HLA-DR3/DR4 alleles were also analysed. All T1AD patients satisfied the American Diabetes Association (ADA) classification criteria for type 1A diabetes [37]. This project was approved by the Ethics Committee for Research Project Analysis of Hospital das Clínicas, University of São Paulo School of Medicine. All the Lumacaftor samples were collected after the patients were provided with guidance and had signed a consent form. Autoantibodies against insulin

(IAA), glutamic acid decarboxylase (GAD65), tyrosine phosphatase (IA2) and 21-hydroxylase (21-OH) were assessed by radioimmunoassay (RSR Limited, Cardiff, UK). Autoantibodies against thyroid peroxidase (TPO) and thyroglobulin (TG) were evaluated by fluorometry (AutoDELPHIA, Turku, Finland). Anti-nuclear antibody (ANA), anti-liver/kidney microsomal

type 1 antibody (LKM1) and anti-smooth muscle (ASM) antibody were quantified using indirect immunofluorescence. Rheumatoid factor (RF) was evaluated using nephelometry, and TSH receptor autoantibody (TRAb) was assessed using iodine radioreceptor assay (RSR Limited). Genomic DNA was extracted by salting-out in blood leucocytes. The region encompassing −448 to +83 base pairs (bp) of the IL-21 gene was amplified and sequenced from samples of 309 Brazilian T1AD patients and 189 control individuals. The following Adriamycin in vivo primers were used for the IL-21 gene: (−448) forward: 5′-CCTTATGACTGTCAGAGAGAACA-3′ and (+83) reverse: 5′-CTTGATTTGTGGACCAGTGTC-3′. Direct sequencing of polymerase chain

reaction (PCR)-amplified products was performed using an ABI 3100 capillary sequencer (Applied Biosystems, Tokyo, selleck kinase inhibitor Japan) with the ABI PRISM BigDye Terminator version 3·1 cycle sequencing kit (Applied Biosystems) and analysed using an ABI PRISM 3730 genetic analyser (Applied Biosystems). The following PCR amplification reaction primers were used: PTPN22 forward: 5′-TCACCAGCTTCCTCAACCACA-3′ and PTPN22 reverse: 5′-GATAATGTTGCTTCAACGGAATTT-3′. PCR amplification products were digested enzymatically using the Xcml restriction enzyme (Uniscience-New England BioLabs, Inc., Ipswich, MA, USA), which resulted in a 215-bp product for the CC variant (wild-type); 215-bp, 169-bp and 46-bp products for the CT variant; and 169-bp and 46-bp products for the TT variant. PTPN22 genotyping was performed in 689 controls and 434 T1AD patients. All results were confirmed using an RsaI restriction enzyme assay (Uniscience). HLA class II typing for DRB1 was performed using PCR with One Lambda’s SSP™ Generic HLA class II (DRB) DNA typing trays (One Lambda, Canoga Park, CA, USA).

There is a possibility that SEB contributes to SSTI, and therefore to MRSA spread in the community. To our knowledge, this is the first isolation of SEB-positive ST5 MRSA. Although the New York/Japan ST5 clone was occasionally positive for the arginine catabolic mobile element (ACME)-arcA (data not shown), two ST5 strains were negative for the arcA gene. The New York/Japan clone has been isolated not only in hospitals, but also from children in the community (14, 15). In Japan, children are frequently treated as outpatients at hospitals near their homes, so it is conceivable that some such children carry the New York/Japan clone to their

homes from hospitals and that transmission of such MRSA occurs among their family members, because MRSA colonizing their nares has also been detected on their hands (2). Probably reflecting such situations, we detected the New York/Japan clone (and its variant) Akt inhibitor in samples from the straps and handrails of trains in this study. MRSA with genotype ST8/spa606(t1767)/SCCmecIVx (unknown subtype)/CoaIII is a major CA-MRSA that is associated not only with SSTI, but also with invasive infections in the community in Japan (2). This clone with the typical genotype (strain PT5) and its variants with spaNew (t986) (strains PT3 and PT4) were isolated in this study (Table 1, Fig. 1). Similarly to clinical isolates (e.g., strain NN4): (i) they were positive for SaPIm1/n1; (ii) they exhibited low degrees

