Recent progress in understanding the interaction between immune/inflammatory cell subsets via interleukins, particularly reciprocal regulation and counter balance between Th1, Th2, Th9, Th17, Th22 and T regulatory cells, as well as B-cell subsets, bring new possibilities for immune intervention. With regard to allergic diseases, the process of developing selleck chemical such diseases is characterized by effector Th2 cells that produce IL-4, IL-5, IL-9 and IL-13 1–4. In addition, recently defined cytokines, such as IL-25, IL-31, IL-32 and IL-33 that contribute to Th2

responses, tissue inflammation, allergen-specific IgE production, eosinophilia, mucous production, and the activation and cell death of the epithelium represent newly emerging and essential players in the pathgogenesis of allergic inflammatory disease 5–9. In the context of tissue-related allergy-driving factors, the IL-1 family member cytokine IL-33 is becoming a key player in the initiation and exacerbation of inflammatory responses. Its effects are exerted via its heterodimeric receptor that consists of ST2 and the ubiquitously expressed IL-1 receptor accessory protein (ILRAcP) Natural Product Library clinical trial 10. IL-33 integrates both innate and adaptive immunity in a unique manner. It affects basophils, mast cells, eosinophils, innate lymphoid cells, NK and NKT cells and Th2 lymphocytes 2, 11. In addition, IL-33 impacts CD34pos precursor cell populations 12 and is involved

in the activation of a cell subpopulation called nuocytes that are crucial for Idelalisib manufacturer parasite repulsion. This nuocyte population was defined as lineageneg ICOSpos ST2pos IL-17RBpos and IL17Rapos

cells and is considered to be an upstream Th2 inducer/amplifier, whose properties still remain to be defined in detail 7. The actions of IL-33 seem to be particularly evident when looking at models of mucosal inflammation. In this issue of the European Journal of Immunology, an article by Besnard et al. adds significant information regarding the role of IL-33 in the context of a mouse model of asthma-like lung inflammation 13. The authors demonstrate that IL-33 acts, in an ST2-dependent manner, as a maturation factor for BM-derived DCs via up-regulation of CD80, CD40 and OX40L. This process is accompanied by the release of pro-inflammatory cytokines, such as IL-6, IL-1β, TNF-α and TARC/CCL17. IL-33-pre-treated DCs were significantly more potent than non-treated DCs at inducing allergen-specific proliferation in naïve T-cells, and the generated T-cell responses were of a Th2 type with IL-5 and IL-13 production. This activation/maturation of lung resident DCs was also confirmed in vivo via local application of IL-33, inducing up-regulation of the homing receptor CCR7 in the CD11cpos fraction. The activated DC phenotype was observed in the draining LN, and PBMCs from the LN displayed a Th2 phenotype upon re-stimulation with anti-CD3/CD28.

Low concentration of CpG-ODN type A, in GM-CSF-pretreated cells, suppressed the production of TGF-β by neutrophils. However, at higher concentrations of CpG-ODN type A or B in the presence of GM-CSF, the TGF-β levels were maintained (Figure 1b). As shown, neither GM-CSF (Figure 1c) nor CpG-ODN class A (Table 2) elicited TNF-α production by neutrophils on their

own. On the other hand, CpG-ODN type A, at concentrations of 15 and 40 μg/mL, significantly stimulated GM-CSF-pretreated cells to secrete selleck compound TNF-α whereas control ODN did not (P < 0·05). Consequently, the production of TNF-α by neutrophils depends on the co-stimulation with GM-CSF and CpG-ODN type A. The secretion of IL-8 and TNF-α was not observed in the presence BAY 80-6946 of different concentrations of CpG-ODN class B regardless

of GM-CSF treatment (Figure 1a,c). Dose–response assessment showed that the optimum concentration of CpG-ODN to stimulate neutrophils was 40 μg/mL. In addition, with regard to TNF-α production, GM-CSF at concentration of 50 ng/mL possesses a co-stimulatory action on neutrophils in the presence of CpG-ODN. Ten healthy individuals were, therefore, tested simultaneously at these concentrations. As shown in Table 3, the obtained results from these experiments confirmed previous data. That is, the level of TNF-α in cells co-stimulated with the combination of these agents increased threefold and fivefold as compared to that in cells stimulated only with GM-CSF and CpG-ODN class A, respectively. The results obtained in healthy donors were followed up by assessment of the same parameters in asymptomatic and nonhealing CL individuals. IL-8, TNF-α and TGF-β were measured in cell supernatants PRKACG after 18 h (Figure 2a). Neutrophils

