Glycocalyx thickness is reduced, glomerular endothelial cell pore size is increased, glomerular charge selectivity is reduced and

podocyte cell foot processes are fused. These changes are associated with reductions in glomerular cell production PI3K inhibitor of proteoglycans and glycosaminoglycans contained within the glycocalyx produced by the glomerular endothelial cells.11 Further evidence for a direct effect of Adriamycin on the kidney comes from a study in which clipping of the renal artery of one kidney protects it from injury.20 Additional studies have examined the molecular mechanisms for Adriamycin-induced renal injury. Increased free radical production has been proposed as a pathogenetic mechanism. This is supported by isolation perfusion studies of hagfish (Myxine glutinosa) glomeruli Autophagy Compound Library in which

Adriamycin was found to reduce glomerular ATPase activity in association with a reduction in water permeability, an effect reversed by the sulfhydryl donor N-acetylcysteine. In addition, depleted levels of glutathione (an anti-oxidant) and elevated levels of lipid peroxide levels in liver, kidney and heart developed after Adriamycin administration.60 Evidence for the role of advanced glycation end products comes from studies of receptor for advanced glycation end product (RAGE)-null mice. These mice are protected from Adriamycin-induced podocyte damage and proteinuria. Adriamycin induced generation of RAGE ligands, an effect reversed by treatment Prostatic acid phosphatase with soluble RAGE. The mechanism for RAGE ligand-induced renal injury involved the activation of nicotinamide adenine dinucleotide phosphate-oxidase and p44/p42 MAP kinase signalling, and upregulation of pro-fibrotic growth factors.61 The changes associated with the slit diaphragm in Adriamycin-induced nephropathy have been studied by Otaki, Kawachi and colleagues.62 Early after Adriamycin administration (day 7), expression of the slit diaphragm

molecules nephrin, podocin and NEPH1 (but not ZO-1- and CD-associated protein) is altered from a continuous to a discontinuous dot-like pattern consistent with podocyte injury. In particular, NEPH1 was disproportionately affected. Using immunoprecipitation and western blot studies of glomerular lysates from animals 7 days after Adriamycin injection, Kawachi’s laboratory found that a large proportion of nephrin lost its affinity with NEPH1. While these data are observational in nature, they do point to slit diaphragm abnormalities as critical early events in the pathogenesis of Adriamycin-induced proteinuric renal injury. Gene profiling using microarray chip technology has identified gene networks that are potential drivers of tubulointerstitial fibrosis in AN.

Supernatants of T-cell proliferation cultures were harvested at 48 h after initiation of culture. Concentrations of IFN-γ, TNF-α, IL-2, IL-4, and IL-5 were measured by mouse Th1/Th2 cytokine bead array assay (BD Bioscience) according to manufacturer’s recommendation. Thymus was obtained from 6-wk-old C57BL/6 mice, and cell suspensions

were stained for 15 min in PBS+0.5% BSA with specific mAbs against CD4-PE and CD8-FITC (BD Bioscience). After staining, cell suspensions were washed and resuspended for analysis. Flow cytometric analysis was performed by a FACScan (BD Bioscience). Splenic T cells from C57BL/6J mice Gamma-secretase inhibitor were starved in RPMI1640 + 0.1% FBS at 37°C and 5% CO2 for 4 h at the cell concentration of 1 × 106/mL. After the starvation, cells were resuspended in RPMI1640 + 0.1% bovine serum albumin (BSA), and seeded in the plates precoated with anti-CD3/ephrin-Bs

