Distinct progenitor origin distinguishes a lineage of dendritic-like cells in spleen

The dendritic cell (DC) compartment comprises subsets of cells with distinct phenotypes. Previously this lab reported methodology for hematopoiesis of dendritic-like cells in vitro dependent on a murine splenic stromal cell line (5G3). Co-cultures of lineage-depleted bone marrow (Lin(-) BM) over 5G3 continuously produced a distinct population of dendritic-like "L-DC" for up to 35 days. Here the progenitor of L-DC is investigated in relation to known BM-derived hematopoietic progenitors. It is shown here that L-DC-like cells also derive from the CD150(+)Flt3(-) long-term reconstituting-hematopoietic stem cells (HSC), and also from the Flt3(+) multipotential progenitor subset in BM. Lin(-) BM co-cultures also produce a transient population of cells resembling conventional (c) DC. Production of cDC-like cells is shown here to be transient and M-CSF dependent, and also appears following co-culture of described common dendritic progenitors or monocyte dendritic progenitors over 5G3. BM cells from C57BL/6-flt3L(tm1lmx) and C57BL/6-Csf2(tm1Ard) mice which lack cDC precursors and monocytes, are shown here to contain L-DC progenitors which can seed 5G3 co-cultures. L-DC are functionally distinct cells, in that they arise independently of M-CSF, and by direct differentiation from HSC.This work was supported by project grant #585443 to Helen Christine O’Neill from the National Health and Medical Research Council of Australia. Sawang Petvises was supported by a graduate scholarship from the Royal Thai Government

Characterisation of dendritic cells arising from progenitors endogenous to murine spleen

Heterogeneity amongst dendritic cell (DC) subsets leads to a spectrum of immune response capacity against pathogens. Several DC subsets in spleen have been described which differ in terms of phenotype and function. We have previously reported a distinct population of CD11c(lo)CD11b(hi)MHC-II(-)CD8(-) dendritic-like "L-DC" in murine spleen, which can also be generated in splenic stromal longterm cultures. Here, the ontogeny of L-DC development in perinatal mice has been compared with other known splenic DC subsets. Flow cytometric analysis has revealed the presence of L-DC at embryonic age (E)18.5 spleen, while plasmacytoid (p)DC and conventional (c)DC appear at 2 and 4 days following birth. Co-cultures of E18.5 spleen above splenic stroma also showed production of only L-DC, while spleen cells from D0 through D5 neonates showed production of both L-DC and cDC-like cells. Addition of an M-CSFR inhibitor to co-cultures revealed that while the development of cDC-like cells depended on M-CSF, many L-DC developed independently of M-CSF. Furthermore, purified hematopoietic stem cells (HSC) and multipotential progenitors (MPP) isolated from neonatal D1 spleen are capable of developing into L-DC in co-cultures. These studies reveal a lineage of dendritic-like cells developing in the spleen microenvironment, and which appear to arise from endogenous progenitors laid down in spleen during embryogenesis.This work was supported by project grant #585443 to H.C.O. from the National Health and Medical Research Council of Australia. S.P. was supported by a graduate scholarship from the Royal Thai Government.

Listeria monocytogenes Biofilms in Food-Associated Environments:A Persistent Enigma

Listeria monocytogenes (LM) is a bacterial pathogen responsible for listeriosis, a foodborne illness associated with high rates of mortality (20–30%) and hospitalisation. It is particularly dangerous among vulnerable groups, such as newborns, pregnant women and the elderly. The persistence of this organism in food-associated environments for months to years has been linked to several devastating listeriosis outbreaks. It may also result in significant costs to food businesses and economies. Currently, the mechanisms that facilitate LM persistence are poorly understood. Unravelling the enigma of what drives listerial persistence will be critical for developing more targeted control and prevention strategies. One prevailing hypothesis is that persistent strains exhibit stronger biofilm production on abiotic surfaces in food-associated environments. This review aims to (i) provide a comprehensive overview of the research on the relationship between listerial persistence and biofilm formation from phenotypic and whole-genome sequencing (WGS) studies; (ii) to highlight the ongoing challenges in determining the role biofilm development plays in persistence, if any; and (iii) to propose future research directions for overcoming these challenges

