Between clinically affected and healthy sheep, no differences were found in the protein levels of mGluR1, while phospholipase

Cβ1 expression in terminally ill Metformin purchase sheep was increased in some brain areas but decreased in others. Adenyl cyclase 1 and A1R levels were significantly lower in various brain areas of affected sheep. No abnormal biochemical expression levels of these markers were found in preclinically infected sheep. Conclusions: These findings point towards an involvement of mGluR1 and A1R downstream pathways in natural scrapie. While classical prion disease lesions and neuromodulatory responses converge in some affected regions, they do not do so in others suggesting that there are independent regulatory

factors for distinct degenerative and neuroprotective responses. “
“Since the first description of the classical presentation of progressive supranuclear palsy (PSP) in 1963, now known as Richardson’s syndrome (PSP-RS), several distinct clinical syndromes have been associated with PSP-tau pathology. Like other neurodegenerative disorders, the severity and distribution of phosphorylated tau pathology are closely associated with the clinical heterogeneity of PSP variants. PSP with corticobasal syndrome presentation (PSP-CBS) was reported to have more tau load in the mid-frontal and inferior-parietal cortices CH5424802 ic50 than in PSP-RS. However, it is uncertain if differences exist in the distribution of tau pathology in other brain regions or if the overall tau load is increased in the brains of PSP-CBS. We sought

to compare the clinical and pathological features of PSP-CBS and PSP-RS including quantitative assessment of tau load in 15 cortical, basal ganglia and cerebellar regions. In addition to the similar age PLEKHM2 of onset and disease duration, we demonstrated that the overall severity of tau pathology was the same between PSP-CBS and PSP-RS. We identified that there was a shift of tau burden towards the cortical regions away from the basal ganglia; supporting the notion that PSP-CBS is a ‘cortical’ PSP variant. PSP-CBS also had less severe neuronal loss in the dorsolateral and ventrolateral subregions of the substantia nigra and more severe microglial response in the corticospinal tract than in PSP-RS; however, neuronal loss in subthalamic nucleus was equally severe in both groups. A better understanding of the factors that influence the selective pathological vulnerability in different PSP variants will provide further insights into the neurodegenerative process underlying tauopathies. “
“Y. Chiba, S. Takei, N. Kawamura, Y. Kawaguchi, K. Sasaki, S. Hasegawa-Ishii, A. Furukawa, M. Hosokawa and A.

275 RENAL (AND HERPETIC) RE-TRANSPLANTATION S SETYAPRANATA1,

KJ WIGGINS1, SG HOLT1,2, WR MULLEY3, PG KERR3, AJ LANDGREN1, A YOUNG4, H OPDAM4, A ROBERTSON1, PD HUGHES1 1Royal Melbourne Hospital, Melbourne, Victoria; selleck chemical 2The University of Melbourne, Melbourne, Victoria; 3Monash Medical Centre, Melbourne, Victoria; 4Donate Life Victoria, Melbourne, Victoria, Australia Aim: Case report of renal re-transplantation, reported only once previously. Report: A middle aged recipient received a kidney transplant from a deceased multi-organ donor. After initially doing well, the patient suffered cardiac arrest several days post-operatively and sustained hypoxic brain injury and was declared brain dead. Following the family’s consent, the allograft INK 128 ic50 kidney was retrieved and re-transplanted into a man with end-stage renal failure secondary to reflux nephropathy. The lungs were used in a separate recipient but the liver was not transplanted due to suspicion of fatty

changes based on macroscopic appearance. Histological analysis of the liver more than 24 hours after transplantation of the other organs revealed coagulative parenchymal necrosis with nuclear inclusions and moderate parenchymal cholestasis, suggestive of herpes viral hepatitis. Examination of the renal implantation biopsy showed histiocytes with enlarged nuclei containing viral inclusions in the capsular fibrous tissue, with positive immunostaining for herpes simplex virus (HSV). Valaciclovir was started immediately after obtaining histological evidence of donor HSV infection and this was subsequently converted to intravenous ganciclovir. Our recipient had pre-formed IgG antibodies to HSV-1 and HSV-2, and was IgM negative pre-transplant. HSV viraemia was detected day 5 post-transplant with a viral load of 7688 copies/mL by

