54 To test if STIM2 and/or ORAI3 activity could be responsible for the differences in costimulation, we compared the effect of 10 μm 2-APB on Ca2+ signals

in CD4+ T-cells (Fig. 8). The application of 10 μm 2-APB increased Ca2+ signals to similar values for both conditions indicating that a difference in the store-independent mode of CRAC channel activation might be the reason for the observed differences between stimulation with dscFv anti-CD33/anti-CD3 in combination with sc CD86/anti-CD33 when compared with dscFv anti-CD33/anti-CD3 in combination with sc CD80/anti-CD33. Tigecycline ic50 100 μm 2-APB decreased Ca2+ influx as previously reported.54 The costimulation effect on Ca2+ influx and the effect of 2-APB were independent of TG because we obtained similar results in the absence of TG (Fig. 8). We conclude that store-independent

Ca2+ entry mediated by STIM2 and/or ORAI3 is likely to be involved in the costimulation-dependent regulation of CRAC channel activity. We show evidence that T-cell costimulation by CD80 or CD86 ligand binding causes differences in net Ca2+ entry depending on the activation state of the T-cell. The differences of Ca2+ entry are not linked to Ca2+ store depletion, offering a potential physiological function for store-independent Ca2+ entry. Store-independent Ca2+ entry by CRAC channels has recently been proposed;21,53 however, so far, no physiological function has been assigned. Our data reveal that the store-independent mode https://www.selleckchem.com/JAK.html of CRAC may be important

to distinguish different modes of costimulation. The interaction of CD80 or CD86 with CD28 Interleukin-2 receptor or CTLA-4 has been established in the early 1990s as the first pathway of T-cell costimulation and co-inhibition and has since been the subject of intense studies.55 The initial work using CD80 or CD86 transfected cell lines was replaced in many studies by CD28-specific monoclonal antibodies because they showed adequate T-cell proliferation in the presence of suboptimal stimulation by TCR cross-linkage. However, anti-CD28 antibodies provide a rather simplistic model for costimulation because they have a different binding pattern on the CD28 molecule and affinity when compared with the natural CD80 or CD86 ligand,33,34,56,57 More importantly, CD28-specific antibodies do not provide any information on the subtle differences between CD80- and CD86-mediated costimulation and cannot mimic the spatial and temporal differences involved in CD28 and CTLA-4 signalling. CD28 is recruited to the IS even in the absence of CD80 or CD86 costimulation and its localization at the IS can be disrupted by CTLA-4, which needs ligand binding to be recruited to the IS.37 Costimulation should, therefore, influence effector T-cell signalling more severely than signalling in naïve T cells because only effector cells express both CD28 and CTLA-4 at high levels. We have linked these findings with our Ca2+ data and developed the following hypothesis (Fig. 9).

Quantitative measures from this second set of simulations were found to correlate extremely well with experimental data obtained from animals treated with an agent that targets

endothelial proliferation (TNP-470). buy R788 Conclusion:  Our direct combination and comparison of in vivo longitudinal analysis (over time in the same animal) and mathematical modeling employed in this study establishes a useful new paradigm. The virtual wound created in this study can be used to investigate a wide range of experimental hypotheses associated with wound healing, including disorders characterized by aberrant angiogenesis (e.g., diabetic models) and the effects of vascular enhancing/disrupting agents or therapeutic interventions such as hyperbaric oxygen. “
“We sought to test the hypothesis that turmeric-derived curcuminoids limit reperfusion brain injury in an experimental model of stroke via blockade of early microvascular inflammation during reperfusion. Male Sprague Dawley rats subjected to MCAO/R were treated with turmeric-derived curcuminoids (vs. vehicle) 1 hour prior to reperfusion (300 mg/kg ip). Neutrophil adhesion to the cerebral microcirculation and measures of neutrophil and endothelial https://www.selleckchem.com/products/Temsirolimus.html activation were assayed during

early reperfusion (0–4 hours); cerebral infarct size, edema, and neurological function were assessed at 24 hours. Curcuminoid effects on TNFα-stimulated human brain microvascular endothelial cell (HBMVEC) were assessed. Early during reperfusion following MCAO, curcuminoid treatment decreased neutrophil rolling and adhesion to the cerebrovascular endothelium by 76% and 67% and prevented >50% of the fall in shear rate. The increased number and activation state (CD11b and ROS) of neutrophils were unchanged by curcuminoid treatment, while increased cerebral expression of TNFα and ICAM-1, a marker of endothelial activation, were blocked by >30%. Curcuminoids inhibited NF-κB activation and subsequent ICAM-1 gene expression in HBMVEC. Turmeric-derived curcuminoids limit reperfusion injury in stroke by preventing

