On the other hand, RIG-I interacted with V and Vcys but not with P/V, Vu, and Vu cys (Fig. 2B), suggesting that the interaction requires the entire V protein and that cysteine mutations did not affect

the interaction. Similar results were obtained in binding of the V protein with IKKɛ and IRF3 (data not shown). These results show that only MDA5 interacts with the V unique region and that RIG-I, IKKɛ and IRF3 interact with the V protein in a mode different from MDA5. We next investigated whether the V and MDA5 interaction was related to inhibition of IRF3 activation. 293T cells were transfected with an IRF3-dependent reporter plasmid, p-55C1B-EGFP, together with FL-MDA5 and one of the viral proteins. Cells were further learn more transfected with poly(I:C), and IRF3 transcription activation was investigated. EGFP expression showed that the V protein significantly suppressed IRF3 activation induced by overexpression of FL-MDA5 (Fig. 3A). Expression of EGFP as well as FL-MDA5 selleck chemical and a viral protein was confirmed by western blotting (Fig. 3B), and light intensity of EGFP protein bands was quantitated and plotted in a graph (Fig. 3C).

The N-terminal part of the V protein lacking the V unique region (P/V) and the C protein (C) did not suppress IRF3 activation. The V protein with a single point mutation of a cysteine to alanine at the V unique region (Vcys2A: C341A and Vcys7A: C365A) also did not suppress IRF3 activation. Amino acid substitutions of the SeV V protein at those positions ameliorated viral load and pathogenicity in a mouse model (11). Influenza virus NS1 protein (NS1) did not suppress IRF3

activation in this condition. SeV C protein and influenza virus NS1 protein are known to inhibit IRF3 activation and IFN-β production (27, Depsipeptide research buy 28, 29). The inhibition is thought to be due to reduction of RNA species that belongs to pathogen-associated molecular patterns. These two proteins did not inhibit IRF3 activation induced by overexpression of MDA5 and poly(I:C) treatment. A similar experiment using an IRF3-dependent GFP and luciferase reporter plasmids showed that V and Vu suppressed IRF3 activation and that the Vcys and Vu cys, which have two point mutations at the cysteine residues in the V unique region, and P/V did not suppress the IRF3 activation (Fig. 4). These results indicate that the V protein suppressed MDA5-induced IRF3 activation in a Vu-dependent and cysteine-dependent manner, corresponding to the mode of interaction of V proteins with MDA5. We previously reported SeV V mutants with attenuated pathogenicity (11, 12). SeV V-H318N, R319W, R320G, and W336G were highly attenuated in virulence by more than 25-fold in 50% mouse lethal dose, and SeV V-E321K and P339T were mildly attenuated (12). We infected FL-MDA5-transfected cells with V mutant viruses, and interaction of the V mutant protein with FL-MDA5 was then investigated by immunoprecipitation and western blotting.

Solomon and colleagues assessed the relationship among the initial haemoglobin response to darbepoetin after two weight-based doses, the haemoglobin level achieved after 4 weeks, the subsequent darbepoetin dose and outcomes in 1872 patients from the TREAT trial who were randomized to darbepoetin.15 The initial dose of darbepoetin was 0.75 µg/kg Y-27632 concentration of body weight and was repeated after 2 weeks if haemoglobin values did not exceed 140 g/L. Poor initial response to darbepoetin was defined as the lowest quartile of per cent change in haemoglobin level (<2%) after the first two standardized doses of the drug. Patients in the lowest quartile of haemoglobin responsiveness were more likely to have cardiovascular

disease, high CRP levels and low ferritin and transferrin saturation levels. The average haemoglobin level after 12 weeks remained marginally but statistically significantly lower among patients with a poor initial response (122 ± 9 g/L) than among those with a better initial response (124 ± 7 g/L, P < 0.001). The average monthly dose of darbepoetin after 12 weeks and throughout the remainder of the trial was substantially higher among patients a poor initial response (median dose, 232 µg; interquartile range, 126 to 390) than those with a better initial response (167 µg; interquartile

range, 95 to 310; P < 0.001). There was significant difference CP-690550 chemical structure in the use of intravenous iron or blood transfusion throughout the trial. Compared with patients with a better initial response, those with a poor initial response were at increased risk of a cardiovascular composite event (HR 1.31, 95% CI 1.09–1.59) and all-cause death (HR 1.41, 95% CI 1.12–1.78). The event rates for the cardiovascular composite outcome and all-cause death in the better initial response group

