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Osteosarcoma stem cells (OSCs) contribute to the pathogenesis of osteosarcoma (OS), which is the most common malignant primary bone tumor. The significance and underlying mechanisms of action of proteoglycans (PGs) and glycosaminoglycans (GAGs) in OSC phenotypes and OS malignancy are largely unknown. This study aimed to investigate the role of PG/GAG biosynthesis and the corresponding candidate genes in OSCs and poor clinical outcomes in OS using scRNA-seq and bulk RNA-seq datasets of clinical OS specimens, accompanied by biological validation by in vitro genetic and pharmacological analyses. The expression of b-1,3-glucuronyltransferase 3 (B3GAT3), one of the genes responsible for the biosynthesis of the common core tetrasaccharide linker region of PGs, was significantly upregulated in both OSC populations and OS tissues and was associatedwith poor survival in patientswithOSwith high stemcell properties. Moreover, the genetic inactivation of B3GAT3 by RNA interference and pharmacological inhibition of PGbiosynthesis abrogated the self-renewal potential of OSCs. Collectively, these findings suggest a pivotal role for B3GAT3 and PG/GAG biosynthesis in the regulation of OSC phenotypes and OS malignancy, thereby providing a potential target for OSC-directed therapy. [ABSTRACT FROM AUTHOR]

T and B lymphocytes are crucial players in cellular and humoral immune responses. The development, activation and differentiation of T and B lymphocytes are regulated by the best characterized PI3K‐PI (3,4,5) P3‐AKT phosphoinositide signalling pathway. As a branch of the phosphoinositide signalling pathway, the lipid phosphatase INPP4B inhibits AKT activation through degrading the phosphoinositide signalling messenger PI (3,4) P2. However, the role of Inpp4b in T and B lymphocytes remains elusive. Here, we reported that Inpp4b was highly expressed in human and murine T‐ and B‐1 lymphocytes. Despite its higher expression in T lymphocytes, neither T cell development and homeostasis nor in vitro T cell activation and CD4+ T cell differentiation were altered upon loss of Inpp4b. Interestingly, combined direct phenotype analysis of Inpp4b conventional knockout mice and adoptive transfer studies revealed that ablation of Inpp4b intrinsically reduced peritoneal B‐1 cells rather B‐2 cells. Moreover, Inpp4b deficiency led to impaired thymus independent (TI) and thymus dependent (TD) antigens‐induced antibody production. Further in vitro analysis revealed that CD40‐mediated B cell proliferation was impaired upon ablation of Inpp4b. Our findings reveal that Inpp4b is required in regulating B‐1 cell numbers and B cell‐mediated antibody production. [ABSTRACT FROM AUTHOR]

Gray mold, caused by Botrytis cinerea, is one of the most devastating agricultural diseases during the pre and postharvest stages. This study aims to investigate the application of Bacillus subtilis as a bioagent against B. cinerea and reducing pre and postharvest disease as a replacement method to chemical pesticides. Five Botrytis isolates were molecularly confirmed and the virulent isolate BC2 was registered for the first time in the National Center of Biotechnology Information (NCBI) with accession number OP257237. The mycelial growth of B. cinerea was inhibited by up to 84% when treated with the bio agent bacterium using the double drawing method in Petri plates. Pre-harvest application of B. subtilis at a concentration of 108/ml CFU reduced postharvest -gray mold of tomato fruit during ripening stages in the greenhouse. Preharvest application of bioagent bacterium B. subtilis induced the levels of B-1, 3-glucanase and Polyphenol oxidase, indicating enhanced plant resistance to gray mold. Disease severity was decreased compared with control and extended the shelf life at 25 °C.The field application of the bio agent bacterium at the maturity of ripening in preharvest had a significant effect on increasing the level of polyphenol oxidase PPO and B-1, 3-glucanase, which are related to the enhancement of plant defenses and resistance. Additionally, it led to an increase in storage life and a reduction in post-harvest losses. The results demonstrated that preharvest application of bio agent bacterium Bacillus subtilis improved the quality of postharvest tomato fruit and significantly reduced gray mold incidence, all without the use of chemical pesticides. [ABSTRACT FROM AUTHOR]