neutrophil adhesion to the cerebrovascular microcirculation and improving shear rate by targeting the endothelium. PIK3C2G “
“Angiotensin II causes potent increases in systemic and local pressure through its vasoconstrictive effect. Despite the importance of angiotensin II for local blood flow regulation, whether angiotensin II regulates the pancreatic islet microcirculation remains incompletely understood. We hypothesized that angiotensin II directly regulates the pancreatic islet microcirculation and thereby regulates insulin secretion. The aims of this study were to develop a new technique to visualize pancreatic islet hemodynamic changes in vivo and to analyze changes in islet circulation induced by angiotensin II or an angiotensin type 1 receptor blocker.

Acinetobacter baumannii has recently emerged as an important Gram-negative pathogen that is reported to account for up to 10% of hospital-acquired

infections and 8.4% of hospital-acquired pneumonia (Hidron et al., 2008; Kallen et al., 2010). The organism’s success as a pathogen can be, in part, attributed to its ability to tolerate desiccation and disinfectants and form biofilms on abiotic surfaces commonly found in healthcare settings (Getchell-White et al., 1989; Musa et al., 1990; Hirai, 1991; Wendt et al., 1997). Colonization of hospital surfaces is thought to provide a reservoir Buparlisib concentration for the transmission and subsequent infection of patients with deficient immune systems. Septicemia and pneumonia, which result in mortality rates of approximately 50% (Seifert et al., 1995; Sunenshine et al., 2007), are the two most severe consequences of A. baumannii infection. Therapeutic intervention of A. baumannii infections has been compromised by an

alarming increase in the organism’s resistance to front-line therapies. Indeed, multidrug resistance in Acinetobacter spp. increased from 6.7% in 1993 to 29.9% in 2004, more than twice that observed in any other Gram-negative bacillus causing nosocomial click here intensive care unit infections (Lockhart et al., 2007). Moreover, strains that are resistant to all currently available antibiotics have been isolated from patients both in the United States and abroad (Siegel, 2008; Doi et al., 2009). Numerous mechanisms

are thought to contribute to the organism’s propensity to circumvent antibacterial agents. Acinetobacter baumannii exhibits an extraordinary ability to acquire antibiotic resistance determinants, which include enzymatic functions such as β-lactamases and aminoglycoside-modifying enzymes (Hujer et al., 2006). Additionally, the organism harbors a repertoire of efflux pumps that have also been hypothesized to Diflunisal contribute to clinical antibiotic failure (Hujer et al., 2006; Peleg et al.,2007a, b). While progress has been made in characterizing the organism’s antibiotic resistance determinants, little is known about their expression patterns or the mechanism(s) by which they are acquired or controlled. Similarly, little is known about the organism’s virulence factors or their regulation. For instance, while it is well recognized that many bacterial virulence factors are expressed in a cell density-dependent manner, we do not yet have a comprehensive assessment of these properties in A. baumannii cells (van Delden et al., 2001; Thompson et al., 2003). Nevertheless, advances in virulence factor identification are being made; using a proteomics approach, Soares and colleagues recently identified 67 proteins that are differentially expressed as A. baumannii ATCC 17978 cells transition from exponential to stationary phase of growth and hypothesized that a subset of these proteins are virulence factors (Soares et al., 2010).

Triggering of these TLRs in human gingival epithelial cells (HGECs) with their specific ligands leads to production of mediators such as IL-8 and antimicrobial β-defensin-2 [[9]], highlighting the critical role of periodontal tissue in innate immunity. To date, there is relatively little

available information regarding periodontal innate antiviral immunity. In addition to TLR ZD1839 in vivo expression, the gingival epithelium and gingival fibroblasts express retinoic acid-inducible gene (RIG)-like receptors (RLRs), including RIG-I and melanoma differentiation associated gene 5 (MDA5) (unpublished observation; [[11, 12]]) which recognize viral ssRNA and dsRNA. Activation via these RLRs results in expression of inflammatory cytokines and type I interferon (IFN) [[13]]. Type I IFN is a key mediator Pexidartinib in defense against viral infection. It eliminates viruses by enhancing the transcription of many IFN-inducible genes such as myxovirus resistance A (MxA) [[14]]. It also enhances dendritic cell maturation, antibody production, and differentiation of virus-specific cytotoxic T lymphocytes, resulting in effective adaptive

