted in significantly higher Root Mass Densities of grassland seed mixture and rye in the sub-soil layer as compared with the other blends. The innovation in this work is the detailed physical, chemical and biological characterisation of silt:overburden blends and effects on root development of plants commonly used in early restoration to bio-engineer soil structural improvements.

Correction: T cell activation and differentiation is modulated by a CD6 domain 1 antibody Itolizumab.

[This corrects the article DOI: 10.1371/journal.pone.0180088.].