polymerase chain reaction (PCR) assay. He completed a 30-day course of intravenous ganciclovir before switching to valganciclovir as standard cytomegalovirus prophylaxis. The HSV PCR became undetectable on day 7 of IV ganciclovir and has remained undetectable. The patient remains well with an estimated glomerular filtration rate of 61 mL/min/1.73 m2 and further investigation of the apparent viral transmission is underway. Conclusions: We report good short term results Obatoclax Mesylate (GX15-070) of renal re-transplantation and HSV transmission by transplantation. 276 ACUTE KIDNEY INJURY DUE TO DECOMPRESSION ILLNESS A VIECELLI, J JAMBOTI, P FERRARI Department of Nephrology, Fremantle Hospital, Perth, Western Australia, Australia Background: Decompression illness is a rare but serious complication of diving caused by intravascular or extravascular gas bubble formation. Case Report: We report the first case of acute kidney injury in a 27-year-old diver caused by arterial gas emboli formation following three rapid uncontrolled ascents.

Kenilworth, NJ, USA), voriconazole (VOR; Pfizer Central Research). In vitro susceptibility testing was performed using the broth microdilution method for filamentous fungi, according to CLSI document Sorafenib M38-A2.15

Stock solutions of antifungal drugs had a concentration of 3200 μg ml−1, while pure substance (powder) of AMB, ISA, ITR, POS, VOR, and ANI were dissolved in dimethyl sulfoxide; for stock solutions of caspofungin and micafungin, sterile distilled water was used. Test concentration solutions were produced using filter-sterilised (0.22 μm filter) RPMI 1640 medium with l-glutamine (Difco, Breda, The Netherlands). For susceptibility testing, strains were re-grown from cryo-preserved cultures on SGA tubes at 30 °C, until colonies revealed strong sporulation (up to 14 days). Inocula were produced by streaking with a sterile cotton swab wetted with 0.9% NaCl + 0.05% Tween 20 solution over the sporulating fungal colonies. Spores were transferred

in a 0.9% NaCl solution + 0.05% Tween 20 to reach a turbidity of approximately 0.5 McFarland. Afterwards, inoculum was adjusted to a light transmission of 68–71% at 530 nm, using a spectrophotometer. Spore solutions were then diluted 1 : 50 in sterile RPMI 1640. Candida parapsilosis (ATCC 22019) and C. BAY 80-6946 krusei (ATCC 6258) were included as quality control strains. Results were read after an incubation time of 72 h at 37 °C. MIC Edoxaban for AMB, ITC, ISA, POS, and VOR was read visually, whereas MEC for ANI, CAS, and MICA was read microscopically. When susceptible to the antifungal agent, hyphae were shorter, more rounded and compact, deformed than those in control wells, and the cell walls of susceptible

hyphae were thickened and the hyphae appeared deformed. Geometric mean MICs and MECs was computed using Microsoft® Office Excel 2003 SP3. For MIC geometric mean calculations, concentrations ≤0.125 μg ml−1 were set as 0.062 μg ml−1 and concentrations ≥16 μg ml−1 were set to 32 μg ml−1. For MEC geometric mean calculations, concentrations ≤0.062 μg ml−1 were set as 0.031 μg ml−1 and concentrations ≥8 μg ml−1 were set to 16 μg ml−1. For MIC50 and MIC90 calculation, MIC data of each antifungal and for all strains belonging to the same species were sorted in ascending order, then median and 90th percentile were determined. The AFLP-electropherograms of clinical isolates (n = 60) were compared with those of the included type strains (Fig. 1). Based on this analysis, they were identified as: P. apiosperma (n = 6), S. aurantiacum (n = 1), P. boydii (n = 15), S. dehoogii (n = 1), P. ellipsoidea (n = 3), S. prolificans (n = 34). No P. angusta, P. minutispora, and P.