Investigation into the prevalence of a novel dendritic-like cell subset in vivo

A novel dendritic-like cell subset termed L-DC was recently identified in murine spleen based on marker expression of a homogeneous cell population derived from long-term culture of neonatal spleen. The function of L-DC is distinct from other splenic dendritic and myeloid cell subsets because of their high endocytic capacity and their ability to cross-present antigen to CD8+ T cells. This paper shows the subset to be unique to spleen and blood, with a similar, but possibly functionally distinct subset also present in bone marrow. The prevalence of the subset is low; ~6% of all dendritic and myeloid cells in the spleen and ~5% in blood. However, they are a distinct cell type on the basis of marker expression, and endocytic and T-cell stimulatory capacity. Attempts to identify an enriched population of these cells in mutant mouse strains with reported increases in myelopoiesis showed either a lack of L-DC or an altered phenotype reflective of the phenotype of the mouse strain

Understanding the direct and indirect mechanisms of xylanase action on starch digestion in broilers

The objective of the current study was to investigate the mechanisms of xylanase action in a maize-soya diet and its effect on starch digestion. A total of 60 broilers were divided into 6 treatment groups; a control group without xylanase, and five other groups supplemented with xylanase (Econase XT 25; 100 g/t) from 1, 2, 3, 4 or 5 weeks before slaughter. At the end of the experiment, digesta was collected from the gizzard, upper and lower small intestine, and both caeca. Digesta pH ranged from pH 2.2-4.4, 5.9-6.6, 6.7-7.8 and 5.7-7.3 in the gizzard, upper small intestine, lower small intestine, and both caeca, respectively, with no effect of xylanase (P > 0.05). Scanning Electron Microscope (SEM) images along with total starch measurements showed the progression of starch digestion through the tract. The SEM did not show any greater disruption to cell wall material with xylanase supplementation. This suggests that xylanase was not working directly on the cell wall and provides evidence for the hypothesis that xylanase works through an indirect mechanism. Peptide YY (PYY) concentration in the blood was higher during the first few weeks of supplementation, with longer periods of supplementation nulling this effect, implying that xylanase may be acting through a prebiotic mechanism. The RT-q PCR results revealed a numerical increase in glucose transporter (GLUT2 and SGLT1) expression at 2 and 3 weeks of xylanase supplementation, respectively, which might suggest a greater absorption capacity of birds. From these results, a potential mechanism of xylanase action in maize-based diets has been proposed

Xylanase, and the role of digestibility and hindgut fermentation in pigs on energetic differences among high and low energy corn samples

The experimental objective was to evaluate the digestibility and fermentation differences between high and low energy corn samples and their response to xylanase supplementation. Four corn samples, 2 with higher DE content (HE-1 and HE-2; 3.74 and 3.75 Mcal DE/kg DM, respectively) and 2 with a lower DE content (LE-1 and LE-2; 3.63 and 3.56 Mcal DE/kg DM, respectively) were selected based upon a previous digestibility trial. Sixteen individually housed barrows (PIC 359 × C29; initial BW = 34.8 ± 0.23kg) were surgically fitted with an ileal T-cannula and randomly allotted to treatments in an 8 × 4 Youden square design. Dietary treatments were arranged in a 4 × 2 factorial: HE-1, HE-2, LE-1, and LE-2, with and without xylanase supplementation. Diets were formulated using one of the 4 corn samples, casein, vitamins, minerals, and 0.4% chromic oxide as an indigestible marker. Feed intake was established at approximately 3 times the estimated energy required for maintenance (NRC 2012) based upon the average initial BW of the pigs at the start of each collection period, which consisted of 9 d adaptation, 2 d of fecal, and 3 d of ileal collections. Diets, ileal, and fecal samples were analyzed for DM, GE, and total dietary fiber (TDF), to determine apparent total tract (ATTD), hindgut fermentation (HF), apparent ileal digestibility (AID) coefficients. A diet × enzyme interaction was not observed for any of the measured variables (P \u3e 0.10). The HE-1 and HE-2 diets had greater ATTD of GE, and HE-2 diet had greater ATTD of DM (P \u3c 0.001 and P = 0.007, respectively). Xylanase, independent of diet, improved the ATTD of GE and DM (84.8 vs. 83.6% for GE with and without enzyme, respectively, P = 0.008; and 84.2 and 83.0% with and without enzyme, respectively, P = 0.007). The energetic differences among these corn samples appeared to be driven by fermentability in the hindgut. Supplementing xylanase improves digestibility irrespective of the digestibility energy content of corn