And when a new stimulus is presented

during the posthabituation test phase, looking time should rebound to reflect a discrepancy with the template. While this “novelty” response during the test phase is the typical outcome, it is not universal; under some circumstances the posthabituation looking times are longer to the familiar stimulus. For example, although almost all of the findings on infant statistical learning report novelty preferences Inhibitor Library (i.e., longer looking to the less frequent or less predictable stimuli), there are exceptions (Fiser & Aslin, 2002; Pelucchi, Hay, & Saffran, 2009). In fact, in looking-time measures of infants’ preferences for their native language, when there is no immediately preceding habituation phase (but only the long-term exposure prior to visiting the laboratory for testing), infants typically listen longer to highly familiar stimuli rather than to novel stimuli (Jusczyk & Aslin, 1995). The foregoing results across literally hundreds of experiments raise the possibility that there is at least one additional variable that is unaccounted for by the canonical reactive view of looking times. Kidd, Piantadosi, and Aslin

(2012) hypothesized that if infants also take an active role in sampling their visual environment, then looking times should vary by how much information infants are able to extract Neratinib clinical trial on a moment-by-moment basis. To be clear, this does not deny the importance of stimulus salience and memory for repeated events as factors that influence infant looking times. Rather, Kidd et al. asked whether this third factor—the ability to estimate the information content of stimulus events—also plays a role in infant looking Pregnenolone times. The logic of the design employed by Kidd et al. (2012) was to create a quantitatively well-defined family of stimulus events whose salience was randomized (to wash

out that effect). Each stimulus event varied in its predictability or surprisal given all previous events in a given sequence. Thus, the goal was to determine, at each stimulus event, whether the infant would continue to look at the display or to terminate fixation and end the trial. Notice that this is quite different from previous studies that ask how long infants will maintain their looking. Kidd et al. asked whether on each stimulus event infants will or will not make an implicit binary decision to stay or go. To achieve this, they created very brief (2 sec) events from an inventory of three possibilities on each trial that varied in information complexity from simple (e.g., AAAAAAA) to complex (e.g., ABACCBBBACAA). The hypothesis was that if infants are active samplers, they will terminate their fixation whenever the sequence of events is either too simple or too complex.

Anderson and co-workers established an innovative approach that allows the detection BAY 80-6946 in vitro of gluten-specific T cells in the peripheral blood of CD patients after a short period of gluten-containing food consumption [4,5]. Basically, gluten-sensitized

CD4+ T cells, normally scarcely detectable in the blood of coeliac patients, circulate transiently in the peripheral blood after 3 days of wheat challenge, and can be detected by a sensitive interferon (IFN)-γ enzyme-linked inmmunospot (ELISPOT) assay. Using this in-vivo procedure, the authors further screened large libraries of prolamin peptides and assessed the hierarchy and immunodominance of gluten T cell epitopes [6]. More recently, T cells reactive to DQ2-α-I and DQ2-α-II epitopes were monitored in the peripheral blood of bread-challenged coeliacs with specific DQ2-tetramer constructs [7,8]. Of BAY 73-4506 clinical trial note, both Australian and Norwegian studies enrolled adult coeliac volunteers, with an average age of 43 years. To our knowledge, no information is available on the responsiveness to short gluten challenge in very young coeliac patients. Furthermore, very little is known about the in-vivo challenge reproducibility

in the same subject cohort, with the exception of a few cases of coeliacs who underwent two separate gluten consumptions described in the above-mentioned studies [7,8]. In the present study we have validated the in-vivo short gluten challenge in a cohort of

14 young CD patients of Italian origin. In particular, we analysed the peripheral blood response against whole gliadin and the immunodominant 33-mer peptide (α-gliadin 57–89). We also assessed the feasibility of exposing the patient cohort to a second in-vivo challenge after a period of 3–10 months of wash-out, in order to estimate the reproducibility of the procedure in the same study population and the intra-individual variations. If replicated successfully in other studies, the short wheat challenge could Fluorouracil concentration represent a strategic tool to evaluate non-invasively the individual’s response to gluten, and could be applied to intervention studies. In fact, the evaluation of small bowel mucosa damage after long-term wheat challenge has been used since the 1950s to assess cereal toxicity or to define the toxic peptides [9–11]. Such studies required repeated endoscopies, before and after treatment, which are always not well accepted by participants. To detect a dysregulated response to gluten, other functional markers, such as faecal fat and xylose malabsorption, resulted in low specificity and sensitivity [12–15]. Furthermore, recent studies have indicated that a short gluten challenge could be used to support diagnosis in doubtful cases of CD [16–18]. Fourteen DQ2-positive volunteers with CD (mean age 18·6, range 15–24 years) participated in the study (Table 1).