from all three groups produced similar levels of IL-8 upon stimulation with L. major. Moreover, in all groups, in comparison with infected neutrophils, IL-8 secretion by infected neutrophils, pretreated with GM-CSF and stimulated with CpG-ODN class A, decreased approximately 1·3-folds. TGF-β was detected, but not induced by either stimulation or infection in both healthy donors and nonhealing individuals. However, neutrophils from asymptomatic subjects did not produce measurable levels of TGF-β regardless of stimulation (Figure 2b). TNF-α production by neutrophils was induced by L. major infection (P < 0·05) in asymptomatic and nonhealing individuals (Figure 2c). Co-stimulation with GM-CSF and CpG-ODN class A did not increase TNF-α levels induced by L. major in nonhealing donors, but did so in asymptomatic individuals (P < 0·05). There was no difference in the level of TNF-α between unstimulated and stimulated infected neutrophils in normal and nonhealing individuals (Figure 2c). High-quality RNAs were isolated from all individuals (healthy, nonhealing and asymptomatic) for further analysis using real-time PCR.

The overall effect of these changes is to reduce PLX 4720 the inflammatory response in the target tissue. This was shown as a marked seasonal reduction in mucosal eosinophil recruitment and an increase in IFN-γ and IL-10 production in nasal mucosal biopsy samples after hay fever immunotherapy [126].

Many of the mechanisms described for conventional weekly up-dosing regimens of immunotherapy cannot apply to the initial phase of rush desensitization, where tolerance is induced within days. While the changes described above may eventually supervene, the initial rapid induction of tolerance to the allergen is likely to represent tachyphylaxis, where repeated doses of allergen induce a progressively weaker mediator response. Changes in histamine release, cytokine production by T cells and monocytes and even antibody binding activity have been described within the first days of rush immunotherapy. The tolerant state is maintained by continued administration of allergen, and a long-lasting immune tolerance develops as maintenance therapy continues. Allergen immunotherapy is a unique treatment, one of only a few that can truly be said to fundamentally alter a disease BIBW2992 clinical trial state. Therefore,

we approach advances in immunotherapy with caution: what can we improve without losing the core benefits? Clearly, we focus on the disadvantages of standard subcutaneous immunotherapy. It is time-consuming both in frequency of treatments and total duration of therapy, it needs to be administered by trained professionals (and is therefore expensive), it requires injections, which are not acceptable to all patients and it is potentially life-threatening. These factors severely restrict the number of individuals who can take advantage of this treatment. If we are to realize the tantalizing

prospect of altering the natural history of allergy in a substantial proportion of allergy patients, and even in the population as a whole, then immunotherapy will need to be dramatically different from what is used routinely today. Allergens extracted from their natural source have been in routine use since the inception Tenofovir solubility dmso of SCIT. Standardization of the potency of these biologically variable products represented a major advance and has led to improved safety and efficacy. Various modifications of the allergen have been attempted to increase potency and specificity and to reduce the risk of acute reactions. Allergoid production by formaldehyde treatment of native antigen has long been used, but is associated with reduced efficacy in allergen immunotherapy. Short peptides, unable to cross-link IgE and induce mast cell degranulation, but able to activate T cells through presentation on human leucocyte antigen (HLA) class II, were shown to induce Th1 reactivity.

Results were expressed as μmol/l of nitrites

synthesized during 48 h in the co-cultures performed in the presence of RSA PBMCs or fertile PBMCs. Co-culture recovered cells were analysed by Western blot for FoxP3, transforming growth factor (TGF)-β, and T-bet expression. Cells were washed extensively with phosphate-buffered saline (PBS), then the cell pellet was mixed gently with 1 ml ice-cold lysis buffer [PBS containing 5 mM ethylenediamine tetraacetic acid (EDTA), 1% NP-40, 0·5% sodium deoxycholate, 0·1% sodium dodecyl sulphate (SDS), 142·5 mM KCl, 5 mM MgCl2, 10 mM HEPES, pH 7·2] with freshly added protease inhibitor cocktail [0·2 mM phenylmethanesulphonyl fluoride (PMSF), 0·1% aprotinin, 0·7 μg/ml pepstatin Deforolimus find protocol and 1 μg/ml leupeptin] and incubated for 1 h on ice. Samples were finally centrifuged at 12 000 g for 20 min at 4°C and the supernatant fluids, representing the whole cell protein lysates, were stored at −70°C until use. Protein concentration was estimated using the micro-BCATM Protein Assay reagent kit (Pierce, Rockford, IL, USA). Equal amounts of proteins were diluted in sample buffer and resolved on SDS-polyacrylamide gels (10% for FoxP3 and T-bet or 15% for TGF-β). After electrophoresis, the separated proteins were transferred onto nitrocellulose membranes and probed with a