immunity against viral infection [[15, 16]]. Saliva and gingival crevicular fluids, which bathe the perio-dontal tissue, contain a variety of innate immune mediators against bacteria, including human α-defensins (commonly known as human

neutrophil peptides) [[17]], β-defensins [[18]], cathelicidin (LL-37) [[19]], thrombospondins [[20]], lactoferrin [[21]], and secretory leukocyte protease inhibitor (SLPI) [[21]]. Some of these molecules have also demonstrated antiviral properties [[22]]. To further gain insight into innate antiviral immunity, we investigated expression of antiviral proteins in periodontal tissue focusing on MxA, a potent antiviral protein against both RNA and DNA viruses [[23-25]]. SLPI has been reported in relation to antiviral defense in perio-dontal tissue [[26]]. In this study, we evaluated the expression of other antiviral molecules, including MxA, oligoadenylate synthetase (OAS), and protein kinase R (PKR) from both healthy periodontal tissue and periodontitis specimens. Using real-time RT-PCR, we found Protein tyrosine phosphatase mRNA expression of MxA, OAS, PKR, and SLPI in all examined periodontal tissues. As compared with healthy periodontal tissues, the mean fold increase of relative quantification of MxA, OAS, PKR, and SLPI in periodontitis tissues was 0.83 ± 0.24, 1.06 ± 0.30, 1.20 ± 0.34, and 2.74 ± 1.37, respectively (Fig. 1). These differences between healthy and periodontitis tissues were not statistically significant (p > 0.05). MxA protein is well known to have antiviral activity against both RNA and DNA viruses [[24, 25]]. We focused on MxA protein throughout our study.

While the levels of circulating CFH in subjects with altered glucose tolerance are usually increased [24], our study showed that the upregulation of CFH in T1D relatives was independent of their metabolic status. However, no evidence of association KPT330 of CFH polymorphisms with T1D has been reported so far [25]. The other category of immune responses where differences observed on the level of a single gene upregulation

were also paralleled on the level of entire pathway represents cytokine and/or chemokine signalling. Namely, when DRLN was compared to the control group, we found the upregulation of genes encoding IL-21 receptor, IL-13 receptor (alpha1) and IL-28 receptor (alpha, IL-28RA). So far, the functional link to T1D and other T cell-mediated diseases was reported only for IL-21 [26, 27]. The analysis on a transcriptome level also revealed differences in the expression of proinflammatory IL-1 as well as of IL-7 and IL-15 cytokines. The recognition of buy Cobimetinib IL-1 signalling as the highest-scored differentially activated pathway in DRLN versus DV comparison is an important outcome of this analysis. IL-1 signalling scored high even when the whole DRL group was compared to controls without consideration of the autoantibody status.

It is necessary to emphasize that none of the participants suffered from any apparent infection at the time of sampling. Several scientific reports described the relationship between IL-1 signalling and the type 1 as well as type 2 diabetes [28]. In this context, our finding suggests that enhanced proinflammatory activity in the group of relatives reflects an inherently increased basal level of signalling status rather than stimulus-mediated activation. The second highest-scored pathway in DRL (whole group) versus DV comparison was IL-7 signalling in B lymphocytes. Common genetic variants of IL-7 receptor alpha (IL-7RA) have been recently shown to affect susceptibility to multiple sclerosis and T1D. While the relationship between IL-7RA signalling and the regulation of T cell homeostasis is well established [29], the mechanistic link between IL-7 signalling in B lymphocytes and

development of T1D is still elusive. IL-15 signalling Tau-protein kinase was recognized in DRL but not in DRLN versus controls comparison. This interleukin is crucial for NK-cell differentiation. Qin and co-workers observed reduced cell numbers and diminished responses of NK cells to IL-2 and IL-15 stimulation in children suffering from T1D [30–32]. It is of note that we have also identified differences in NKG2D signalling between DRL as well as DRLN and the control group. Changes in the activation of two chemokine cascades, CCR3 and CXCR4, were also revealed. CCR3 signalling in eosinophiles scored the highest in DRL versus patients with T1D. The protein encoded by CCR3 gene is highly expressed in eosinophils and basophils and is also detectable in Th1 and Th2 cells [33].

