C57BL/6J (B6) mice were purchased from the Jackson Laboratory (Bar Harbor, ME). B6.129P-Hrh1tm1Wat (H1RKO) [[51]], B6.129P-Hrh2tm1Wat (H2RKO) [[52]], B6.129P2-Hrh3tm1Twl (H3RKO) [[53]], and B6.129P-Hrh4tm1Thr (H4RKO)

mice (generated by Lexicon Genetics, Woodlands Park, TX) [[54]] were maintained at the University of Vermont (Burlington, VT). All strains were backcrossed to the C57BL/6J background for at least 10 generations. Individual HRKO mice were interbred and the resulting F1 mice were intercrossed together to generate H1H2RKO and H3H4RKO mice. The experimental procedures used in this study were approved by the Animal Care and Use Committee of the University of Vermont. Mice were immunized for the induction of EAE using a 2× immunization protocol. The animals were injected subcutaneously in the posterior right and left flank with a sonicated phosphate-buffered saline (PBS)/oil emulsion containing 100 μg of MOG35–55 and

Fer-1 datasheet CFA (Sigma-Aldrich, St. Louis, MO) supplemented with 200 μg of Mycobacterium tuberculosis H37Ra (Difco Laboratories, Detroit, MI). One week later, all mice received an identical injection of MOG35–55-CFA [[31]]. Mice were ranked scored daily for clinical quantitative trait variables beginning at day 5 after injection as follows: 0, no clinical expression of disease; 1, flaccid tail without hind limb weakness; 2, hind limb weakness; 3, complete hind limb paralysis and floppy tail; 4, hind leg paralysis accompanied Idasanutlin solubility dmso by a floppy tail and urinary or fecal incontinence; 5, moribund. Assessments of clinical quantitative trait variables were performed as previously described [[31]].

Histopathological evaluations were done as previously described [[55]]. Briefly, brains and spinal cords were dissected on 30th day postimmunization, from calvaria and vertebral columns, respectively, and fixed by immersion in 10% phosphate-buffered formalin (pH 7.2). After fixation, trimmed and representative transverse section-embedded in paraffin and mounted on glass slides. Sections were stained with hematoxylin and eosin for routine evaluation and Luxol fast blue-periodic STK38 acid-Schiff reagent for demyelination. Representative areas of the brain and spinal cords were selected for histopathological evaluation. The following components of the lesions were assessed: (i) severity and extent of the lesion; (ii) extent and degree of myelin loss and tissue injury (swollen axon sheaths, swollen axons, and reactive gliosis); (iii) severity of the acute inflammatory response (predominantly neutrophils); and (iv) severity of the chronic inflammatory response (lymphocytes/macrophages). Lesions in the brain and spinal cord (SC) were evaluated separately and assigned a numerical score based on a subjective scale ranging from 0 to 5. A score of 0 indicates no lesions; 1 indicates minimal; 2, mild; 3, moderate; 4, marked; and 5, severe lesions. BBB permeability was assessed as previously described [[56]].

Three of the five ‘classical’ HIES patients

had known STAT3 mutations (R382W twice and V463del) [5] (Table 1). Two of the patients with ‘classical’ HIES had no STAT3 mutation. To investigate the immunological functional properties Birinapant chemical structure with respect to Th17 responses in HIES patients with different mutations, PBMC from healthy volunteers, ‘classical’ HIES patients and three members from a HIES family with ‘variant’ HIES were assessed for the capacity to mount IL-17 responses. We have developed a new methodology of Th17 generation using human PBMC stimulated with whole microbial stimuli relevant for HIES: S. aureus and C. albicans[18]. HIES patients had a defective response to C. albicans, although IL-17 was measurable in all patients (Fig. 2a). Interestingly, IL-17 production was completely absent in PBMC stimulated with S. aureus in all ‘classical’ HIES patients (Fig. 2b). In contrast, PBMC isolated from the variant HIES patients, bearing the STAT3 mutations in the linker domain, were able to produce IL-17 in response to S. aureus, albeit at lower concentrations when compared to healthy volunteers (Fig. 2b and c). IFN-γ production was distorted in HIES patients when compared to healthy controls, while IL-10 was found to be elevated in HIES patients when stimulated with

