We report a progressive lower extremity lymphedema (LEL) case successfully treated with a ladder-shaped LVA. A 67-year-old female with secondary LEL refractory to conservative treatments underwent LVA. A ladder-shaped

LVA was performed at the left ankle. In the CAL-101 research buy ladder-shaped LVA, 3 lymphatic vessels and 1 vein were anastomosed in a side-to-side fashion; 2 lymphatic vessels next to the vein were anastomosed to the vein, and the other lymphatic vessel was anastomosed to the nearby lymphatic vessel. Using ladder-shaped LVA, 6 lymph flows of 3 lymphatic vessels could be bypassed into a vein. Six months after the LVA operation, her left LEL index decreased from 212 to 195, indicating edematous volume reduction. Ladder-shaped ACP-196 molecular weight LVA may be a useful option when there are 3 lymphatic vessels and 1 vein in a surgical field. © 2013 Wiley Periodicals, Inc. Microsurgery 34:404–408, 2014. “
“This study evaluated the results of repair of the radius defect with a vascularized tissue

engineered bone graft composed by implanting mesenchymal stem cells (MSCs) and a vascular bundle into the xenogeneic deproteinized cancellous bone (XDCB) scaffold in a rabbit model. Sixty-four rabbits were used in the study. Among them, four rabbits were used as the MSCs donor. Other 57 rabbits were divided into five groups. In group one (n = 9), a 1.5 cm bone defect was created with no repair. In group two (n = 12), the bone defect was repaired by a XDCB graft alone. In group three (n = 12), the defect was repaired by a XDCB graft that included a vascular bundle. In group four (n = 12), the defect was repaired by a XDCB graft seeded with MSCs. In group Palbociclib five (n = 12), the defect was repaired by a XDCB graft including a vascular bundle and MSCs implantation. The rest three rabbits were used as the normal control for the biomechanical test. The results of X-ray and histology at postoperative intervals (4, 8, and 12 weeks) and biomechanical examinations at 12 weeks showed that combining MSCs and a vascular bundle implantation

resulted in promoting vascularization and osteogenesis in the XDCB graft, and improving new bone formation and mechanical property in repair of radius defect with this tissue engineered bone graft. These findings suggested that the vascularized tissue engineered bone graft may be a valuable alternative for repair of large bone defect and deserves further investigations. © 2011 Wiley-Liss, Inc. Microsurgery, 2011. “
“The comparisons of two different methods of donor nephrectomy were performed in this study. Fisher inbred rats were used as donors and recipients of kidneys. In method A (the conventional technique), meticulous blunt dissection of the abdominal aorta, inferior vena cava, and renal arteries/veins, was followed by ligating and cutting the superior mesenteric artery and small vessels entering the above vessels. Both donor kidneys were irrigated after the suprarenal aorta and inferior vena cava were cross-clamped (n = 10).

Genomic profiling

can be used as a powerful tool to identify novel differences and separate out these subpopulations in a more detailed manner. The early stages of human lymphopoiesis are poorly characterized. Common lymphoid progenitors commit to either the NK-cell or the B/T-cell lineages. Two subsets of CD34+ hematopoietic progenitor cells (HPCs) have been proposed as candidate common lymphoid progenitors: CD45RA+CD38–CD7+ cells from the umbilical cord blood and CD45RAhiLin–CD10+ cells from the BM [39, 40]. In vitro experiments showed that umbilical cord blood derived CD34+CD45RAhiCD7+ HPCs skew toward generating T/NK lineages in vitro, while CD34+CD45RAhiLin–CD10+ BM-derived HPCs predominantly exhibit a B-cell potential [39]. Gene expression profiling by DNA microarrays confirmed that CD34+CD45RAhiCD7+ HPCs selectively express NK and T lineage committed genes while Navitoclax mw retaining expression of genes related to the granulomonocytic lineage, whereas CD34+CD45RAhiLin–CD10+ HPCs exhibit a typical pro-B-cell transcriptional profile and generally lack genes unrelated