B-1 lymphocytes exhibit specialized roles in host defense against multiple pathogens. Despite the fact that CD19+CD93+B220lo/- B cells have been identified as B-1 progenitors, the definition for B-1 progenitors remains to be elucidated as CD19+CD93+B220+ B cells are capable to give rise to B-1 cells. Given that transcription factor Bhlhe41 is highly and preferentially expressed in B-1 cells and regulates B-1a cell development, we generated a transgenic mouse model, Bhlhe41dTomato-Cre, for fate mapping and functional analysis of B-1 cells. Bhlhe41dTomato-Cre mice efficiently traced Bhlhe41 expression, which was mainly restricted to B-1 cells in B-cell lineage. We showed an efficient and specific Cremediated DNA recombination in adult B-1 cells and neonatal B-1 progenitors rather than B-2 cells by flow cytometric analysis of Bhlhe41dTomato-Cre/+Rosa26EYFP mice. Treatment of Bhlhe41dTomato-Cre/+Rosa26iDTR mice with diphtheria toxin revealed a robust efficacy of B-1 cell depletion. Interestingly, using Bhlhe41dTomato-Cre mice, we demonstrated that neonatal B-1 progenitors (CD19+CD93+B220lo/-) expressed Bhlhe41 andwere identical to well-defined transitional B-1a progenitors (CD19+CD93+B220lo/-CD5+), which only gave rise to peritoneal B-1a cells. Moreover, we identified a novel population of neonatal splenic CD19hidTomato+B220hiCD43loCD5lo B cells, which differentiated to peritoneal B-1a and B-1b cells. Bhlhe41 deficiency impaired the balance between CD19hidTomato+B220lo/-CD5hi and CD19hidTomato+B220hiCD5lo cells. Hence, we identified neonatal CD19hidTomato+B220hiCD43loCD5lo B cells as novel transitional B-1 progenitors. Bhlhe41dTomato-Cre/+ mouse can be used for fate mapping and functional studies of B-1 cells in host-immune responses. [ABSTRACT FROM AUTHOR]

B-1 lymphocytes exhibit specialized roles in host defense against multiple pathogens. Despite the fact that CD19+CD93+B220lo/- B cells have been identified as B-1 progenitors, the definition for B-1 progenitors remains to be elucidated as CD19+CD93+B220+ B cells are capable to give rise to B-1 cells. Given that transcription factor Bhlhe41 is highly and preferentially expressed in B-1 cells and regulates B-1a cell development, we generated a transgenic mouse model, Bhlhe41dTomato-Cre, for fate mapping and functional analysis of B-1 cells. Bhlhe41dTomato-Cre mice efficiently traced Bhlhe41 expression, which was mainly restricted to B-1 cells in B-cell lineage. We showed an efficient and specific Cre-mediated DNA recombination in adult B-1 cells and neonatal B-1 progenitors rather than B-2 cells by flow cytometric analysis of Bhlhe41dTomato-Cre/+Rosa26EYFP mice. Treatment of Bhlhe41dTomato-Cre/+Rosa26iDTR mice with diphtheria toxin revealed a robust efficacy of B-1 cell depletion. Interestingly, using Bhlhe41dTomato-Cre mice, we demonstrated that neonatal B-1 progenitors (CD19+CD93+B220lo/-) expressed Bhlhe41 and were identical to well-defined transitional B-1a progenitors (CD19+CD93+B220lo/-CD5+), which only gave rise to peritoneal B-1a cells. Moreover, we identified a novel population of neonatal splenic CD19hidTomato+B220hiCD43loCD5lo B cells, which differentiated to peritoneal B-1a and B-1b cells. Bhlhe41 deficiency impaired the balance between CD19hidTomato+B220lo/-CD5hi and CD19hidTomato+B220hiCD5lo cells. Hence, we identified neonatal CD19hidTomato+B220hiCD43loCD5lo B cells as novel transitional B-1 progenitors. Bhlhe41dTomato-Cre/+ mouse can be used for fate mapping and functional studies of B-1 cells in host-immune responses.