of oxacillin and imipenem resistance (MICs, 64

and <  2  μg/mL, respectively); and (iii) they were resistant to a limited number of antimicrobial agents, such as gentamicin (many CA-MRSA strains are resistant to gentamicin Cobimetinib in Japan [2]). Since the three strains (PT3 to PT5) were isolated from different trains, we concluded that either ST8 CA-MRSA is circulating in trains or that very ST8 CA-MRSA spreading in the community has appeared in trains. One ST8 MRSA (strain PT6) was slightly divergent from previously described clinical isolates and not closely related to the ST8 reference strain (NN4) (Table 1, Fig. 1). Similarly to CA-MRSA (consistent with NN4): (i) it exhibited the genotype ST8/spa606/agr1/CoaIII; (ii) it exhibited low degrees of oxacillin and imipenem resistance (MICs, 4 and <  0.06  μg/mL, respectively); and (iii) it was resistant to a limited number of antimicrobial agents (including chloramphenicol, which is rarely used in humans); however, (iv) it exhibited SCCmecI, which is generally associated with HA-MRSA (3, 10). Therefore, bacteriological assignment as CA- or HA-MRSA was impossible for strain PT6. ST88 MRSA and ST89 MRSA are representative CA-MRSA and are isolated from bullous impetigo and positive for the causative toxin, exfoliative toxin (A for ST88, and B for ST89) (2). Although ST88 MRSA (strain PT7) and ST89 MRSA (strain PT8) respectively resembled ST88 and ST89 clinical isolates from bullous impetigo (Table 1 and Fig. 1), they lacked exfoliative toxin.

We have recently generated a high-throughput, homogenous version of this assay, based upon a scintillation proximity principle allowing online, real-time monitoring of the dissociation of 125I-labeled β2m from recombinant MHC-I heavy Romidepsin chains [[14]]. Here, we have used this assay to address

the stability of immunogenic and nonimmunogenic pMHC-I complexes. Using panels of affinity-balanced peptides, we could demonstrate that the stabilities of pMHC-I complexes involving known T-cell epitopes are significantly more stable than pMHC-I complexes involving peptides of similar-binding affinity that are not known to be immunogenic. Our results also suggest that HLA-A*02:01-binding peptides become destabilized if the P2 anchor residue,

and to a lesser extend the P9 anchor residue, are not optimal; and that anchor optimization increases both affinity and stability. In conclusion, our results suggest that some peptides, despite exhibiting high-affinity binding to HLA class I molecules, may fail to become immunogenic because Akt inhibitor they fail to form stable complexes with HLA class I molecules. We used high-throughput homogenous biochemical assays to measure the affinity and stability of pMHC-I complexes [[14, 15]]. To generate pMHC-I complexes, biotinylated MHC-I heavy chain molecules were diluted more than 100-fold into a folding buffer containing β2m and peptide; and incubated to reach steady-state pMHC-I complex formation. All in vitro biochemical peptide-MHC-I affinity measurements (and all complex formation Tyrosine-protein kinase BLK for subsequent dissociation experiments)

were done at 18°C to avoid the confounding loss of complexes due to temperature instability [[16]]. In contrast, the dissociation phase of dissociation experiments was conducted at 37°C. To measure the affinity of peptide-MHC-I interactions, dose–response experiments were done in order to determine the peptide concentration (EC50) resulting in half-saturation of folded pMHC-I complexes (Fig. 1A). A homogenous luminescence oxygen channeling immunoassay (LOCI) was used to measure the resulting formation of folded pMHC-I complexes [[15]]. Under conditions of limited receptor concentration ([MHC-I HC] ≤ KD), the EC50 is a reasonable approximation of the equilibrium dissociation constant, KD. To measure the rate of peptide dissociation, we exploited an observation made initially by Parker et al. [[13]] showing that dissociation of 125I-labeled β2m is an accurate measurement of peptide dissociation. We recently showed that pMHC-I dissociation can conveniently be monitored in real time using a scintillation proximity assay (SPA) [[14]]. To this end, pMHC-I complexes were generated under conditions that led to optimal incorporation of 125I-labeled β2m.

