As well as these new developments, there also appears to be a protective role for women taking progestogen-only birth control pills, particularly those with anti-gonadotrophic activity such as norethisterone [25]. In summary, it is hoped that this and future audits will serve to help inform the decision-making process in planning future care for patients with bradykinin-mediated angioedema. The British Society for Immunology Clinical Immunology and Allergy Section (BSI-CIAS) received an unrestricted grant Everolimus cost of £5000 from Shire to support data entry. S. J. is supported by an NISCHR Fellowship. S Jolles – Consulting, speaker, meeting support from Shire, CSL Behring, Viropharma and SOBI. P Williams – No disclosure

E Carne – Meeting support CSL Behring and Shire. H Mian – No Disclosure A Huissoon – Meeting support CSL Behring, Shire and Viropharma. Consulting Viropharma. G Wong – No Disclosure S Hackett – Meeting support CSL Behring J Lortan – No disclosure V Platts – No Disclosure H Longhurst and S Grigoriadou and members of their department have received funding to attend conferences and other educational events, have acted as medical advisor or speaker, have received donations to her departmental fund, have received financial and other assistance with patient care projects and/or have participated in clinical trials with the following companies: CSL Behring, Pharming/Swedish

Orphan, Jerini/Shire, buy GPCR Compound Library Dyax, Viropharma, Baxter and Grifols. J Dempster – Performed consultancy work for Virophrama,

Selleckchem C225 Shire and CSL Behring S Deacock – No disclosure S Kahn – No Disclosure J Darroch – Meeting support Shire C Simon – No Disclosure M Thomas–No Disclosure V Pavaladurai – No disclosure H Alachkar – No Disclosure A Herwadkar – No Disclosure M Abinun – No Disclosure P Arkwright – No Disclosure M Tarzi – Speaker and travel support CSL Behring and Shire. M Helbert – Speaker, consulting, conference support CSL Behring, consulting and conference support Shire and consulting Viropharma. C Bangs – No Disclosure C Pastacaldi – No Disclosure C Phillips – Consulting for Viropharma H Bennett – Consulting for Viropharma T El-Shanawany – Consulting and meeting support from Shire, CSL Behring and Viropharma. “
“The present authors have previously reported that Vibrio mimicus expresses 77-kDa and 80-kDa outer membrane proteins in response to iron-limited conditions, and that the 77-kDa protein serves as the receptor for ferriaerobactin. In this study, it was found that V. mimicus can use heme and hemoglobin as iron sources. FURTA was then applied to V. mimicus 7PT to obtain candidate gene fragments involved in utilization of heme and hemoglobin. One FURTA-positive clone was shown to contain a partial gene, whose predicted amino acid sequence correlated with the N-terminal amino acid sequence determined for the 80-kDa outer membrane protein and also shared homology with heme/hemoglobin receptors of Gram-negative bacteria.

01; group P vs MR, P < 0.05). Nephrin, podocin and podocalyxin staining was preserved in the glomeruli of groups MR and AR compared with group P. Conclusion:  The MR and AR blockers decreased proteinuria in the acute model of nephrotic syndrome with preserved expression of glomerular podocyte protein independently of blood pressure. "
“Aim:  Adverse cardiovascular events resulting from accelerated atherosclerosis are the leading cause of mortality in uraemic

patients on maintenance haemodialysis (MHD). Chronic inflammation due to antigen-specific selleck chemicals llc responses is an important factor in the acceleration of atherosclerosis. The balance between CD4+ CD25+ forkhead/winged helix transcription factor (Foxp3)+ regulatory T cells (Treg) and T helper (Th)17 cells has been reported to play an important role in the development of inflammatory and autoimmune diseases. The aim of the present

study was to assess the Treg/Th17 pattern in uraemic patients on MHD and to explore the significance of Treg/Th17 imbalance in the development and outcome of acute cardiovascular events. Methods:  A total of 42 uraemic patients on MHD were evaluated. Of the 42, 22 patients with a history of acute cardiovascular events served as the MHD1 group and 20 patients without acute cardiovascular events served as the MHD2 group. Thirty patients with advanced chronic kidney disease (CKD) without acute cardiovascular events just before haemodialysis LEE011 ic50 therapy served as the CKD control group and 30 healthy volunteers as the normal control group. The Treg and Th17 frequencies were measured by flow cytometry. The retinoic acid receptor-related orphan receptor γt (RORγt) and Foxp3 expressions Ergoloid were measured by real-time reverse transcription polymerase chain reaction. Serum cytokines and C-reactive protein were detected by enzyme-linked immunosorbent assay and immunoturbidimetry. Results:  Patients with uraemia exhibited an obvious imbalance