B cell receptor signaling and downstream NF-κB activity are crucial for the maturation and functionality of all major B cell subsets, yet the molecular players in these signaling events are not fully understood. Here we use several genetically modified mouse models to demonstrate that expression of the multifunctional BRCT (BRCA1 C-terminal) domain-containing PTIP (Pax transactivation domain-interacting protein) chromatin regulator is controlled by B cell activation and potentiates steady-state and postimmune antibody production in vivo. By examining the effects of PTIP deficiency in mice at various ages during ontogeny, we demonstrate that PTIP promotes bone marrow B cell development as well as the neonatal establishment and subsequent long-term maintenance of self-reactive B-1 B cells. Furthermore, we find that PTIP is required for B cell receptor and T:B interaction-induced proliferation, differentiation of follicular B cells during germinal center formation, and normal signaling through the classical NF-κB pathway. Together with the previously identified role for PTIP in promoting sterile transcription at the Igh locus, the present results establish PTIP as a licensing factor for humoral immunity that acts at several junctures of B lineage maturation and effector cell differentiation by controlling B cell activation. [ABSTRACT FROM AUTHOR]

In the developing mouse embryo, there are several waves of hematopoiesis. Primitive and definitive erythromyeloid lineages appear prior to hematopoietic stem cell (HSC) emergence, and these waves are considered to be transient and support embryonic homeostasis until HSC-derived hematopoiesis is established. However, recent evidence strongly suggests that HSC-independent immune cells, such as tissue macrophages and some innate lymphoid cells, develop in the mouse embryo and persist into postnatal life. Innate type B-1 cells have also been reported to emerge from hemogenic endothelial cells in the extraembryonic yolk sac and para-aortic splanchnopleura, and continue to develop in the fetal liver, even in HSC-deficient mouse embryos. Here, this review discusses B-1 cell development in the context of the layered immune system hypothesis of B lymphopoiesis and the emergence of B-1 cells independent of HSCs. [ABSTRACT FROM AUTHOR]

B-1 lymphocytes produce natural immunoglobulin ( Ig)M, among which a large proportion is directed against apoptotic cells and altered self-antigens, such as modified low-density lipoprotein ( LDL). Thereby, natural IgM maintains homeostasis in the body and is also protective against atherosclerosis. Diabetic patients have an increased risk of developing certain infections as well as atherosclerosis compared with healthy subjects, but the underlying reason is not known. The aim of this study was to investigate whether diabetes and insulin resistance affects B-1 lymphocytes and their production of natural IgM. We found that diabetic db/db mice had lower levels of peritoneal B-1a cells in the steady state-condition compared to controls. Also, activation of B-1 cells with the Toll-like receptor ( TLR)-4 agonist Kdo2- Lipid A or immunization against Streptococcus pneumoniae led to a blunted IgM response in the diabetic db/db mice. In-vitro experiments with isolated B-1 cells showed that high concentrations of glucose, but not insulin or leptin, caused a reduced secretion of total IgM and copper-oxidized ( CuOx)- LDL- and malondialdehyde ( MDA)- LDL-specific IgM from B-1 cells in addition to a decreased differentiation into antibody-producing cells, proliferation arrest and increased apoptosis. These results suggest that metabolic regulation of B-1 cells is of importance for the understanding of the role of this cell type in life-style-related conditions. [ABSTRACT FROM AUTHOR]

