Specific B-cell subsets can regulate T-cell immune responses, and are termed regulatory B cells (Breg). The majority of Breg cells described in mouse and man have been identified by IL-10 production and are known to suppress allergy and autoimmunity. However, Breg cell mediated immune suppression, independent of IL-10, also occurs. Here we show that Breg cells play a critical role in regulating humoral immunity mediated by CD4 þ CXCR5 þ PD-1 þ follicular helper T cells, and can suppress inflammation in autoimmune disease through elevated expression of PD-L1. We have also identified that these B cells are resistant to aCD20 B-cell depletion. This work describes how Breg cells are critical in humoral homoeostasis and may have implications for the regulation of autoimmune diseases.

Ion-sensitive responsive polymers are prepared under fully aqueous conditions using controlled radical polymerization. Variations in comonomer content and sequence lead to temperature and salt-dependent solution behavior, with cloud-points ranging by +/-40 degrees C following addition of Hofmeister series salts. A "hybrid" block copolymer, composed of a statistical sequence of monomers tipped with a hydrophilic block, formed stable micelle-like assemblies that exhibited burst release of an encapsulated model drug in response to addition of a kosmotrope, Na2SO4, at room temperature.

ILC2s interact with CD4 T cells during immune responses against parasitic helminths. Schwartz et al. describe that PD-L1–expressing pulmonary ILC2s stimulate CD4 T cells via PD-1 to up-regulate the type 2 master transcription factor GATA3 and thereby promote IL-13 production from Th2 cells.

BackgroundNeuroblastoma is responsible for 15% of all childhood cancer deaths. Despite advances in treatment and disease management, the overall 5-year survival rates remain poor in high-risk disease (25-40%). MiR-497 was previously identified by our laboratory as a member of a miRNA expression signature, predictive of neuroblastoma patient survival and has been reported as a tumor suppressor in a variety of other cancers. WEE1, a tyrosine kinase regulator of the cell cycle and predicted target of miR-497, has emerged as an oncogene in several cancer types and therefore represents an attractive potential target for novel therapy approaches in high-risk neuroblastoma. Our aim was to investigate the potential tumor suppressive role of miR-497 in high-risk neuroblastoma.MethodsExpression levels of miR-497 and WEE1 in tissues and cells were determined using RT-PCR. The effect of miR-497 and siWEE1 on cell viability was evaluated using MTS assays, apoptosis levels were determined using FACS analysis of Annexin V/PI stained cells, and target protein expression was determined using western blot. Luciferase reporter plasmids were constructed to confirm direct targeting. Results were reported as mean±S.E.M and differences were tested for significance using 2-tailed Students t-test.ResultsWe determined that miR-497 expression was significantly lower in high-risk MYCN amplified (MNA) tumors and that low miR-497 expression was associated with worse EFS and OS in our cohort. Over-expression of miR-497 reduced cell viability and increased apoptosis in MNA cells. We identified WEE1 as a novel target for miR-497 in neuroblastoma. Furthermore, our analysis showed that high WEE1 levels are significantly associated with poor EFS and OS in neuroblastoma and that siRNA knockdown of WEE1 in MNA cell lines results in significant levels of apoptosis, supporting an oncogenic role of WEE1 in neuroblastoma. Cisplatin (CDDP) treatment of both miR-497 over-expressing cells and WEE1 inhibited cells, resulted in a significant increase in apoptosis in MNA cells, describing a synergistic effect and therefore a potential therapeutic for high-risk neuroblastoma.ConclusionOur study’s results are consistent with miR-497 being a candidate tumor suppressor in neuroblastoma, through the direct targeting of WEE1. These findings re-enforce the proposal of WEE1 as a therapeutic target in neuroblastoma.

The use of live helminth infections is currently in clinical trials as a novel approach for the treatment of a range of allergic and autoimmune diseases. This rapid progression from observational studies some 20 years ago to helminth clinical trials can be attributed to huge advances in not just pre-clinical and clinical evidence, pertaining to the efficacy of these parasites in unrelated diseases, but also a greater understanding of the complex immunological mechanisms that underpin these effects. Helminths have exerted significant evolutionary selective pressures on the host immune genome or "immunome". Studies on helminths were pivotal in a paradigm shift in immunology with recent discoveries of a number of novel immune cell populations. Critically, these new discoveries highlight the need to further understand the underlying mechanism behind the desirable therapeutic effects that helminths offer. With these unknown unknowns there is the distinct possibility that a true, fundamental modus operandi for helminth therapy will arrive long after it has been established in the clinic.

