S. G. M. received honoraria for lecturing and travel expenses for attending meetings and has received financial research support from Bayer, Biogen

Idec, Sanofi-Aventis, Bayer Schering, Merck Serono, Novo Nordisk, Genzyme, MSD and Teva. All authors declare no relevant conflicts of interest. “
“Apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) is an important component of the inflammasome, INK 128 mouse functioning as an adaptor protein that facilitates the recruitment and activation of procaspases that in turn promote the maturation of interleukin-1β (IL-1β) and IL-18. Despite initial focus on the inflammatory properties of ASC there is emerging evidence that highlights the Fulvestrant concentration importance of ASC in facilitating adaptive immune

responses. However, the cellular and molecular basis for the involvement of ASC in adaptive immunity remains largely unexplored. We have previously demonstrated that activated ASC-deficient T cells have dampened proliferative responses. We have therefore explored the underlying cellular mechanism(s) by which ASC regulates T-cell proliferation. We show that under activating conditions (anti-CD3/CD28 stimulation) in bulk T-cell cultures the presence of ASC−/− CD4+ T cells is sufficient to suppress the proliferative responses of neighbouring T cells. Furthermore, ASC−/− CD4+ T cells upon activation exhibit a suppressive cytokine profile, with elevated production of IL-10 and reduced secretion of T helper type 1 cytokines, interferon-γ and IL-2. This increase in IL-10 secretion within the activated ASC−/− CD4+ T-cell compartment Phospholipase D1 was not associated with a proportional increase in conventional Foxp3+ regulatory T (Treg) cells. Interestingly, when equal numbers of fluorescence-activated cell sorted ASC+/+ and ASC−/− Treg cells (CD4+ CD44intermediate/high CD25+) were activated in vitro, the ASC−/− fraction produced significantly more IL-10 than their wild-type counterparts, suggesting that ASC−/− Treg cells have greater suppressive capacity. Collectively,

these results imply that the ASC may influence the development and functioning of Treg cells. Apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) is an integral component of the inflammasome, a cytosolic multiprotein platform that facilitates the activation of pro-inflammatory caspases, which in turn promote the maturation and subsequent secretion of interleukin-1β (IL-1β) and IL-18.1,2 ASC is a simple adaptor protein with two linked protein–protein interaction domains of the death domain superfamily: an N-terminal pyrin/PAAD death domain and a C-terminal caspase recruitment domain, which interact with the different NOD-like receptors, the sensory elements of the inflammasome and pro-caspase-1, respectively.3–5 These two domains enable ASC to function as an essential link between the sensor protein and effector molecules during inflammasome assembly.

Brucella melitensis is the first intracellular pathogen in which a QS system was described. Although no acyl-homoserine lactone (AHL) synthase has been found as yet, this bacterium produces two AHLs detectable in culture supernatants: a dodecanoyl-homoserine lactone (C12-HSL) and a putative 3-oxo-dodecanoyl-homoserinelactone (3-oxo-C12-HSL) (Taminiau et al., 2002), and possesses two LuxR-type regulators, called VjbR and BabR (Delrue et al., 2005). We demonstrated previously that QS, through VjbR, is a major regulatory system of important cell surface structures of Brucella (Delrue et al., 2005; Uzureau et al., 2007). Moreover,

we showed Protein Tyrosine Kinase inhibitor that vjbR-deficient strains, all unresponsive to C12-HSL, display a clumping phenotype in liquid culture and that these aggregates contain an unknown exopolysaccharide(s) (Uzureau et al., 2007). Clumping development is a complex process that is initiated when bacteria attach to a surface using exopolysaccharide polymers or other adhesins and develop into microcolonies. Bacteria can undergo an additional maturation step

in which they develop as complex three-dimensional (3D) structures called biofilms (O’Toole et al., 2000). These structures are classically defined as matrix-enclosed bacterial populations adherent to each other and/or to surfaces or interfaces (Costerton et al., 1995). The biofilm Ivacaftor in vitro development process requires complex cellular regulatory mechanisms in which QS is often involved (Davies et al., 1998; Hammer & Bassler, 2003; Rice et al., 2005). Aggregates of bacteria not attached to a surface are commonly termed