B-1 cells play critical roles in defending against microbial invasion and in housekeeping removal of cellular debris. B-1 cells secrete natural antibody and manifest functions that influence T cell expansion and differentiation and in these and other ways differ from conventional B-2 cells. B-1 cells were originally studied in mice where they are easily distinguished from B-2 cells, but their identity in the human system remained poorly defined for many years. Recently, functional criteria for human B-1 cells were established on the basis of murine findings, and reverse engineering resulted in identification of the phenotypic profile, CD20+CD27+CD43+CD70−, for B-1 cells found in both umbilical cord blood and adult peripheral blood. Human B-1 cells may contribute to multiple disease states through production of autoantibody and stimulation/modulation of T cell activity. Human B-1 cells could be a rich source of antibodies useful in treating diseases present in elderly populations where natural antibody protection may have eroded. Manipulation of human B-1 cell numbers and/or activity may be a new avenue for altering T cell function and treating immune dyscrasias. [ABSTRACT FROM AUTHOR]

Banana fruit allergy is well known, but neither immunoglobulin E recognition patterns to purified plant food allergens nor true prevalences of putative banana allergens have been established. This study aimed to characterize β-1,3-glucanase and thaumatin-like protein (TLP) as banana allergens, testing them, together with other plant food allergens, in 51 children with allergic reactions after banana ingestion and both positive specific IgE and skin prick test (SPT) to banana. Banana β-1,3-glucanase and TLP were isolated and characterized. Both banana allergens, together with kiwifruit TLP Act d 2, avocado class I chitinase Pers a 1, palm pollen profilin Pho d 2 and peach fruit lipid transfer protein (LTP) Pru p 3, were tested by in vitro and in vivo assays. Banana β-1,3-glucanase (Mus a 5) was glycosylated, whereas banana TLP (Mus a 4) was not, in contrast with its homologous kiwi allergen Act d 2. Specific IgE to both banana allergens, as well as to peach Pru p 3, was found in over 70% of sera from banana-allergic children, and Mus a 4 and Pru p 3 provoked positive SPT responses in 6 of the 12 tested patients, whereas Mus a 5 in only one of them. Both peptidic epitopes and cross-reactive carbohydrate determinants were involved in the IgE-binding to Mus a 5, whereas cross-reactivity between Mus a 4 and Act d 2 was only based on common IgE protein epitopes. Profilin Pho d 2 elicited a relevant proportion of positive responses on in vitro (41%) and in vivo (58%) tests. Therefore, Mus a 4 and LTP behave as major banana allergens in the study population, and profilin seems to be also a relevant allergen. Mus a 5 is an equivocal allergenic protein, showing high IgE-binding to its attached complex glycan, and low in vivo potency. [ABSTRACT FROM AUTHOR]

Aims: To elucidate the roles of the β-1,3-endoglucanase EngA in autolysis of the filamentous fungus Aspergillus nidulans and to identify the common regulatory elements of autolytic hydrolases. Methods and Results: A β-1,3-endoglucanase was purified from carbon-starving cultures of A. nidulans. This enzyme is found to be encoded by the engA gene (locus ID: AN0472.3). Functional and gene-expression studies demonstrated that EngA is involved in the autolytic cell wall degradation resulting from carbon starvation of the fungus. Moreover, regulation of engA is found to be dependent on the FluG/BrlA asexual sporulation signalling pathway in submerged culture. The deletion of either engA or chiB (encoding an endochitinase) caused highly reduced production of hydrolases in general. Conclusions: The β-1,3-endoglucanase EngA plays a pivotal role in fungal autolysis, and activities of both EngA and ChiB are necessary to orchestrate the expression of autolytic hydrolases. The production of cell wall-degrading enzymes was coordinately controlled in a highly sophisticated and complex manner. Significance and Impact of the Study: No information was available on the autolytic glucanase(s) of the euascomycete A. nidulans. This study demonstrates that EngA is a key element in fungal autolysis, and normal activities of both EngA and ChiB are crucial for balanced production of hydrolases. [ABSTRACT FROM AUTHOR]