of Treg/Th17 function when compared to the normal control group, displaying increased peripheral Th17 frequency, Th17-related cytokines (interleukin [IL]-17, IL-6 and IL-23) and RORγt mRNA levels. These patients also displayed decreased Treg frequency, Treg-related cytokines (IL-10, transforming growth factor-β1) and Foxp3 mRNA levels. This imbalance was more pronounced in the MHD2 group, while there was no significant difference between the MHD1 and CKD control group (P < 0.01 between normal control and uraemic patients; P > 0.05 between CKD control and MHD1; and P < 0.05 between MHD1 and MHD2). It was also observed that the imbalance of Treg/Th17 was not only consistent with the cardiovascular disease but also correlated with a microinflammatory state. Conclusion:  The Treg/Th17 balance was disturbed by uraemia, especially in patients with adverse cardiovascular events.

Some of these organs, such as the pineal gland (PG), subcommissural organ (SCO), and organum vasculosum of the lamina terminalis, might be the sites of origin of

periventricular tumors, notably pineal parenchymal tumors, papillary tumor of the pineal region and chordoid glioma. In contrast to the situation in humans, CVOs are present in the adult rat and can be dissected by laser capture microdissection (LCM). In this study, we used LCM and microarrays to analyze the transcriptomes of three CVOs, the SCO, the subfornical organ (SFO), and the PG and the third ventricle ependyma check details in the adult rat, in order to better characterize these organs at the molecular level. Several genes were expressed only, or mainly, in one of these structures, for example, Erbb2 and Col11a1 in the ependyma, Epcam and Claudin-3 (CLDN3) in the SCO, Ren1 and Slc22a3 in the SFO and Tph, Aanat and Asmt in the PG. The expression of these genes in periventricular tumors should be examined as evidence for a possible origin from the CVOs. Furthermore, we performed an immunohistochemical study

of CLDN3, a membrane protein involved in forming learn more cellular tight junctions and found that CLDN3 expression was restricted to the apical pole of ependymocytes in the SCO. This microarray study provides new evidence regarding the possible origin Carbohydrate of some rare periventricular tumors. “
“Formation of cytoplasmic aggregates in neuronal and glial cells is one of the pathological hallmarks of amyotrophic lateral sclerosis (ALS). Mutations in two genes encoding transactivation response (TAR) DNA-binding protein 43 (TDP-43)

and fused in sarcoma (FUS), both of which are main constituents of cytoplasmic aggregates, have been identified in patients with familial and sporadic ALS. Impairment of protein degradation machineries has also been recognized to participate in motoneuron degeneration in ALS. In the present study, we produced recombinant adenovirus vectors encoding wild type and mutant TDP-43 and FUS, and those encoding short hairpin RNAs (shRNAs) for proteasome (PSMC1), autophagy (ATG5), and endosome (VPS24) systems to investigate whether the coupled gene transductions in motoneurons by these adenoviruses elicit ALS pathology. Cultured neurons, astrocytes and oligodendrocytes differentiated from adult rat neural stem cells and motoneurons derived from mouse embryonic stem cells were successfully infected with these adenoviruses showing cytoplasmic aggregate formation. When these adenoviruses were injected into the facial nerves of adult rats, exogenous TDP-43 and FUS proteins were strongly expressed in facial motoneurons by a retrograde axonal transport of the adenoviruses.