The number of intestinal intraepithelial lymphocytes (IEL) expressing the αβ T cell receptor (TCR) is greatly reduced in axenic mice in addition to a reduced cytotoxic ability of these cells, although no difference was found in the number of γδ TCR-positive IELs [16–18]. While the intestinal microflora has essential beneficial functions, this same endogenous non-pathogenic microflora and/or its antigens are also implicated in the pathogenesis of chronic intestinal inflammation during inflammatory bowel diseases [19]. Several axenic rodent models of chronic intestinal buy Vemurafenib inflammation

have demonstrated that disease development is dependent upon bacterial colonization [6,7,20]. While healthy wild-type animals have developed tolerance to their endogenous intestinal microflora, animals that are genetically prone to develop chronic intestinal inflammation lack

this tolerance and mount an uncontrolled immune response to enteric bacteria and/or their components. This response is apparent locally in the mucosal, gastrointestinal compartment as well as systemically and involves both humoral and cellular immune responses [21,22]. Our results indicate that acquisition of the normal faecal endogenous flora later in life can induce a transient intestinal inflammation. Mice that are kept in axenic conditions while their immune system matures without exposure to bacterial antigens lack tolerance to endogenous microflora. Thus, without previous exposure to luminal U0126 purchase microflora, if faecal and bacterial antigens are encountered in the presence of a mature immune system a rapid-onset mucosal and systemic immune response ensues. The first response appears to be dominated by a local intestinal innate response that is skewed towards T helper type 1 (Th1) proinflammatory cytokine production. Early transient activation of proinflammatory gene expression and innate signal transduction has been demonstrated in intestinal epithelial cell lines and naive epithelial cells isolated following monoassociation of axenic Florfenicol rats with probiotic Bifidobacterium lactis, suggesting a role for

activation of proinflammatory transcription factors in initiating epithelial cell homeostasis at an early stage of bacterial colonization [23]. Here we show that the initial proinflammatory response is followed by a response that appears to be dominated by the adaptive immune system characterized by systemic activation of antigen-specific lymphocytes and a subsequent infiltration of immune cells in the intestinal tissue. The latter may be facilitated by the increase in intestinal G-CSF. The initial relative abundance of mucosal proinflammatory cytokines instigates a transient colonic inflammation that then resolves, in conjunction with a subsequent anti-inflammatory response and establishment of a homeostatic cytokine balance.

3C) and spontaneous (data not shown) capability of BMDMs to repair a wound generated by scratching a confluent cell monolayer. Our results show that Abl is a component of podosomes in myeloid leukocytes and its expression and function is essential for podosome formation, cell migration in 2D and 3D and trans-endothelial migration. These check details findings have a particular significance in the context of two aspects of leukocyte biology. The first one concerns the implication of podosome protrusive

activ-ities in trans-endothelial migration of leukocytes from blood to the interstitium during inflammation [[3, 17]]. Notably, the Abl kinase inhibitor imatinib mesylate has been reported to prevent and treat murine collagen-induced NVP-AUY922 datasheet arthritis [[18]] although a possible effect of the drug on leukocyte migration was not specifically addressed in this study. The second one concerns the decrease in osteoclast activity in patients treated with imatinib [[19, 20]]. In fact, although targeting of c-fms and other growth

factor receptors by imatinib may affect osteoclast differentiation [[20]] our findings point to an additional more direct role of the drug on podosome organization to explain its ability to inhibit bone resorption. Previous studies on carcinoma cells [[15, 16]] and this one highlight that targeting of Abl may result in reduction of cancer cell invasive capacity but also of myeloid leukocyte recruitment into the tumor. Notably, tumor-induced inflammation has emerged as one of the hallmarks of cancer [[21, 22]] thus pointing to the exciting possibility that Abl targeting