As BAFF is able to induce CSR, the intestinal immunoglobulins may well be of another isotype than IgE. The results indicate that BAFF might be particularly involved in non-IgE-mediated reactions. Determination of BAFF levels in different body fluids, as in gut lavage fluid in our study, thus

supports the notion that BAFF is produced locally in different compartments of the body, not only in joints and airways but also in the gut, in response to inflammation and allergic reactions. In addition, our study raises the possibility that Hydroxychloroquine ic50 delayed-type hypersensitivity reactions to food may result from a unique immunoglobulin class switching in the intestine. Enhanced BAFF expression has been noted in several viral infections such as in human immunodeficiency virus (HIV), Epstein–Barr virus (EBV) and hepatitis C virus (HCV) infections [45–47]. Studies in patients with HIV suggest that these patients have increased levels of BAFF and IL-10 in their serum, and BAFF concentration increased with disease progression [45, 46]. EBV-infected

B cells have been shown to express BAFF [4, 47]. In patients with HCV, increased BAFF levels in serum were associated with the presence of arthritis/arthralgia and/or vasculitis, and high values at onset of acute HCV infection can predict its evolution to chronic infection [48]. Another Selleckchem Palbociclib significant association was found between increased serum BAFF levels and liver fibrosis in HCV-infected patients, showing that patients with cirrhosis have more BAFF expression than non-cirrhotic patients [49, 50]. B-cell expansion and lymphoproliferation are common features in patients chronically infected with HCV [51]. Induction of BAFF expression during HIV, EBV Cediranib (AZD2171) and HCV infections may explain the connection

between viral infections and the occasional development of autoimmunity. Persistent viral infection may enhance cell apoptosis and the release of various nuclear antigens including heat shock proteins and the binding of toll-like receptors (TLRs) [52, 53]. Following such activation, dendritic cells become overactivated and increase their production of proinflammatory cytokines, one of which is BAFF, which may terminate B-cell tolerance and stimulate autoreactive B cells to produce autoantibodies. Neoplastic B cells express one or more of the receptors for BAFF on their surface, and impaired TACI upregulation contributes to hyperactive B cells and cancer development [3, 4]. In addition to autoimmune and allergic diseases, high BAFF levels were demonstrated in the serum of patients with B-cell chronic lymphocytic leukaemia (CLL), multiple myeloma and non-Hodgkin’s lymphoma [54–57]. One study showed that many patients had increased levels of BAFF on circulating CLL compared with non-transformed B cells [54]. In different types of non-Hodgkin’s lymphoma, BAFF concentrations were at least threefold higher in serum of patients with follicular lymphoma [56, 58].

Therefore, the defect in ovalbumin (OVA) -specific IgA production is unlikely to be linked to the reduced frequency of CD11b+ DC but rather would be linked to the lack of CD47 expression by non-haematopoietic

cells. CD47−/− BALB/c (back-crossed for 16 generations) and DO11.10 mice were bred in specific pathogen-free conditions at the Experimental Biomedicine Animal Facility, University of Gothenburg. BALB/c (WT) mice were purchased from Taconic, Ry, Denmark. To generate bone marrow (BM) chimeric mice, BM cells from donor WT mice were filtered, red blood cells were lysed, and the remaining cells Ibrutinib were resuspended in PBS. Recipient WT or CD47−/− mice were irradiated (1000 rad) before 2 × 106 to 5 × 106 donor BM cells were transferred intravenously to generate WT  CD47−/− (WT/CD47) chimeras or CD47−/−  CD47−/− irradiation controls (CD47/CD47) and WT  WT (WT/WT). Irradiated mice and mice which underwent mesenteric lymphadenectomy were left to recover for 6 weeks before being included in experiments. The

chimerism was confirmed by flow cytometry. All experiments performed SCH772984 were approved by the Swedish government’s Animal Ethics Committee and followed institutional animal use and care guidelines. Cells were isolated from LN and spleen by mechanical disruption. For DC isolation, tissues were pre-treated with liberase (0·4 mg/ml; Roche, Indianapolis, IN) in Hank’s buffered saline solution (HBSS, GIBCO/Invitrogen, Leek, The Netherlands) supplemented with 2% fetal bovine serum (FBS) FER at 37° for 30 min. Small intestines were flushed with calcium-free and magnesium-free HBSS (GIBCO/Invitrogen) and cut into smaller pieces. The PP were excised from intestinal tissue and washed. For removal of epithelial cells, tissues were incubated at 37° for 15 min with HBSS containing EDTA (5 mm),