both S. aureus and C. albicans. The in vitro stimulations described above suggest that HIES patients have a significant defect in the generation of Th17 cells. This was selleck compound indeed the case for the patients with ‘classical’ HIES, either bearing STAT3 mutations or not (Fig. 3). Surprisingly, when the familial variant HIES patients were challenged with disease-related microorganisms, they showed a clear induction

of single IL-17-positive and IL-17/IFN-γ-positive CD4+ cells compared to normal controls (Fig. 3). IL-6 augmented IL-17 production induced by GBA3 C. albicans and S. aureus in cells isolated from healthy controls (Fig. 4a). No effect was apparent in the HIES patients, independently of the type of STA3 mutation. In contrast to IL-6, IL-10 reduced the amount of IL-17, and this effect was observed both in healthy controls and HIES patients (Fig. 4b). Mutations in the SH2 and DNA-binding domain of STAT3 have been reported to be the cause of disease in a large proportion of HIES patients [4]. These mutations function as dominant-negative mutations [4] and result in a defective Th17 response in these patients [9,10], explaining many of the clinical features of HIES. In the present study we confirm, on one hand, the relationship between HIES and defective Th17 responses; on the other hand, we also refine this notion to include the relationships between the type of STAT3 mutation, immunological response to relevant microbial stimulation and clinical phenotype of the patients.

111) and TNF-α-PECy7 (MAb11; all from BD Biosciences), IL-17-AlexaFluor647 (eBio64CAP17, eBiosciences) and CD4-QDot605 (SK3, Invitrogen). For the 24 children, GM-CSF-PE (BVD2-21C11; BD Biosciences) was also included in the antibody panel. For adolescents an additional set of rAg85A-, BCG-stimulated and unstimulated cells was available and the surface phenotype of cytokine-producing CD4+ T cells was determined

with the following panel: CD3-Pacific Blue, CD4-QDot605, IFN-γ-AlexaFluor700, IL-2-FITC, TNF-α-PECy7, IL-17-AlexaFluor647, CD45RA-PerCPCy5.5 (HI100, eBiosciences) and CCR7-PE (150503, R&D Systems). At least 1 million total cells were acquired on an LSR II flow cytometer (BD Biosciences). Cell doublets were excluded using forward scatter-area versus forward scatter-height parameters. Unstained cells and single-stained mouse κ beads were used to calculate compensations for every run. Data analysis buy DAPT was performed with FlowJo software version 8.5.3 (TreeStar). The boolean gate platform was used with individual cytokine gates to create all possible response pattern combinations. For the IFN-γ ELISpot assay, the cut-off for positive responses was 17 spot forming cells per million

PBMC. The cut-off for positive response measured by the intracellular cytokine detection assay was 0.01% of gated cells. A minimum of 20 cytokine-positive cells were selleck required for surface phenotypic analysis. The data analysis programs PESTLE (version 1.5.4) and SPICE (Simplified Presentation of Incredibly Complex Evaluations; version 4.1.6)

were used to analyse flow cytometry data and generate graphical representations of T-cell responses using background-deducted flow cytometric data (both kindly provided by Mario Roederer, Vaccine Research Center, NIAID, NIH). Statistical tests were performed with Prism 4.03 (GraphPad). The distributions of the T-cell frequency data were extremely skewed, and log transformations did not result in symmetrical distributions. As a result, normal-base linear regression-type models could not be used to model the frequency data. These measurements were thus summarized by time point, by use of medians and interquartile ranges, and were compared at each timepoint by use of the Kruskal–Wallis (for overall effect) and Mann–Whitney U tests. Resulting p values should be interpreted conservatively because http://www.selleck.co.jp/products/BAY-73-4506.html of the increased chance of false-positive findings resulting from multiple testing. The authors thank all the participants who took part in this trial. They thank Tom Ottenhoff and Kees Franklin from Leiden for the recombinant Ag85A protein and Zia Sherrell for administrative support and project management. This work was supported by the Wellcome Trust (081122/Z/06/Z) and Europe AID (SANTE/2006/105–066). T. J. S. is a Wellcome Trust Research Training Fellow (080929/Z/06/Z), H.M. is a Wellcome Trust Senior Clinical Fellow, A. V. S. H. is a Wellcome Trust Principle Research Fellow. W.A.H.