7). Collectively, these data suggest that EphB4 may contribute to the unique biphasic modulatory effect by ephrin-B1/B2 through

the recruitment of SHP1 (Fig. 6C). In contrast to ephrin-B1/B2, the phospho-EphB4 induced by ephrin-B3 could not couple with SHP1, which has the inhibitory effect of Lck phosphorylation. In this study, we elucidated that ephrin-B1 and ephrin-B2 belong to a novel class of costimulatory molecules with unique action, namely, a concentration-dependent switch from costimulation to inhibition; whereas, ephrin-B3 simply exerts a steadily increasing stimulatory effect in TCR-mediated regulation of primary T cells via Eph receptors other than EphB1/B2/B3/B6. The unique inhibitory effects selleck chemical by the high concentrations of ephrin-B1/B2 occur buy RG7204 as a consequence of cross-talk of EphB4 signaling on TCR cascade, most likely targeting Lck. Although Eph receptors/ephrin ligands were initially recognized as mediators of repulsive signals in growing axons, it is now clear that their functions

are versatile, including attractive and adhesive property. In vivo, ephrin-Bs have been shown to act as both attractants and repellents for retinal axons during the developmental stage of the visual system [[24]]. However, it remains unclear whether the reciprocal effects in vivo are mediated by the same ephrin ligand in the same cell since these effects are dependent in time and space where the expression of Eph receptors/ephrin ligands would

be variable. Definitive demonstration of biphasic action of this system can be done in in vitro system. Recently, Hansen et al. elegantly demonstrated that ephrin-As induced the biphasic retinal axon growth [[21]]. Alfaro et al. [[7]] demonstrated that immobilized Eph-B2-Fc and ephrin-B1-Fc modulated the anti-CD3 antibody-induced apoptosis of CD4+CD8+ thymocytes in a concentration-dependent 5-Fluoracil in vivo manner. Our present study has addressed the direct proof of biphasic effect of ephrin-Bs on the proliferation of the primary immune cells. In addition to the function in cell positioning including attraction, adhesion, and repulsion which have been mainly investigated in the nervous system, our study demonstrated for the first time that this biphasic regulation is functional in cell proliferation, as well. According to the studies for functional determination, Eph receptors can promote adhesion/attraction in a kinase-independent manner; whereas, repulsive function requires tyrosine kinase activity and receptor phosphorylation [[26, 38, 39]]. Eph receptors may possess two distinct functional sites, (i) adhesion by extracellular kinase-independent domain and (ii) repulsive/inhibitory signaling by intracellular kinase-dependent domain. The concentration of ephrin ligands would be one of the factors to determine the balance between these two functions.

The LTα1β2 then signals via LTβR to drive mesenchymal stromal cells to differentiate into lymphoid tissue organizer cells (LTos),[9] accompanied by the up-regulation of chemokine (e.g. CXCL13, CCL19 and CCL21) and adhesion molecule (e.g. vascular cell adhesion molecule-1, intercellular adhesion molecule-1, mucosal addressin cell adhesion molecule-1)[12] expression in the LN anlagen. Chemokines, as well as the up-regulated expression of RANKL PLX4032 chemical structure and interleukin-7 (IL-7) by LTos,[9, 10] induce the recruitment and survival of further cells to the expanding LN anlagen.[13] The arrival of more LTα1β2-expressing cells, which includes few LTis[14] but after birth is dominated by lymphocytes