at 2 × 105 cells/well. The plates were centrifuged at 350 rpm for 3 min to achieve rapid contact between the cells and the bottom of the culture wells. The cells were incubated at 37°C and 5% CO2 for 2 h. Then, the cells were harvested and washed with ice-cold PBS. Cell lysis and subsequent Western blotting were performed learn more as previously described [[58]] with minor modifications. Briefly, cells were lysed in cell lysis buffer containing 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 1% Triton X-100, 1 mM sodium vanadate, 50 mM sodium fluoride, and protease inhibitor cocktail (Sigma Aldrich). For immunoprecipitation, RIPA lysing buffer (50 mM Tris-HCl, pH 7.5, 137 mM NaCl, 2 mM EDTA, 1% NP-40, 0.1% SDS, 0.5% sodium deoxycholate, 1 mM sodium vanadate, 50 mM sodium fluoride, and protease inhibitor cocktail) was used. The lysates were boiled with SDS-loading buffer. Equal amount of sample proteins (35 μg) were separated on 7.5–16% SDS-PAGE and transferred onto PVDF membranes (Immobilon, Millipore, Billerica, MA, USA). The membranes were first incubated with TBST (20 mM Tris-HCl,

Ergoloid pH 7.5, 137 mM NaCl, 0.1% Tween20) containing 5% nonfat dried milk and probed with specific antibodies using primary and horseradish peroxidase-conjugated goat anti-rabbit secondary antibodies (Cell Signaling Technologies, Danvers, MA, USA). Immune complexes were detected by chemiluminescence (Immobillon Western, Millipore). For immunoprecipitation, total cell lysates were incubated with anti-PY antibody (clone 4G10, Millipore) and protein G-sepharose (GE Healthcare Bio-Sciences AB, Sweden) for 18 h at 4°C. The immunoprecipitates were washed with lysis buffer and then with PBS. The blotting membranes were incubated with biotinylated rabbit anti-goat IgG (BA-5000, Vector Laboratories, Burlingame, CA, USA) followed by the amplification with ABC system (Vectastain Elite ABC Reagent, Vector Laboratories).

By contrast, LASV- and MOPV-infected macrophages activated NK cells, as shown by the upregulation of CD69, NKp30, and NKp44, the downregulation of CXCR3, and an increase in NK-cell proliferation. NK cells acquired enhanced cytotoxicity, as illustrated by the increase in granzyme B (GrzB) expression and killing of K562 targets, but did not produce IFN-γ. Contact between NK cells and infected

macrophages and type I IFNs were essential for activation; however, NK cells could not kill infected cells and control infection. Overall, these findings show that MOPV- as well as pathogenic LASV-infected macrophages mediate NK-cell activation. Lassa fever (LF) is a viral hemorrhagic fever caused by Lassa virus (LASV). It is endemic in West

Africa and causes 100,000–300,000 cases and 5000–6000 deaths each year [1]. find more The absence of a vaccine and the limited use of ribavirin, the only antiviral drug licensed, in endemic countries, render LF a public health problem. https://www.selleckchem.com/products/Fludarabine(Fludara).html LASV and Mopeia virus (MOPV) are very closely related Old-World Arenaviruses with a common animal reservoir, Mastomys natalensis, a peridomestic rodent [2]. Unlike LASV, MOPV is not pathogenic to nonhuman primates (NHPs), in which this virus has even shown to confer protection against challenge with LASV [3]. The immune responses to LASV and MOPV are poorly understood. The control of LASV seems to involve the induction of T cells, rather than Staurosporine ic50 humoral responses [4]. Indeed, cellular immune responses specific for viral glycoproteins appear to protect NHPs against lethal challenge [5]. By contrast, severe LASV infections seem to be associated with immunosuppression and structural changes to secondary lymphoid organs. LASV and MOPV display tropism for APCs, such as DCs and macrophages (MΦs) [6-8]. These cells are the first targets of the viruses and they release large numbers of viral particles without cytopathic effects. APCs display only very low levels of activation or maturation after LASV infection

[6] and produce only small amounts of type I IFN [9]. By contrast, MOPV infection results in type I IFN production by MΦs and, to a lesser extent, by DCs, and triggers the early and strong activation of MΦs [8]. The different responses of APCs to LASV and MOPV infections are probably involved in the difference in pathogenicity between the two viruses. It has been shown that CD4+ and CD8+ T cells are strongly and rapidly activated in response to MOPV-infected DCs, resulting in proliferation, differentiation into effector, cytotoxic, and memory cells. By contrast, LASV-infected DCs can induce only weak and delayed T-cell responses in vitro [10]. Like APCs, NK cells are at the crossroads between the innate and adaptive responses. They have effector functions in innate immunity, through their cytotoxic properties, and also produce cytokines involved in the induction of T-cell responses.