flocs or clumps and have many of the characteristics of a biofilm (Hall-Stoodley et al., 2004). Because bacterial clumping is one of the initial steps of biofilm formation, the clumping phenotype in B. melitensis 16M described previously was the first evidence that this alphaproteobacterium could form biofilms during its lifecycle. Biofilm or clump formation constitutes the natural behavior of numerous environmental and pathogenic bacteria. The most distinctive feature of these aggregative structures is the extracellular matrix that plays a structural role, benefiting the bacterium by enabling attachment to surfaces, improving nutrient acquisition RAS p21 protein activator 1 or providing protection from environmental stresses and host defenses (Sutherland, 2001; Branda et al., 2005). Matrix polymers of bacterial biofilms are predominantly exopolysaccharide, whose compositions vary between strains and can be affected by the growth conditions and the age of the biofilm (Sutherland, 2001). In addition to exopolysaccharide, the matrices generally contain nucleic acids, proteins, lipids and outer membrane vesicles (OMVs) in the case of Gram-negative bacteria (Tsuneda et al., 2003; Schooling & Beveridge, 2006).

3E). These data indicated that the activated phenotype of NK cells was determined by MHC class I down-regulation

rather than by NKG2D-L levels expressed on early-stage tumors. Nevertheless, the higher MHC class I and lower NKG2D-L expression levels found in late tumor stages suggested that both, MHC class I PD-1 inhibitor recovery and loss of NKG2D-L, may provide mechanisms of immune escape. To directly test the role of NKG2D-L loss in immune escape, we established cell lines from lymphoma-bearing mice with reduced MHC class I expression and selected variants with different NKG2D-L levels. Cell line myc-E showed background levels of NKG2D-L. In contrast, cell line myc-B had a 20-fold enhanced expression of NKG2D-L (Fig. 4A). After transfer into naïve WT mice, the myc-B line grew out slowly www.selleckchem.com/products/Sunitinib-Malate-(Sutent).html and was even rejected in 50% of the animals. In contrast, all mice injected with myc-E cells rapidly succumbed to tumor growth (Fig. 4B). Importantly, when NKG2D-L on myc-B cells were blocked with NKG2D multimers

prior to injection, protection was lost, and mice died as rapidly as those receiving the myc-E line. Protection against myc-B was also abrogated by NK-cell depletion (Fig. 4B). The data show that in these cell lines showing low MHC class I levels, expression of NKG2D-L is a signal that is required for NK cell-mediated elimination of tumor cells. To test the hypothesis that tumor escape from NK-cell surveillance results from re-expression of MHC class I and from suppression of NKG2D-L, we analyzed the outgrowing lymphomas in mice having received cell line myc-B. Indeed, the tumor cells that grew out after challenge with MHC

class Ilow/NKG2D-Lhigh myc-B cells were converted to MHC class Ihigh/NKG2D-Llow cells (Fig. 4C). NK cells isolated upon growth of myc-B also showed an activated status and decreased NKG2D expression (data not shown). The data demonstrate that tumor progression is not only due to exhaustion or paralysis of NK cells following their initial activation, as described above. In addition, loss of NKG2D-L as well as recovery of MHC class I on tumor cells contribute to escape from NK-cell surveillance. Protection from NK-cell attack and the NKG2D modulation observed on NK cells from tumor-bearing mice Niclosamide might be an effect of NKG2D-L shedded from tumor cells. In two different assay systems (See the Materials and methods section), there was no evidence for the presence of soluble NKG2D-L in sera from tumor mice (Supporting Information), but a clear NKG2D down-regulation was seen when WT NK cells were incubated with ligand-expressing lymphoma cells in vitro (Fig. 4D). If NKG2D-L expression is needed for tumor elimination (Fig. 4B, C) although it was not correlated with the expression of NK-cell activation markers (Fig.

As a result of its speed and potential sensitivity, nucleic acid amplification via polymerase chain reaction (PCR)-based protocols appear as an attractive alternative.33 However, as Bennett pointed out,34 the lack of a reference standard other than blood culture is a significant impediment to the development of standardised assay. Specifically, it is hard to decide if the detection of Candida nucleic acids in blood culture-negative samples is a false-positive result or reflects a lower threshold

of detection. In addition, the substantial resource requirements and costs of a high-quality PCR laboratory limit the immediate use of PCR in the individual patient, thus diminishing its time advantage AZD6244 concentration when compared with culture-based diagnostics. Evidence built up in the last couple of years unequivocally indicates that the time point of initiation