β-aminobutyric acid (BABA) was assessed for the ability to protect two artichoke cultivars, C3 and Exploter, against white mould caused by Sclerotinia sclerotiorum, which represents a major problem in the cultivation of this crop in many growing areas of Central Italy. Changes in the activity and isoenzymatic profiles of the pathogenesis-related (PR) proteins β-1,3-glucanase, chitinase and peroxidase in plantlets upon BABA treatment and following inoculation of the pathogen in plantlets and leaves detached from adult plants were also investigated as molecular markers of induced resistance and priming. BABA treatments by soil drenching induced a high level of resistance against S. sclerotiorum in artichoke plantlets of both cultivars C3 and Exploter with a similar level of protection and determined a consistent increase in peroxidase activity paralleled with the differential induction of alkaline isoenzyme with a pI 8.6. A consistent change was found in Exploter in the peroxidase activity following BABA treatments and pathogen inoculation and was paralleled with the expression of an anionic band in plantlets and both anionic and cationic bands in leaves. Our results showed a correlation between BABA-induced resistance (BABA-IR) and a augmented capacity to express basal defence responses, more pronounced in cultivar C3 and associated β-1,3-glucanase accumulation in both plantlets and leaves inoculated with the pathogen, whereas chitinase resulted affected only at plantlet stage. The present results represent the first one showing the effect of BABA in inducing resistance in artichoke and associated accumulation of selected PRs. If confirmed in field tests, the use of BABA at early plant stages may represent a promising approach to the control soilborne pathogens, such as the early infection of S. sclerotiorum. [ABSTRACT FROM AUTHOR]

A wheat β-1,3-glucanase gene ( TaGluD) was identified as a fungal defense candidate. Its transcript induction was more than 60-fold higher in a resistant wheat line, Shannong0431, than in a susceptible wheat line, Wenmai6, after infection with Rhizoctonia cerealis. The TaGluD protein was overexpressed as inclusion bodies in Escherichia coli. After refolding and purification, TaGluD with 1 unit of β-1,3-glucanase showed antifungal activity in vitro against Rhizoctonia solani, R. cerealis, Phytophthora capsici and Alternaria longipes with inhibition rates of 42%, 43%, 32% and 30%, respectively. Thus TaGluD may be useful for enhancing fungal resistance in several crop species. [ABSTRACT FROM AUTHOR]

Previously, we isolated a strain of Bacillus that had antifungal activity and produced lytic enzymes with fungicidal potential. In the present study, we identified the bacterium as Paenibacillus ehimensis and further explored its antifungal properties. In liquid co-cultivation assays, P. ehimensis IB-X-b decreased biomass production of several pathogenic fungi by 45%-75%. The inhibition was accompanied by degradation of fungal cell walls and alterations in hyphal morphology. Residual medium from cultures of P. ehimensis IB-X-b inhibited fungal growth, indicating the inhibitors were secreted into the medium. Of the 2 major lytic enzymes, chitinases were only induced by chitin-containing substrates, whereas β-1,3-glucanase showed steady levels in all carbon sources. Both purified chitinase and β-1,3-glucanase degraded cell walls of macerated fungal mycelia, whereas only the latter also degraded cell walls of intact mycelia. The results indicate synergism between the antifungal action mechanisms of these enzymes in which β-1,3-glucanase is the initiator of the cell wall hydrolysis, whereas the degradation process is reinforced by chitinases. Paenibacillus ehimensis IB-X-b has pronounced antifungal activity with a wide range of fungi and has potential as a biological control agent against plant pathogenic fungi. Nous avons précédemment isolé une souche de Bacillus qui possédait une activité antifongique et qui produisait des enzymes lytiques à potentiel fongicide. Dans la présente étude, nous avons identifié la bactérie comme Paenibacillus ehimensis et nous avons exploré davantage ses propriétés antifongiques. Lors d’essais de co-culture en milieu liquide, la présence de P. ehmensis IB-X-b diminuait la production de biomasse de champignons pathogènes de 45 % à 75 %. L’inhibition était accompagnée par la dégradation de la paroi cellulaire des champignons et modifiait la morphologie des hyphes. Le milieu de culture résiduel de P. ehmensis IB-X-b inhibait la croissance des champignons, ce qui signifie que des inhibiteurs étaient secrétés dans le milieu. Parmi les deux enzymes lytiques majeures, les chitinases n’étaient induites que par des substrats contenant de la chitine, alors que la β-1,3-glucanase était exprimée à des niveaux constants sur toutes les sources de carbone. La chitinase et la β-1,3-glucanase purifiées dégradaient toutes deux la paroi cellulaire des mycéliums macérés alors que seule la dernière dégradait aussi la paroi cellulaire de mycéliums intacts. Les résultats indiquent qu’il existe une synergie entre les mécanismes d’action antifongiques de ces enzymes dans lesquels la β-1,3-glucanase est l’initiatrice de l’hydrolyse de la paroi cellulaire alors que le processus de dégradation est renforcé par les chitinases. Paenibacillusehimensis IB-X-b possède une activité antifongique importante sur une vaste gamme de champignons et démontre un potentiel en tant qu’agent de contrôle biologique des champignons pathogènes. [ABSTRACT FROM AUTHOR]