The LTα1β2 then signals via LTβR to drive mesenchymal stromal cells to differentiate into lymphoid tissue organizer cells (LTos),[9] accompanied by the up-regulation of chemokine (e.g. CXCL13, CCL19 and CCL21) and adhesion molecule (e.g. vascular cell adhesion molecule-1, intercellular adhesion molecule-1, mucosal addressin cell adhesion molecule-1)[12] expression in the LN anlagen. Chemokines, as well as the up-regulated expression of RANKL Midostaurin cost and interleukin-7 (IL-7) by LTos,[9, 10] induce the recruitment and survival of further cells to the expanding LN anlagen.[13] The arrival of more LTα1β2-expressing cells, which includes few LTis[14] but after birth is dominated by lymphocytes

(both T and B cells),[15, 16] creates a positive feedback loop EPZ-6438 molecular weight (Fig. 1), further increasing signalling through the LTβR and

the subsequent expression of LTo-derived factors. Using conditional ablation of the Ltbr gene exclusively in VE-Cadherin+ endothelial stromal cells, Onder et al.[17] recently revealed that the development of multiple peripheral LNs required LT signalling specifically into this LTβR+ stromal compartment. Interestingly, not all LNs required endothelial sensitivity to LTα1β2, as the mesenteric LNs of the intestine were fully intact in these mice, hinting at a requirement for distinct LTβR+ stromal cell populations in the development of anatomically disparate peripheral LNs in vivo. Other homeostatic SLOs develop in a fundamentally similar way to the LN with only minor differences between tissues. For instance in the Peyer’s patches of the small intestine, although ligands of the receptor tyrosine kinase RET acting on a distinct population of CD45+ IL-7Rα− CD11c+ cells contributes to stromal activation in the developing anlagen,[18] LTis and LTα1β2 are still important in this developmental process,[4] although it is not clear if LTα1β2 expression is induced by RANK as in early LN development. However,

the earliest steps in homeostatic intestinal SLO development are still under intense investigation.[19] Lymphoid tissue organizers differentiate into the various non-haematopoietic stromal subtypes present in the adult SLO via LTβR signalling,[20] Bay 11-7085 although the ontogeny and lineage relationships of the various stromal cell subsets within the LN is still under investigation.[21, 22] Mesenchyme-derived stromal cells can be divided into several subsets including follicular dendritic cells (FDCs), marginal reticular cells and populations of fibroblastic reticular cells (FRCs). Lymph node stromal endothelial cells can be divided into blood endothelial cells and lymphatic endothelial cells,[23] and all SLOs contain high endothelial venules composed of endothelial cells with distinct morphology and phenotype. Four CD45− stromal subsets can therefore be identified by a dual CD31 (PECAM-1) and Podoplanin (gp38) stain.[23] Identification of further subsets can be achieved using a range of different surface markers (Table 1).

We note, however, that the proportion of inter-population variation differs depending on the genetic system: it is around 15% for allozymes,24 most DNA markers,22,23 and HLA-DPB1,25,49 and is slightly lower for the other HLA loci (∼ 10% on average), but is notably higher for GM (∼ 46%, including ∼ 39% among geographic groups and ∼ 7% among populations within geographic groups).12 This may be the result, in the

case of GM, of a bias in frequency estimation because of serological typing (as discussed above), although the effect of positive selection cannot be totally ruled out. In the case of HLA, we can conclude that balancing selection lowers inter-population variation although this effect is not Compound Library very pronounced. Immunogenetics is therefore an informative tool in anthropology, despite the effect of natural selection, which is clearly demonstrated for HLA but appears to be weak. Moreover, the study of immunogenetic markers may provide important novel information for anthropological studies. Indeed, what is often considered to be a disadvantage in anthropological studies – a non-neutral mode of evolution of the studied polymorphisms – may

be highly relevant to understanding Roxadustat ic50 complementary aspects of human evolution, like environmental changes. Relevant results obtained through computer simulation have recently been obtained by Currat et al.,91 who estimated an unequal coefficient of selection for HLA-DRB1 in Southwest European (0·7%) and Northwest African (1·9%) populations separated by the Strait of Gibraltar. This difference can be seen as a genetic signature of heterogeneous environments in the past, i.e. different pathogen richness or prevalence of specific infectious diseases in the two regions. Also, the case of Amerindians would deserve deeper investigation to understand Methisazone the evolution of their peculiar HLA genetic profiles. This could also be carried out by simulating different

scenarios taking into account both the initial settlement of America and its recent history marked by European colonization, which brought many new pathogens to this continent. The study of polymorphisms of important molecules for immune responses opens crucial areas of research in the field of human evolution, such as gene–pathogen co-evolution. This work received financial support from the Swiss National Science Foundation (SNF, Switzerland) grants no. 3100A0—112651 and 31003A—127465 (A.S.M.), the ESF (Europe) COST grant of Action BM0803 ‘HLA-NET’ (A.S.M.), the Oslo University Hospital Rikshospitalet, and Medinova (E.T.), and the US National Institute of Health Grant no. AI067068 (J.A.H. and S.J.M.). The authors declare no conflicts of interests.