were comparable with 4-Aminobutyrate aminotransferase the placebo group. These findings indicate that requirement of high-dose ESA to achieve target haemoglobin rather than achieved haemoglobin may be responsible for the poor outcome. Interestingly, the event rates for stroke were comparable in the two response groups, but higher in both groups than in the placebo group. It still remains unclear whether the use of ESA or high haemoglobin target contributed to increased risk of stroke in patients treated with darbepoetin. A summary of the observational studies is provided in Table 2. In a US Medicare study of 75 283 prevalent patients receiving haemodialysis between July–December 1993, a haematocrit level of 33–36% was associated with a similar risk of mortality (adjusted RR 0.96, 95% CI 0.91–1.01) compared with a reference haematocrit level of 30–33%.16 In contrast, lower haematocrit levels were associated with increased risk of mortality (haematocrit <27% RR 1.33, 95% CI 1.26–1.40; haematocrit 27–30% RR 1.12, 95% CI 1.08–1.17). The pattern of higher mortality with lower haematocrit was similar in diabetic and non-diabetic patients.

Results: Twenty-three studies (n≥4675 respondents) were included. The studies were conducted in the United Kingdom, United States, Australia, Sweden, Netherlands, and Pexidartinib concentration Iran. Four (17%) were multinational

studies. Nephrologists’ preferences varied with respect to: medical suitability – some indicated lower likelihood of recommending transplantation for patients with cardiovascular disease, diabetes, obesity, and infection; non-adherence was regarded by some as a contraindication for transplantation; and socio-demographic characteristics – patients of older age, ethnic minorities, or low socio-economic status were less likely to be recommended. Six major themes underpinned nephrologists’ perspectives: prioritising individual benefit and safety, maximising efficiency, patient accountability, justifying gains, protecting unit outcomes, and reluctance to raise patients’ expectations. Conclusions: Variability in nephrologists’ preferences may be contributing to disparities in access to transplantation. Evidence-based guidelines supplemented with pragmatic tools for determining selleck chemicals medical and psychosocial criteria for referral and waitlisting may support more systematic and equitable decision-making.

Continuing medical education informed by current evidence on transplant outcomes, and psychosocial and educational interventions, particularly for high-risk or disadvantaged patient populations, could help to reduce overall disparities in access to transplantation. 259 POLYCYSTIC KIDNEY DISEASE AS A RISK FACTOR FOR NEW ONSET DIABETES AFTER RENAL TRANSPLANTATION: A META-ANALYSIS J JANARDAN1, R WALKER2,3 1Department of General Medicine, The Alfred hospital, Melbourne, Victoria; 2Department of Renal Medicine, The Alfred hospital, CHIR-99021 chemical structure Melbourne; 3Monash University, Melbourne, Victoria, Australia Aim: A systematic review of published medical literature on autosomal dominant polycystic kidney disease (ADPKD) as a risk factor for new onset diabetes after transplantation

(NODAT) in renal transplant recipients. Background: NODAT is an important complication of renal transplantation with reported rates varying from 3% to 46%, depending on the diagnostic criteria and length of follow-up. There is conflicting data regarding the increased incidence of NODAT in patients with ADPKD. Methods: We searched the PUBMED database for studies published before February 2014. Out of 129 citations, 12 suitable studies were selected for analysis. The incidence of NODAT in patients with ADPKD was compared to patients with alternative renal pathology using odds ratio (OR) and respective 95% confidence interval (CI). Results: The analysis revealed a higher incidence of NODAT in the ADPKD population (OR: 1.15, 95% CI: 1.06–1.25).

Some of the text is canted towards the generalist and will be useful to early stage trainees. There is a brief discussion see more on the use of squash/smear preparations in which the authors discuss the pros and cons vs. frozen section. They conclude that relative usage depends on the technical availability of quality frozen sections and by which method the pathologist was trained.