of adequate antifungal therapy greatly impacts the outcome of Candida bloodstream infections in terms of hospital mortality. This was most impressively demonstrated in patients with septic shock: in a large sample, Kumar et al. [35] retrospectively found a crude hospital mortality of 87% in patients with Candida spp. as the causative agent compared to 52% in patients with bacterial pathogens learn more in cohorts with similar baseline APACHE II score and age. They showed that the median time to effective therapy was 35 h in fungal septic shock compared to only 6 h in bacterial septic shock. If the data were adjusted for time from onset of hypotension to start of appropriate antimicrobial therapy, there was no difference in mortality of the two cohorts. This clearly demonstrates that the excess mortality in fungal septic shock is attributable to delays of effective antifungal therapy. In patients receiving antifungal therapy within 2 h after onset of hypotension, the mortality rate was only 19% were compared

to 94% if antifungal therapy was delayed by 12 h. Mortality increased by approximately 8% STK38 per hour of delay. By the way, these data may serve as a strong indicator of the close correlation between the time of initiation of antifungal treatment and mortality rates of severe Candida sepsis. Similarly, the same group showed that appropriateness of initial therapy, i.e. coverage of the causative pathogen by the first administered drug, was associated with increased survival in Candida septic shock with 5–10-fold reductions in hospital mortality for both C. albicans and C. non-albicans infections.36 As pointed out, blood cultures remain the backbone of diagnosis of fungal bloodstream infection. Incubation times to positivity tend to be substantially longer with Candida spp. when compared with bacterial pathogens because of the generation times of several hours in contrast to <1 h for bacteria commonly involved in septic infections.

As detailed in the section entitled Novel protein synthesis is required to maintain a prolonged

IL-10-mediated STAT3 activation, activation of IL-10 signaling via STAT3 rapidly culminates in a reduction in the transcriptional rate of LPS-induced pro-inflammatory genes, whereas simultaneously enhancing the transcription of LPS-induced anti-inflammatory genes. A working model that could explain both actions of IL-10 is one in which STAT3 is directly recruited to the promoter of target genes and, in turn, modifies the composition of the transcriptional complexes or the accessibility of the promoter to the transcriptional machinery. Consistent with this depiction, adenoviral gene delivery of a constitutively active STAT3 in bone marrow-derived DC blocks LPS-induced IL-12p40 gene expression and c-Rel recruitment to the IL-12p40 promoter 43. In this context, IL-1ra

represents a Decitabine chemical structure potential target gene of IL-10. Previous studies had indeed shown that IL-10 strongly potentiates the production of IL-1ra in LPS-stimulated phagocytes 12, 14, 44, whereas concomitantly inhibiting pro-inflammatory gene expression. IL-1ra was first isolated in 1986 as a soluble factor that competed with IL-1α and IL-1β for binding to their receptor 45, thus inhibiting their biological activities. It is now well established that the balance between IL-1 and IL-1ra may determine whether the outcome of a given response to damage will be pro- or anti-inflammatory. Indeed, in a variety of experimental animal models, Nutlin-3 concentration an imbalance between IL-1 and IL-1ra in favor of IL-1 has been shown to predispose to the development of diseases such as arthritis, inflammatory bowel disease, granulomatous and fibrotic lung disorders, kidney disease, diseases of the liver and pancreas, graft-versus-host disease, leukemia and cancer, osteoporosis and diabetes, central nervous system diseases, infectious diseases and arterial disease 46. Consistent with the anti-inflammatory

role of IL-1ra, mice lacking a functional IL-1ra gene develop spontaneous signs of polyarthritis, vasculitis or skin inflammation 47–49. Moreover, the use of conditional knockout mice in which IL-1ra production has been selectively Sorafenib chemical structure deleted in myeloid cells has suggested that IL-1ra derived from monocyte/macrophages and/or neutrophils plays a critical role in controlling the development and severity of collagen-induced arthritis, by modulating Th1 and Th17 responses in lymphoid organs 50. More recently, homozygous germ-line mutations of the sequence encoding the IL-1ra gene (IL1RN) have been demonstrated to cause a human syndrome, named deficiency of IL-1ra, characterized by a striking IL-1-mediated systemic and local inflammation that is apparent soon after birth 51, 52.