to the B-cell lineage [41]. https://www.selleckchem.com/products/CAL-101.html Human NK cells account for a small fraction of total lymphocytes (∼10%) in the peripheral blood and are composed of two different subpopulations: the predominant CD56dimCD16+ mature subset (∼95%) and the much smaller CD56brightCD16– immature subset (∼5%) [29]. CD56dim and CD56bright pNK cells have differential expression patterns for cell receptors, adhesion molecules, cytokines, chemokines, TFs, and cytolytic molecules [29, 42, 43]; three studies to date have characterized these two NK-cell subpopulations using genomic profiling (Table 4). All three studies revealed that, compared with CD56bright pNK cells, CD56dim pNK cells upregulate killer cell Ig-like

receptors (KIRs) (including Kir2dl1 and Kir2d2), cytolytic molecules (including Prf1, Gzma, and Gzmb), and chemokines (including Cxcl8, Mip-1b, and Mip-1b) [42-44]. Additionally, Koopman et al. [43] compared CD56bright dNK cells with CD56bright or CD56dim pNK cells and found that CD56bright pNK cells were more similar to the CD56dim pNK-cell subset than they were to the CD56bright dNK cells. Hanna et al. [42] analyzed ∼20 000 genes among purified CD56brightCD16+, CD56dimCD16–, check and in vitro activated CD16+ pNK cells to find that overexpression of certain tetraspanin family receptors (CD9, CD53, CD81) on activated NK cells might enhance or alter their migration to, and retention in, inflamed tissues. Wendt et al. [44] analyzed ∼33 000 genes in resting CD56bright and CD56dim pNK cells, and verified the observed changes in cytokine and chemokine genes at the protein level using cytometric bead array and protein arrays. While GM-CSF, TARC, and TGF-β3 were exclusively expressed in CD56bright pNK-cell supernatants, CD56dim pNK cells were the main producers of IGF-1 and IGFBP-3. GDNF, IGFBP-1, EGF, and TIMP-2 were detected in both CD56bright and CD56dim pNK subsets [44].

In addition to documenting the safety of this

approach, we found that patients treated with OK432-stimulated DCs displayed unique cytokine and chemokine Kinase Inhibitor Library high throughput profiles and, most importantly, experienced prolonged recurrence-free survival. Inclusion criteria were a radiological diagnosis of primary HCC by computed tomography (CT) angiography, hepatitis C virus (HCV)-related HCC, a Karnofsky score of ≥ 70%, an age of ≥ 20 years, informed consent and the following normal baseline haematological parameters (within 1 week before DC administration): haemoglobin ≥ 8·5 g/dl; white cell count ≥ 2000/µl; platelet count ≥ 50 000/µl; creatinine < 1·5 mg/dl and liver damage A or B [23]. Exclusion criteria included severe cardiac, renal, pulmonary, haematological or other

systemic disease associated with a discontinuation risk; human immunodeficiency virus (HIV) infection; prior history of other malignancies; history of surgery, chemotherapy or radiation therapy within 4 weeks; immunological disorders including splenectomy and radiation to the spleen; corticosteroid or anti-histamine therapy; current lactation; pregnancy; history of organ transplantation; or difficulty in follow-up. Thirteen patients (four women and nine men) presenting at Kanazawa www.selleckchem.com/products/sorafenib.html University Hospital between March 2004 and June 2006 were enrolled into the study, with an age range from 56 to Tryptophan synthase 83 years (Table 1). Patients with verified radiological diagnoses of HCC stage II or more were eligible and enrolled in this study. In addition, a group of 22 historical controls (nine women and 13 men) treated with TAE without DC administration between July 2000 and September 2007 was included in this study. All patients received RFA therapy to increase the locoregional effects 1 week later [24]. They underwent ultrasound, computed tomography (CT) scan or magnetic resonance imaging (MRI) of the abdomen about 1 month after treatment and at a minimum of

