Relatively little financial support has been given in Australia and New Zealand for studies of immunity to parasitic nematodes in rodent hosts and as a consequence, few laboratories have consistently published on the topic. There has been some interest in using natural parasitic nematodes to prevent the sometimes devastating house mouse (Mus domesticus) plagues that engulf large areas of agricultural land in Australia, but experimental field studies have so far been disappointing (14,15). The remainder of this review will address the last 30 years

of research in Australia and New Zealand on parasitic nematode infections in mice. These and other studies have been informative in modelling natural infections in human and agricultural host species. Such investigations have repeatedly provided novel data on immunity to parasitic https://www.selleckchem.com/products/BAY-73-4506.html worms and more importantly have identified or tested central paradigms of relevance to the widest facets of immunology. A number of parasitic nematode species commonly used internationally in murine models have not been studied in Australasia,

either because of quarantine restrictions (Trichinella spiralis) or because of limited local experience and opportunity (Trichuris muris, Strongyloides venezuelensis, Angiostrongylus cantonensis, Litomosoides sigmodontis). The nematodes most frequently studied in murine hosts in Australasia are Heligmosomoides bakeri (previously known as Heligmosomoides polygyrus or Nematospiroides dubius), Strongyloides ratti, Nippostrongylus brasiliensis and Toxocara canis. Many of these parasites were not initially discovered in murine species, though host specificity VX-809 research buy in the wild has not necessarily been well-defined, especially given the diverse geographical locations in which the parasites might OSBPL9 be endemic. However, in each case, mice are permissive and convenient laboratory hosts. Most of the Australasian investigations reviewed were directed at understanding protective immunity, immune regulation and

immunopathology in the host and immune evasion strategies used by parasites. Murine models have also been used to test vaccines and anthelmintics and are increasingly being applied to identifying parasite-derived molecules that might have therapeutic applications in controlling inflammatory and allergic diseases. N. brasiliensis has been widely studied internationally, both as a model for hookworm and Strongyloides stercoralis infections and because it has proven useful in understanding broadly applicable concepts in both innate and adaptive immunity. Studies of N. brasiliensis infections have added to our understanding of IgE production (16), Type 2 cytokines and helper T cell regulation (17–19), the importance of alternatively activated macrophages (20), as well as the interplay between the immune system and epithelial and smooth muscle cells to control gastric motility and the expulsion of worms (21,22). New regulatory and effector leucocytes have also been identified in studies of N.

024). Based on these

findings, it seems that individuals with the genotype AE, AG or Tel-B/B, or haplotypes 1 and 6 are susceptible to syphilis, whereas individuals with genotype P or haplotype 17 are protective from syphilis in the Chinese Han population. Killer immunoglobulin-like receptor (KIR) molecules are encoded by the KIR gene family that clusters within the leucocyte receptor complex on chromosome 19q13.4. KIR genes exhibit learn more allelic, haplotypic and gene content variability [1–4]. The haplotypes have a framework of four conserved blocks containing KIR3DL3, KIR3DP1, KIR2DL4 and KIR3DL2 and differ in the number and type of KIR genes. In general, most KIR haplotypes belong to one of two broad groups, termed A and B. Haplotype A is composed of KIR3DL3, KIR2DL3, KIR2DP1, KIR2DL1, KIR3DP1, KIR2DL4, KIR3DL1, KIR2DS4 and KIR3DL2 genes, while all the other haplotypes are described as haplotype B. The genes encoding KIR are found in two adjacent clusters, where framework genes flank each cluster: KIR3DL3 Selleck EPZ-6438 and KIR3DP1 flank the centromeric (Cen) cluster, and KIR2DL4 and KIR3DL2 flank the telomeric (Tel) cluster. KIR haplotypes A and B have distinctive Cen and Tel gene content motifs [5]. Both groups of haplotypes