Background and Aims: Two aspects of the competence of abscission zone cells as a specific class of hormone target cell are examined. The first is the competence of these target cells to respond to a remote stele-generated signal, and whether ethylene acts in concert with this signal to initiate abscission of the primary leaf in Phaseolus vulgaris. The second is to extend the concept of dual control of abscission cell competence. Can the concept of developmental memory that is retained by abscission cell of Phaseolus vulgaris post-separation in terms of the inductive/repressive control of β-1,4-glucan endohydrolase (cellulase) activity exerted by ethylene/auxin be extended to the rachis abscission zone cells of Sambucus nigra? [ABSTRACT FROM PUBLISHER]

An extracellular β-1,3-glucanase with antifungal properties was secreted by the novel mycoparasite, Periconia byssoides. The glucanase has a molecular mass of 35 kDa estimated by SDS-PAGE. Its optimum activity was at pH 6.0 and 50°C (over 2 h). The purified β-1,3-glucanase was capable of degrading cell walls, and inhibiting mycelia growth and spore germination of plant pathogenic fungi including Fulvia fulva, Fusarium sp. and Rhizoctonia solani. The N-terminal amino acid residues of the purified β -1,3-glucanase are LKNGGPSFGA, which do not have any homology with previously described glucanases, suggesting it may be a novel member of the fungal β -1,3-glucanases. [ABSTRACT FROM AUTHOR]

B-1a cells reside in both the peritoneal cavity and the spleen. LPS stimulates splenic B-1a to differentiate to plasma cells producing natural IgM specific for microbial and self antigens. However, there are conflicting views as to whether the B-1a cells divide before this differentiation occurs, and hence how the resident B-1a population is maintained in the spleen. Studies here resolve this dispute in favor of both sides: we show that (some or all) B-1a cells resident in the spleen respond to LPS by differentiating to plasma cells immediately, without dividing; however, we also show that additional B-1a cells immigrate into the spleen after LPS stimulation and divide at least once before differentiating. Importantly, the studies we presently describe reveal the complex cell migration and differentiation events that collectively underlie the rapid production of natural antibodies in response to in vivo LPS stimulation. Thus, the studies present a different view of the roles that B-1a cells play in the early phases of the innate immune response. [ABSTRACT FROM AUTHOR]