We established that systemic treatment of mice with PI inhibited TNBS-induced colitis, a widely used murine model for

Crohn’s disease. The efficacy of anti-IL-12 treatment and studies of TNBS colitis in mouse models that are deficient at certain checkpoints of T-cell activation have unequivocally established a contributive role for T cells in this disease and its respective models 16–20. We show that PI treatment dramatically reduced disease severity of TNBS colitis as exhibited by a large decrease in weight loss and the absence of severe gastro-intestinal inflammation on ICG-001 histological evaluation. The effect of PI was mediated by T-cell inhibition as T cells derived from colon-draining selleck chemicals lymph nodes of PI-treated mice secreted much less of the hallmark inflammatory T-cell cytokines IL-17 and IFN-γ 3. These results were the first indication of PI as a potential T-cell inhibitor in a clinical setting. Next to exerting inhibition on the adaptive immune system, PI may affect innate immunity in TNBS colitis. Previously, it has been shown that TNBS colitis involves the innate immune system 21. Moreover, local mucosal application

of PI has been shown to have restorative effects on inflamed mucosa in a rat model for acetic acid-induced intestinal inflammation 22. It is unclear whether i.p. application of PI may affect mucosal innate immune cells in tetracosactide a similar degree although no effect on epithelial proliferation rate was observed (Supporting Information Fig. 1.). Additionally, in vitro, PI did not affect TNF-α release by LPS-activated peritoneal macrophages (Supporting Information Fig. 2). Under physiological conditions, clearance of immune cells may be achieved through apoptosis associated with the release of various tissue-derived molecules, amongst which phospholipids. In turn, these cell components have been suggested

to possess anti-inflammatory capacities. In this regard, other phospholipids such as phosphatidylcholine and phosphatidylserine have been identified as anti-inflammatory 8, 9. As such, future application of PI in human inflammatory disease may be explored. Current immunosuppressants are accompanied by a wide range of side effects and complications. These properties severely limit the application of these drugs. For example, steroids can only be prescribed for a limited period of time. Other immunosuppressants such as azathioprine are not to be used at high dosages 6, 20, 23. Finally, many novel drugs are only efficacious in a subset of patients. Therefore, treatment with this novel class of anti-inflammatory agents may be particularly interesting as long-term maintenance therapy.

“
“Please cite this paper as: Gaynes B, Teng P-Y, Wanek J, Shahidi M. Feasibility of conjunctival hemodynamic measurements in rabbits: reproducibility, Panobinostat in vitro validity, and response to acute hypotension. Microcirculation 19: 521–529, 2012. Objective:  To evaluate the feasibility of conjunctival hemodynamic measurements based on assessment of reproducibility, validity, and response to acute hypotension. Methods:  Image sequences of the conjunctival microvasculature of rabbits were captured using a slit lamp biomicroscope under a steady-state condition, after topical administration of phenylephrine, and after intravenous administration of esmolol. Venous hemodynamic parameters (diameter, blood velocity,

blood flow, and wall shear stress) were derived. Results:  Conjunctival venous diameters ranged from 9 to 34 μm and blood velocities ranged selleckchem from 0.08 to 0.95 mm/s. Coefficients of variation of venous diameter and blood velocity measurements were, on average, 6% and 14%, respectively. Automated and manual measurements of venous diameter and velocity were highly correlated (R = 0.97; p < 0.001; n = 16). With phenylephrine administration, diameter and velocity were reduced by 21% and 69%, respectively. Following esmolol administration, blood pressure was reduced with a concomitant decrease in velocity, followed by recovery to baseline. Venous blood velocity, flow, and WSS were correlated with blood pressure (R ≥ 0.52; p ≤ 0.01). Conclusions: 

The feasibility of quantifying alterations in microvascular hemodynamics in the bulbar conjunctiva was established. The method is of potential value in evaluating microcirculatory hemodynamics related to cardiovascular function. “
“Please cite this paper as: Adderley, Sridharan, Bowles, Stephenson, Sprague and Ellsworth (2011). Inhibition of

ATP Release from Erythrocytes: A Role for EPACs and PKC. Microcirculation18(2), 128–135. Objective:  Here we demonstrate that, in human erythrocytes, increases in cAMP that are Docetaxel manufacturer not localized to a specific receptor-mediated signaling pathway for ATP release can activate effector proteins resulting in inhibition of ATP release. Specifically we sought to establish that exchange proteins activated by cAMP (EPACs) inhibit ATP release via activation of protein kinase C (PKC). Methods:  ATP release stimulated by iloprost (ILO), or isoproterenol (ISO), was determined in the absence and presence of selective phosphodiesterase inhibitors and/or the EPAC activator, 8CPT2OMecAMP (8CPT). To determine whether EPACs inhibit ATP release via activation of PKC, erythrocytes were incubated with phorbol 12-myristate 13-acetate (PMA) prior to either forskolin or ILO in the absence and presence of a PKC inhibitor, calphostin C (CALC). Results:  Selective inhibition of PDEs in one pathway inhibited ATP release in response to activation of the other cAMP-dependent pathway. 8CPT and PMA inhibited both ILO- and ISO-induced ATP release.