(both T and B cells),[15, 16] creates a positive feedback loop Daporinad (Fig. 1), further increasing signalling through the LTβR and

the subsequent expression of LTo-derived factors. Using conditional ablation of the Ltbr gene exclusively in VE-Cadherin+ endothelial stromal cells, Onder et al.[17] recently revealed that the development of multiple peripheral LNs required LT signalling specifically into this LTβR+ stromal compartment. Interestingly, not all LNs required endothelial sensitivity to LTα1β2, as the mesenteric LNs of the intestine were fully intact in these mice, hinting at a requirement for distinct LTβR+ stromal cell populations in the development of anatomically disparate peripheral LNs in vivo. Other homeostatic SLOs develop in a fundamentally similar way to the LN with only minor differences between tissues. For instance in the Peyer’s patches of the small intestine, although ligands of the receptor tyrosine kinase RET acting on a distinct population of CD45+ IL-7Rα− CD11c+ cells contributes to stromal activation in the developing anlagen,[18] LTis and LTα1β2 are still important in this developmental process,[4] although it is not clear if LTα1β2 expression is induced by RANK as in early LN development. However,

the earliest steps in homeostatic intestinal SLO development are still under intense investigation.[19] Lymphoid tissue organizers differentiate into the various non-haematopoietic stromal subtypes present in the adult SLO via LTβR signalling,[20] Parvulin although the ontogeny and lineage relationships of the various stromal cell subsets within the LN is still under investigation.[21, 22] Mesenchyme-derived stromal cells can be divided into several subsets including follicular dendritic cells (FDCs), marginal reticular cells and populations of fibroblastic reticular cells (FRCs). Lymph node stromal endothelial cells can be divided into blood endothelial cells and lymphatic endothelial cells,[23] and all SLOs contain high endothelial venules composed of endothelial cells with distinct morphology and phenotype. Four CD45− stromal subsets can therefore be identified by a dual CD31 (PECAM-1) and Podoplanin (gp38) stain.[23] Identification of further subsets can be achieved using a range of different surface markers (Table 1).

Findings are discussed in relation to parenting roles and family dynamics. “
“The interactions between attention and stimulus encoding in infancy were examined using heart rate (HR) and visual habituation measures. At 3, 6, and 9 months of age, infants (N = 119) were habituated to an adult face; longest look (LL) duration was measured as an indicator of encoding speed. Three groups were formed based on LL change from 3 to 9 months: Large Decrease, Small Decrease, and Increase. Using concurrent electrocardiograph

recordings, attention was measured through the percentage of looking time in orienting, sustained attention, and attention termination. We partially replicated previous findings regarding developmental patterns of attention in these three Akt inhibitor groups, notably that these patterns were different for the Increase group. Looks away from the stimulus were also assessed in each attentional phase and, as predicted, HR acceleration

phases showed less visual engagement than HR deceleration phases. We also found anomalous behavior for the LL Increase group. In general, this small but distinct group showed similarities at 3 months to the presumably more mature behavior of typical 9 month olds, but by 9 months, they behaved more like typical 3 month olds regarding some, but not all, cognitive Rapamycin datasheet measures. These results are discussed in the context of the development of endogenous attention. “
“We investigated the effects of distraction on attention and task performance during toddlerhood. Thirty toddlers (24- to 26-month-olds) completed different tasks (2 of each: categorization, problem solving, memory, free play) in one of two conditions: No Distraction or Distraction. The results revealed that the distractor had varying effects on performance scores depending on the task: The problem solving and memory tasks were more susceptible to distraction. In addition, the two conditions click here showed different patterns of attention over time.