once every 3 months thereafter, and tumour recurrences were followed for up to 360 days. The Institutional Review Board reviewed and approved the study protocol. This study complied with ethical standards outlined in the Declaration of Helsinki. Adverse events were monitored for 1 month after the DC infusion in terms of fever, vomiting, abdominal pain, encephalopathy, myalgia, ascites, gastrointestinal disorder, bleeding, hepatic abscess and autoimmune diseases. DCs were generated from blood monocyte precursors, as reported previously [25]. Briefly, peripheral blood mononuclear cells (PBMCs) were isolated by centrifugation in LymphoprepTM Tubes (Nycomed, Roskilde, Denmark). For generating DCs, PBMCs were plated in six-well tissue culture dishes (Costar, Cambridge, MA, USA) at 1·4 × 107 cells in 2 ml per well and allowed to adhere to plastic for 2 h.

Preparation of cell suspensions from liver, lung and bone marrow was as described

previously 13. To enumerate cell number, cytometric bead-based counting assays were performed as described previously 13. Intracellular cytokine staining was performed according to standard procedures 4 using stimulation with OVA323-339 (16–18 h) or PMA/ionomycin (3 h, Merck Biosciences, Darmstadt, Germany; Fluka, Switzerland). Cytometric data were collected using a FACSCalibur or FACSCanto cytometer (BD Biosciences, San Jose, CA, USA) and analyzed with CellQuest or FACSDiva software. The total number of IFN-γ-producing OT-II cells was calculated based on the intracellular cytokine staining and absolute OT-II cell number determined using a bead-based counting assay. To assess proliferation in vivo, OVA-specific OT-II T cells were labeled with CFSE as described previously 46 and used for selleck compound culture or injected i.v. Three days later, recipient spleens or LN were harvested and CFSE dilution (CD45.1+/CD4+ gated cells)

assessed by flow cytometry. To determine responsiveness to antigen challenge, mice were immunized s.c. at the tail base with OVA (100μg) emulsified in complete Freund’s adjuvant. In vitro peptide restimulations were performed on splenocyte single-cell suspensions plated at 2×106/mL (1 mL, 24-well plates) in learn more complete RPMI with or without added OVA323–339 (10 μg/mL). Culture

3-mercaptopyruvate sulfurtransferase supernatants were harvested after 3 days and ELISA assays were performed using standard procedures with the following capture and detection antibodies (capture/detection IFN-γ: R4-6A2/XMG1.2, IL-2: JES6-1A12, JES6-5H4, IL-4:11B11/BVD6-24G2) or kits purchased from eBioscience and used in accordance with the manufacturer’s instructions (IL-10 and TGF-β). Comparison of means was performed using Student’s t-test and multiple comparisons were performed using one-way ANOVA followed by Newman–Keuls post-test (GraphPad Prism). This study was supported by the National Health and Medical Research Council (R. J. S.) and Juvenile Diabetes Research Foundation (R. J. S.). Conflict of interest: The authors declare no financial or commercial conflict of interest. “
“Staphylococcal enterotoxin A (SEA) is one of the bacterial products tested for modulation of unwanted immune responses. Of all the staphylococcal enterotoxins, SEA is the most potent stimulator of T cells. When administered orally, SEA acts as a superantigen (SA), producing unspecific stimulation of intra-epithelial lymphocytes (IELs) in the intestinal mucosa. This stimulation results in amplification of the normal local immunologic responses, which are mainly regulatory. This amplification is based on increased local production of IFN-γ by IELs, which acts on the nearby enterocytes.