have been found in all populations analysed so far, but their distributions vary considerably among ethnic groups [1–3]. Syphilis is caused by the sexually transmitted spirochetal pathogen Treponema pallidum (T. pallidum), which is a worldwide public health problem. The World Health Organization (WHO) estimates that there are 12 million new cases of syphilis each year, with more than 90% occurring in developing nations [6]. In China,

a total of 217,473 syphilis cases were reported in 2007 with the incidence rate of 15.88/100,000 population, which was 5.17-folds more than that in 1998 [7]. In a study of the sexual contacts of patients with syphilis, 48.5–62.1% of contacts at risk developed syphilis [8]. Syphilis has primary and secondary clinical stages with large numbers of T. pallidum organisms found in mucous membrane and skin lesions. Once spirochetes persist in the host, signs and symptoms of late or tertiary syphilis ensue and even lead to death. Without anti-microbial therapy after infection, approximately one-third of patients mafosfamide with syphilis will eventually develop symptomatic late syphilis; the remaining two-thirds seem to clear the infection [9]. The immunological response of host has long been suggested to play a critical role in the occurrence and development of syphilis [10]. However, because of the inability to cultivate T. pallidum in vitro and the lack of a suitable inbred animal model for immunological studies [11], many questions remain obscure regarding the basic immunobiological aspects of syphilis, for example, why do some contacts not contract T.

The findings presented in this study should also be relevant for researchers using rats to study obesity and/or inflammatory processes such as arteriosclerosis, where the importance of iNKT cells has emerged over the last years [34, 35]. Moreover, our results are also of high relevance in the fields of pharmacology, physiology, and surgery in which the rat is the major selleck inhibitor model organism and where iNKT cells have been ignored so far. Altogether, we hope that the current study will help and motivate researchers to analyze iNKT cells in the rat model, which shows some promising

similarities to humans, and we anticipate that such studies will greatly enhance our understanding of iNKT-cell biology. F344/DuCrl

and LEW/Crl inbred rats and C57BL/6J/Crl inbred mice originally obtained from Charles River were kept and bred in the animal facilities of the Institute for Virology and Immunobiology, University of Würzburg, Würzburg, Germany. The procedures for performing animal experiments as well as animal care were in accordance with the principles of the German law. Permission to keep and breed the animals was given by the city of Würzburg, Germany (OA/he-wa07.12.1987). All animals were maintained under specific pathogen-free I-BET-762 datasheet conditions and were used at 6–18 weeks of age. Thymocytes and splenocytes were prepared by mechanical disruption using a stainless steel mesh. Erythrocytes were eliminated by lysis with TAC buffer (20 mM Tris, pH 7.2, and 0.82% NH4Cl). Rat and mouse IHLs were isolated as described previously [36]. Rat and mouse CD1d dimers were produced in our laboratory as previously described for mouse CD1d dimer [37, 38]. Modifications such as the use of rat-β-2 microglobulin transduced

J558L cells for rat CD1d-dimer production and construction of the CD1d dimer expression vectors have been performed unless as described in [36]. The dimers (at a final concentration of 250 ng/μl) were loaded with 40× molar excess of α-GalCer in the presence of 0.05% Triton X-100 for 16 to 24 h at 37°C. As previously shown by Porcelli and colleagues [39], the presence of Triton X-100 was crucial for appropriate loading of α-GalCer into the CD1d molecules. The vehicle used for dilution of α-GalCer was DMSO, thus as control, the dimers were loaded only with the corresponding amount of DMSO. Nonspecific binding of the Ab/dimers to mouse Fc receptors were blocked by incubating the cells first with anti-mouse Fc receptor mAb (2.4G2). CD1d dimer stainings were carried out at room temperature for 30 min with 1 μg of dimers (4 μl) per 100 μl of sample containing up to 106 cells suspended in FACS buffer (PBS pH 7.4, BSA 0.1%, 0.01% NaN3). Bound CD1d dimers were detected with a fluorophore-labeled donkey F(ab′)2 fragment anti-mouse IgG (H+L) with minimal cross-reactivity to rat and other species serum proteins (Dianova), referred hereafter as DαM.