8 T-cell differentiation occurs by a complex transcriptional programme initiated by TCR and environmental signals but it is also accompanied by epigenetic changes at specific loci.9 We first review the transcription factors that are activated downstream of TCR signalling and then explore certain principles that might operate in regulating them. Signalling through the TCR activates at least three families of transcription factors: nuclear factor of activated T cells (NFAT), activating protein 1 (AP-1) and nuclear factor-κB (NF-κB) (see Fig. 1). Gene expression Deforolimus by these transcription factors is not restricted to

T cells but rather is found in almost every cell type in the body. As a result, extensive biochemical analysis has been performed over the years describing

the network of interacting proteins that activate them. We will briefly review the regulation of these factors in T cells. The NFAT family consists of five members: NFAT1 (NFATp or NFAT c2), NFAT2 (NFATc or NFATc1), NFAT3 (NFATc4), NFAT4 (NFATc3) and NFAT5; NFAT3 is not expressed in immune cells. All NFAT proteins contain a conserved Rel homology domain (regulatory domain) and an NFAT homology domain (DNA-binding domain). All except NFAT5 are regulated by calcium.10 NFAT is a transcription factor that is normally resident in the cytoplasm and is de-phosphorylated by a calcium-dependent phosphatase, calcineurin. This de-phosphorylation activates it and causes its translocation into the nucleus.11 Nuclear export of NFAT is mediated by phosphorylation. Glycogen 17-AAG synthase kinase 3 (GSK-3) is known to phosphorylate conserved serine residues necessary for nuclear export.12 In peripheral lymphocytes, antigen receptor signalling leads to the rapid inactivation of GSK-3. Activators of PKA suppress interleukin-2 (IL-2) production and T-cell activation, consistent with the possibility that NFAT is a substrate for protein kinase A (PKA).12 NFAT4 is known to be negatively regulated through phosphorylation by casein kinase 1 in the cytoplasm.13 Another mechanism of negative regulation of NFAT involves calcipressin, a target of NFAT that

binds to and inhibits calcineurin.10 Members of the homer family have been shown to bind to NFAT and compete with calcineurin, hence negatively regulating NFAT Flucloronide activation.14 Nuclear retention of NFAT can also be achieved by sumoylation, adding another level of complexity in its regulation.15 Unlike NFATc2, which is constitutively transcribed in T cells, transcription of the NFATc1 gene in effector T cells is strongly induced within 3–4 hr of TCR and co-receptor stimulation.16 Members of the NFAT family are redundant, as the mice lacking individual NFAT proteins show mild alterations in immune function whereas more severe defects are observed when more than one member is knocked out.10 NFAT plays a crucial role in T-cell differentiation.

PolyI:C was purchased from GE Healthcare company, and solved in milliQ water. For polyI:C treatment, polyI:C (50 μg/mL)