Nukuzuma, unpublished data). Proliferation characteristics of COS-tat cells may provide important background information for studies using these cell lines. Thus, we first compared the cell proliferation of three COS-tat cell lines

with those of parental check details COS-7 cells. COS-7 cells (ATCC CRL 1651) and COS-tat cell clones (8) were cultivated in EMEM containing 10% FBS (hereafter called culture medium). Cell cultures were maintained at 37°C in a humidified incubator containing 5% CO2 in air. The relative number of live cells was determined by measuring mitochondrial succinate dehydrogenase activity using MTT assay. COS-7 cells and COS-tat cell clones were each plated in five wells of 96-well culture plates at a concentration of 2 × 103 cells/well in 100 μL culture medium and incubated at 37°C in a CO2 incubator. MTT assay was performed using a Cell Proliferation Kit I (MTT) (Roche, Penzberg, Germany) according to

the manufacturer’s instructions. After an incubation period of 5 days, 10 μL MTT solution was added to each well to a final concentration of 0.5 mg/mL, and the plates incubated for 4 hr. Then, click here 100 μL solubilization solution was added to each well, and the plates placed in an incubator overnight. The formazan products were solubilized, and spectrophotometric data were measured using an enzyme-linked immunosorbent assay reader (Bio-Rad, Hercules, CA, USA) at a wavelength of 550 nm with a reference wavelength of 650 nm. The significance of inter-group differences was statistically determined by Student’s t-test. As shown in Table 1, the enzyme activity of COS-tat7 and COS-tat15, and COS-tat22 cells was lower than that of parental COS-7 cells and this difference PI-1840 was statistically significant (P < 0.01). Of note, the enzyme activity of COS-tat22 cells was lower than that of COS-tat7 and COS-tat15 cells (P < 0.01). To measure the doubling time, COS-7 cells and COS-tat cell clones were plated in 6-well culture plates at a concentration of 4 × 104 cells/well in 2 mL culture medium. After an incubation period of 72 hr, cell numbers were counted. The

doubling time of COS-7, COS-tat7, COS-tat15, and COS-tat22 were 21.6, 24.6, 22.8, and 30.8 hr, respectively. The doubling time of COS-7 COS-tat cells were in agreement with the proliferation characteristics of the cells as judged by MTT assay. Taken together, these results indicate that stable expression of Tat leads to down-regulation of cell proliferation. We next compared the production of PML-type JCV in COS-tat cell clones with that in parental COS-7 cells. Since JCV capsids have the property of agglutinating human type O erythrocytes, HA assay has been traditionally employed to determine the virus titer (12). COS-7 and COS-tat cell clones were cultured in 35-mm dishes containing 2 mL culture medium until the cells were 50–80% confluent.

1). A 3D reconstruction of the contact zone confirmed that LPL localized outside the cSMAC and thus in the p/dSMAC of the BYL719 research buy IS (Fig. 1C). A quantification of the contact zone of 150 cell couples from three experiments

revealed that LPL was found in 62% in and around (i.e. distal) the LFA-1 staining (p/dSMAC) (Fig. 1D). The mechanism by which LPL is targeted to the IS was so far unknown. To scrutinize the mechanisms that promoted the relocalization and stabilization of LPL in the contact zone, we created LPL cDNA constructs lacking the potential calmodulin-binding domain (ΔCBD-LPL), actin-binding domain (ΔABD-LPL) or EF-hand calcium-binding domains (ΔEF1/2-LPL). In addition, we mutated the phosphorylation site at Ser5 to alanine (5A-LPL) 17. The expression of theses mutants in T cells was at a similar level as wt-LPL (Fig. 1E). After incubation with superantigen-loaded APC, the localization of these

mutants in the IS was analyzed and judged as enriched if at least 50% of the protein was found in the cell interface (Fig. 1F). Compared to wt-LPL the number of cells that contained ΔCBD-LPL in the contact zone was reduced by about 50% as analyzed by LSM. Hardly any cells displayed ΔABD-LPL in the contact zone. In contrast, mutation of the phosphorylation site (5A-LPL) or deletion of the calcium-binding sites (ΔEF1/2-LPL) had no effect on the relocalization of LPL. Thus, the binding sites of LPL for calmodulin and F-actin GW-572016 ic50 are important for the redistribution of LPL to the T-cell/APC contact zone. MIFC allows quantification of the F-actin content of T cells that form a contact with APC (Supporting Information Fig. 1). These analyses revealed