Accordingly, IL-9 production by Th9 cells strictly correlated with IRF4 expression, and Irf4–/– CD4+ T cells failed to differentiate into IL-9 producers under Th9-inducing conditions [44]. Conversely, transient deletion of IRF4 in wild-type (WT) CD4+ T cells prevented the differentiation of Th9 cells. At the molecular level, IRF4 directly induced IL-9 expression by binding to and activating the Il9 promoter. The importance of IRF4 for Th9 development in vivo was shown in a mouse model for allergic asthma, in which Irf4–/– mice were totally resistant to the induction of allergic airway disease. Importantly, reconstitution of the mice with WT Th9 cells restored asthma symptoms, demonstrating not only

the importance of IRF4 for this website Th9 development in vivo, but also APO866 clinical trial a role for Th9 cells during allergic airway disease [44]. Consistent with the finding that AICEs are present in the upstream regulatory elements of the Il9 and Il10 genes [16], BATF cooperates with IRF4 for the induction

of IL-9 [42]. Accordingly, mouse and human Th9 cells depend on BATF for IL-9 production. Similarly to IRF4, BATF expression in Th cells promotes allergic airway inflammation [42]. As Th9-cell differentiation was in addition described to depend on the ETS transcription factor PU.1 [45], IRF4 might also regulate Th9-cell differentiation via EICE binding in concert with PU.1. Finally, for the induction of IL-9 production, IRF4 cooperates with SMAD2 and SMAD3 proteins, which are induced by TGF-β signaling [21], indicating multiple mechanisms and interaction partners utilized by IRF4 during Th9-cell differentiation (Fig. 1A). The relevance of IRF4 for the in vivo development of Th17 cells has been demonstrated in several

autoimmune disease models, in which pathogenic Th17 cells play a central role. Irf4–/– mice have been shown to be totally resistant to the induction of experimental autoimmune encephalomyelitis (EAE), which is a mouse model for multiple sclerosis (MS), and this resistance correlated with lack of Th17-cell differentiation [46]. Reconstitution of Irf4–/– mice with WT CD4+ T cells restored PLEK2 their susceptibility to the disease and the transferred cells developed a Th17 phenotype, again pointing to a T-cell intrinsic defect of Irf4–/–CD4+ T cells [46]. Furthermore, IRF4 deficiency was protective in T-cell-dependent colitis models, such as transfer colitis and oxazolone-induced as well as trinitrobenzene sulfonic acid induced colitis [47]. Again, resistance to colitis induction correlated with defective differentiation of naïve Irf4–/–CD45RBhighCD4+ T cells into Th17 cells, along with reduced IL-6 production by Irf4–/– mucosal T cells. Consistent with these findings, IRF4 levels were augmented in patients with inflammatory bowel disease and correlated with enhanced production of IL17 and IL22 mRNA [47, 48]. Thus, lack of IRF4 seems to cause resistance to Th17-mediated autoimmune diseases.

The role of Sorafenib research buy D. massiliensis in tick natural history and its influence on tick’s

fitness are unknown. However, a recent study suggests that this bacterium is pathogenic towards humans (Subramanian et al., 2011). Spiroplasmas (class Mollicutes) are helical, motile, wall-less prokaryotes associated with a variety of insects, other arthropods and some plant hosts (Tully et al., 1981). They are usually considered as commensal organisms in their arthropod hosts, but several are pathogenic for insects and plants. Several species were associated with a male-killing phenomenon (Jiggins et al., 2000). Spiroplasmas have been identified in ticks (Haemaphysalis leporispalustris and Ixodes pacificus) and blood-sucking members of the Diptera, including horseflies