1:500 anti- FoxP3 Ab (eBioscience, San Diego, CA, USA) or 1:500 TGF-β (R&D Systems) or 1:500 T-bet (Santa Cruz Biotechnology, Santa Cruz, CA, USA). Blots were then incubated with a 1:3000 dilution of a horseradish peroxidase (HRP)-conjugated anti-goat immunoglobulin (Ig)G for FoxP3 and T-bet or anti-rabbit for TGF-β and developed using an enhanced chemoluminiscence detection kit (Amersham). Equal selleck products loading and absence of protein degradation were checked by Ponceau S staining (Sigma, St Louis, MO, USA). The immunoreactive protein bands were

analysed with a Fotodyne Image Analyzer® (Fotodyne, Inc., Hartland, WI, USA). Results were expressed as relative densitometric values by means of the Image Quant software normalized to β-actin expression. Flow cytometric analysis was performed according to the manufacturer’s instructions (human regulatory T cell staining kit; eBioscience). Briefly, 1 × 106 cells were stained with a CD4/CD25 cocktail. After 30 min cells were washed with staining buffer and then incubated with the fixation/permeabilization buffer for 1 h. After washing, unspecific sites were blocked by adding 2 μl (2% final) normal rat serum in approximately 100 μl for 15 min. Cells were then incubated with the anti-human FoxP3 (PCH101) antibody or rat IgG2a isotype control for at least 30 min at 4°C. Finally, cells were washed with permeabilization buffer and analysed.

In addition, patients with fibrosis had lower FCRN mRNA levels compared to patients without fibrosis (P = 0·041). No relationship between FCRN mRNA levels and other phenotypical features of CVID (presence of chronic diarrhoea, splenomegaly, granulomas, lymphadenopathy or autoimmune phenomena) INK 128 price was documented. No correlation was found between FCRN mRNA level and pre-infusion IgG and also serum albumin levels

in CVID patients. However, a correlation was demonstrated between FCRN mRNA level and the decline in serum IgG concentration during the second week after IVIg infusion (D14/D7 ratio) (P = 0·045 Spearman’s correlation coefficient). The higher the FCRN mRNA expression, the less pronounced the decrease in IgG concentration in the tracked period after IVIg infusion was observed [6].

We also showed a significant positive correlation between FCRN mRNA expression and the ‘efficiency index’ defined as: [IgG trough level – IgG residual level (g/l)]/IgG dose (g/kg/week [7]; P = 0·05). check details We did not document any correlation between FCRN mRNA expression and serum albumin levels in our CVID patients (P = 0·258). Our findings show that FcRn may play a role in the development of lung structural abnormalities, which are the principal life-threatening complications in patients with CVID, as well as in the catabolism of therapeutically administered IVIg. However, our results were obtained in a limited number of patients and show borderline statistical significance, and

need to be interpreted carefully. This study was supported by grant NT 111414-5/2010 of the Czech Ministry of Health. J. L. has received consultation fees from Baxter and LFB Biotechnologies; research ZD1839 chemical structure support from Shire and Baxter; honoraria for lectures from Biotest and Baxter; and support for clinical studies from Octapharma and CSL Behring. “
“Our and others’ previous studies have shown that Schistosoma japonicum (SJ) infection can inhibit allergic reactions. We recently reported that DCs played an important role in SJ infection-mediated inhibition of allergy, which was associated with enhanced IL-10 and T regulatory cell responses. Here, we further compared the role of CD8α+ DC and CD8α− DC subsets for the inhibitory effect. We sorted CD8α+ DC (SJCD8α+ DC) and CD8α− DC (SJCD8α− DC) from SJ-infected mice and tested their ability to modulate allergic responses in vivo. The data showed that the adoptive transfer of SJCD8α− DC was much more efficient than SJCD8α+ DC for the suppression of allergic airway eosinophilia, mucus overproduction, antigen-specific IgE responses, and Th2 cytokines (IL-4 and IL-5).