FBS (2%) and antibiotics, and then shaken vigorously. The procedure was repeated twice for small intestinal lamina propria (LP) and once for PP. The LP was then digested with collagenase D (100 U/ml; Roche) in RPMI-1640 medium supplemented with FBS (10%), HEPES (15 mm) and antibiotics during two 1 hr incubations. The PP were digested with liberase (0·4 mg/ml) in HBSS containing polymycin B (10 U/ml) at 37° for 27 min. Remaining tissue was disrupted over nylon mesh and counted using a cell counter (Sysmex, Kungsbacka, Sweden) or manually using trypan blue to exclude dead cells. Mesenteric lymph nodes and small intestines were frozen in OCT compound, then 8-μm cryosections were collected on gelatin-coated slides, air-dried and fixed in 1% paraformaldehyde for 5 min.

Here, we demonstrate that CD22 is efficiently activated in trans by complexes of Ag and soluble IgM (sIgM) due to the presence of glycan ligands on sIgM. This result strongly suggests sIgM as a natural trans ligand for CD22. Also, CD22 appears to serve as a receptor for

sIgM, which induces a negative feedback loop for B-cell activation similar to the Fc receptor for IgG (FcγRIIB). CD22 is a 140 kDa glycoprotein on the surface of B cells that negatively regulates signaling through the B-cell Ag receptor (BCR) 1–3. There are six tyrosine residues within the cytoplasmic portion of CD22, four of which are located within ITIMs 4. These tyrosine residues are phosphorylated upon BCR cross-linking, leading to recruitment of SHP-1 4, 5. SHP-1 subsequently dephosphorylates the BCR-proximal signaling molecules, resulting in downmodulation of BCR signaling. Consistent with this, B cells BGB324 cost from CD22-deficient mice are hyperactive 6–9. The extracellular portion of CD22 is composed of seven immunoglobulin (Ig)-like domains, the most distal of which is a V-set Ig-like domain that recognizes α2,6-linked sialic acid (α2,6Sia)-containing glycoconjugates 3, 10. α2,6Sia is common at the terminal of N-linked glycans and is abundantly expressed

on various kinds of cells, including erythrocytes, monocytes, B cells, and T cells. α2,6Sia also exists on soluble plasma proteins such as serum-soluble IgM (sIgM) 11. CD22 is a member of the sialic Luminespib supplier acid-binding Ig-like lectin (Siglec) family, and is also referred to as Siglec-2. CD22 appears to interact with various ligands in cis and in trans to modulate B-cell activity 10. Potential CD22 ligands, including IgM, CD45, and CD22 itself, have been identified 12. Among them, only CD22 has been identified as a natural

glycan ligand for CD22 in cis 13. Furthermore, CD22 regulates BCR signaling induced by Ags expressed on other cells in an α2,6Sia-dependent manner 14. It has recently been reported that sialylated multivalent DOCK10 Ags engage CD22 in trans and inhibit B-cell activation 15. Thus, various interactions between CD22 and its ligands have been shown. However, the overall interactions and the subsequent effects on B-cell activation are not fully understood. In this study, we further evaluated the role of CD22 ligand binding in trans in B-cell activation and propose a novel model of CD22 function. Since sIgM has been shown to bind to recombinant CD22 fusion protein (CD22-Fc) 11, we tested whether sIgM binds to CD22-expressing cells. The mouse myeloma line J558L fails to express the CD22 glycan ligand α2,6Sia at the terminal of N-glycan due to a lack of β-galactoside α2,6-sialyltransferase I (ST6GalI) expression. Introduction of a ST6GalI expression vector can restore α2,6Sia on cell-surface glycoproteins and we showed previously that the soluble CD22 fusion protein (CD22-Fc) bound to J558L cells expressing ST6GalI (J558L/ST6) but not to J558L cells 16.