(Tabanus spp.), deerflies (Chrysops spp.) and mosquitoes (Aedes spp., Culex spp.). Spiroplasma ixodetis was first isolated from I. pacificus, a principal vector of Lyme disease on the west coast of the USA (Tully et al., 1981). Later, a nearly identical bacterium PLX-4720 chemical structure was isolated from a pool of Ixodes ticks in North Rhine-Westphalia (Germany) (Henning et al., 2006), and another strain of Spiroplasma spp. (genetically very close to S. ixodetis) was recently isolated on the XTC cell line from a I. ricinus tick sampled in Slovakia, where prevalence of tick infection by Spiroplasma was 2.5% (Subramanian et al., in press). Virtually nothing is known about the relationship between Spiroplasma and ticks. However, several publications support the pathogenic role RVX-208 of this bacterium towards humans. Thus, Lorenz et al. (2002) found a Spiroplasma

sp. infection causing a unilateral cataract in a premature human baby. Spiroplasmas have been reported to be involved in neurodegenerative diseases such as scrapie or Creutzfeldt–Jakob disease (Bastian et al., 2004, 2011). In addition to the four potential tick endosymbionts discussed above and to established human pathogens known to be transmitted by ticks (Table 3), several other fastidious intracellular bacteria have been shown to be closely associated with ticks, including Candidatus Midichloria mitochondrii (Sassera et al., 2006), Francisella-like bacteria (Sun et al., 2000), Wolbachia spp., and different Rickettsiales. More studies are needed in this emerging field, whose results may have many applications, including the control of vectorborne diseases of humans and animals. Indeed, the concept of targeting endosymbionts as a mean to control ticks and tickborne diseases has been tested using a chemotherapeutic approach (Ghosh et al., 2007). Novel methods for the isolation and characterization of tick-associated bacteria will likely promote new approaches to control ticks by targeting their endosymbionts.

Additionally, one set of samples was pretreated with vehicle Alectinib purchase or 10 mM dimedone for 1 h prior to stimulation and sulfenic acid labeling. For immunoprecipitation experiments, 2–4 × 106 purified B cells were stimulated with 10 μg/mL anti-IgM and lysates were prepared as previously described by Michalek et al.

[14]. Briefly, cells were washed with PBS prior to lysis in the presence of DCP-Bio1 and lysates were precleared for 1 h at 4°C with protein G beads (Dynal). Following magnetic bead removal, lysates were incubated with 2.5 μg/mL anti-SHP-2 (BD Pharmingen), anti-SHP-1, or anti-actin (Santa Cruz Biotechnology) at 4°C with constant rotation overnight. The following day, protein G beads were added and the lysates were rotated at 4°C for 3 h. After discarding the supernatant, the magnetic beads were washed three times, resuspended in lysis buffer, and protein was eluted by boiling in reducing sample buffer (Pierce). Affinity capture of biotinylated proteins was performed according to Nelson et al. [47]. Samples were boiled with reducing sample buffer, separated on a 10% precast SDS denaturing gel, and transferred to a PVDF (polyvinylidene fluoride) membrane. The Y-27632 mw membrane was blocked and probed with anti-PTEN (Cell Signaling) or anti-CD45 (Santa Cruz Biotechnology) according to the manufacturer’s protocol. For sulfenic acid

detection, samples lysed in the presence of 1 mM dimedone were separated on a 10–12% precast SDS denaturing gel and transferred to a PVDF membrane. The membrane was blocked and incubated with anti-dimedone antibody (Millipore) according to the manufacturer’s protocol. Proteins were visualized as previously described [14]. The blot was stripped and probed with anti-actin.