In malaria endemic settings, adults develop LY2157299 protective immunity after repeated exposures (2), but women are more

susceptible to malarial infection when they become pregnant (1). Severe clinical manifestations associated with this infection include premature delivery and intrauterine growth restriction, contributing to low birth weight (LBW), stillbirth, abortion and maternal mortality (3–5). Increased synthesis of inflammatory cytokines like tumour necrosis factor (TNF), interleukin (IL)-2 and interferon (IFN)-γ (6–8) has been shown in malaria during pregnancy, and levels of TNF in particular have been associated with maternal anaemia and LBW (6,9). Interestingly, such pathogenic immune responses to malaria appear to be influenced by host genetic factors. For example, infants homozygous for TNF2, a polymorphism in the TNF promoter region that is associated with increased TNF production (10), are at increased risk of preterm birth and mortality, suggesting that poor birth outcomes in malaria endemic areas are precipitated by a genetically determined maternal tendency to produce high levels of this inflammatory cytokine (11). In mice, the immune response to malaria is complex and varies as a function

of mouse genetic background (12) and anatomical sites analysed. Moreover, it is dependent on parasite species and strain as well as on route of infection. In B6 mice, early production of TNF, IFN-γ, IL-12 (13) and granulocyte–macrophage colony-stimulating factor (14) is required for resistance to Montelukast Sodium blood-stage selleck products P. chabaudi AS infection. In contrast, susceptible A/J mice mount early, predominantly Th2-biased cytokine responses (15) and succumb to infection (16). Treatment of these mice with recombinant IL-12 early in infection results in increased production of IFN-γ and TNF and facilitates elimination of parasites and survival (17). Interestingly, A/J mice overcome their Th2-cytokine bias later in infection, exhibiting increased TNF expression in the liver and high serum TNF levels coincident with the time that they begin to succumb to infection (18). Recently, we initiated studies of malaria

during pregnancy using P. chabaudi AS infection in B6 mice as a model platform (19–21). This model recapitulates the severe pregnancy outcomes, namely foetal loss, seen in low endemic areas and in some heavily exposed primigravidae (1). Importantly, similar to human malaria during pregnancy (6,9), TNF plays a critical role in embryo loss in this model; antibody-mediated neutralization of this cytokine rescues mid-gestational pregnancy (21). Because TNF is well known to have a negative impact on pregnancy outcomes even in the absence of infection (22,23), the tendency to produce this factor in response to malarial infection may represent a common pathogenic factor that relative to other host elements is central to disease pathogenesis.

We have used similar conditional immortalization techniques to generate human glomerular endothelial cell lines.19 We are particularly interested to study the interactions of these cells with normal and mutant podocytes, also to develop three-dimensional culture systems including flow, so as to mimic as closely as possible the in vivo situation.20 Human podocyte cell lines can now be reliably propagated for study in vitro. We believe that VX-809 in vitro conditional immortalization provides the most reliable and representative

cell lines: we are proud of the fact that podocyte cell lines originally developed in Bristol are now in widespread use across the world and we would like to encourage other workers to reproduce our results. “
“Immunoglobulin A nephropathy (IgAN), characterized by predominant or exclusive deposition of IgA1 in glomerular mesangium, is the most common primary glomerulonephritis worldwide. At present, the treatment is always limited due to the incomplete understanding of the pathogenesis of check details IgAN. Mesangial deposited IgA1 is the common final pathway leading to glomerulonephritis and renal injury. IgA1 protease, a proteolytic enzyme with strict substrate specificity for human IgA1, may be an effective therapeutic candidate for IgAN by removing the mesangial deposited IgA1. “
“Aim:  Overseas kidney transplantation has often been reported to have unsatisfactory outcomes.

This study aims to compare post-transplantation outcomes between overseas and domestic kidney transplant (KT) recipients in Taiwan. Methods:  The Taiwanese National Health Insurance Research Database was used to identify 310 domestic and 643 overseas KT recipients, who survived for longer than 1 month after