Having touched upon these matters, the authors are entirely clear that this book is solely focussed on frozen section diagnosis and readers expecting to learn something of smear diagnosis interpretation should look elsewhere – there is only one smear micrograph in the whole book. Chapter 3 is dedicated to identifying non-neoplastic disease and avoiding the pathologist’s nightmare of a false positive tumour diagnosis. As with the initial chapters, FK866 purchase this is approached in a structured manner, directing the reader to observe the presence or absence of specific features (‘flags’) and leading them through a diagnostic algorithm suggesting suitable differential diagnoses that are conveniently summarized in a couple of tables. Chapters 4 and 5 are a logical extension to Chapter 3 and deal with tumours of the cerebral parenchyma, addressing first the metastatic lesions (Chapter 4) and then the primary brain tumours (Chapter 5).

There is more of a descriptive approach to these chapters and the major histological features of intrinsic tumours and their sub-types, as detailed in the current WHO manual, are rehearsed in brief. The authors interestingly advocate providing the surgeons with a WHO grade in this provisional assessment. The subsequent chapters follow this general format and cover dural based tumours, intraventricular lesions, cerebellar based lesions, pituitary gland and sellar lesions, pineal U0126 in vivo gland lesions and spinal cord lesions. Each chapter adequately addresses the range of possibilities one might reasonably expect to encounter, en route indicating pitfalls and providing differential diagnoses. Overall

the writing style is clear and concise but some readers may find it possibly a little too narrative for ‘flick and find’ rapid reference as the publishers intend. Most chapters have an introductory paragraph to set the scene. Presumably owing to the volume’s compact size, the print size is slightly smaller than the usual text book (I estimate around 11 point) and the presbyopic will need their reading glasses. The micrographs (c. 164 in number) are generally of good print quality and colour balance and as large as the format allows with a maximum of two per page covering the available width. Most of the frozen section material from which these micrographs derive are of outstanding quality and can easily be taken for paraffin embedded H&E’s.

α-GalCer (Alexis Biochemicals) and β-GalCer (Galactocerobroside, Sigma) were dissolved in DMSO. The anti-CD1d mAb WTH-1 [13] was added to the cultures 30 min before the addition of any stimuli. Spots were analyzed

and enumerated using the CTLImmunoSpot S5 Versa analyzer ELISPOT reader and the ImmunoSpot 4.0 Software (both from CTL). Small spots (smaller than 0.096 mm2) obtained in cultures with medium only were considered nonspecific background and were subtracted from all the samples. Single cell suspensions prepared from spleens and livers were plated at a density of 106 cells per 1 ml of RPMI 1640 supplemented as aforementioned. Cells were cultured with 20 ng/ml α-GalCer during the first 7 days. During the second week, the cells were cultured 5-Fluoracil with 10 ng/ml α-GalCer and 10% of T-cell

growth factor-containing medium (supernatant from Con A-stimulated rat splenocytes blocked with α-methylmannoside) usually adding fresh media at day 13. We would like to especially thank T. Hünig for his continuous support to this project and N. Beyersdorf for critical reading of the manuscript and helpful comments. This selleck screening library work was supported by the Deutsche Forschungsgemeinschaft Graduate College 520 Immunomodulation and HE 2346/6-1. EMC was also supported by a STIBET Doktoranden grant of the Deutsche Akademische Auslandsdienst. DBS was supported by NIH NIAID R01 AI083988 and AI059739 and by the Robert Wood Johnson Foundation (grant no. 67038) to the Child Health Institute. The authors declare no financial or commercial conflict of interest. As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. Technical

support issues arising from supporting information Leukotriene-A4 hydrolase (other than missing files) should be addressed to the authors. Figure S1. Figure S2. Table S1. Table S2. Table S3. “
“Dendritic cells (DC) are key factors in regulating immune responses, and they induce immune response or tolerance depends on its maturation states. Previous studies demonstrated that blocking IKK2 in bone marrow-derived dendritic cells (BMDC) by adenoviral transfection with a kinase-defective dominant negative form of IKK2 (IKK2dn) could inhibit NF-κB activation and impair DC maturation. Here, we transfected IKK2dn into recipient rat (Lewis) BMDC by adenovirus vector (Adv-IKK2dn-DC) and found that Adv-IKK2dn-DC had reduced B7-2 and B7-1 expression under alloantigen stimulation. Their ability to induce allogeneic T-cell proliferation was markedly reduced in comparison with uninfected DC. A higher IL-10 secretion and a lower IFN-γ secretion were detected in Adv-IKK2dn-DC-stimulated allogenic T cells. Furthermore, we showed that Adv-IKK2dn-DC pulsed with BN (Brown Norway rats) splenocyte lysates markedly prolonged the survival of renal allografts in an antigen-specific manner.