To quantify sulfenic acid, actin and cysteine sulfenic acid levels were normalized between samples using a Kodak Image Station 4000R and Carestream Montelukast Sodium Molecular Imaging Software. The entire length of the gel lane was used to determine sulfenic acid levels. Only the protein band was used for actin. The sulfenic acid signal was then normalized to actin protein levels. Detection of sulfenic acid during immunoprecipitation experiments was performed as previously described [14]. Briefly, samples lysed in the presence of 5 mM DCP-Bio1 were separated on a 7.5–15% SDS denaturing gel and transferred to a PVDF membrane. The membrane was blocked overnight at 4°C with 5% FCS in tris buffered saline supplemented with 0.1% Tween-20 (TBS-T). The following day, the membrane was washed three times and incubated with 1:50,000 dilution Streptavidin-HRP (Southern Biotech) in 5% FCS in TBS-T for 1 h at room temperature. After washing, the membrane was developed as previously described [14]. CFSE (5–6-carboxyfluorescein diacetate, succinimidyl ester, Molecular Probes) was resuspended in DMSO at a 5 mM stock and stored at −20°C. Purified B cells were washed with cold PBS three times and resuspended in PBS at 20 × 106 cells/mL. The CFSE stock solution was diluted in PBS to 6.

Dry weight (normotension without the need for Panobinostat order antihypertensive medications) is targeted in the same way for patients on SDHD and NHD as for those on conventional HD. However, patients are more likely to achieve their dry weight with more frequent HD regimens. Despite generally lower ultrafiltration rates and better volume control, patients at home can have a tendency to achieve excessive interdialytic weight gains given the increased flexibility of dialysis regimens and liberalization of diet and fluids. Patients on SDHD and NHD should still be encouraged to reduce fluid accumulation and limit gains <2 L

in between sessions. With improved volume control, blood pressure may drop over time in both SDHD and NHD requiring reduction or even discontinuation of antihypertensive medications.34 Generally, non-cardioprotective antihypertensive medications should be stopped first. As with conventional HD, good vascular access is crucial for successful dialysis with alternative HD regimens. Difficult learn more access means difficult needling, longer training time and an unhappy patient. An arteriovenous fistula

(AVF) is the preferred vascular access for alternative HD regimens. NHD can be delivered successfully with an AVF using double-needle or even single-needle cannulation techniques; and patients at home are usually self-needling. Single-needle cannulation may potentially increase safety in case of accidental needle dislodgement and theoretically could increase access survival because of fewer cannulations. Although this technique Thalidomide reduces the dose of dialysis by decreasing effective dialysis time and potentially increasing the degree of access recirculation, this problem is less of a concern with

NHD. Central venous catheter (CVC) use at home is also possible but not encouraged. In the most recent IQDR, 63% of patients undertaking NHD at home in Australia and New Zealand were dialysing through a native AVF and 32% were dialysing though a CVC.6 These proportions are similar to those for the conventional HD population in both countries as well as for alternative HD patients in Canada undertaking NHD at home. In the Australian cohort alone however, the prevalence rates for CVC were between 0% and 9% according to the IQDR report in 2008, much better than the HD population in Australia as a whole.35 The reasons for the higher AVF rates in NHD patients at home in Australia are not known but may include patient characteristics that increase the likelihood of having an AVF created in the first place. There are several methods of AVF cannulation for alternative HD regimens. The ‘buttonhole technique’ involves creation of a subcutaneous tract (composed of scar tissue between the skin and the access) allowing for repeated cannulation at the same arterial and venous sites.

1A). NF1 site is located between positions -3992/–3982 from the ATG (A corresponding to position +1 of isoform 1). This site has been previously described and characterized in human airway epithelial cells [16]. NF2 site is located between positions