Toddlers in the No Distraction condition were more attentive, and their attention remained consistently high across the session. Toddlers in the Distraction condition increased their attention to the task and decreased their attention to the distractor in the second half of the session. This study demonstrates how the presence of distraction influences toddlers’ performance on individual cognitive tasks and contributes to our understanding of distractibility and endogenous attention during toddlerhood. This work also has implications for how environmental noise, such as background television, may influence cognitive development. “
“Behavioral and electrophysiological indices of memory were examined in 12-month-old typically developing control infants (CON) and infants with history of perinatal hypoxic-ischemic injury (HII) across 2 days.

parapertussis lipopolysaccharides stimulates the TLR4 response inefficiently, allowing the organism to avoid the robust inflammatory response involved in rapid antibody-mediated clearance (Wolfe et al., 2009). This is in contrast to the lipopolysaccharides of B. bronchiseptica and B. pertussis, which are relatively stimulatory of the TLR4 response in mice (Mann et al., 2005; Wolfe et al., 2009). Wolfe et al. (2009) observed that coinfection of C57BL/6 mice with B. bronchiseptica and B. parapertussis

resulted in more efficient control of B. parapertussis infection by the host, concluding that increased neutrophil recruitment due to the presence of B. bronchiseptica lipopolysaccharides led to the more efficient clearance of B. parapertussis. However, these observations are in conflict with those made in our study, where coinfection of Balb/c mice with B. pertussis Panobinostat and B. parapertussis did not result Daporinad cost in increased clearance of B. parapertussis,

but rather an increase in B. parapertussis numbers. It may be that PT produced by B. pertussis provides B. parapertussis with protection against the TLR4-mediated responses, because PT can inhibit cytokine production and neutrophil recruitment in response to an intranasal administration of lipopolysaccharides (Andreasen & Carbonetti, 2008). Alternatively, the effects may be mouse strain-dependent. Previous studies with 2-week-old suckling mice demonstrated that when infected with a mixed inoculum of B. pertussis 18-323 and B. parapertussis strain 422, persistent colonization with B. parapertussis was observed (Kawai et al., 1996). However, when mice were inoculated with B. parapertussis alone, this organism failed to colonize mice, suggesting a relationship between B. pertussis and B. parapertussis, where the former facilitates colonization

by the latter in a mixed infection (Kawai et al., 1996). This group hypothesized that for B. parapertussis to adhere to lung epithelia cells and consequently establish infection, these epithelial cells must first be damaged by B. pertussis infection. In our infection studies, we observed a similar relationship between these two species whereby B. pertussis facilitates infection by B. parapertussis. However, unlike in the 2-week-old mice, B. parapertussis alone is able to establish infection in 6-week-old Balb/c mice. Staurosporine clinical trial Our study examined the effects of coinfection on early events in naïve hosts. Several reports have examined the effect of immunity to one of these Bordetella pathogens (from vaccination or infection) on infection by the other in mouse models. Current pertussis vaccines do not provide protective immunity against B. parapertussis (Komatsu et al., 2010) and can confer this organism with an advantage in a mixed infection (Long et al., 2010), although a novel live-attenuated pertussis vaccine was found to protect against B. parapertussis by a T-cell-mediated mechanism (Feunou et al., 2010).

Akt2 and Akt3 seem not to play a major role in placental angiogenesis because Akt2-null mice display a type-II diabetes-like syndrome and mild growth retardation and age-dependent loss of adipose tissue [121] and Akt3 has been shown to be important in postnatal brain development [31]. The potent vasodilator NO is generated during the conversion of l-arginine to l-citrulline by a family of NO synthases (NOS), including eNOS, inducible NOS (iNOS) and neuronal NOS (nNOS) [106]. Placental

NO production increases during pregnancy, which Selleck Alvelestat is highly correlated with eNOS, but neither iNOS nor nNOS expression [127, 88], suggesting that eNOS is the major NOS isoform responsible for the increased NO in the placenta. During normal sheep pregnancy placental NO production increases [127, 69] in association with elevated local expression of VEGF and FGF2, vascular density, and blood flow to the placentas [128, 9], suggesting that eNOS-derived NO is important in placental angiogenesis. Indeed, the eNOS-derived NO is critical for the VEGF and FGF2- stimulated angiogenesis in vitro [76, 24] and in vivo [44]. The eNOS-derived