3–5,44 Hence, the diurnal suppression of Tres cytokine secretion by nTreg might, in part, be driven by the cellular circadian clock of nTreg via yet-unknown pathways. Therefore, the analysis of the circadian BMN-673 clock in T cells should be addressed in future studies. Besides the cellular circadian clock, the hormonal priming of T cells in vivo could be another mechanism

for the diurnal rhythm of cytokine secretion by Tres.13 To investigate this possible mechanism we analyzed the hormone levels from all subjects and performed a multiple linear regression analysis. We found a negative correlation between cortisol serum levels and T-cell cytokine secretion. Furthermore, we demonstrated in vitro that a 2 hr pre-incubation with physiological daytime levels of cortisol decreased cytokine secretion. This Trametinib supplier is in line with in vitro data published by other investigators demonstrating an immunosuppressive effect of cortisol.8,26,30,45–47 A positive correlation

was found between melatonin and prolactin serum levels and T-cell cytokine secretion. Whereas we could show in vitro that pre-incubation of Tres with prolactin increased the secretion of IL-10 but decreased that of IL-2 by Tres, we were unable to demonstrate this effect for melatonin. Prolactin was described to display immune-stimulatory functions in vitro, whereas conflicting data are published for melatonin.27,30,48,49 We also observed increased IFN-γ after prolactin pre-incubation AMP deaminase but this effect was not significant, as previously described by Matera et al. and Dimitrov et al.29,30 However, Matera et al. investigated unstimulated T cells while we used polyclonally stimulated Tres. Dimitrov et al. studied the percentage of IFN-γ-producing T cells in whole blood which were stimulated with PMA/ionomycin in the presence of prolactin. By contrast, we pre-incubated T cells with prolactin, performed

the assays (αCD3 stimulated) without prolactin and measured the concentration of IFN-γ in the supernatant. Despite these different approaches, our observations are broadly similar to these other reports.29,30 Our findings on the effect of melatonin are in line with other investigators who did not observe stimulatory effects of melatonin in vitro.49 We could not confirm the proposed Th1-enhancing effect of melatonin in vitro but these published data are from in vivo experiments in mice and conflicting data have also been published.50 In any case, one can speculate, from the effects of cortisol and prolactin, that the hormonal milieu could be one mechanism of the diurnal rhythm of cytokine secretion by Tres. The suppressive activity of nTreg on cytokine secretion by Tres did not correlate with the serum levels of any of the hormones.

rubrum-specific primers. Of the scale samples, 16% were positive for T. rubrum in the culture and PCR as well, 9% were positive in the PCR only and 3% in the culture only, whereas 5% were only KOH-positive. The corresponding results for nail samples were 17%, 20%, 3% and 7%. PCR results were available after 2–5 days, culture results after 2–3 weeks. Our results show that a specific PCR assay can successfully be used to detect T. rubrum directly in samples collected from superficial skin lesions and nails under routine

conditions. Compared with conventional methods, it is faster and more sensitive. We recommend its complementary use. Superficial tinea including onychomycosis is the most frequent cutaneous fungal infection in Germany with Trichophyton rubrum as the causative agent in about 80–90% of all cases.1,2 However, the

clinical picture of tinea caused by T. rubrum is not diagnostic because a multitude of other diseases can cause phenotypic changes find more identical to those induced by various dermatophytes, including T. rubrum. Therefore, a definite diagnosis of T. rubrum-tinea needs a positive proof of T. rubrum within the tissue. The most common Venetoclax mw and approved methods to detect dermatophytes in skin samples are KOH-mounts that allow a rapid demonstration of fungal elements, but no species identification and mycological cultures for species recognition. However, for various reasons, cultures can remain false negative, a positive culture can easily take 3 weeks and occasionally even a positive culture may not allow a definite identification. On the other hand, T. rubrum can nowadays unambiguously be identified by molecular analysis3,4 and modern PCR-based genetic methods to detect dermatophytes reliably and rapidly in infected skin and nails are currently proposed.5–11 In our study, we systematically analysed unselected skin samples collected under routine conditions from suspected tinea lesions by KOH-mounts, dermatophyte cultures and a

T. rubrum-specific PCR to check the Astemizole applicability and benefit of the latter method in the daily routine. Unselected samples of skin scales and nail scrapings obtained from dermatological patients that were submitted to our laboratory for mycological testing were employed. No particular instructions had been given for the collection of these samples and all samples had been taken under routine conditions from skin lesions or nails to prove or exclude a fungal infection. The samples included scrapings from lesional stratum corneum and from nails (almost exclusively toe nails) and were submitted in glass tubes without any additives. Samples from nails were taken by scraping off material from the destructed nail plate and/or subungual debris at a site as closely as possible to the proximal margin of the lesional area by use of a curette. The time period of collection was from April 2007 to November 2008 and all submitted samples with a sufficient amount of material were included.