that T cells had an elevated total F-actin content if they formed a contact Buspirone HCl with their APC compared to solitaire T cells within the same sample (Supporting Information Fig. 1D). Actin polymerization could account for the enrichment of LPL at the T-cell APC contact zone since plastins bind to F-actin during actin polymerization 14, 15 and LPL mutants lacking the actin-binding domain did not localize to the IS (See Fig. 1D). Vice versa, LPL is an actin-bundling protein that stabilizes F-actin. Therefore, one would assume that lack of LPL affects the F-actin content of T cells. We knocked down LPL expression with small interference RNA (siRNA) to test this hypothesis. T cells that were transfected with LPL-specific siRNA (LP) displayed a reduction of LPL-expression by at least 90% compared to cells that were transfected with a non-targeting control siRNA (con) (Fig. 2A). The mean F-actin content in unstimulated LPL knock-down T cells (T-cell singlets) was only slightly lower compared to unstimulated control siRNA-treated cells (Fig. 2B). However, the rise in the F-actin content observed upon APC encounter was significantly lower than in control siRNA-treated T cells (Fig. 2B).

We found that LPG induced opposing effects on the PMA-induced oxidative burst of macrophages from both mouse strains. Whereas in macrophages

of BALB/c mice, LPG inhibited the oxidative burst by 11·33%, compared with control values (P < 0·002), in C57BL/6 macrophages, LPG enhanced the oxidative burst by 13·7% (P < 0·017) over the controls not incubated with LPG. When the macrophages were pre-incubated with the PKCα inhibitor Gö6976, either alone or in combination with LPG, the oxidative burst of macrophages of both mouse strains was inhibited in relation to the macrophages in the absence of Gö6976, albeit the degree of inhibition was higher in the BALB/c macrophages (P < 0·002) (Figure 3a). We also analysed the effect of L. mexicana promastigotes on the PMA-induced oxidative burst Acalabrutinib clinical trial of peritoneal macrophages obtained from both mouse strains. The assay was performed in the absence or presence of the PKCα inhibitor Gö6976. L. mexicana promastigotes significantly diminished the oxidative burst induced by PMA on macrophages of both mouse strains, yet the degree of inhibition was significantly higher in BALB/c macrophages (46·4%, P < 0·002) than in C57BL/6 macrophages (19·4%P < 0·00013) as compared with controls not incubated with the parasite. In the presence of Gö6976, the degree of inhibition exerted

by L. mexicana promastigotes on the oxidative burst of BALB/c macrophages was similar to that achieved without Gö6976. In ADP ribosylation factor C57BL/6 macrophages, Gö6976 was able to increase the Bcl-2 inhibitor degree of inhibition of the oxidative burst exerted by L. mexicana promastigotes, as compared with the oxidative burst in the absence of Gö6976 (P < 0·00013) (Figure 3b). The purity of peritoneal macrophages ranged between 80% and 85% (data not shown). The levels of oxidative burst in cells not stimulated

by PMA, but treated only with LPG or L. mexicana promastigotes, were also measured. In both cases, the levels of oxidative burst were below the detection level of the apparatus. To determine a possible correlation between PKCα activity and the effectiveness of burst oxidation with the intracellular survival of the parasite, peritoneal macrophages from both mouse strains were infected with L. mexicana promastigotes. The oxidative burst was then induced by PMA and the parasite survival was analysed. Results show that in C75BL/6 macrophages stimulated with PMA, only 66% (259 parasites/100 macrophages) of the parasites survived as compared with parasite survival in macrophages not stimulated with PMA (390 parasites/100 macrophages). In contrast, 92% (280 parasites/100 macrophages) survived in BALB/c macrophages as compared with parasite survival in nonstimulated macrophages (304 parasites/100 macrophages).