This claim is far from being

uncontroversial. According to the social cognitive perspective, the ability to be jointly engaged with a partner is brought about by a strong reorganization of infant mind—the so-called 9-month cognitive revolution (Tomasello, 1995a, 1995b, 1999)—occurring at around the end of the first year of life, owing to the emergence of the infant’s understanding of other persons as intentional agents. Therefore, that ability is viewed as a sudden achievement that appears in quite an abrupt way and pushes infants from the dyadic to the triadic period. Recent research has XL765 molecular weight challenged this view. Infants younger than 9 months of age actively

coordinate their attention between people and objects (Flom & Pick, 2005; Striano & Bertin, 2005; Striano, Stahl, & Cleveland, 2009) and even 3-month-olds can appreciate the triadic situation if they are provided with a facilitated condition, such as when the adult’s gaze on an object is coordinated with the infant’s gaze (Striano & Stahl, 2005). The few neurophysiological data available so far are also consistent with the above findings, as 5-month-olds’ attention to an object, measured as activation of neural correlates, was higher in joint attention GDC-0068 condition, where the experimenter alternated her gaze from the object to the infant’s eyes, than in nonjoint attention condition (Parise, Reid, Stets, & Striano, 2007), and 4-month-olds exhibited enhanced neural processing when looking at an object at which the adult did not look compared with the

object the adult looked at, suggesting that the cued object is perceived as more familiar than the uncued one (Reid, Striano, Kaufman, & Johnson, 2004). Overall, infants appear to be sensitive to key components of triadic interaction very early in development. It is thus hard to argue for a sharp discontinuity between the dyadic and triadic L-NAME HCl period owing to the alleged sociocognitive shift. Instead, infants’ earlier appreciation of rudimentary aspects of triadic interactions in the dyadic period could represent the first step in joint attention development (Moore, 1996; Striano & Rochat, 1999; Striano & Stahl, 2005), giving it the nature of a process that is “nurtured during the early period of face-to-face play and expands during the emergence of the triadic interactive system” (Bakeman & Adamson, 1984, p. 1288). Indeed, recent literature supports the continuity perspective (Müller, Carpendale, Budwig, & Sokol, 2008).

RNA isolation, cDNA synthesis and quantitative PCR.  The samples frozen in liquid nitrogen were homogenized in Tri Reagent (Applied Biosystems, Foster City, CA, USA), using the Ultra-Turrax apparatus (Janke&Kunkel, Staufen, Germany) or VWR Pellet Mixer (VWR, West Chester, PA, USA) for the solid organs. Total RNA was isolated after chloroform phase separation using the RNeasy kit (Qiagen, Düsseldorf, Germany), as instructed by the manufacturer.

First-strand HIF inhibitor cDNA was synthesized using oligo-dT primer (Sigma Aldrich, St. Louis, MO, USA) and avian myeloblastosis virus reverse transcriptase (Finnzymes, Espoo, Finland). Real-time quantitative PCR (qPCR) was performed using the TaqMan Gene Expression Master Mix (Applied Biosystems) and the appropriate primer-probe sets, and the samples were analysed using the iCycler iQ instrument (Bio-Rad, Hercules, CA, USA). All samples were run in duplicate and unless otherwise stated, were normalized against Hprt expression