Mizoribine (MZR) is a selective inhibitor of the inosine monophosphate dehydrogenase – a key enzyme in the de novo pathway of guanine nucleotides – that was developed in Japan.[1] Clinically, MZR has been successfully used without any serious adverse effects

for the long-term treatment of young patients with lupus nephritis.[1-3] Besides its immunosuppressive effects, MZR has recently been reported to suppress the progression of histologic chronicity in selected patients with lupus nephritis and immunoglobulin A (IgA) nephropathy.[1-4] Moreover, some experimental reports described that MZR attenuates tubulointerstitial fibrosis in Dinaciclib ic50 rat models of unilateral ureteral obstruction, non-insulin-dependent diabetes and peritoneal fibrosis via suppression of macrophage infiltration of the interstitium.[5-7] Also, we recently confirmed a significant suppression of intraglomerular macrophage infiltration accompanied with significant suppression of the chronicity indices following MZR treatment in a patient with proliferative lupus nephritis.[8] These laboratory

and clinical observations suggest another beneficial mechanism of action of MZR from the histologic standpoint in the treatment of lupus nephritis. Since most of the oral dose of MZR is excreted unchanged in urine,[9] the CB-839 ic50 drug is thought to expose directly to residual renal cells. Thus, it is important to examine the direct effects of MZR against inflamed residual renal cells.[10] Glomerular mesangial cells (MCs) have been reported to produce a wide variety of proinflammatory molecules that play an important role in immune and inflammatory reactions in the kidney, and MCs itself are thought to play a pivotal role in the pathogenesis of renal diseases.[11]

Interestingly, it has been reported that the implication of ‘psuedoviral’ immunity as a novel disease concept of lupus Adenosine triphosphate nephritis, that is, the detection of self-nucleic acid particles resembling viral particles by toll-like receptors (TLRs) results in the activations of the downstream signalling cascades and subsequent type I interferons (IFNs) production.[12] In this context, we have examined the TLR3 signalling cascades treated with polyinosinic-polycytidylic acid (poly IC), a synthetic analogue of viral dsRNA, that makes ‘pseudoviral’ infection in cultured human MCs, and found that the activation of mesangial TLR3 upregulated the expression of functional molecules including monocyte/macrophage chemoattractants: CC chemokine ligand (CCL) 2 (or monocyte chemoattractant protein-1 [MCP-1]), CCL5 (or regulated on activation, normal T-cell expression and secretion [RANTES]), CXC ligand 10 (CXCL10) (or IFN-γ-induced protein 10 [IP-10]), fractalkine (or CX3CL1), and neutrophil chemoattractant: interleukin (IL)-8 (or CXCL8), in cultured human MCs.

Although renal prognosis and mortality is different among the underlying glomerulonephritides, corticosteroid-based immunosuppressive therapy is their main treatment modality and, therefore, they face the same clinical target, how to maximize the benefit of immunosuppressive therapy and minimize their disadvantages. The aims of the multicenter prospective cohort study, Japan Nephrotic Syndrome Cohort Study (JNSCS), are to provide the basic epidemiological date in primary CFTR modulator nephrotic syndrome in Japan, including the renal

prognosis and all-cause mortality, the response to the modern immunosuppressive practice patterns, and adverse events associated with these immunosuppressive therapy. JNSCS started in 2008 and 396 patients with primary nephrotic syndrome in 57 hospitals were enrolled during 3 years’ entry

period between 2008 and 2010. Diagnosis of glomerular diseases are minor change disease (MCD, n = 165 [41.6%]) and membranous nephropathy (MN, n = 158 [39.9%]), Rapamycin focal segmental glomerulosclerosis (FSGS, n = 38 [9.6%]), IgA nephropathy (n = 15 [3.8%]), membranoproliferative glomerulonephritis (n = 9 [2.3%]), non-IgAN mesangial proliferative glomerulonephritis (n = 7 [1.8%]), extracapillary proliferative glomerulonephritis (n = 2 [0.5%]) and intracapillary proliferative glomerulonephritis (n = 2 [0.5%]). Median age was 42 (interquartile range 26, 61) years in MCD, 66 (59, 75) years in MN, 62 (29, 73) in FSGS, and 58 (45, 71) in others. Male gender was 57.6%, 53.8%, 65.8%, and 57.1% in MCD, MN, FSGS, and others, respectively. Until December 2012, 359 (90.7%) patients received immunosuppressive therapy, including 162 MCD patients (98.2%), 136 MN patients (86.1%), 35 FSGS patients (92.1%), and 26 other patients (74.3%). Besides oral prednisolone (PSL), major initial immunosuppressive agents within 1 month of the immunosuppressive therapy were intravenous methylprednisolone (29.0%, 18.5%, 28.6%, and 50.0% in MCD, MN, FSGS, and others, respectively) Olopatadine and cyclosporin (14.8%, 45.2%, 42.9%, and 23.1% in MCD, MN, FSGS, and others, respectively). In contrast, only a few patients received cyclophosphamide