may represent a double-edged sword, acting simultaneously on tumor cells and cancer-related inflammation. Anti-Abl, anti-Arg, and anti-CrkL antibodies from Santa Cruz Biotechnology Inc. (Santa Cruz, CA) and anti-pCrkL antibody from Cell Signaling Technology (Beverly, MA) were used for immunoblotting experiments. Anti-Vinculin antibody (clone hVin-1) from Sigma Aldrich (St. Louis, MO) anti-Abl antibody from Millipore (Billerica, MA) anti-Cortactin (phosphoY466), anti-Arg and anti-Cortactin from Abcam (Cambridge, UK) were used for immunofluorescence experiments. Secondary antibodies from Invitrogen (Carlsbad, CA) were goat-anti mouse Methane monooxygenase IgG1 FITC conjugated, goat anti-mouse Alexa 647 conjugated and goat anti-rabbit Alexa 647 conjugated. Rhodamine phalloidin from Cytoskeleton (Denver, CO) was used to label F-actin. Imatinib/Gleevec/STI-571 was from Santa Cruz. LPA was from Sigma Aldrich. BMDMs were isolated from femurs and tibias of 8-week-old wild-type C57BL/6J or fgr–/– and hck–/–fgr–/– mice as previously described [[12]]. Macrophages differentiated from the bone marrow, nonadherent, cell population for 7–8 days [[12, 13]] were detached by scraping and then plated for 24 h on fibronectin- or gelatin-FITC-coated coverslips in the above medium with a FCS concentration of 1%.

However, HDAC inhibitors can impact a wide array of cellular functions through alterations in gene expression or post-translational protein modification. The functional unresponsiveness characteristic of CD4+ T cell anergy was initially demonstrated in CD4+ T cells stimulated in the absence of co-stimulatory signals [8]. Subsequent work established that these anergized CD4+ T cells were sequestered in the G1 phase of

the cell cycle [9]. This finding inspired a search of various pharmacological agents known to block cell cycle progression, with the aim of locating an agent that could induce CD4+ T cell anergy, even in the presence of co-stimulation selleck screening library [10]. Known G1 blocker and HDAC inhibitor n-butyrate was shown to be such an agent. n-Butyrate induced anergy in murine CD4+ T cells stimulated with antigen in the presence of co-stimulation, DZNeP order but not in un-stimulated CD4+ T cells [11]. This observation suggests that short-term exposure to an HDAC inhibitor such as n-butyrate could control unwanted immune responses through deactivation of activated effector CD4+ T cells while allowing naïve T cells to respond to future challenge. One of the functions attributed to HDAC inhibitors is the capacity to enhance the generation and/or activity of Treg cells [12]. Murine Treg cells express transcription factor FoxP3 and have been shown to suppress activated effector

CD4+ T cells [13]. Treg cells may arise naturally from the thymus as part of immune tolerance or can be induced experimentally as a means of inhibiting unwanted T cell-mediated immune responses [14]. If the only mechanism for HDAC inhibitor–induced anergy requires Treg cell activity, the therapeutic potential of this class of drugs might be limited. Studies suggest a correlation between autoimmune disease and an increased risk for development of cancer in the lungs, liver,

skin and pancreas [15, 16]. Positively skewing an autoimmune patient’s Treg cell profile may be harmful, as it is known that the suppressive properties of Treg cells present an obstacle to immune clearance of tumours [17]. Documenting Treg cell-independent Galeterone CD4+ T cell anergy induced by HDAC inhibitors would underline the functional significance of this class of drugs for patients in which increased Treg cell activity may not be helpful. The Gilbert lab has previously reported that n-butyrate induced anergy in Th1 CD4+ T cell clones [10, 11, 18, 19]. Those CD4+ T cells were highly differentiated and unlikely to exhibit the plasticity needed to convert into Treg cells. However, a direct role for Treg cells in n-butyrate-induced CD4+ T cell anergy was not examined in those studies. We extended those studies to determine if n-butyrate-induced CD4+ T cell anergy requires the generation of suppressive CD4+FoxP3+ Treg cells. Mice.

5). This observation may appear contradictory to the result that cultured K5-PLCε-TG keratinocytes autonomously exhibit elevated selleckchem expression of IL-23 and Camp (Fig. 7). One of the possible explanations for this phenomenon is that cytokines with anti-inflammatory activity, such as IL-10 5, whose expression is elevated at P26 in the K5-PLCε-TG mouse skin along with the Treg marker Foxp3 (Fig. 5), may result in downregulation of the cytokine expression in PLCε-overexpressing keratinocytes. We also find that the relapse of the symptoms occurring in ∼5% of aged K5-PLCε-TG mice is accompanied by a vast increase in the IL-23 mRNA level (data