5D). To address whether compensation by the upregulated HRs in H1H2RKO and H3H4RKO CD4+ T cells affects the expression of HDC or the production of HA, anti-CD3, and soluble anti-CD28 mAb stimulated CD4+ T cells

were analyzed for HDC expression (Fig. 5E) by qRT-PCR and screened for HA production by enzyme immunoassay (EIA) (Fig. 5F) at 24, 48, and 72 h. Surprisingly, we did not detect a significant strain effect or (strain × time) interaction for HA production or HDC expression. These Small molecule library cell assay data therefore do not support the existence of a compensatory loop with respect to HA production and HDC expression by H1H2RKO and H3H4RKO CD4+ T cells. However, the HR expression studies clearly indicate that disease severity in H1H2RKO and H3H4RKO mice is associated with compensatory upregulation of the corresponding receptors. Here, we have assessed the overall contribution to EAE susceptibility imposed by H1R and H2R (couple via stimulatory G proteins) and the H3R and H4R (couple via inhibitory

G proteins). The results of our study demonstrate that H3H4RKO mice develop a significantly more severe mTOR inhibitor clinical disease course compared with B6 and H1H2RKO mice in association with greater pathology in the brain but not the spinal cord. In contrast, despite a significant difference in the severity of the clinical disease courses between B6 and H1H2RKO, a significant difference in pathology was not detected in either the brain or spinal cord, suggesting as in H3RKO mice [[18]], H1R and/or H2R may also play a significant role in central functions related to the severity of clinical signs. Increased susceptibility to EAE in H3H4RKO mice is associated with significantly higher production of IFN-γ and IL-17 in MOG35–55 specific ex vivo recall assays. In contrast, H1H2RKO mice exhibit decreased susceptibility to EAE and decreased BBB permeability. We have also shown that CD4+ T cells from H1H2RKO mice, upon in vitro activation, have an intrinsic immune deviation toward the Th2 phenotype, while activated T cells from H3H4RKO mice have an intrinsic PTK6 immune bias toward Th1 type cells. The results of our current study

indicate that HA signaling through H1R and H2R augments EAE susceptibility by influencing antigen-specific T-cell effector responses, immune deviation, and BBB permeability. It is well known that HA and HRs modulate the innate and adaptive immune systems [[4]]. Previously, we have shown that H1RKO mice develop less severe EAE that was associated with an immune deviation of the CD4+ T-cell population from an encephalitogenic Th1 response to a protective Th2 response [[27, 31]]. In addition, mice deficient for H2R are significantly less susceptible to acute early phase EAE and T cells from immunized H2RKO mice exhibit blunted Th1 effector cell responses [[32]]. Similar to H1R, HA acting through H2R determines T-cell effector functions and their polarization.

38 Regular updates of the numbers of alleles observed at each HLA locus (current numbers are given in Table 2) are recorded in the IMGT/HLA database (http://www.ebi.ac.uk/imgt/hla/), which also provides DNA and amino acid sequences and alignments of HLA alleles and molecules, and nomenclature information.39 This nomenclature has recently been modified substantially according to the allele naming system shown in Fig. 2. The high level of diversity

found at the HLA loci is principally located in exons 2 and 3 for class I genes, and in exon 2 for class II genes. Such exons correspond, at the protein level, to the peptide-binding region (PBR) of the HLA molecules. The mean pairwise DNA sequence differences between HLA alleles are between Wnt inhibitor ∼ 10 and 26 nucleotides, depending on the locus (Table 2 and ref. 40), suggesting a functional relevance. Analysis of the amino acid sequence of HLA molecules shows that allelic variants differ from each other mainly by substitutions in residues contributing to the PBR, in particular in some pockets in the PBR that accommodate side chains of the bound peptides. Hence, peptides eluted from different HLA class I molecules show distinctive amino acid patterns at certain positions, in particular corresponding to Hydroxychloroquine molecular weight pockets 2 and 9 of the HLA molecules.41 It is therefore assumed that the polymorphism of