Likewise blockade of PD-1 signaling reverses tumor-induced T-cell exhaustion and enhances the functions of CD8+ T cells [37, 38] In the present work, PDL-1 and PDL-2 do not seem to be involved in the regulation of CD4+ T cells as we could not observe an effect on responding CD4+ T cells after neutralizing of these ligands on DX5+CD4+-modulated DCs (data not shown). Nonetheless, their high expression levels on DCs after modulation with DX5+CD4+ supernatants, combined with the phenomena described above, also point Sorafenib nmr to the multiple pathways implemented DX5+CD4+ T cells to steer the outcome of T-cell responses. These pathways do not only involve

the modulation of cytokine secretion but also the expression of molecules known to affect T-cell responses. Whether the action selleck chemicals llc of DX5+CD4+ T cells on DC function and phenotype is responsible for the effects observed in disease models is not known. Nonetheless, our findings are in line with data obtained in vivo indicating a preferential reduction of Th-1-associated IgG2a against collagen type II in CIA model following adoptive transfer of DX5+CD4+ T cells [19]. Likewise, some of our findings resemble the findings observed in studies focusing on FoxP3+ CD25+CD4+ Treg-cell-mediated immune regulation. Like DX5+CD4+ T cells, CD25+CD4+

Treg cells can exert their suppressive effect directly and indirectly via suppressing T-cell responses and altering the phenotype and the function of DCs, respectively. In addition, human Treg cells inhibit the maturation and antigen presentation of monocyte-derived DCs to become poor APCs [7, 10, 13-17]. Together our results point to the plethora of pathways

affected by DX5+CD4+ T cells that could be involved mafosfamide in the control of autoimmune responses. Understanding of these pathways might be instrumental to further define potential immune modulating strategies with the aim to counteract autoimmune diseases. D011.10 (OVA-specific TCR Tg) mice were used for the generation of bone marrow DCs and for the isolation of CD4+ T cells and C57BL/6 mice were used for the generation of DX5+CD4+ T cells and DX5−CD4+ T cells. D011.10 mice were housed and bred in the animal facility of the Leiden University Medical Center. C57BL/6 mice were purchased from Charles River. Experiments were performed in accordance with our institutional guidelines on animal use in research. Splenocytes were isolated from spleens of mice that were injected three times (7, 5, and 3 days before purification of DX5+CD4+ T cells) intraperitoneally with 1 × 106 immature DCs in PBS. RBCs were lysed and CD4+ T cells were purified by positive selection using Dynal CD4 positive isolation kit (Invitrogen). Afterwards, DX5+ and DX5− cells were isolated from CD4+ T cells derived from the same mice using CD49 (DX5) MicroBeads (Miltenyi Biotec). The purity was 80–90%. DX5+CD4+ and DX5−CD4+ T cells were isolated from the same mice.

Although the invasion and inflammatory phenotypes are the best studied pathogenic mechanisms of Shigella infection, clinical data show that a considerable number of patients develop a self-limiting watery diarrhea (Keusch et al., 1986; Vargas et al., 1999). These clinical observations led to the description of two candidate enterotoxins

in Shigella flexneri, called ShET-1 and ShET-2, encoded on the chromosome and the Inv virulence plasmid, respectively (Fasano et al., 1995; Nataro et al., 1995). ShET-2 was initially described in enteroinvasive Escherichia coli strain EI-34, but was also found in most isolates of the FK506 in vitro genus Shigella (Nataro et al., 1995; Vargas et al., 1999). The protein was purified after recombinant gene expression and was found to induce rises in short-circuit current in rabbit intestinal tissue mounted in the Ussing chamber (Nataro et al., 1995). Recently, vaccine trials using live attenuated Shigella strains with deletions in the genes encoding ShET-1 and ShET-2 suggested that one or both of these toxins contribute to virulence in humans (Kotloff et al.,