not shown). To understand the molecular basis of these phenomena, further clarification of the PLCε-regulated signaling in keratinocytes Osimertinib is required. The development of the skin phenotype of K5-PLCε-TG mice seems to be driven by aberrant expression of proinflammatory molecules represented by IL-23 and IL-22. These molecules are implicated in the pathogenesis of a variety of human inflammatory diseases including psoriasis, rheumatoid arthritis, and inflammatory bowel disease 4. Indeed, the characteristic features, such as acanthosis, keratinocyte STAT3 activation, aberrant infiltration of leukocytes, and elevated expression

of Th cytokines, which are found in the symptomatic K5-PLCε-TG mouse skin, are evident in the psoriatic skin 7, 32. Therefore, K5-PLCε-TG

mice could be used for the Histidine ammonia-lyase study of the immunopathogenesis of inflammatory diseases. The full-length mouse PLCε cDNA 33 was inserted into the Pme I site of pCAG-XstopX-IRES-NLLacZ, a derivative of pCAG-XstopX-polyA 34, to derive pCAG-XstopX-mPLCε-IRES-NLLacZ. Founders of CAG-XstopX-PLCε mice were produced by pronuclear injection of the linearized pCAG-XstopX-mPLCε-IRES-NLLacZ into fertilized eggs of L7-Cre mice, which had been backcrossed to C57BL/6J mice for at least eight generations 34, 35. After backcrossing to C57BL/6J mice for more than five generations, CAG-XstopX-PLCε mice (Lines A, G, and H) were crossed to K5-Cre transgenic mice 19 to yield K5-PLCε-TG mice and control WT littermates. For global overexpression of PLCε, CAG-PLCε transgenic mice were generated by germline excision of the XstopX cassette from CAG-XstopX-PLCε mice (Line E) by mating with CAG-Cre transgenic mice 36. Genotypes were determined by PCR. All the animals were maintained at the animal facilities of Kobe University Graduate School of Medicine. The use and care of the animals were reviewed and approved by the Institutional Animal Care and Use Committee of Kobe University.

The same antibody was unfortunately not efficacious in treating MS [45], perhaps due to the fact that IL-23 may be important prior to the appearance of clinical symptoms and not in subsequent LY2157299 manufacturer disease stages when patients appear with MS-associated neurological impairments.

Alternatively, it is possible that the neutralizing antibody will have limited access to the inflamed CNS where IL-23 has been shown to perpetuate the immune response [46]. Lastly, Ustekinumab also blocks IL-12, which has been proposed to have a regulatory function in autoimmunity [24, 25]. Hence, a more specific blockage of IL-23 without simultaneously neutralizing IL-12 might have been a more efficacious approach for the treatment of MS. The rationale behind blockade of IL-23 in vivo stems

from the idea that IL-23 is the major inducer of IL-17, a cytokine linked to many autoimmune diseases including multiple sclerosis and Crohn’s disease [47-52]. However, the attempts to block IL-17A itself have shown limited efficacy in some systems, implying that inflammatory mediators other than IL-17 are important in these diseases. Some early experimental studies indicated that blockade of IL-17 may not be efficacious in human Crohn’s disease patients, as neutralization of IL-17 was shown to exacerbate colitis in a mouse model [53]. Nonetheless, neutralization of IL-17A is now achievable in humans using Secukinumab (AIN457), and is shaping up after Phase II clinical trials to be a successful therapy in the pathogenesis of psoriasis, LY2606368 concentration rheumatoid arthritis, and uveitis [54]. In fact, neutralization of IL-17A in human psoriasis patients was linked to a simultaneous downregulation of upstream

signaling molecules important for IL-17A expression itself, including IL-12p40. Taken together, Th17 cells appear to be present in a number of autoimmune diseases, but their hallmark cytokine, IL-17, is not necessarily responsible for the symptoms associated with the diseases themselves. The clear correlation between many autoimmune diseases and the presence of cytokine-expressing effector T cells at the sites of inflammation should Chlormezanone allow us (in theory) to recognize the proteins secreted and make educated guesses at those proteins responsible for the tissue damage. However, a classical example of how this logic may fail is illustrated in the case of EAE, for which Th1 cells were thought to be ultimately responsible. Yet treating animals that had been immunized with the appropriate antigens to induce EAE with the hallmark Th1 cytokine IFN-γ surprisingly alleviated clinical disease. Conversely, blocking IFN-γ enhanced disease severity [55, 56]. Prior to this finding, administration of IFN-γ had been tested as a potential treatment for MS in the clinic. Deleterious effects had been reported in patients receiving this cytokine, and IFN-γ was subsequently deemed an unsuitable treatment for MS [57].