Abstract: Mature B cells expressing surface IgM as B cell receptor (BCR) consist of two B-cell subpopulations, B-1 and B-2 cells that regulate innate and acquired immunity, respectively. B-1 cells become natural Ig-producing plasma cells in response to natural ligands or self-antigens. Mature B-2 cells proliferate and differentiate into Ig-producing plasma cells under the influence of T helper (Th) cells. Antigen stimulation of B-2 cells induces class switch recombination (CSR) from IgM to other isotypes that is regulated by cytokines and activation-induced cytidine deaminase (AID). IL-5 activates B cells and eosinophils and regulates the innate and acquired immune response. IL-5 stimulates innate B-1 cells and induces the homeostatic proliferation, survival and differentiation of B cells leading to increased secretion of natural antibody. IL-5Rα-deficient mice show the impaired contact sensitivity that is mediated by activated T cells and IgM antibody produced by B-1 cells. IL-5 induces AID and Blimp-1 expression in activated B-2 cells leading to μ to γ1 CSR and IgG1 production. In contrast, IL-4 does not induce μ to γ1 CSR in anti-CD38-activated B-2 cells. Intriguingly, the costimulation of anti-CD38-activated B-2 cells with 8-mercaptoguanosine and IL-4 induces μ to γ1 CSR and IgG1 production, suggesting that unidentified factor(s) is/are required together with AID to complete CSR. We will discuss the role of IL-5 in the innate immune system and disease control, particularly the regulation of contact sensitivity and chronic inflammation through B-1 cell activation and IgM production. [Copyright &y& Elsevier]

An air-dry developmental state with low-hydrated tissues is a characteristic of most plant seeds. Seed dormancy is an intrinsic block of germination and can be released during after-ripening, that is air-dry storage of mature seeds. Both seed-covering layers, testa and endosperm, cause the coat-imposed dormancy of tobacco (Nicotiana tabacum). After-ripening and over-expression of class Iβ-1,3-glucanase (βGlu I) confer maternal effects on testa rupture and dormancy release. Very little is known about the molecular mechanisms of after-ripening and whether gene expression is possible in low-hydrated seeds. Transient, low-levelβGlu I transcription and translation was detected during tobacco seed after-ripening.1H NMR 2D micro-imaging showed uneven distribution of proton mobility in seeds.βGlu I gene expression is associated spatially with the inner testa and temporally with the promotion of testa rupture. Local elevation in moisture content seems to permit local, low-levelβGlu I gene transcription and translation in the maternal tissues of air-dry, low-hydrated seeds.De novogene expression is therefore proposed to be a novel molecular mechanism for the release of coat-imposed dormancy during oilseed after-ripening. [ABSTRACT FROM AUTHOR]

Using cord blood samples obtained from fetuses between 16 and 40 weeks gestation, we have used a lysed whole blood flow cytometric technique to study the natural history of lymphocyte phenotypes known to be highly represented in cord blood at birth. The majority (51.0 ± 14.7%) of lymphocytes expressed CD45RA+ a marker of "virgin' cells and there was a correlation between incrca.sing percentages of CD45RA+ lymphocytes and gestational age (r = 0 44, P < .001). Few cells (8.5 ± 4.2%) expressed the CD45RO marker of primed lymphocytes and very few (1.0 ± 0.7%) co-expressed CD45RA and RO, indicating little traffic between the two maturation markers, The percentage of B lymphocytes co-expressing CD5 was high in the fetal circulation (55.5 ± 10.5%) compared with healthy adults (23.2 ± 14.3%; P < 0.001) and the level of CD5+ B cells declined with gestational age In an exponential manner (r= -0.45, P < 0.05). Similarly, levels of T lymphocytes expressing the γδ T cell receptor (TCR) declined exponentially (r = -0.59, P < 0.005). These results demonstrate that lymphocytes remain almost entirely unprimed before birth. In addition, CD5+ B lymphocytes and TCR-γδ + T lymphocytes decline exponentially towards birth, in a manner suggesting that they may be seeding peripheral sites such as the spleen, skin and mucosae. [ABSTRACT FROM AUTHOR]