HLA alleles is to a large extent functional because different HLA molecules bind different sets of peptides. A high sequence diversity is therefore required in the PBR of the HLA molecules to bind a high variety of pathogen-derived peptides that are subsequently presented to T-cell receptors. The distribution of HLA alleles in different populations may be a consequence of this functional polymorphism. In many instances the immune response to a particular peptide epitope of a pathogen may depend on the HLA alleles carried by the individual. Individuals heterozygous for HLA alleles may have a wider

peptide binding repertoire and therefore a capability to respond to more pathogen variants, causing selection of heterozygotes. On the other hand, the existence of several different loci both within Histamine H2 receptor the class I (A, B and C) and II series (DR, DQ and DP) of molecules may to some extent compensate for the deficits of homozygosity. It should also be noted that there exists a very strong linkage disequilibrium (LD), or non-random association, between HLA alleles at different loci; i.e. some HLA alleles are found together in populations more frequently than expected based on their gene frequencies. For example some alleles of the DRB1 locus demonstrate strong LD with specific alleles at the DQA1 and DQB1 loci. Furthermore, in many populations HLA alleles at one locus with high sequence homology, i.e. DRB1, are in LD with the same alleles at other loci, i.e. DQA1 and DQB1, which may indicate an evolutionary relationship between some alleles, i.e. DRB1.

In this Fulvestrant way, females differed from males, which showed no

significant differences in IgE levels when immunized with different doses and at different ages. Other studies, too, have demonstrated clearly higher IgE, cytokine and/or airway inflammatory responses in females compared with males in i.p. sensitization models using young adult mice (6–8 week old) [27–30]. These studies were performed in the BALB/c, C57Bl/6 and NIH/OlaHsd strains. In line with these previous studies, obvious differences related to sex were found in our i.n. sensitization model. Sex differences were most pronounced for antibody production and influx of inflammatory cells into the airways (BALF) and into the lung tissue (histopathology), where females had higher responses than males. Cytokine secretion and MLN cell numbers were marginally influenced by the sex of the animals. The same was recently observed for cytokines in lung tissue in an i.n. house dust mite sensitization model with adult BALB/c mice [29] DMXAA solubility dmso and for cytokines in BALF following OVA inhalation [26].

Further, 1-week-old female mice also appeared to have stronger IgE and inflammatory responses than male mice, which is different from the i.p. model, where no sex differences were observed in 1-week-old mice. This discrepancy between the i.p. and i.n. sensitization studies may be ascribed to the route of immunization and OVA dose. It could, however, also be because of the fact that the importance of allergen dose was examined in the i.p., but not in the i.n. mouse models, and as more factors are investigated a higher power is needed to detect significant effects. During the i.n. model development, the 10 μg OVA and 120 μg Al(OH)3 doses were found to be optimal for IgE responses. A 0.1-μg OVA dose did not stimulate IgE production in BALB/c mice (unpublished data). It cannot be ruled out that a dose–response relationship could be found comparably to the i.p. Resminostat model, but higher

doses were not investigated in our i.n. model. Table 3 summarizes the findings of age-related effects for the i.p. study (using 0.1 or 10 μg OVA in 1 mg Al(OH)3 for sensitization) and for the i.n. study (10 μg OVA in 120 μg Al(OH)3 for sensitization). Compared to the low or high dose i.p. model, the outcomes of the i.n. model did not resemble one of these more than the other. Overall, the OVA-specific IgE and IgG1 production were unaffected or increased with age. Importantly for both models, the BALF eosinophil pattern was followed by IL-5 and IL-13, which regulates eosinophil inflammation and airway hyperresponsiveness [31, 32]. Histopathology was only performed in the i.n. sensitization model. When comparing trends in the three age groups, it appeared that the perivascular and partly the peribronchial inflammation followed the IgE/IgG1 response, while eosinophil numbers in BALF followed the IL-5/IL-13 response.