–369/–359 of I-BET-762 the human TSLP promoter. Two additional putative NF-κB sites, named NF3 and NF4 are located, respectively, at positions –1528 and –3421 of TSLP promoter. A search of the relevant vertebrate databases revealed that the region of human TSLP promoter containing the NF2 site, is conserved in numerous mammals namely Pongo abelii, Pan troglodytes, Mus musculus, Rattus norvegicus, Equus caballus, and Bos taurus (Fig. 1B). Within these species, no sequence corresponding to human NF1 was found in M. musculus and R. norvegicus. A sequence similar but not identical to human NF1 was found in E. caballus and B. taurus. As expected, both NF1 and NF2 sites, as well as NF3 and NF4 sites, were

conserved in primates. The latters were also found in E. caballus EGFR inhibitor and M. musculus but not in B. Taurus or R. norvegicus. Three putative AP-1 binding sites (AP1–1, AP1–2, and AP1–3), are located at positions –3942, –1255, and –263, respectively (Fig. 1). Like NF1 binding site, AP1–1 site has been described in human airway epithelial cells [16]. Moreover, AP1–2 and AP1–3 are conserved between human and mice but not AP1–1. Since NF-κB and AP-1 are key transcription factors involved in various inflammatory pathologies in both humans and mice and several reports suggest TSLP to be regulated by NF-κB [16, 19] we focused our work on a number of inflammatory tuclazepam agonists including IL-1, TNF-α, and PMA as well as TLRs ligands to evaluate, at the transcriptional level, TSLP regulation

in human IECs. For this purpose, we used a luciferase reporter assay where the luciferase gene was cloned under the control of a 4-kb-long fragment of TSLP promoter. Among the TLRs ligands used Flagellin and FSL1 were able to stimulate the reporter gene activity in HT-29 and Caco-2 cells, respectively (Supporting Information Fig. 1). When Caco-2 cells were stimulated with IL-1, a 12-fold increase in luciferase activity was measured at 24-h poststimulation, whereas a weaker activation was observed in cells stimulated with TNF (twofold) (Fig. 2A). PMA, a diacyglycerol analog that activates PKC and butyric acid, is an end-product of bacterial fermentation, that strongly regulates gene expression in IECs [20-22]. We found that PMA also strongly induced TSLP-dependent luciferase activity (ninefold). Moreover, when Caco-2 cells were co-incubated with PMA and butyric acid a dramatic stimulation (100-fold) of luciferase activity was noted (Fig. 2A). Similar results were obtained with HT-29 cells, however as expected, HT-29 cells were less sensitive to PMA and much more to TNF (data not shown).

CD19 can also associate with the BCR in

the absence of CD21 to promote BCR ICG-001 mw signalosome assembly upon recognition of membrane-associated antigens 4. The cytoplasmic tail of CD19 contains two canonical motifs for recruitment of PI3K (YXXM), and these are required for CD19 function 5. Genetic evidence supports a functional role for AKT downstream of CD19, in that combined deletion of two AKT genes (Akt1 and Akt2) in mouse B cells confers a defect in marginal zone (MZ) B-cell development 6 similar to the phenotype of CD19-deficient mice 5, 7. However, it is not yet clear which AKT substrates regulate MZ-cell development. Forkhead box subgroup O (Foxo) transcription factors activate or suppress target genes in a cell type-specific and context-dependent manner 8, 9. In resting lymphocytes, Foxo proteins are localized to the nucleus and activate genes that maintain quiescence as well as proper homing and recirculation 1. Phosphorylation by AKT causes cytoplasmic sequestration and degradation of Foxo factors, inhibiting the Foxo gene expression program. The Foxo1 family member has been studied in lymphocytes by conditional deletion using Cre-lox systems. This work has identified unique roles

for Foxo1 Gefitinib supplier in several aspects of B-cell function 10. Deletion of the Foxo1 gene in early B-cell progenitors using Mb1Cre caused a block at the pro-B cell stage. Deletion at a later stage with Cd19Cre caused a partial block at the pre-B-cell stage. Deletion