Such observations are consistent with previous reports where a relatively high APOE ε4 allele frequency was also found among AD (and DLB) cases with capillary involvement compared with those without capillary involvement [11, 14, 22, 23]. The type 4 phenotype was regarded as the CAA-predominant phenotype in which a heavy Aβ deposition was observed in leptomeningeal vessels, click here cortical vessels and capillaries with abundant perivascular deposition of Aβ (dyshoric change). Plaques were either absent or relatively sparse. This phenotype was observed in four (3%) patients,

where at least one region (occipital cortex, but usually all three regions) of the brain was involved. Other workers have reported similar cases, and termed them the ‘vascular variant of Alzheimer’s disease’ or ‘sporadic amyloid angiopathy’. A similar pathology has been described in inherited forms of AD associated with APP692 (Flemish) mutation where Aβ

deposition was referred to as ‘vasculocentric’ [24]. Vidal et al. [25] reported on two sporadic AD cases, both homozygous for APOE ε4 allele, PLX3397 research buy without mutations in APP or PSEN-1 genes, whose main pathological feature was diffuse amyloid angiopathy without evidence of SP. They hypothesized that APOE ε4 allele homozygosity could have been a contributing factor favouring vascular amyloid deposition in leptomeningeal and cortical vessels. APOE genotypes were only available for three of the patients in our cohort, two were APOE ε4 allele carriers (one being APOE ε4 homozygous and one being heterozygous), but the other was a non-APOE ε4-allele carrier (APOE ε3/ε3). Therefore, it cannot be presumed that APOE ε4 allele homozygosity is the sole driving force underlying this phenotype. Interestingly, while the clinical phenotype was available for only one of the present type 4 cases (‘memory’ predominant), selleck chemical one of the other patients had been diagnosed with Frontotemporal

dementia, and thereby was likely to have presented as the ‘frontal’ variant of AD. Vidal et al. [25] further reported that both of their patients had markedly impaired short term verbal recall memory. It is possible therefore that the type 4 pathological phenotype may be more associated with a focal variant of AD, than presenting as ‘typical’ AD. Curiously comparisons of plaque density across the four phenotypes failed to bear out visual impressions of a difference between this group and the other three groups. This is probably due to the low number of type 4 cases available for analysis. Although Thal et al. [11] reported an increased frequency of APOE ε2 allele among their type 2 compared with type 1 (our type 3), CAA cases, we were unable to formally demonstrate such an association in the present study, probably due to the small number of cases of any histological type possessing APOE ε2 allele.

Our results demonstrate that CD4+ T cells from Irf5+/+ mice produce negligible amounts of IL-4;

in contrast, a fraction of CD4+ T cells from Irf5−/− mice produced IL-4 (Fig. 4A). Both Th1 (IFN-γ+IL-4−) and Th2 (IFN-γ−IL-4+) cells, but not Th0 (IFN-γ+IL-4+), exist in Irf5−/− mice, whereas only Th1 cells could be detected in Irf5+/+ mice. The frequency of IFN-γ producing CD4+ T cells from Irf5−/− mice was comparable Lumacaftor supplier with those from Irf5+/+ (Fig. 4A). Together, these data support a critical role for IRF5 in the regulation of Th1/Th2 polarization contributing to pristane-induced lupus pathogenesis. Since IFN-γ production was not impaired in T cells from Irf5−/− mice, the emergence of Th2 cells in Irf5−/− mice is not solely due to a lack of Th1 polarization in this model. In SLE, activated T and B cells can infiltrate tissues to cause organ damage. Recent data in the Yaa murine lupus model [[23]] indicated that IRF5 was critical for T-cell activation. In a similar manner, we investigated whether loss of Irf5 affects lymphocyte activation. At 6 months postpristane, Adriamycin molecular weight we found that expression of the early activation marker