was mixed with DEAE-dextran (0.5 mg/mL) (Sigma) in the culture medium, and the cell culture supernatant was replaced with the medium containing polyI:C and DEAE-dextran. Using DEAE-dextran, polyI:C is incorporated into the cytoplasm to activate RIG-I/MDA5. VSV Indiana strain or poliovirus type 1 Mahoney strain were used for virus assay. SB203580 ic50 Vero derived cell (Vero-SLAM) was used for propagation and plaque assay for VSV indiana strain or poliovirus type 1 Mahoney strain. HEK293 cells were infected with viruses at MOI=0.001 in a 24-well plate. The virus titers of culture media at indicated hours post infection in the figures were determined by plaque assay using Vero-SLAM cells. In some experiments that require rapid virus propagation, high MOI (0.1∼1) was used for infection. HEK293FT cells were transfected in a 6-well plate with plasmids encoding DDX3, IPS-1, RIG-I or MDA5 as indicated in the figures. Twenty-four hours after tranfection, the total cell lysate

was prepared by lysis buffer (20 mM Tris-HCl (pH 7.5) containing 125 mM NaCl, 1 mM EDTA, 10% glycerol, 1% NP-40, 30 mM NaF, 5 mM Na3Vo4, 20 mM IAA and 2 mM PMSF), and the protein was immunoprecipitated R788 concentration with anti-HA polyclonal (Sigma) or anti-FLAG M2 mAb (Sigma). The precipitated samples were resolved on SDS-PAGE, blotted onto a nitrocellulose sheet and stained with anti-HA (HA1.1) monoclonal (Sigma), anti-HA polyclonal or anti-FLAG M2 mAb. HeLa cells were plated onto cover glass in a 24-well plate. In the Tyrosine-protein kinase BLK following day, cells were transfected with indicated plasmids using Fugene HD (Roch). The amount of DNA was kept constant by adding empty vector. After 24 h, cells were fixed with 3% of paraformaldehyde in PBS for 30 min, and then permeabilized with PBS containing 0.2% of Triton X-100 for 15 min. For the polyI:C stimulation, 100 ng of polyI:C were transfected into HeLa cell in 24-well plates together with IPS-1 or DDX3 expressing vectors, and 24 h after

the transfection, the cells were fixed and stained for confocal microscopic analysis. Permeabilized cells were blocked with PBS containing 1% BSA and were labeled with anti-Flag M2 mAb (Sigma), anti-HA polyclonal Ab (Sigma) or Mitotracker in 1% BSA/PBS for 1 h at room temperature. The cells were then washed with 1% BSA/PBS and treated for 30 min at room temperature with Alexa-conjugated Ab (Molecular Probes). Thereafter, micro-cover glass was mounted onto slide glass using PBS containing 2.3% DABCO and 50% of glycerol. The stained cells were visualized at ×60 magnification under a FLUOVIEW (Olympus, Tokyo, Japan). The authors thank Dr. M. Sasai, Dr. T. Ebihara, Dr. K. Funami, Dr. A. Matsuo, Dr. A. Ishii, Dr. A. Watanabe and Dr. M. Shingai in our laboratory for their critical discussions.

The ATF6 branch of UPR also plays a role in plasma cell function [97]. Murine B cells transduced with a dominant-negative form of ATF6 had diminished IgM secretion after treatment with LPS. Expression of Ig transcripts in these cells happened

at the same levels U0126 manufacturer as in control cells, while protein levels were diminished. This suggests that protein synthesis is impaired and/or degradation of nascent chains is enhanced in the presence of ATF6 dominant-negative mutant [97]. Most of what we know about the UPR pathway refers to C. elegans and mice studies. A few years ago, we got involved with studying the UPR pathway based on the hypothesis that the hypogammaglobulinemia observed in Common Variable Immunodeficiency (CVID) was a

result of defective activation of the UPR pathway [98]. CVID is the most prevalent immunodeficiency of adult humans and it is a syndrome diagnosed by the loss of at least two immunoglobulin isotypes. Several defects have been identified as causes of CVID, but a large number of patients still have unknown underlying causes for their phenotype (reviewed by [99]). We identified one CVID patient whose activation of the IRE1/XBP-1 pathway occurs at a slower rate as compared to a matched healthy control. CH5424802 manufacturer Ex vivo and EBV-immortalized B cells were treated with LPS or brefeldin A (ER stressor) and the levels of transcripts for XBP-1s, IRE1α, and BiP were quantified over time. XBP-1 splicing was performed at a much slower rate in this patient, as well as transcription of BiP and IRE1Α. Peripheral blood B cells were enlarged and did not present typical membrane-bound IgM. Instead, filipin chains of IgM co-localized with BiP inside the ER. Both the XBP-1 and endonuclease/kinase domains of IRE1α were sequenced, and had no mutations that could explain the defective activation. Because the defect(s) resulted in deficient BiP transcription,