2000, 2004, 2007). More thorough characterization of these two factors is therefore warranted. Multiple virulence factors of Shigella spp. are secreted by type III secretion systems (T3SS) or by the autotransporter (type V) mechanisms. However, no experimental data have been published implicating BYL719 either of these mechanisms for ShET-1 or ShET-2 secretion. Inositol oxygenase Notably, neither putative toxin exhibits a typical Gram-negative signal sequence (Nataro et al., 1995) and no signature suggesting T3SS-dependent translocation has been reported. The Shigella T3SS, encoded on the 31-kb Inv plasmid-encoded entry region, comprises a multiprotein bacterial complex that forms a needle-like structure, termed the injectosome; this nanomachine mediates the translocation

of bacterial effector proteins directly to the eukaryotic cytoplasm (Mota & Cornelis, 2005). In Shigella, the T3SS is induced upon contact of the bacteria with epithelial cells (Watarai et al., 1995) or by adding Congo red (CR) dye to the growth medium (Bahrani et al., 1997). Constitutive secretion of T3SS effectors is observed after inactivation of the ipaB or the ipaD genes (Menard et al., 1994). In an S. flexneriΔipaBCDA mutant, 14 other type III effectors encoded on the Inv virulence plasmid were identified and designated as outer Shigella proteins (Osp proteins). These proteins were organized in groups OspB to OspG according to similarities in their amino-acid sequence (Buchrieser et al., 2000). The OspD group includes three members: OspD1 (a proven type III effector) (Parsot et al., 2005), OspD2 (of unknown function) and OspD3 (also known as ShET-2). Notably, this first report did not directly document dependence of OspD3 secretion on the T3SS.

However, some bacteria are resistant to the microbicidal effectors of amoebae (1) by being either true symbionts, that are

living in close association during a specific period of their lifetime with amoebae, or (2) by being true amoebal pathogens able to lyse the amoebae before or after completing an intra-amoebal replication cycle (Birtles et al., 2000; Greub et al., 2003). Amoebae may thus be considered as a replicative niche for both amoebal symbionts and amoebal pathogens. However, amoebae are not a neutral replicative site, but a potent evolutionary crib that promotes the selection of virulence traits leading to survival against phagocytic cells (Steenbergen et al., 2001; Greub & Raoult, 2004; Molmeret et al., 2005; Greub, 2009). This supports the use of amoebae as a model PD98059 to assess the bacterial virulence of amoebae-resisting microorganisms (Goy et al., 2007). Amoebae also represent protective armour for the internalized bacteria when encysted, and at least for some symbionts, a source of energy and nutrients. The evidence of the importance of amoebae as a reservoir of Legionella spp. led T. Rowbotham to use amoebae as cells in a cell culture system Compound Library to culture Legionella species (Rowbotham, 1983). Since that time, this amoebal co-culture method (see reference Lienard et al., 2011 for an up-to-date protocol) has

proven successful for the recovery by culture of a large biodiversity of amoebae-resisting bacteria (reviewed in Winiecka-Krusnell & Linder, 2001; Greub & Raoult, 2004; Lamoth & Greub, 2010; Lienard et al., 2011). Amoebae are also increasingly considered as an Agora where gene exchanges take place (Greub, 2009; Moliner & Raoult, 2010; Thomas & Greub, 2010). This intra-amoebal cross-talk has been corroborated by a recent analysis of gene exchanges occurring between amoebae-resisting microorganisms,

Adenosine triphosphate whereby as many as nine horizontal gene transfer events between Legionella species, Chlamydia-related bacteria and members of the Order Rickettsiales (Gimenez et al., 2011) were identified. Moreover, the genome of amoebae-resisting bacteria are commonly encoding proteins sharing a domain conserved in eukaryotic proteins (Schmitz-Esser et al., 2010; Gimenez et al., 2011), suggesting that horizontal transfer may also be at play between the bacterial symbiont and the amoebal host. Three major groups of amoebae-resisting bacteria have been extensively investigated, the Legionella, mycobacteria and Chlamydia-related organisms (Fig. 2), and several relatively recent reviews are already available (Horn, 2008; Greub, 2009; Lamoth & Greub, 2010). Here, we thus focus on rickettsial symbionts and on two other Candidatus species for which recently available genomic data illuminate the biology and their interactions with amoebae: Odysella thessalonicensis and Amoebophilus asiaticus.