the transplantation, in a cohort of 45 453 chronic haemodialysis patients in 1997–2002. Cox proportional hazards models were used to assess Morin Hydrate risks of mortality and graft failure. Results:  The 1, 3 and 5 year survival rates for domestic KT recipients were 96.5%, 93.3% and 91.6%, respectively, while those for overseas KT recipients were 94.9%, 87.9% and 77.1%, respectively (P = 0.015). For the overseas group, those who received a KT before 2001 had significantly higher hazard ratios of mortality and graft failure (2.85 and 1.71, respectively). However, for those receiving a KT in 2001–2002, no significant outcome difference could be found between overseas and domestic recipients. Conclusion:  The risk disparity between overseas and domestic KT recipients is mainly attributable to when the transplantation was performed. In attempting to dissuade potential recipients from organ trafficking, merely emphasizing the previously acknowledged poor outcomes no longer suffices as a valid reason. “
“Aim:  Nestin, an intermediate filament originally identified as a marker of neural progenitor cells, is transiently expressed in endothelial cells and tubuloepithelial cells during kidney development.

“
“Somatic hypermutation (SHM) is an important

step in antigen-driven B cell development creating B lymphocytes expressing high-affinity antibody receptors. It is known that the peripheral B lymphocyte compartments of healthy children and adults differ considerably. However, the development of SHM with age has not been studied in detail previously. Therefore, we used the immunoglobulin (Ig)κ-restriction enzyme hot-spot mutation assay (Igκ-REHMA) to gain an estimation of SHM levels in different age groups in order to relate this to the size of the memory B lymphocyte subpopulations. We show that the level of SHM increases rapidly during the first 2 years of life. This reflects the changes of the memory B cell subpopulations, but also changes in the SHM within memory Ceritinib mw B cell subsets, probably reflecting an increase of secondary memory B cell responses. “
“Toxoplasmosis is a world-wide zoonosis that causes significant public health and veterinary problems. The study of vaccines remains the most promising method for the future prevention and control of toxoplasmosis. Recombinant

Toxoplasma gondii cyclophilin has been shown to have potent PPIase and IL-12-inducing activities, thus promoting the stabilization of T. gondii’s Z-VAD-FMK cost life cycle and maintaining the survival of its host during evolution. In this study, the T. gondii cyclophilin gene was used to construct a DNA vaccine (pVAX1-TgCyP). The immune response and protective efficacy of the vaccine against T. gondii infection in BALB/c mice were evaluated. All BALB/c mice that were vaccinated with pVAX1-TgCyP developed a high response CYTH4 with TgCyP-specific antibodies, and significant splenocyte proliferation (P < 0·05) compared with pVAX1 vector and PBS groups. pVAX1-TgCyP also induced a significant Th1 type immune response, indicated by the higher production of IL-2 and IFN-γ (P < 0·05). The survival rate of BALB/c mice increased significantly after vaccination with pVAX1-TgCyP (37·5%) (P < 0·05). These results indicate that TgCyP is a highly efficacious vaccine candidate that can generate protective immunity against

T. gondii infection in BALB/c mice. Toxoplasma gondii (T. gondii), the aetiological agent of toxoplasmosis, is an apicomplexan protozoan parasite that infects wide variety of cell types in humans and other warm-blooded animals [1, 2]. A variety of clinical syndromes can develop following T. gondii infection, especially in immune-compromised patients (such as AIDS patients), pregnant women and congenitally infected children [3]. T. gondii can cause severe or lethal toxoplasmosis that leads to significance economic losses in the veterinary industry, due to abortion, neonatal loss, foetal death, stillbirths and various other problems in livestock, which are mostly associated with sheep. [4, 5]. Treatment of toxoplasmosis is difficult due to the toxicities of available drugs, and re-infection occurs rapidly.

We previously identified

the adapter protein HS1 as a putative Nck-interacting protein. We now demonstrate that the SH2 domain of Nck specifically interacts with HS1 upon phosphorylation of its tyrosine residue 378. We report that in human T cells, ligation of the chemokine receptor CXCR4 by stromal cell-derived factor 1α (SDF1α) induces a rapid and transient phosphorylation selleck compound of tyrosine 378 of HS1 resulting in an increased association with Nck. Consequently, siRNA-mediated downregulation of HS1 and/or Nck impairs SDF1α-induced actin polymerization and T-cell migration. “
“The neonatal Fc receptor (FcRn) was first described as a receptor-mediating transplacental immunoglobulin (Ig)G transfer from mother to fetus, but it has other significant biological functions. It plays a key role in IgG and albumin homeostasis by efficient protection from catabolism [1]. It binds endocytosed IgG at acidic pH (< 6·5) within endosomes, Lenvatinib diverts it from degradation in lysosomes and instead transports the IgG–FcRn complexes back to the cell surface where, at neutral pH (> 7·0), IgG is released [1]. This process is highly efficient; FcRn recycles an equivalent amount of albumin and even four times as much IgG as can be produced