we hypothesized that a rescue of function could be achieved by providing these cells with chemical chaperones. Indeed, in vitro treatment of the cells with DMSO rescued secretion of IgM and IgG, suggesting that there is no defect on the secretory pathway of the cells [98]. More recently, we started analyzing ex vivo cells from CVID patients to check whether the differentiation programme of their B cells is completed by the time these cells reach periphery. It is conceivable to hypothesize that the UPR pathway will be properly activated only when the cell has reached a certain developmental stage. Our preliminary data suggest that B cells from CVID patients represent a heterogeneous group, where cells at different stages of differentiation can be found based on expression of FMC7, CD5, CD19, CD23, CD38 and CD45.

“
“Sustained research efforts over the last 50 years have revealed

a considerable amount of information about immunity to taeniid cestode infections in the parasites’ intermediate hosts. As a product of this research, a series of effective recombinant vaccines have been developed which have no parallel in any other group of parasitic organisms. There are, however, many important aspects relating to immunity that remain to be elucidated. Some concepts have come to be firmly held as selleck facts and yet the supportive data are either conflicting or unconfirmed. This review considers the phenomenon of immunity to re-infection with taeniid cestodes in their intermediate hosts, examining carefully the nature of the evidence that is available to support conclusions that have been drawn in this area. “
“Replacement therapy with exogenous factor VIII (FVIII) to treat haemorrhages or used in prophylaxis induces inhibitory anti-FVIII immunoglobulin G (IgG) in some patients with haemophilia A. Therapeutic strategies to prevent

the onset of the deleterious anti-FVIII immune response are still lacking. Maternal IgG is transferred to the offspring during fetal and neonatal life. While protecting the offspring from bacterial and viral infections, maternal IgG may alter the repertoires of T and B lymphocytes, and may impair vaccination in early infancy. Using selleck screening library haemophilic mice, we demonstrate that the transfer of maternal anti-FVIII IgG modulates the onset of anti-FVIII inhibitory IgG in early adulthood. The protective effect is reproduced upon reconstitution of naive mice with anti-FVIII IgG, PLEK2 suggesting that the reduced ability to mount an anti-FVIII immune response is the result of an interference between circulating anti-FVIII IgG and the administered FVIII rather than to a profound remodelling of lymphocyte repertoires occurring during the ontogeny of the immune system. Administration of exogenous factor VIII (FVIII) to patients with haemophilia A leads, in up to 30%

of the cases, to the development of neutralizing anti-FVIII alloantibodies that inhibit the pro-coagulant activity of FVIII. Different therapeutic strategies are being used to eliminate FVIII inhibitors, such as the administration of B-cell-depleting anti-CD20 antibodies (Rituximab®, Genentech Inc, South San Francisco, CA, USA) or the induction of immune tolerance upon repeated injection of high doses of FVIII.1 In patients, prophylaxis has been proposed as one of the rare solutions towards a reduction of the risk for the onset of the deleterious anti-FVIII immune response.2,3 During fetal life, maternal immunoglobulin G (IgG) of the IgG1 subclass is delivered through the placenta to the fetus via interactions with the neonatal Fc receptor (FcRn).

A statistical test based on measures of central tendency comparison was not applicable to the particular case of anti-IgM combined with IL-21. A P-value less than 0·05 was considered statistically significant. B cells die from apoptosis if maintained unstimulated in culture [31]. After 3 days, spontaneous apoptosis was higher in CD27+ than in CD27– B cells (79·2 versus 57·6%, P < 0·001) (Fig. 2a). When B cells are stimulated, they are rescued from apoptosis.