(0.6%, 4.4%, 0.0%, and 11.5% in MCD, MN, FSGS, and others, respectively), which KDIGO guideline 2012 recommended as the first-line immunosuppressive agent for MN. Interestingly, use of immunosuppressive agents were substantially different geographically. During median 2.3 years (interquartile range, 1.9–3.0) of observational period, cumulative probabilities of complete remission of proteinuria defined as <0.3 g/day of urinary protein, <0.3 urinary protein/urinary creatinine ratio, or negative or trace of dipstick urinary protein after initiation of immunosuppressive therapy (n = 359 [90.7%]) or kidney biopsy if no immunosuppressive therapy (n = 39 [9.3%]) were 0.85, 0.89, 0.93, and 0.95 at 2, 6, 12, and 24 months in MCD, 0.08, 0.27, 0.53, and 0.68 in MN, 0.32, 0.46, 0.58, and 0.65 in FSGS, and 0.09, 0.21, 0.42, and 0.

[29] Recognition of RSV though PRR is schematized in Fig. 1. Among the pro-inflammatory cytokines described below, IL-8 is a key molecule produced by epithelial cells and macrophages during the early response to hRSV and works as a chemoattractant in the recruitment of neutrophils, which infiltrate the site of infection.[35] Another important molecule of the innate response against hRSV infection FDA approved Drug Library cell assay is

IL-1β, a pro-inflammatory cytokine involved in the antiviral response. First, hRSV stimulates PRR to induce the expression of pro-IL-1β (IL-1β precursor) and inflammasome components, trigged by TLR2/MyD88 that activates the NF-κB pathway.[33, 35] Second, the assembly of the inflammasome www.selleckchem.com/products/azd-1208.html complex takes place and caspase-1 cleaves pro-IL-1β into IL-1β in response to the production of reactive oxygen species, cellular potassium efflux, or cathepsin leakage into the cytosol after lysosomal disintegration.[34,

36] The NF-κB pathway is important for the activation of an innate response against hRSV, not only for the cytokine response, but also for the formation of tight junctions between nasal epithelial cells.[37] Infection with hRSV induces the up-regulation of genes encoding structural components of tight junctions, including claudin-2, -4, -7, -9, -14, -19, occludin, ZO-2, cingulin and MAG-1, mediated by the protein kinase Cδ signalling.[37] This phenomenon seems to be beneficial for the replication of the virus, because inhibition of NF-κB and protein kinase Cδ

activation leads to an impairment of viral replication and formation of virus filaments.[37] In addition, the induction of tight junctions could increase the cell polarity necessary for viral budding.[13] Human RSV infection has been associated with an inefficient adaptive immune response, characterized by an excessive T helper type 2 (Th2) and a deficient Teicoplanin antiviral Th1 response.[15, 36, 38] The Th1 responses usually involve the production of IFN-γ, IL-2 and tumour necrosis factor-α, whereas IL-4, IL-5, IL-10 and IL-13 secretion characterize Th2 responses. Further, a Th17 response has been associated with hRSV pathogenesis because it contributes to the development of asthma in infected children.[15, 39] Studies using an in vitro model comprising both human airway epithelial cells (A549 cells) and human immune cells (peripheral blood mononuclear cells) have shown that hRSV infection induces the production of IFN-γ,IL-4 and IL-17, suggesting that the three subsets (Th1, Th2 and Th17) can be activated upon viral infection.[40] Assays performed with peripheral blood mononuclear cells co-cultured with hRSV-infected A549 cells have also shown a Th2 and Th17 differentiation and the suppression of the generation of regulatory T cells.[8, 41] Indeed, as shown in Fig.