Bullous pemphigoid is an autoimmune blistering skin disease often associated with chronic inflammation and malignancies [40]. Increased serum BAFF levels in patients with bullous pemphigoid may be because of inflammation-enhanced production of inflammatory cytokines (INFγ) triggering BAFF secretion in affected tissues, which in turn stimulates B-cell and

T-cell responses locally as well as in neighbouring lymph nodes contributing to the development of bullous pemphigoid in susceptible individuals [26]. The clinical relevance of BAFF in allergic diseases has been pointed out in a number of papers [10, 41, 42]. Allergy can be antibody-mediated (IgE) or cell-mediated (non-IgE). Mast cells and MS 275 basophils

are key effector cells for IgE-mediated allergy whereas the reaction/inflammation is mediated by allergen-specific T lymphocytes in non-IgE-mediated allergy [43, 44]. A possible pathogenetic role of BAFF and its association with delayed-type hypersensitivity reactions and atopy were investigated in patients with asthma, rhinitis and self-reported food hypersensitivity. A study by Kang et al. [41] showed markedly an increased serum levels of BAFF in patients with non-IgE-mediated asthma, apparently related to the degree of airway hyperresponsiveness. Decreased serum levels of BAFF after improvement with anti-asthmatic therapy suggest that BAFF could be AZD2014 solubility dmso a novel parameter for monitoring the severity of asthma symptoms. Another study reported

that BAFF is upregulated in the airways of allergic subjects after allergen exposure [10]. In 12 patients with allergic asthma and four patients with allergic rhinitis, bronchoalveolar lavage fluids were collected after challenge with allergen or saline. Significantly, increased concentrations of BAFF in lavage fluids after segmental challenge (20–24 h after Selleck Decitabine challenge) suggest that BAFF production occurs locally within the airways. Recently, we demonstrated for the first time that BAFF concentrations in serum and gut lavage fluid were significantly increased in patients with non-IgE-mediated hypersensitivity reactions to food [42]. Non-atopic patients had significantly higher levels of BAFF in serum than both atopic patients and controls, and there was no significant correlation either between serum BAFF concentration and total IgE levels or between BAFF concentration in gut lavage fluids and serum total IgE levels. To eliminate the possibility that high concentrations of BAFF in gut lavage merely reflect vascular leakage, BAFF values in gut lavage fluids and blood were normalized to respective concentrations of albumin. The ratio of BAFF to albumin in gut lavage fluids was much higher than the ratio in blood suggesting that BAFF is produced locally in the intestines.

[123, 124] Therefore, IL-22 is likely to be an important factor in the pathogenesis and clinical

outcome Vorinostat of hepatitis B virus and hepatitis C virus infections, where the liver is a major target organ such as in DHF/DSS.[64, 125] Recently, it has been shown that acute DENV-2 infection elicited high levels of IL-17 in patients with severe disease (DHF).[126] However, other studies found a correlation between IL-17 levels and mild infection (DF).[127] Malavige et al.[128] found no differences for IL-17 levels in patients with DHF who developed shock and those who did not. Furthermore, Talarico et al.[33] demonstrated age-related differences in the primary response to DENV, characterized by an immature Th2 polarization and MK-2206 chemical structure Th17 suppression in infants. Hence, the ultimate role of Th17 cytokines in the pathogenesis of dengue is yet to be unveiled. In the experimental model of DENV-2 infection, using the P23085 adapted strain, we showed that mice deficient for the cytokine IL-22 were more susceptible to experimental DENV infection, presenting increased inflammation and severe tissue injury, especially in the hepatic parenchyma.[68] This was associated with increased mortality, levels of AST/ALT in serum, greater neutrophil accumulation and/or activation and a small increase in viral load