Pain (NRS) 7 NSAIDs; AED; narcotics. Heart disease. CRPS24 F/42 Motor vehicle accident (MVA); right BPTI;

disk at C6-C7; surgery with fusion/5·5 years Neurogenic oedema; autonomic dysregulation; positive Tinel signs; generalized mechano allodynia; hyperalgesia. Pain (NRS) 8 NSAIDs; AED; antidepressants; spasmolytics; narcotics. Depression; hypertension; hypercholesterolemia. CRPS25 F/49 L5-S1 disc; fall with BPTI/18 years Dynamic and static mechano allodynia; thermal allodynia; hyperalgesia; Wnt inhibitor spread from leg to brachial plexus; generalized weakness; decreased initiation of movement. Pain (NRS) 7·5 NSAIDs; AED; antidepressants; narcotics; intravenous ketamine; intravenous lidocaine. Hypertension; hypercholesterolemia; L5-S1 radiculopathy; migraine. “
“The tapeworm Echinococcus granulosus is the causative agent of hydatid disease and affects sheep, cattle, dogs and humans worldwide. It has a two-stage

life cycle existing as worms click here in the gut of infected dogs (definitive host) and as cysts in herbivores and humans (intermediate host). The disease is debilitating and can be life threatening where the cysts interfere with organ function. Interruption of the hydatid life cycle in the intermediate host by vaccination may be a way to control the disease, and a protective oncosphere antigen EG95 has been shown to protect animals against challenge with E. granulosus eggs. We explored the use of recombinant vaccinia virus as a delivery vehicle for EG95. Mice and sheep were immunized with the recombinant vector, and the result monitored at the circulating antibody level. In addition, sera from immunized mice were assayed for the ability to kill E. granulosus oncospheres in vitro. Mice immunized once intranasally developed effective oncosphere-killing antibody by day 42 post-infection. Antibody responses and oncosphere killing were correlated and were significantly enhanced by boosting mice with either EG95 protein or recombinant vector. Sheep antibody responses to the recombinant vector or to EG95 protein mirrored those in mice. Hydatid disease is a parasitic infection that affects

sheep, cattle, dogs and humans (1,2). The disease Glutathione peroxidase is endemic in many countries and is a worldwide problem (3). A vaccine approach to control this parasite may offer a cost-effective strategy (4,5). The tapeworm Echinococcus granulosus is the causative agent of hydatid disease. It has a two-stage life cycle, existing as worms in the gut of infected dogs and other canids (definitive host), and as fluid-filled cysts containing immature tapeworm heads in sheep and cattle and other herbivores, including humans (intermediate hosts) (2). Prevention of hydatids using anthelmintic treatment of dogs and the prohibition of feeding uncooked offal to dogs are the ways in which several countries including New Zealand, Tasmania and Iceland managed hydatid disease (5).

2% and 10.3%. The S-Cr level did not increase further and was stable at 2.8 mg/dL. The patient was discharged from our hospital on day 58. After leaving hospital, in spite of the above therapy, his S-Cr level was not decreased less than 2.7 mg/dL. The additional biopsy was performed 2 years after kidney transplantation and found the obstinate mild peritubular capillaritis and mild capillary basement membrane thickening. Further analysis showed de novo anti-DQ4 antibodies increased to 14 315 on MFI values. Again, for treatment of the