levels. The primer-probe sets for Hprt, Foxp3 and CD19 were commercially available assays-by-demand (Applied Biosystems). The primer-probe set for T cell receptor (TCR) Cα was an assay-by-design (Applied Biosystems), consisting of the primers 5′-CAA AGA GAC CAA CGC CAC CTA and 5′- CGG TCA ACG TGG CAT CAC, and probe 5′-6FAM- CCA GTT CAG ACG TTC CC-quencher. All assays were intron spanning. Statistical analysis.  The data are shown as mean ± SD. Comparison of means was carried out by Student’s two-tailed t-test, with P < 0.05 as the limit for statistical significance. To test how cells from Aire−/− C646 mice behave in a situation strongly biased in favour of autoreactivity, but with normally functioning Aire protein, we transferred

lymph node cells from Aire−/− and control Aire+/+ mice into lymphopenic Aire+/+ recipients, thus triggering LIP. In the first Methocarbamol group of recipients (hereafter Aire-group), the proliferating cells have developed in the absence of Aire, but the proliferation takes place in an Aire-sufficient environment. The second group of recipients will hereafter be denoted the control group. In this way, we can separate the central effects of Aire from its peripheral functions, and all the differences we note between the Aire and control group will be consequences of the defective thymic development in Aire−/− donors. The major lymphocyte populations in the donors were analysed prior to the adoptive transfer, and no significant differences were found in the frequency of T or B cells, CD4+ cells, CD8+ cells, Foxp3+ Treg cells, or in the fraction of Ki-67+ cells within these subsets (data not shown). Each lymphopenic recipient was injected with 1 million mononuclear cells to tail vein. After the transfer, we allowed the lymphocytes to proliferate for 2 months in order to reach the plateau phase of LIP.

01 when compared with mice pre-sensitized with FITC and treated with control rat IgG). The results indicated that CD4+CD25+ T-cell-mediated negative regulation induced by FITC sensitization suppressed the subsequent activation of DNFB-specific CD8+ T cells in the skin-draining LN. this website Consistent with the results of this report, CD4+CD25+ T-cell-mediated negative regulation of the activation of CD8+ T cells specific to a second hapten (FITC) correlated with decreased total numbers of LC presenting this hapten in the LN of mice pre-sensitized with DNFB and treated

with control rat IgG at the time of first sensitization (Fig. 6B). The numbers of FITC-presenting LC were increased to the level observed in the control group when mice were given anti-CD25 mAb at the time of the first sensitization with DNFB. Antigen-specific CD8+ T cells undergo rapid expansion within the lymphoid priming site in response to pathogen infection and the number of these effector cells rapidly declines following antigen clearance 17, 18. One critical factor regulating antigen-specific CD8+ T-cell expansion is the duration of CD8+ T-cell exposure to antigen and co-stimulatory signals provided by the APC. In vitro models have indicated apoptosis of APC during culture with antigen-specific

effector CD4+ T cells suggesting this elimination as a mechanism affecting the Selleck Trichostatin A magnitude and duration of T-cell-mediated immune responses these 2, 19. In vivo studies have identified Fas-dependent elimination of APC as a mechanism restricting systemic autoimmune disorders such as lymphoproliferation and production of autoimmune Ab 4. LC resistant to apoptosis through a deficiency in Bid or Fas induced stronger CD4+ T-cell-mediated immune responses than WT DC 2, 3. The increased lifespan of DC and B cells in mice with a targeted FasL gene deletion

in T cells suggests that FasL-expressing T cells down-regulate autoimmune responses by controlling APC numbers 20. It remains unclear, however, whether the same mechanism down-regulates CD8+ T-cell-mediated immune responses to antigens deposited in the skin as well as the identity of the cells mediating this negative regulation. Our previous studies suggested that FasL-expressing CD4+ T cells regulate hapten-presenting DC activation of effector CD8+ T cells for CHS 1. We had also reported that attenuation of the regulatory CD4+CD25+ T-cell compartment by anti-CD25 mAb treatment during initiation of CHS responses enhanced the magnitude of hapten-specific CD8+ T-cell expansion and subsequently increased the magnitude and duration of CHS responses mediated by these effector CD8+ T cells 13. This suggested that CD4+CD25+ T cells might negatively regulate CD8+ T-cell-mediated CHS responses through FasL-dependent mechanisms.