NO is also a potent vasodilator in the perfused human muscularized fetoplacental vessels [87], which might be critical for the maintenance of low vascular resistance in the fetoplacental circulation in pregnant sheep in vivo [18]. Early studies have shown that pharmacological NOS inhibition by l-NG-nitroarginine methyl ester results in preeclampsia-like symptoms and reduced litter size in rats [11]. This has been confirmed in eNOS-null mice whose dams develop proteinuria

[68] and fetuses Selleckchem PF2341066 are growth restricted [68, 67, 66]. In eNOS-null pregnant mice, uteroplacental remodeling is impaired and their vascular adaptations to pregnancy are dysregulated [66, 114], resulting in decreased uterine and placental blood flows and greatly reduced vascularization in the placenta [67, 66]. These selleck inhibitor studies suggest that eNOS is critical for both vasodilation and angiogenesis, that is, the two rate-limiting mechanisms for blood flow regulation at the maternal–fetal interface. Numerous studies have shown that activation of the MAPK (ERK1/2, JNK1/2, and p38MAPK), PI3K/Akt1, and eNOS/NO pathways is critical for VEGF- and FGF2-stimulated angiogenesis in various endothelial cells. In placental endothelial cells, we have shown that activation of the MAPK pathways are important for the differential regulation of placental endothelial cell proliferation, migration, and tube formation (i.e., in vitro angiogenesis) in response to VEGF and FGF2 stimulation in vitro [130, 82, 35, 36]. Inhibition of the ERK1/2 pathway partially attenuates the FGF2-stimulated cell proliferation, whereas it completely blocks the VEGF-stimulated cell proliferation as well as the VEGF- and FGF2-stimulated cell migration [75, 76, 130, 35, 36].

[1] Donor-derived T cells are considered the main effector cells mediating acute GVHD because they recognize MHC disparities (allo-antigen) between the donor and recipient, which are presented by antigen-presenting cells (APC). T-cell activation in response to allo-antigen Adriamycin requires two stimulatory signals.[1] The primary signal is delivered through the T-cell receptor (TCR), which recognizes antigens on MHC molecules. This signal is necessary but not sufficient to induce full T-cell activation, which also requires co-stimulation that drives T cells to proliferate and produce cytokines. The co-stimulation signal is mediated by a number of ligand–receptor pairs expressed

on APC and T cells, and is a composite or net effect of stimulatory and inhibitory signals mediated between these two

cells. The inhibitory TCR include cytotoxic T-lymphocyte antigen-4 (CTLA-4),[2] programmed cell death-1 (PD-1)[3] and B- and T-lymphocyte attenuator Ivacaftor in vitro (BTLA).[4] Studies using experimental models of acute GVHD have shown that co-stimulatory molecules play a pivotal role in initiating acute GVHD.[5] By contrast, much less is known about co-inhibitory pathways in this process, better understanding of which would make them useful therapeutic targets. Recently, we discovered a new co-inhibitory pathway composed of DC-HIL on APC and syndecan-4 (SD-4) on activated T cells.[6, 7] DC-HIL is a highly-glycosylated type I transmembrane receptor (95 000–120 000 molecular weight) expressed constitutively by many APC sub-sets including