obstinate refractory AMR, we performed an additional three sessions of PEX and IVIG. In addition, we administered rituximab (200 mg/body) because his CD19/20 level increased to 1.5% and 2%. His S-Cr beta-catenin pathway level was still high at the S-Cr level

of 2.8 mg/dL 30 months after kidney transplantation. In this study, we report a refractory case of JNK inhibitor purchase PCAR accompanied by acute AMR. This case report helps to inform at least two debates: (1) the difficulties of diagnosis and management of PCAR when it is accompanied by AMR; and (2) the difficulties of diagnosis of AMR when it is resultant of anti-HLA-DQ antibody in ABO-incompatible kidney transplantation, because HLA-DQ antigen screening is not always required. PCAR is characterized by the presence of mature plasma cells that comprise more than 10% of the inflammatory cell infiltration in a renal graft.[1] This pathologic finding is noted in approximately 5–14% of patients with biopsy-proven acute rejection. Although therapy for this condition has not been generally established, graft survival is poor.[2] To diagnose PCAR, physicians should pay attention to PTLD

caused by Epstein-Barr (EB) viral infection, because the treatment for PTLD is contrary to that for PCAR.[4] In our case, we confirmed that there was no monoclonality for kappa and lambda by immunohistochemistry. In addition, EBER staining was negative by in situ hybridization. Authorities stated that there could be an AMR variant of PCAR. C4d-positive PCAR with circulating DSAbs responds adequately to treatment aimed at AMR, such as rituximab and IVIG combination of therapy. On the other hand, C4d-negative PCAR is intractable to treatment. In our case, treatment aimed at AMR showed good response. Current anti-humoral therapies in transplantation and autoimmune disease do not target the mature antibody-producing plasma cells. Matthew et al. reported that bortezomib therapy may be effective for treating mixed rejection (AMR and acute T cell-mediated rejection) with minimal toxicity and for sustaining reduction of DSAb and non-DSAb levels.[5] In this context, a strategy for treating PCAR needs to be established in the future. The importance of HLA matching in kidney transplantation is well recognized, with HLA-DR compatibility having the greatest influence on outcome.

In a 1964 review lecture, Renkin [15] analyzed the available data on the transport of macromolecules Everolimus nmr between plasma and lymph and considered how well they could be accounted for by ultrafiltration through Grotte’s large pores and by transcytosis by vesicles. By so doing, he showed that if vesicular transport were responsible for macromolecular permeability, it could be described in quantitative terms and these terms placed restrictions on the numbers and behavior of the vesicles. Renkin’s review stimulated considerable experimental work by both physiologists and electron microscopists in the late 1960s and throughout the 1970s. Trans-endothelial channels were reported to be formed by a chain of fused

vesicles [23], and some analyses GPCR Compound Library suggested both convective and non-convective mechanisms of macromolecular transport

operated in parallel. Convective transport and non-convective transport were interpreted in terms of large pores and transcytosis, respectively. In 1979, however, Rippe et al. [16], working on isolated perfused rat hind limb preparations, published a definitive set of experiments providing strong evidence that, in this preparation, the movement of serum albumin from plasma to tissue occurred entirely by convection. In the same year, Bundgaard et al. [3] published the first of a series of papers in which electron micrographs showed that all the vesicles in capillary endothelium were arranged in fused clusters, which communicated with caveolae at either the luminal or abluminal surface of the cells, but never at both. In their later papers, they [9] reconstructed three-dimensional models of the vesicle clusters from TEMs of ultra-thin C225 serial sections. It was argued [2,6] that the vesicle clusters

were static structures incompatible with transcytosis because single unattached vesicles were never present, and this was inconsistent with the simple model of transcytosis. It was not, however, inconsistent with the later fusion–fission model [5]. Furthermore, they found no evidence of channels formed as connections between chains or clusters of vesicles opening on to both luminal and abluminal cell surfaces. To account for the appearance of a blood-borne label in abluminal vesicles, it was proposed that the label had entered the abluminal vesicles from the interstitial fluid, having crossed the endothelium by a nearby intercellular cleft, which lay just out of the plane of section. A few years later, direct evidence rebutting this last argument was reported. Wagner and Chen [24] used terbium as a tracer of transport from blood to tissue in the rete mirabile of the eel. By making TEMs from serial sections, they showed that the tracer reached the abluminal surface via vesicles when no intercellular clefts were in the vicinity. Furthermore, the terbium density decreased with distance from a discharging caveola.