4). We compared the performance of gene sets with their constituent genes in profiles from high versus low HAI responders to influenza vaccination. We found that the top-scoring gene sets in TIV responders were more strongly correlated with the high antibody response phenotype than any constituent GSK-3 activity gene in either gene set (Supporting Information Fig. 5A). Moreover,

although both complement and antibody genes were present in gene sets enriching in responders, the antibody genes were among those most upregulated (Supporting Information Fig. 5A and B). Thus a gene set based analytic approach identifies signatures of proliferation and immunoglobulin genes that are strongly correlated ABT199 with high antibody response. We next sought

to determine if enrichment of the immunoglobulin and/or proliferation gene sets could be used as a predictor of vaccine response, using high or low HAI titers as an outcome. To do this, we selected the most differentially enriched gene set from each of the two clusters, and fitted them into logistic regression models. Both models closely fit the data and yielded an AUC of ∼0.9 (Fig. 3A and B), suggesting that each independent gene set could provide a strongly predictive model of vaccine response. To integrate both biological processes into a single model, we applied Bayes’ rule, and found that the integrated model achieved an AUC of 0.94 (Fig. 3C). To compare our integrated gene set based model with the single-gene level model previously described for this dataset [16], we tested our model in a validation dataset comprised of PBMC samples Oxymatrine from an independent trial of TIV vaccination. We found that our predictive model yielded an accuracy of 88% in the test set, comparable

to the performance of the single-gene level predictor [16]. This indicates that gene set based analysis of expression profiles provide accurate predictors of response to vaccination. An advantage of a gene set enrichment analysis is that it can capture subtle changes in gene expression distributed across transcriptional networks. We therefore compared the degree of differential expression of genes in the predictive gene sets (proliferation and immunoglobulin gene sets) with that of the genes selected in the single-gene level predictor originally applied to this dataset (Fig. 4). Predictive genes selected in the study by Nakaya et al. [16] were all highly differentially expressed in day seven PBMC expression profiles from responders compared to nonresponders, as expected (mean fold change 3.36). In contrast, the gene sets identified in our analysis included many genes that were much less differentially expressed (mean fold change of proliferation cluster 2.13; mean fold change of immunoglobulin cluster 2.53) (Fig. 4).

Incidentally, Fujii et al. 8 reported a phenomenon describing NKT cell turnover, a decrease in the NKT cell population on day 1 after α-GalCer administration later found to be due to TCR down-regulation, after administration of free α-GalCer that was “less rapid and severe” when DCs pulsed with α-GalCer were administered. Antigen-specific cellular click here immune responses were measured after each dose of the α-GalCer adjuvant and OVA antigen mixture, similar to our previously reported studies with a different antigen 7. Both these studies demonstrate that multiple doses of α-GalCer, administered by the intranasal route, are necessary to induce

efficient antigen-specific cellular immune responses, regardless of the mouse strain used. In addition to the antigen-specific cellular immune responses, effectiveness of α-GalCer as an adjuvant after intranasal immunization to induce humoral immune responses, in terms of antigen-specific IgA and IgG responses has been described in the literature

24 and also observed in other unrelated studies in our laboratory (data not shown). Thus, our studies provide mechanistic support for mucosal delivery of α-GalCer adjuvant as an attractive strategy for vaccination regimens. It is also important to note potential inflammatory effects from the intranasal administration of α-GalCer. Different mouse model studies revealed that intranasal administration of α-GalCer can induce airway infiltration of a combination of eosiniphils, neutrophils, Neratinib mouse and/or monocytes 25, 26. Preliminary studies in our lab showed increase in the percentages of eosinophils but not neutrophils or monocytes (data not shown). However, clinical trials performed by Kunii et al. 4 showed that administration of α-GalCer by a nasal sub-mucosal route was safe. Overall, this investigation has shown that α-GalCer can be administered by the intranasal route for primary and booster immunizations to induce cellular immune responses to co-administered antigens, without inducing NKT cell anergy. This is in striking contrast to α-GalCer administration

by the intravenous route, in which a single dose leads to NKT cell anergy and a reduction in the ability of the adjuvant to boost adaptive immune responses to co-administered antigen. Thus, old our data support the intranasal route of immunization as an attractive route for immunization especially because the ability to deliver multiple doses of the vaccine is essential for most therapeutic applications against infectious diseases and cancer. Female C57Bl/6 mice aged 6–10 wk were purchased from the National Cancer Institute. All procedures on the animals were carried out in accordance with institutionally approved protocols. The animals were housed in microisolator cages and provided with sterile food and water. The animal facility is fully accredited by the Association for Assessment and Accreditation of Laboratory Animals Care International.