of Foxo1 in late transitional B cells with Cd21Cre blocked class-switch recombination. We have examined in more detail the phenotype of mature B cells in mice with Cd19Cre-mediated deletion of Foxo1. We find that these mice have fewer FO B cells and a higher percentage of MZ cells. In mice homozygous for the Cd19Cre knock-in allele, which lack CD19 protein, MZ cells are absent as reported previously 5, 7 but this defect is reversed by the concomitant Metformin order deletion of Foxo1. This genetic epitasis analysis suggests the possibility that CD19 negatively regulates Foxo1 to promote MZ B-cell development. We generated a conditional Foxo1 allele by inserting LoxP sites flanking the first exon of Foxo111. Mice homozygous for the Foxo1-flox allele are denoted Foxo1f/f herein. We bred Foxo1f/f mice with Cd19Cre mice in which the Cre recombinase is knocked into the Cd19 locus 12. Splenic B cells from Foxo1f/fCd19Cre mice expressed no detectable Foxo1 protein as determined by immunoblot, whereas Foxo3a expression was unchanged (Supporting Information Fig. 1A). Several aspects of B-cell development in these mice were altered in a manner similar to the phenotype of another strain of Foxo1f/fCd19Cre mice reported by Dengler et al.10. In particular, our Foxo1f/fCd19Cre mice had fewer IgM+ bone marrow B cells (Supporting Information Fig. 1B), and a population of peripheral B220+ cells lacking surface expression of IgM or IgD (Supporting Information Fig.

2A). In contrast, claudin-18 Selleckchem Napabucasin and 23 mRNAs were not increased upon IL-4-stimulation, while claudin-8 and 9 mRNAs were not detected at all in C57BL6 thio-PEM. Thus, besides E-cadherin, claudin-1, 2 and 11 are members of the junction protein family whose mRNAs are induced in IL-4-stimulated thioglycollate-elicited

peritoneal macrophages. For this reason, we confined our further analysis to this limited set of claudin genes. Of note, the IL-4-mediated induction of these three claudins is largely STAT-6 dependent, as demonstrated by their significantly reduced upregulation in STAT-6-deficient thio-PEM (Fig. 2B). Finally, to extrapolate these findings to other macrophages, BALB/c bone marrow-derived macrophages (BMDM) were treated with IL-4 and assessed for Cdh1, Cldn1, Cldn2 and Cldn11 gene expression. Although Cdh1 and Cldn2 are still inducible by IL-4 in BMDM, the level of induction is less pronounced as compared to thio-PEM (Fig. 2C). As opposed to this, Cldn11 mRNA is strongly induced in BMDM. Of note, the differences in IL-4-mediated Cldn1, 2 and 11 gene inductions between BALB/c thio-PEM and BALB/c BMDM are not because of significant differences in the basal

expression levels of these genes in the DNA Damage inhibitor respective macrophage populations (Table S1). Together, these data confirm the IL-4-induced, STAT6-dependent gene expression of Cldn1, Cldn2 and in particular Cdh1 and Cldn11 in AAMs, but the extent of gene induction depends on the macrophage type. E-cadherin regulation in AAMs was already studied in detail before [8] and is therefore not included in the remainder of this manuscript. To evaluate whether enhanced gene expression of the selected claudins resulted in increased protein levels, we performed a FACS staining on naive and IL-4-stimulated BALB/c thio-PEM. While E-cadherin expression was clearly detected at the cell surface of IL-4-treated,

but not naive thio-PEM as documented before [8], claudin-1, 2 and 11 were below the detection limit (data not shown). Furthermore, no claudin-1, 2 and 11 proteins were detected by Western blot in complete cell lysates of (-)-p-Bromotetramisole Oxalate IL-4-stimulated BALB/c thio-PEM. Complete brain and kidney homogenates were used as controls and scored positive for the tested claudins, validating the experimental procedure for detecting these proteins (Fig. S1). The effect of IL-4 on Cldn1 gene expression in macrophages was rather limited. Besides IL-4, other cytokines such as IL-10 and TGF-β have been reported to induce an M2 macrophage activation state. Therefore, BALB/c and C57BL/6 thio-PEM and BALB/c BMDM were treated with IL-4, IL-10 and TGF-β, and Cldn1 induction was assessed.