Substantial differences in the analyte profiles were notable, with the group demonstrating the highest level of periodontitis showing elevated levels of IL-6, IL-8 and LBP and significantly decreased levels of PGE2 and BPI. By the time of delivery, and following ligation of teeth in four quadrants, all animals had a CIPD >20 (not periodontally healthy). Again, the most diseased animals provided a profile of serum analytes that was distinctive from animals expressing primarily gingival inflammation,

with a lower level of destructive disease. These data suggested that the variation in naturally occurring periodontal Doxorubicin clinical trial inflammation and disease in the female baboons was reflected by patterns of systemic inflammation. Moreover, those animals that responded more robustly to the infection burden accompanying ligation generally

demonstrated a unique profile of mediator levels. As we have observed previously, these findings are consistent with a subset of these non-human primates that show an increased susceptibility to dysregulated local responses eliciting greater disease and allowing a more substantial challenge to the systemic inflammatory apparatus. These outcomes would also suggest that animals with a more effective adaptive immune response to the microbial challenge would demonstrate less disease, as we have reported previously [46,55], and less systemic challenge with lower serum inflammatory responses. Examination of the relationships between the inflammatory mediators and antibody in serum showed that elevated or decreased antibody specificities were coincident Pirfenidone solubility dmso with levels of selected mediators. However, identification of a particular pattern of antibodies that best described the systemic inflammatory response profiles was somewhat complex. Generally, the acute phase reactants were delineated by

unique patterns of antibody responses that were observed at specific time-points during the study. The chemokines IL-8 and MCP-1 demonstrated some similarity in the patterns of antibody correlations, particularly at baseline new and mid-pregnancy. IL-6 levels were best described by distinctive antibody specificities during the protocol. However, of the 20 antibody specificities that were evaluated, levels of F. nucleatum, P. gingivalis, A. actinomycetemcomitans and C. rectus showed some consistency in contributing to relationships with the range of inflammatory mediators analysed. However, within the model system, a pattern of the serum analytes provided some insight into describing the expression of disease. We observed a clear association of IL-6, IL-8 and LBP levels across disease expression and throughout pregnancy. When broken down further, we observed that these relationships were related primarily to the characteristics of the disease expression in the individual animal, and generally related less to the stage of pregnancy at which the sample was obtained.

As shown in this study, NFATc2 and c-Jun transcription factors are able to induce an open chromatin conformation at the target locus. Various transcription factors with chromatin remodeling activity were described earlier, including CTCF, GATA-3, NF-κB, and NFAT family members interacting with regulatory elements of the GM-CSF locus [86], members of NFAT family for IL-3 [87, 88], IL-4

[89] and IFN-γ [90] enhancers and IL-2 promoter [91]. Notably, chromatin remodeling at regulatory elements shows different requirements for transcription factor AP-1. learn more GM420 element of the GM-CSF enhancer can bind both NFAT and AP-1, while NFAT motif of GM420 alone is sufficient for the formation of the DH site [92]. In contrast, active (phosphorylated) form of c-Jun alone could maintain open chromatin conformation at the TNF TSS in CsA-independent manner in quiescent Th1 and Th17 cells (Fig. 6B and Supporting Information Fig. 6). Overexpression of c-Jun alone can induce open chromatin conformation at the TNF TSS in cultivated T cells (Fig. 6D). In contrast, chromatin remodeling at the IL-2 promoter is resistant to inhibition

of c-Jun phosphorylation by SP600125, but depends on de novo synthesis of c-Fos [93], indicating that Z-VAD-FMK research buy AP-1 transcription factors required for chromatin remodeling of TNF TSS and IL-2 promoter may have distinct compositions. Pharmacological inhibition of calcineurin/NFAT activity by CsA has a long and successful history of clinical applications for preventing transplant rejections and Ureohydrolase for the treatment of autoimmune pathologies [94-98]. On the other hand, blocking the MAPK/AP-1 cascade has been proposed as a therapeutic approach in various disease conditions including arthritis [99], colitis [100], neurodegenerative

disorders [101, 102], and cancer [103, 104], and new inhibitors of this pathway are being developed [105, 106]. Here, we demonstrated that the NFAT and AP-1 pathways are involved at additional level for TNF expression control in T cells. We also uncovered a distinct role for the AP-1 component c-Jun in the maintenance of open chromatin conformation at TNF TSS in potentially pathogenic Th1 and Th17 cells. C57BL/6 mice were purchased from Charles River Laboratories and FoxP3-IRES-GFP mice were kindly provided by Dr. Bernard Malissen [48]. Animals were bred and maintained under specific pathogen-free conditions. All animal experiments were performed in accordance with institutional, state, and federal guidelines (Landesamt für Gesundheit und Soziales—LAGeSo, Berlin, Germany). All reagents were purchased from Sigma-Aldrich (St.