Aims/hypothesis The aim of this study was to determine whether social deprivation in the presence of diabetes is an independent predictor of developing a foot ulcer and separately of mortality. Methods This was a primary-care-based retrospective analysis of 13,955 adults with type 1 (n = 1370) or type 2 (n = 12,585) diabetes after a median follow-up of 10.5 years. Demographic characteristics, indices of social deprivation and clinical variables were assessed at baseline. The primary outcomes were new foot ulceration (in those without a previous history of foot ulcers) and all-cause mortality. Cox proportional hazard models were used to describe the associations among foot ulceration, social deprivation and mortality. Results The mean age of the population was 69.4 (range: 16-89) years. The incidence of foot ulceration was greater in individuals with type 2 (8.6%) compared with type 1 diabetes (4.8%). Occurrence was similar by sex, but increased with age and deprivation index. Individuals in the highest quintile of deprivation were 77% more likely to develop a foot ulcer compared with those in the lowest quintile (OR 1.77 [95% CI 1.45, 2.14], p < 0.0001). Overall, 2946 (21.1%) deaths were recorded. Compared with individuals without a foot ulcer, the development of a foot ulcer was associated with a higher age-and sex-adjusted mortality rate (25.9% vs 14.0%), and a 72% (HR Conclusions/interpretation This study confirms the high mortality rate in individuals with diabetes-related foot ulcers. In addition, socioeconomic disadvantage was found to be an independent effect modifier, contributing to an increased burden of mortality in people with diabetes who develop foot ulceration. In light of this, and as diabetes service configurations are orientated for the next 5-10 years, modelling of foot ulceration risk needs to take socioeconomic disadvantage into account.

Eur. J. Immunol. 2015Immunol. . 45: 1842Immunol. -1854 Leukocyte signaling 1843 IntroductionA subset of B cells that has garnered intense interest are regulatory B (Breg) cells [1][2][3][4]. These include B cells that produce the anti-inflammatory cytokine IL-10, termed B10 cells [5][6][7], including murine CD19 + CD1d hi CD5 + B cells [8,9]. However, there is still no universally recognized phenotype for these cells. In naïve mice, Breg cells that secrete IL-10 exist as no more than 1-2% of all B220 + cells [8,10] with such IL-10-secreting B cells potently suppressing inflammation in several murine disease models [10][11][12][13][14] and have been shown to have effects in man [14]. Specific factors and mechanisms that induce or regulate Breg cells in vivo have recently been explored. We have previously demonstrated that helminths can expand a Breg-cell population that can suppress in allergic inflammatory conditions in mice in an IL-10-dependent manner [10,11]. Given that helminths can generate IL-10-producing Breg cells in both mouse and man [10,11,15], it is important to understand and define noninfective mechanisms by which to generate such cells both in vitro and in vivo [9]. Paradigm shifts in the role of B cells have described their effects in both innate and adaptive arms of the immune response [2,[16][17][18]. In particular, engagement of Toll-like receptors (TLR) on B cells can produce a plethora of cytokine responses [18].In order to gain further insight into the function of IL-10-secreting B cells in mice, we undertook RNA microarray analysis of Breg cells from helminth-infected mice and compared its gene expression profile with that of B cells from both infected and uninfected animals. On analysis, we discovered that Breg cells from Schistosoma mansoni infected mice contained upregulated expression of genes associated with pattern recognition of bacteria and viruses. Of particular significance was the elevated expression of the endosomal-resident TLR, Tlr7.Previously, we have shown that helminth-derived, IL-10-producing B cells can mediate airway hyperresponsiveness (AHR) through the induction of CD4 + FoxP3 + regulatory T (Treg) cells [10]. Here, we now demonstrate that TLR7 is expressed on CD19 + CD1d hi B cells and that agonism of this receptor can induce IL-10 production, both in vitro and in vivo. Furthermore, using adoptive transfer strategies we describe how these TLR7-elicited CD19 + CD1d hi B cells facilitate the suppression of AHR in mice through the induction of lung-resident Treg cells. We also indicate that these ligands can generate IL-10-secreting B cells in man, thus supplementing the current evidence base that TLR7 has the potential to be a therapeutic target in respiratory disease and providing novel insight into IL-10-producing Breg cell function. ResultsPreferential upregulation of Tlr7 on CD19 + CD1d hi B cells determined using helminth-infected mice Splenic CD19 + CD1d hi B cells are expanded in mice infected with S. mansoni [19]. These helminth-generated B cells have similar...

BackgroundMetformin is the most common oral hypoglycemic used and associated with certain abnormalities. The objective was to evaluate and define the occurrence and bases of vitamin B12 deficiency amongst patients on Metformin for diabetes mellitus type II.MethodsA cross-sectional study was conducted on 209 patients having diabetes type II between January-December 2016. The patients aged > 45 years and who had taken metformin for at least three months were recruited with regular follow-up at the Endocrinology Unit of Hayatabad Medical Complex and Diabetic Center Hayatabad, Peshawar. The patients were included in a survey after which they had their serum B12 levels measured. Serum B12 levels < 150 pg/ml is defined as the B12 deficiency.ResultsAbout 29.66% of diabetic patients had confirmed the B12 insufficiency through laboratory tests. The patients on metformin had statistically lower values of B12 (P = 0.01). For the patients who smoked, vitamin B12 deficiency was significantly higher than those who did not smoke (p= <0.001). Also in patients using multivitamins, vitamin B12 deficiency was lower compared to nonusers (p=0.05).ConclusionOur study shows that for the patients with type 2 diabetes (T2DM), long-term treatment with metformin and smoking are associated with higher chances of developing vitamin B12 deficiency. Clinicians should, therefore, recognize this significant element and should screen diabetics who are on metformin treatment for any B12 insufficiency, which may be hidden, especially patients coming with neurologic symptoms. Additionally, multi vitamins taken daily may have a protective role.