The effectiveness of the rescue depends upon both the kind of stimulus used and the subpopulation of B cells. For CD27– B cells, the strongest rescue effect was induced by anti-CD40 followed by CpG-ODN and to a lesser extent by anti-IgM, whereas for CD27+ B cells, CpG-ODN appeared to be the strongest rescue stimulus (Fig. 2b). Nevertheless, all the stimuli evaluated were more efficient in the CD27– than in the CD27+ buy Enzalutamide population: anti-CD40 (77·9 versus 23·9%, P < 0·001), CpG-ODN (71·4 versus selleckchem 57·3%, P < 0·01) and anti-IgM (52·7 versus 36·9%; P < 0·01) (Fig. 2b). Proliferation was evaluated simultaneously. Anti-CD40 and anti-IgM did not induce proliferation of either CD27– or CD27+ B cells while CpG-ODN induced proliferation of both subpopulations (Table 2). Although CpG-ODN

induced a lower level of proliferation on CD27– than CD27+ B cells (PI = 0·1 versus PI = 1·8, respectively; P < 0·001) (Table 2), it induced higher rescue from apoptosis in the CD27– population (Fig. 2b). These aforementioned results suggest that proliferation and rescue from apoptosis are two independent processes. CD27– B cells from CVID MB0 patients were less sensitive to rescue from apoptosis when stimulated with a T-dependent stimulus (anti-CD40) than control subjects (65·4 versus 77·9%, P < 0·05)

(Fig. 3a). They were also less sensitive to rescue from apoptosis when stimulated with a T-independent stimulus (CpG-ODN) than control subjects or CVID MB1 patients, although differences did not reach statistical significance (58·8 versus 71·4 and 63·0%, respectively, P = 0·075). CD27– B cells from CVID MB1 patients were rescued from apoptosis similarly to controls, regardless of the stimulus used (Fig. 3a). After BCR engagement with anti-IgM CD27– B cells from both CVID MB0 and MB1, patients Methane monooxygenase were rescued equally from apoptosis than healthy controls. CD27+ B cells from CVID MB0 patients, stimulated with either a T-dependent (anti-CD40) or a T-independent stimulus (CpG-ODN), were less sensitive to apoptosis rescue than control subjects (6·0 versus 23·9%, P < 0·01; and 23·2 versus 57·3%, P < 0·05, respectively) and CVID MB1 patients (6·0 versus 30·6%, P < 0·001; and 23·2 versus 65·7%, P < 0·01, respectively). They were also less sensitive to rescue from apoptosis after BCR engagement with anti-IgM than control subjects (19·2 versus 36·9%, P < 0·05) or CVID MB1 patients (19·2 versus 38·2%, P < 0·01) (Fig. 3b).

In fact, it is interesting to observe that in NSCLC patients, who had not been exposed to any antitumor treatment (including radio or chemotherapy), we could not detect cytotoxic anti-NeuGcGM3 antibodies in the conditions used for our study. This behavior was observed even in those patients less than 60 years of age. Only six of the 53 NSCLC patients studied had a low response against NeuGcGM3, and their sera were not able to bind to tumor cells expressing the antigen. The levels of IgG and IgM antibodies did not decrease with the learn more age of the cancer patients, however,

we did detect a significantly lower total IgM concentration in the cancer patients’ sera when compared with healthy Selleckchem Panobinostat donors’. In contrast, the IgG concentrations were similar, suggesting that the IgM reduction is not due to a general state of immunosuppression in these patients. The reduced level of anti-NeuGcGM3 antibodies detected in these patients could be a