in a given time [2, 3]. Another notable function of FcRn is antigen delivery. FcRn was shown to be involved in the transcytosis of monomeric serum IgG from the basolateral to the apical side of the epithelium; immune complexes formed in the lumen are consequently delivered by FcRn to the lamina propria for antigen processing and triggering immune responses Terminal deoxynucleotidyl transferase [4]. Therefore, FcRn in the epithelium is probably able to sense luminal and epithelial infections and transmit evidence of these infections to the local and systemic immune apparatus. In the regulation of FCRN expression, polymorphism in the promoter region of the human gene consisting of a variable number of 37-base

pairs (bp) tandem repeats (VNTR) plays an important role. The allele with two tandem repeats (VNTR2) is associated with decreased promoter activity compared with the most common VNTR3 allele. VNTR2 carriers have been shown to have lower FCRN mRNA levels and decreased binding capacity of monocytes to immobilized IgG than VNTR3/3 homozygotes that predominate in general population [5]. We sought to determine whether FCRN expression influences intravenous immunoglobulin (IVIg) catabolism and clinical phenotype in patients with common variable immunodeficiency (CVID). This effect may be due not only to the role of FcRn in IgG protection from degradation, but also by influencing mucosal antigen presentation.

This inhibitory effect was confirmed in S2 cells stably expressing viral Pellino following lentiviral transduction. Viral Pellino also displayed cytoplasmic localisation upon stable expression (Fig. 2C) and inhibited C106-induced activation of the drosomycin promoter (Fig. 2D). This confirms that the entomopoxviral protein can obstruct a key insect immune–response pathway. The high degree of sequence and mechanistic conservation between insect Toll and mammalian TLR signalling pathways led us to further explore the potential immunomodulatory capabilities of viral Pellino in human cells. Expression of increasing amounts of viral Pellino in HEK293-TLR4 cells (Fig. 3A) showed dose-dependent

inhibition of LPS-induction of an NF-κB-responsive promoter–reporter construct (Fig. 3B). We next confirmed that viral Pellino could block the endogenous NF-κB pathway in a cell Sirolimus in vitro that was naturally responsive to LPS by demonstrating that lentivirally delivered viral Pellino

blocked the LPS-induced phosphorylation of the NF-κB subunit p65 upon stable expression in U373 cells (Fig. 3C). The regulatory effects of viral Pellino on the NF-κB pathway find more have functional consequences for pro-inflammatory gene expression since the transduction of THP-1 monocytic cells with varying titres of lentivirus, conferring stable viral Pellino expression, caused an inhibition of Montelukast Sodium LPS induced expression of the NF-κB-responsive gene IL-8 (Fig. 3D). The highest titre also inhibited LPS induction of TNF

in THP-1 cells (Fig. 3E). These studies confirm the regulatory effects of viral Pellino on TLR4 signalling in a number of cell types. We next investigated the mechanistic basis to the regulatory effects of viral Pellino on TLR signalling. IRAK-1 was an obvious target for viral Pellino, given that the mammalian Pellinos have been shown to associate with IRAK-1 10, 25, probably via their FHA domain, and that the homology modelling studies detailed above suggest the presence of a core FHA domain in viral Pellino. Co-immunprecipitation studies demonstrated that vPellino and IRAK-1 associated upon co-expression. This was observed upon immunoprecipitation of viral Pellino and immunoblotting for IRAK-1 (Fig. 4A). Furthermore, viral Pellino was found to interact with endogenous IRAK-1 upon immunoprecipitation of the latter (Fig. 4B). The IRAK-1-viral Pellino interaction is also apparent under conditions where viral Pellino is expressed at more physiologically relevant levels as facilitated by lentiviral-mediated delivery of viral Pellino into U373 cells (Fig. 4C). Further evidence in support of the IRAK-1-Pellino interaction is provided by co-localisation of IRAK-RFP and viral Pellino-GFP in HEK293 cells (Fig. 4D). Conflicting reports exist on the importance of IRAK-1 kinase activity in the interaction between mammalian Pellinos and IRAK-1 14, 15.