To assess responses

to GAD65 epitopes that could be processed and presented from intact protein, CD4+ T cells were primed by stimulation with GAD65 protein and then screened using tetramers loaded with each of the antigenic peptides identified by tetramer-guided epitope mapping. Briefly, 2·5 × 106 ‘no-touch’ Microbead-enriched CD4+ T cells were stimulated with 1·2 × 105 GAD65 protein loaded monocytes in one well of a 48-well plate. CD14+ monocytes were isolated and pulsed with recombinant GAD65 protein as in the protein-stimulated proliferation assays. At least four replicate wells (of a 48-well plate) were set up for each subject. The T cells were cultured for 14 days, adding fresh media and interleukin-2

as needed starting on day 7. Expanded cells were stained AZD5363 supplier with HLA-DR0401 tetramers loaded with each antigenic selleck chemicals GAD65 peptide. Again, tetramer responses were considered positive when distinct staining that was more than twofold above background (this was set to 0·2% and subtracted) was observed. As described in the Materials and methods section, the tetramer-guided epitope mapping approach was used to comprehensively investigate DR0401-restricted epitopes within GAD65. Peptide pools spanning the entire GAD65 sequence were used to stimulate CD25-depleted T cells from multiple donors with DR0401 haplotypes. Consistent with the representative results shown in Fig. 1(a), a total of 17 different peptides (from 11 peptide pools) elicited a positive response from at least one of the subjects tested. With the exception of pool #6, the antigenic peptides

within each of these peptide pools could be identified using tetramers loaded with individual peptides. The antigenic peptide from pool #6 could not be identified using this approach. However, peptide p26 (GAD201–220) from pool #6 was identified as the antigenic peptide by means of a proliferation assay (Fig. 1b) and was further confirmed by stimulating Selleck Pazopanib of CD4+ T cells with the individual GAD201–220 peptide and staining with the DR0401/GAD#6 pooled tetramer (data not shown). The peptide sequences containing these epitopes are summarized in Table 1. The 17 antigenic peptides identified included five pairs of adjacent, overlapping peptides. It seemed likely that some of these adjacent overlapping peptides contain a single, shared antigenic sequence. To delineate the antigenic sequences within these adjacent overlapping peptides, we generated tetramer-positive T-cell lines for at least one peptide from each pair. As shown in Fig. 2, we assessed the proliferation of these lines in response to each of the adjacent peptides. These results suggested that three pairs of overlapping peptides (GAD105–124 and GAD113–132, GAD265–284 and GAD273–292, GAD545–564 and GAD553–572) appear to contain distinct antigenic sequences, because T-cell lines only proliferated in response to one of the peptides.

01). In conclusion, neurological deteriorations of diabetic rats were alleviated with PGE1, which is associated with inhibition of NGF and enhancement of VEGF at the entrapment site. © 2014 Wiley Periodicals, Inc. Microsurgery 34:568–575, 2014. “
“Medicinal leech therapy (MLT) to salvage venous congestion in native skin and local flaps is commonly practiced. However, the role of MLT in compromised regional and free flaps remains unclear. Leeches were used in 39 patients to treat venous congestion in native skin (n = 5), local flaps (n = 6), regional flaps (n