in the liver. DENV-2-infected HepG2 cells treated with recombinant human IL-22 showed reduced cell death and IL-6 production. These data clearly suggest that IL-22 appears

to play a key role in liver homeostasis in the course of DENV infection. Regarding the main leucocyte subsets that participate in our experimental system, γδ T cells and NK cells were the major sources of IL-17A and IL-22, respectively. Although we had observed a minor production of IL-17 by CD4+ Th17 cells in the spleens of infected WT mice, these populations do not appear to represent the real key players in this experimental setting. Recently, γδ T cells (but not Th17 cells) have been shown to be the primary source of IL-17A production in the early phase of Escherichia coli infection, which is related to an early infiltration of neutrophils such as in our model of DENV-2 in mice.[129] Moreover, γδ T-cell-derived IL-17A is critical for the optimal SB-3CT induction of cytotoxic T lymphocyte responses and protection against primary intracellular Listeria monocytogenes infection in the liver.[130] Interleukin-17A production during experimental DENV-2 infection was strongly correlated with disease severity, which was confirmed by the fact that infected IL-17RA-deficient mice were less susceptible than WT mice.[68] Immature or mature NK cells (CD3− NKp46+) have been identified in the mucosa and found to be capable of producing IL-22 in different models of infection.[121, 131] We have shown here that NK cells (CD3− NK1.1+) are the major producers of IL-22 in the present model.

Although TGF-β can mediate B cell production of IgA in vitro in general, TGF-β alone under the present culture conditions did Selleckchem Navitoclax not alter B cell differentiation, nor did it augment the sCD40L- or IL-10-mediated IgA induction. Rather, IgA production induced by sCD40L and IL-10 was reduced significantly, albeit slightly, by addition of TGF-β (20·93 ± 6·09 µg/ml versus 34·71 ± 7·17 µg/ml, P < 0·05, Fig. 2a). Therefore, TGF-β was not used further in this study in addition to sCD40L and IL-10 as a differentiation/switch factor to induce B cell IgA production. Next, we examined if our culture conditions engaged the intracellular phosphorylation of the classical NF-κB (Fig. 3a) and

STAT3 (Fig. 3b) pathways. We used ELISA to detect pNF-κB p65 and Everolimus pSTAT3 in nuclear extracts from B cells stimulated with sCD40L (50 ng/ml) and/or IL-10 (100 ng/ml) for 30 min. The sCD40L + IL-10 combination and, to a lesser extent, sCD40L

alone, increased the pNF-κB p65 levels significantly in cultured B cells. IL-10 alone gave no signal over the control (Fig. 3a). In sharp contrast, sCD40L addition gave no signal over control signal for STAT3 phosphorylation, of which IL-10 was shown to be a powerful stimulator. No significant gain in pSTAT levels was observed when IL-10 was combined with sCD40L (Fig. 3b). Thus, in the in vitro conditions that initiate purified human blood B cell differentiation into IgA-secreting cells, sCD40L was able to induce the phosphorylation of NF-κB

p65 but not of STAT3, while IL-10 induced the phosphorylation of STAT3 but not of NF-κB p65. Whereas sCD40L and IL-10 did not increase IgA production levels synergistically compared to sCD40L or IL-10 alone (Fig. 2a), IL-10 clearly increased CD40L-mediated activation of NF-κB p65 (Fig. 3a). IL-6 has long been considered to be involved in Ig (particularly IgA) production [29]. Recently, IL-6 was also found to be one the main cytokines that is capable of inducing ROS1 phosphorylation of STAT3 [30]. Moreover, IL-6 is released quickly by B cells after activation. We then asked whether IL-6 could behave as a mediator between IL-10 signalling and STAT3 phosphorylation. We hypothesize that IL-10 (through IL-10R) induces IL-6 release from B cells. This IL-6 could then be recaptured by B cells (through IL-6R) and activates STAT3. To test whether the IL-10-driven activation of the STAT3 pathway is direct or indirect, we measured both B cell production of IL-6 and IgA and also STAT3 phosphorylation in the presence or absence of IL-6R or IL-10R blocking antibodies. B cells were incubated with IL-6R or IL-10R blocking antibodies for 120 min and were then stimulated by IL-6 or IL-10 for 30 min. The level of STAT3 phosphorylation was measured by ELISA (Fig. 4a). In the absence of inhibitors, both IL-6 and IL-10 significantly induced STAT3 phosphorylation.