CD69, in splenic CD4+ T cells of Irf5−/− mice, was significantly reduced (Fig. 4B). Because IRF5 regulates type I IFN production [[15, 42]] and type I IFN signaling is central to the pathogenesis of pristane-induced SLE [[25]], we examined the contribution of IRF5 to pristane-induced type I IFN production. Levels of serum IFN were determined by the type I IFN reporter cell assay that measures the ability of sera to cause IFN-induced gene expression [[43]]. buy Baf-A1 A significant decrease in

mRNA levels of the IFN stimulated gene (ISG) IRF7 was observed in L929 cells stimulated with sera from pristane-injected Irf5−/− mice as compared with Irf5+/+ (Fig. 5A). No increase in surface expression of the ISG Sca-1 [[44, 45]] was observed on CD19+ B cells from the PB of Irf5−/− mice 2 weeks postpristane injection (Fig. 5B). Similar results were found at 6 months postinjection in different cellular compartments of Irf5−/− mice (Fig. 5C) and decreased mRNA expression of the ISGs — MCP-1 (ccl2) and MX1 (myxoma response protein) — was also observed in the bone marrow (BM) of Irf5−/− mice (Supporting Information Fig. 2). Ly6Chi monocytes, which are recruited rapidly to the peritoneal cavity (PC) in response to pristane, are thought to be the major source of type I IFN in this model [[44]]. As such, Ly6Chi monocytes were isolated from the PC [[45]] and quantitative PCR (qPCR) performed to determine IRF7 and MX1 mRNA levels. Expression of IRF7 and MX1 were significantly decreased in Irf5−/− mice (Fig. 5D).

g., diet, physical

activity, and smoking) may affect the morphology of the retinal vasculature. Being easily accessible and non-invasively visualized, the retinal microvasculature therefore can be a clinically useful biomarker of reversible sub-clinical physiologic deviation of the systemic circulation as results of such unfavorable exposures. Importantly, quantitative analysis of the retinal microvasculature may be utilized as a prognostic tool, allowing for targeted vascular therapies before the Selleckchem RAD001 onset of overt cardiovascular and metabolic disorders. This review summarizes the modifiable lifestyle and environmental risk factors that affect retinal microvascular structure and the possible clinical implications of such relationships. The retinal microcirculation may reflect healthy and pathophysiologic processes affecting systemic

circulation [64]. The vascular architecture within the retina, as well as elsewhere in the body, is thought to follow the principles of optimality, which allows the blood distribution to peripheral tissue within the quickest time with the least amount of energy [45,65]. Therefore, deviations from optimal structure of the retinal vasculature (e.g., arteriolar narrowing, venular widening) may represent deviation of the circulation from its optimal state, indicating any pathophysiologic processes. During the last few decades, the retinal vasculature has received increasing attention. With the advancement of retinal imaging, the retinal vasculature may allow non-invasive visualization to examine and monitor human circulation systems in vivo Pifithrin-�� (Figure 1). For example, computer-based analysis techniques from digital retinal images has allowed accurate and reproducible measurement 2-hydroxyphytanoyl-CoA lyase of several parameters of the retinal vasculature (e.g.,

caliber, fractal dimension [complexity of vessel network], and branching angle) [6,11,41,61,62]. A number of large-scale epidemiological studies have demonstrated that subtle changes in these parameters carry important information regarding the future risk of systemic vascular diseases [18,25,30,39,40,50,58,60,62]. Importantly, changes in the retinal vasculature have also been shown to have strong associations with systemic and environmental cardiovascular risk factors in a range of populations (for review see Ref. [51]), even before the clinical manifestation of diseases. These subtle retinal vascular changes have been suggested to mirror preclinical changes in both the cerebral [32] and coronary [53] microcirculations. Although the mechanisms remain questionable, this may indicate that abnormalities in the retinal vasculature incorporate a cumulative effect of systemic damage. Recently, many of the largest determinants of this sub-optimal retinal microvasculature have been found to be modifiable [40], such as diet and medications.

The obtained images were analyzed by particle-tracking software for clot size distributions of removed clot fragments, and for non-lysed blood clot areas as function of time. Based on the experimental results, a probabilistic phenomenological model of blood clot dissolution was developed, in which mechanical forces of streaming plasma are in balance with binding forces of blood cells to the remaining clot. Results:  The clot dissolution rate and maximum size of removed clot fragments were

increased with greater flow rate. AZD6244 nmr A 3.3-fold flow rate increase resulted in a two-fold clot dissolution rate increase, while sizes of the removed fragments were in the range of single blood cells, up to thousand-cell clusters. Our phenomenological microscale model of clot dissolution suggests that thrombolysis is a corrosion–erosion-like process. Conclusions:  The findings of this study provide a possible explanation for the origin of clot fragment formation in the blood clot dissolution process. “
“Microcirculation (2010) 17, 3–20. doi: 10.1111/j.1549-8719.2010.00008.x Peripheral arterial disease is a LY2109761 price major health problem and there is a significant need to develop therapies to prevent its progression to claudication and critical limb ischemia. Promising results in rodent models of arterial occlusion have generally failed to predict clinical success and led to questions of their relevance.