= 14), and free flaps (n = 14). There were no total losses in patients with compromised native skin or local flaps. One patient who had received a radial forearm RAD001 mouse free flap expired before flap outcome could be assessed, and was excluded from analysis. Of the remaining 27 regional and free flaps, 33.3% were salvaged, 33.3% were partially salvaged, and 33.3% were lost. Means of 38.3 ± 34.0, 101.0 ± 11.2, selleck compound and 157.9 ± 224.4 leeches and 1.7 ± 3.6, 3.2 ± 4.4, and 5.6 ± 5.2 units of blood were required for the salvaged, partially salvaged, and lost groups, respectively. Twenty-two patients required blood transfusion (57.9%). No patients developed wound infection with Aeromonas hydrophilia. Two patients developed donor site hematomas, and four patients developed recipient site hematomas. MLT is efficacious in congested native

skin and local flaps. Some regional and free flaps can be totally orpartially salvaged. However, the morbidity of MLT must be weighed against the risks of flap loss. © 2012 Wiley Periodicals, Inc. Microsurgery, Tenofovir concentration 2012. “
“The purpose of this study was to evaluate the effect of direct administration of nerve growth factor (NGF) into an epineural

conduit across a short nerve gap (10 mm) in a rabbit sciatic nerve model. The animals were divided into two groups. In group 1, n = 6, a 10-mm defect was created in the sciatic nerve and bridged with an epineural flap. A dose of 1 μg of NGF was locally administered daily for the first 21 days. NGF administration was made inside the epineural flap using a silicone reservoir connected to a silicone tube. In group 2, n = 6, the 10-mm defect was bridged with a nerve graft. This group did not receive any further treatment. At 13 weeks, all animals, before euthanasia, underwent electromyography (EMG) studies and then specimen sent for histology morphometric analysis. NGF administration ensured a significantly increased average number of myelinated axons per μm2 (P = 0.028) and promoted fiber maturation (P = 0.031) and better EMG results (P = 0.046 for latency P = 0.048 for amplitude), compared with the control group. Although nerve grafts remain the gold standard for peripheral nerve repair, NGF-treated epineural conduits represent a good alternative, particularly when an unfavorable environment for nerve grafts is present. © 2011 Wiley-Liss, Inc.

OHASHI YASUSHI1, TAI REIBIN1, AOKI TOSHIYUKI1, MIZUIRI SONOO2, OGURA TOYOKO3, TANAKA YOSHIHIDE1, OKADA TAKAYUKI1, AIKAWA ATSUSHI1, SAKAI KEN1 1Department of Nephrology, School of Medicine, Faculty of Medicine, Toho University, Tokyo; 2Division of Nephrology, Ichiyokai Harada Hospital, Hiroshima; 3Department of Nutrition, Toho University Omori Medical Center, Tokyo Introduction: Fluid imbalance due to sodium

retention and malnutrition Rucaparib chemical structure can be characterized by the ratio of extracellular water (ECW) to intracellular water (ICW). Our objectives are to investigate whether fluid imbalance between ICW and ECW is a risk factor for adverse outcomes. Methods: Body fluid composition was measured in 149 patients with chronic kidney disease from 2005 to 2009, who were followed until death, loss to follow-up, or August 2013. Patients were categorized according to the ECW/ICW ratio tertile. The ratio of ECW to total body water, calculated by the Watson formula, was used as an indicator of ECW excess. Main outcomes were adverse Talazoparib supplier renal outcomes, as defined by a decline of 50% or more

from baseline glomerular filtration rate or initiation of renal replacement therapy, cardiovascular events, and all-cause mortality. Results: Patients with higher tertile tended to be older and have diabetes mellitus, treatment-resistant hypertension, ECW excess, decreased protein intake per calorie, lower renal function, hypoalbuminemia, and higher proteinuria and furosemide usage (P < 0.01). Compared with patients in the lowest tertile during a median 4.9-year follow-up, those in the highest tertile had the worst adverse renal outcomes (15.9 vs. 5.1 per 100 patient-years, P < 0.001), cardiovascular events (4.1 vs. 0.3 per 100 patient-years, P = 0.002), and mortality (11.2 vs. 1.3 per 100 patient-years, P < 0.001)