Our recent studies on an autoantibody-transgenic mouse line demonstrated that peritoneal B-1 cells are responsible for autoimmune symptoms. However, whether B-1 cells in the peritoneum are generally involved in the pathogenesis of autoimmune disease remains controversial. To test the possible involvement of peritoneal B-1 cells in autoimmune symptoms of autoimmune-prone NZB mice, we eliminated the peritoneal cells by hypotonic shock with repeated I.p. injection of distilled water every 7 days into neonatal or 8-week-old NZB mice. By this treatment, B-1 cells, which self- renew within the peritoneal cavity, are expected to be preferentially eliminated, while other peritoneal cells can be easily supplied from bone marrows after this treatment indeed, in distilled water-treated old NZB mice, the number of B-1 cells decreased in spleen as well as in lamina propria of the gut but the numbers of conventional B cells and T cells did not change. Moreover, the production of autoantibodies against erythrocytes significantly decreased and the occurrence of autoimmune hemolytic anemia was reduced in 12-month-old treated NZB mice. Similarly, the elimination of peritoneal cells of NZB/NZW (NZB/W) F; mice by water injection decreased anti-DNA IgG antibodies in the sera and reduced the pathological changes of the kidney. These results suggest that peritoneal B-1 cells may be a source of autoantibody-producing cells in autoimmune diseases of NZB and NZB/W F; mice. [ABSTRACT FROM PUBLISHER]

The development of nanocarriers (NC) for biomedical applications has gained large interest due to their potential to co-deliver drugs in a cell-type-targeting manner. However, depending on their surface characteristics, NC accumulate serum factors, termed protein corona, which may affect their cellular binding. We have previously shown that NC coated with carbohydrates to enable biocompatibility triggered the lectin-dependent complement pathway, resulting in enhanced binding to B cells via complement receptor (CR)1/2. Here we show that such NC also engaged all types of splenic leukocytes known to express CR3 at a high rate when NC were pre-incubated with native mouse serum resulting in complement opsonization. By focusing on dendritic cells (DC) as an important antigen-presenting cell type, we show that CR3 was essential for binding/uptake of complement-opsonized NC, whereas CR4, which in mouse is specifically expressed by DC, played no role. Further, a minor B cell subpopulation (B-1), which is important for first-line pathogen responses, and co-expressed CR1/2 and CR3, in general, engaged NC to a much higher extent than normal B cells. Here, we identified CR-1/2 as necessary for binding of complement-opsonized NC, whereas CR3 was dispensable. Interestingly, the binding of complement-opsonized NC to both DC and B-1 cells affected the expression of activation markers. Our findings may have important implications for the design of nano-vaccines against infectious diseases, which codeliver pathogen-specific protein antigen and adjuvant, aimed to induce a broad adaptive cellular and humoral immune response by inducing cytotoxic T lymphocytes that kill infected cells and pathogen-neutralizing antibodies, respectively. Decoration of nano-vaccines either with carbohydrates to trigger complement activation in vivo or with active complement may result in concomitant targeting of DC and B cells and thereby may strongly enhance the extent of dual cellular/humoral immune responses. [ABSTRACT FROM AUTHOR]

Over the years research has found an association between B lymphocytes and pathogenesis during Leishmania sp. infections. Recently we demonstrated that B-2 lymphocytes are the main producers of IL-10 during L. amazonensis infection, and that the disease severity in BALB/c mice was attributed to these IL-10-producing B-2 lymphocytes. Here, we aim to understand the role of peritoneal B-1 lymphocytes in the pathogenesis of L. amazonensis infection. We found that infection resulted in a decrease in the number of B-1a lymphocytes and increase in B-1b lymphocytes in the peritoneal cavity of WT BALB/c mice but not in B lymphocyte deficient mice (BALB/Xid) mice. In vitro interaction between B-1 lymphocytes and L. amazonensis showed that the amastigote form of the parasite was able to induce higher levels of IL-10 in B-1 lymphocytes derived from infected BALB/c mice than the promastigote. Moreover, B-1 lymphocytes derived from infected mice produced more IL-10 than B-1 lymphocytes derived from naïve mice under amastigote interaction. However, the repopulation of BALB/Xid mice with B-1 lymphocytes from WT BALB/c mice did not affect the lesion development. Together, these results suggest that although B-1 lymphocytes are able to produce IL-10 during in vitro interaction with L. amazonensis , they are not directly related to pathogenesis in vivo. [ABSTRACT FROM AUTHOR]