While sub-optimal models may have contributed to the lack of progress, we suggest that advancement has also been hindered by misconceptions of the human capacity for compensation and the specific vessels which are of primary importance. We present and summarize new and existing data from humans, Ossabaw miniature pigs, and rodents which provide compelling evidence that natural compensation to occlusion of a major artery (i) may completely restore perfusion, (ii) occurs in specific pre-existing small

arteries, rather than the distal vasculature, via mechanisms involving flow-mediated dilation and remodeling (iii) this website is impaired by cardiovascular risk factors which suppress the flow-mediated mechanisms and (iv) can be restored by reversal of endothelial dysfunction. We propose that restoration of the capacity for flow-mediated dilation and remodeling in small arteries represents a largely unexplored potential therapeutic opportunity to enhance compensation for major arterial occlusion and prevent the progression to critical limb ischemia in the peripheral circulation. “
“This collection of papers is based on talks presented at the IUPS meeting in Birmingham, UK last summer, in a symposium as part of the ESM & EVBO program, sponsored by the British Microcirculation Society and Microcirculation. In this issue we discuss new insights into the control of angiogenesis, including regulation of different aspects of endothelial cell biology by the tissue stroma, during inflammatory disease, and active remodelling of the microcirculation.

Mannose-binding lectin was identified as an important part of natural human immunity and a basic protein activating the lectin complement pathway. Mutations in the promotor and coding area of MBL2 gene are known to lead to significant decreases in serum MBL concentration which might be connected with immunodeficiency manifestation in young age [24]. Infections have been described as a potential trigger of oedema formation in HAE patients [25]. One could speculate that MBL gene mutations could positively influence the HAE phenotype becasue of lower MBL potential to

complement activation, which could mean a lowered MAPK Inhibitor high throughput screening disposition to triggering the complement cascade and oedema development after infectious stimulus in patients with C1 Inh deficiency. However, Cedzynski et al. [26] did not find any relationship between the HAE phenotype and MBL levels and ability of MBL to activate complement, respectively. Our study, which considered a number of varied parameters characterizing a course of the disease, also did not find any evidence on MBL involvement in HAE clinical manifestation. In conclusion, examination of particular functional polymorphisms in BDKR1, BDKR2, ACE and MBL2 genes did not support a hypothesis about the potential

disease-modifying role of these genes on the HAE phenotype. It seems likely that other genetic and/or environmental factors are responsible for HAE clinical variability in Caucasians. However, it has to be emphasized that the study was performed on a limited number of heterogenous patients and therefore

with a limited power of performed this website Etomidate analyses. Becasue of a heterogeneous nature of the disease and its rare occurrence, it seems to be difficult to collect sufficiently large amount of homogeneous HAE patients for powerful analysis in a single country. It is noteworthy that comparable numbers of patients were used also in other studies addressing the influence of genetic factors on HAE clinical manifestation [7, 8, 14, 26]. Evidently, only a large international multicentre study could bring powerful results targeting these topics. We thank Lenka Suchankova for the technical help. The study was supported by the grants Nos. NR7921-3 and NR9192-3 of the Internal grant agency of the Ministry of Health, Czech Republic. Table S1 Frequency of BDKR1, BDKR2, ACE, and MBL2 gene polymorphisms and mutations in HAE patients and control individuals. Table S2 Association of HAE clinical score and gene variants in the BDKR1, BDKR2, ACE, and MBL2 genes in all HAE patients. Table S3 Association of HAE disease severity, frequency of attacks and disease onset with analysed gene variants in the BDKR1, BDKR2, ACE, and MBL2 genes in all HAE patients. “
“OTHER ARTICLES PUBLISHED ON ANCA IN THIS ISSUE Animal models of anti-neutrophil cytoplasmic antibody-associated vasculitis. Clinical and Experimental Immunology 2012, 169: 229–37.