by Kaplan–Meier survival analysis. The adjusted hazard ratio (95% confidence intervals) for adverse renal outcomes, cardiovascular events, and all-cause mortality were 1.15 (1.03–1.26, P = 0.011), 1.12 (0.93–1.31, P = 0.217), and 1.29 (1.11–1.50, P < 0.001), respectively. Conclusion: Fluid Etofibrate imbalance between ICW and ECW, driven by cell volume decrease and ECW excess, was associated with adverse renal outcomes and mortality. These findings emphasize the importance of cell volume retention as well as appropriate extracellular volume. CHEN SZU-CHIA1, HUANG JIUN-CHI1,2, CHANG JER-MING1,2, HWANG SHANG-JYH1, CHEN HUNG-CHUN1 1Division of Nephrology, Department of Internal Medicine, Kaohsiung Medical University Hospital; 2Department of Internal Medicine, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung Medical University Introduction: The P wave parameters measured by 12-lead electrocardiogram (ECG) are commonly used as noninvasive tool to assess for left atrial enlargement.

In contrast, when combined with TGF-β and IL-23, the cytokines IL-6 or IL-21 can induce Th17 cells, which produce IL-17, IL-21, and IL-22, express the lineage-specific transcription factor ROR-γt, and protect from extracellular bacterial and fungal infections. Finally, naïve FOXP3+ Treg cells under Th1 or Th2 inflammatory conditions acquire effector function and have anti-inflammatory properties. Although all T-cell subsets mentioned above have protective

functions under physiological conditions, uncontrolled responses of the respective Th subsets may cause immunopathology. Thus, Th1 and Th17 cells have been implicated in autoimmune tissue inflammation, including autoimmune encephalomyelitis and inflammatory bowel disease, ITF2357 nmr whereas Tfh cells contribute to a lupus-like syndrome, and Th2 as well as Th9 cells to allergy and asthma [32-35]. Although early studies Raf inhibition have demonstrated the T-cell intrinsic importance of IRF4 for Th2-cell differentiation [36-39], its role for Th1-cell development is less clear. Contradictory data show either diminished [36, 38] or normal [37]

IFN-γ production by Irf4–/– Th cells cultured under Th1 conditions in vitro. In an infectious model with the intracellular protozoon Leishmania major, in which Th1 responses promote healing and parasite clearance, whereas Th2-driven responses cause chronic disease [40], Irf4–/– mice failed to control the infection. However, this defect could not solely be explained by impaired Th1-cell differentiation, because the responding T cells also completely failed to develop a Th2-cell phenotype. Furthermore, disease susceptibility correlated with extraordinarily enhanced apoptosis of Irf4–/– Thiamet G CD4+ T cells, which was reflected in almost total loss of cellularity in the draining lymph node (LN) [41]. Th2-cell differentiation can be compromised

in vivo not only as a result of the T-cell intrinsic loss-of-function of IRF4 but also owing to T-cell extrinsic defects in IRF4-controlled functions, such as DC development [5]. Within T cells, IRF4 controls Th2-cell differentiation through several mechanisms (Fig. 1A). First, IRF4 promotes IL-4 production directly by binding to the IL-4 promoter in cooperation with the transcription factors NFATc2 in mouse [36] or NFATc1 in human cells [39]. Second, IRF4 is important for the upregulation of GATA3, and overexpression of GATA3 partially rescued IL-4 production in Irf4–/– Th2 cells, suggesting a crucial role of IRF4-dependent GATA3 expression for Th2-cell differentiation [38]. Third, IRF4 is important for the expression of growth factor independent 1 (Gfi1), a transcription factor that regulates IL-2-mediated Th2-cell expansion [37]. Given that BATF is required for Th2-cell development [42, 43] and that AICEs have been found in Th2 cells [16], it is highly probable that IRF4 also regulates Th2-cell differentiation in cooperation with BATF–JUN heterodimers.