consequence of the low total IgM levels, the isotype of the antibodies that recognize NeuGcGM3. But this specificity could be particularly affected, resembling what we observed for elderly healthy donors. In the case of these cancer patients, the observed behavior could be due to the anti-NeuGcGM3 antibody-secreting B-cell population being affected, or to the capacity of this B-cell population to secrete antibodies with this specificity being inhibited. By idiotypic vaccination, however, we have been able to boost this kind of immune response in cancer patients, which suggests that these cells are not completely deleted [17]. Another possibility is ID-8 that, in NSCLC patients, anti-NeuGcGM3 antibodies form immune complexes with gangliosides released from the tumor cells, which might affect their detection. This phenomenon could also result from the recruitment of such antibodies to the tumors since the presence of NeuGcGM3 in NSCLC tumor samples has been reported [41-43]. To our knowledge this is the first report showing that the levels of anti-NeuGcGM3 antibodies are lower in cancer patients in comparison with

healthy donors. Previous work reported that, depending on the ganglioside and the kind of tumor, higher or lower concentrations of antibodies against gangliosides in the sera of cancer patients with respect to healthy donors, could have a prognostic value [25, 44]. Further studies are needed to evaluate whether this is also the case for the antibody response against NeuGcGM3. Currently, we are carrying out experiments to elucidate the cause of the reduced levels of anti-NeuGcGM3 antibodies in NSCLC patients and extending these determinations to other kinds of tumors. In particular, we are trying to understand if the absence of this kind of response is a consequence of disease, or one of the causes increasing susceptibility to malignant transformation.

Efficient responses to the fungus require a complex network of immunological mechanisms. Together with alveolar macrophages and neutrophils, which constitute a primary line of innate cellular defence against A. fumigatus,1,2 the crucial role of the adaptive immunity has been extensively demonstrated.3 Indeed, besides the well-characterized protective role of T helper type 1 (Th1) lymphocytes,4–7 the newly described regulatory T cells and interleukin-17 (IL-17) -producing cells (Th17) represent important mediators of the inflammatory and anti-inflammatory

FDA-approved Drug Library ic50 host responses against A. fumigatus.8 However, dendritic cells (DCs) also play a fundamental function in initiating and modulating the specific immune responses upon recognition of A. fumigatus.5,9,10 After internalization of A. fumigatus conidia, DCs mature and acquire the capacity to polarize

naive T cells and, in turn, to promote a protective response.9 In keeping with these findings, in vivo results on the migration of lung DCs into lymphoid organs, where they drive an appropriate T-cell response to fungal antigens,11 have brought DCs centre stage as promising targets for intervention in immunotherapy and fungal vaccine development.12 In addition, it is important Sirolimus mw to consider several studies that have recently pointed to DCs and type I interferons (IFNs) as special players in the immune response tailored to combat tumours and infections.13–15 Indeed, although the anti-microbial properties of these cytokines have not been fully characterized yet, type I IFNs represent important immunomodulators of the innate, as well as the adaptive, arm of the immune system. Type I IFN can promote

the differentiation of human blood monocytes into DCs and contribute to their maturation.16,17 This leads to the generation of DCs able to stimulate a primary human antibody response, a Th1 proliferation,18 and a cross-priming of CD8 T cells against viral antigens.19 In addition, one crucial outcome of type I IFN-induced effects is the ability to directly stimulate IFN-γ production in natural killer and T cells,20–22 which in turn promotes the development of a cell-mediated immune response. Based on these immunoregulatory properties, in this work the expression and the MYO10 capacity of type I IFN, namely IFN-β, to modulate the T-cell polarizing capacity of A. fumigatus-infected DCs was investigated in an attempt to evaluate the effects induced by this cytokine on anti-fungal immunity. Although the phagocytosis of the fungus was not affected by IFN-β treatment, the maturation induced by A. fumigatus infection was enhanced in IFN-β-primed DCs, as evaluated by analysing the immunophenotype and the release of pro-inflammatory and regulatory cytokines. Accordingly, IFN-β endowed DCs with potent Th1 polarizing capacity because an enhanced IFN-γ production in T cells co-cultured with A. fumigatus-infected DCs was observed in the presence of IFN-β.