macrophages, monocytes, epidermal Langerhans cells, CD11c+ CD4+ lymphoid dendritic cells (DC), CD11c+ CD8+ myeloid DC and CD11c+ PDCA-1+ plasmacytoid DC.[8] It is also known as glycoprotein nmb (Gpnmb),[9] osteoactivin[10] and haematopoietic growth factor-inducible neurokinin-1 type (HGFIN).[11] DC-HIL binds to heparan sulphate-like structures on SD-4 expressed by activated (but not resting) T cells, Carteolol HCl and their binding inhibits strongly the anti-CD3 response of T cells, resulting in cessation of interleukin-2 (IL-2) production and prevention of T-cell entry into the cell cycle.[6, 12] Consistent with a previous finding that SD-4 is expressed primarily by effector/memory (but not recently activated) T cells,[13] infusion of DC-HIL or SD-4 soluble receptor during the elicitation (but not sensitization) phase of contact hypersensitivity effectively blocked the inhibitory function of the endogenous DC-HIL/SD-4 pathway, thereby enhancing ear-swelling responses in this experimental model.[7] Conversely, depletion of SD-4+ T cells by infusion of toxin-conjugated DC-HIL inhibited elicitation (but not sensitization) of contact hypersensitivity.[13] These findings support the concept that binding of DC-HIL to SD-4 inhibits pre-primed T-cell responses. To determine whether SD-4 is the sole T-cell ligand of DC-HIL and whether its negative regulatory role applies to acute GVHD, we took advantage of SD-4 knockout (KO) mice.

In Fig. 1c it can be seen that a higher amount of fluorescence appeared after incubation with FITC-AGE-OVA

compared with FITC-OVA. Blocking of RAGE by a neutralizing antibody did not inhibit internalization of FITC-OVA or FITC-AGE-OVA. To investigate click here the proliferation of CD4+ T cells induced by OVA or AGE-OVA, CD4+ T cells were co-cultured together with autologous mature DCs that had been loaded with different concentrations of OVA or AGE-OVA. Figure 2(a) shows that both allergens were able to induce a concentration-dependent proliferation of T cells compared with the background proliferation of unloaded DCs (medium) which did not reach the level of the positive control tetanus toxoid (TT). There was no significant difference between OVA- and AGE-OVA-loaded DC-induced T-cell proliferation. To eliminate a possible influence of lipopolysaccharide (LPS) at the highest concentration of OVA or AGE-OVA, polymyxin B sulphate was added together with the allergen during DC culture, without changing the results (data not shown). Next, we wondered whether glycation of OVA induces a change in the cytokine profile of mature DCs and subsequent co-cultures of DCs and CD4+ T cells. Therefore, we measured the secretion of IL-6 and

IL-12p40 by DCs as well as the secretion of the Th2 cytokines IL-4 and IL-5 and the Th1 cytokine IFN-γ by CD4+ T cells. Additionally, we measured the production of the regulatory cytokine IL-10. Figure 2(b) shows that AGE-OVA induced a stronger expression of IL-6 in mature DCs than OVA, while IL-12p40 until production was not affected by OVA or AGE-OVA. In the co-cultures,

RG7204 cell line stimulation of CD4+ T cells with autologous OVA- or AGE-OVA-loaded mature DCs induced concentration-dependent production of all cytokines (Fig 2c). Compared with tetanus toxoid-pulsed DCs, the Th2 cytokines IL-4 and IL-5 were more weakly expressed after stimulation with OVA- or AGE-OVA-pulsed DCs, while the production of IFN-γ and IL-10 almost reached the levels found in the positive control. Interestingly, AGE-OVA-loaded DCs induced greater Th2 cytokine production (P < 0·05 for IL-5), while OVA-loaded DCs induced a significant Th1 or regulatory cytokine profile. This bias towards Th2 cytokine production after stimulation with AGE-OVA-pulsed DCs was confirmed by intracellular staining of the co-cultures for IFN-γ and IL-4. Again, IFN-γ-producing cells were greatly reduced after stimulation of CD4+ T cells with AGE-OVA-pulsed DCs compared with OVA-pulsed DCs, while IL-4-producing cells were slightly increased (Fig. 2d). For analysis of the expression of RAGE on immature and mature DCs, cells were analysed by flow cytometry, and 16·1 ± 5·6% expression of RAGE by immature DCs and 12·8 ± 6·1% expression by mature DCs were found (Fig. 3a,b). As RAGE expression is up-regulated after contact of AGEs with RAGE on monocytes,28,29 we examined whether AGE-OVA also enhances the expression of RAGE on immature and mature DCs.