Natural killer T cells expressing an invariant T cell antigen receptor recognize glycolipid antigens by their invariant TCR; however, natural antigens recognized by this receptor were not identified for many years. Recent studies have shown that iNKT cells recognize glycolipids from microbes such as Sphingomonas spp. (41–43) and B. burgdorferi (49), suggesting that the iNKT TCR detects certain microbes. The crystal structures of two ternary complexes of mouse CD1d-bacterial glycolipid-iNKT TCR have revealed that the iNKT TCR recognizes bacterial glycolipids by inducing conformational

changes in antigens and CD1d to adopt a conserved binding mode (53). We speculate that iNKT TCR recognizes microbial glycolipids whose structures are similar to known microbial antigens. Importantly, iNKT cells also respond to microbes via inflammatory cytokines and/or endogenous antigens in the absence of microbial glycolipids. However, in some cases, this website iNKT cells participate in the pathogenesis of inflammatory diseases (28, 59). Therefore,

it is important to clarify the mechanisms that initiate and regulate iNKT Selisistat in vitro cell mediated inflammatory responses. Furthermore, an important future goal of iNKT cell research is the identification of endogenous antigens for these cells. Although it has been reported that one glycolipid is the endogenous antigen that is responsible for iNKT cell development (66), later studies have disputed this (67–69). More studies are needed Carnitine dehydrogenase to identify the endogenous antigen for iNKT cells. Many mouse studies have shown that glycolipid mediated

iNKT cell activation augments antimicrobial responses in various microbial infections (2, 4, 9, 10). Moreover, recent studies indicate that iNKT cell antigens are useful adjuvants for vaccines against microbial pathogens such as influenza virus (70–74), malaria (75, 76), HIV (76–78) and HSV-2 (79). Positive results have been reported from several clinical trials of tumor immunotherapy with αGalCer pulsed APCs and in vitro expanded iNKT cells (80, 81). These data indicate that iNKT cell glycolipid antigens may also be useful for new antimicrobial therapies and vaccines. This work was supported by grants from the Japan Society for the Promotion of Science and the Japanese Ministry of Education, Culture, Sports, Science and Technology (22689031), the Ministry of Health, Labor and Welfare of Japan (H22seisakusouyakuippan012), and the Uehara Memorial Foundation. “
“Specific cytokines and the costimulatory protein CD40 play role in inducing immunoglobulin (Ig)A production by B cells in the humoral immune response. However, to date, the role of these mediators was not investigated in chronic periodontitis. Therefore, the aim of this study was to assess the local levels of interleukin (IL)-21, IL-21 receptor (IL-21R), IL-4, IL-10 and CD40 ligand (CD40L) on chronic periodontitis subjects and their relationship with the salivary levels of IgA.

As a second approach to test our hypothesis, we compared the capability of cutaneous DC that do or do not express functional Fas to prime Midostaurin purchase effector CD8+ T cells for CHS responses. DC were purified from the skin-draining LN of DNFB-sensitized WT or lpr

mice and were transferred intradermally into naïve WT mice as previously described 15, 16. The magnitude of CHS responses induced by transfer of DC isolated from DNFB-sensitized WT mice decreased to background levels at 120 h post-challenge. In contrast, the magnitude of CHS responses in mice receiving DC from Fas-defective lpr mice was markedly increased and sustained (Fig. 4A, *p<0.05). The characteristics of these CHS responses correlated with the magnitude of hapten-specific CD8+ T-cell development in the skin-draining LN of DC-transferred mice. At day +5 post-transfer, hapten-specific CD8+ T cells producing IFN-γ were easily detectable in mice primed with WT DC, but within 2 days (i.e. day +7 post-transfer), the number of these CD8+ T cells decreased more than three-fold (Fig.

4B). In contrast, considerably higher numbers of hapten-specific CD8+ T cells producing IFN-γ were observed on day +5 in the LN of mice primed with lpr DC (Fig. 4B, WT DC versus lpr DC, *p<0.05), and these numbers continued to increase 3-MA research buy by day +7 post-transfer. Thus, the augmented

and prolonged ear swelling responses observed in mice primed with Fas-defective DC correlated with increased and sustained numbers of hapten-specific CD8+ T cells Tolmetin producing IFN-γ in the LN. These results were consistent with negative regulation of DC priming functions in CHS responses through Fas–FasL. To directly test whether regulatory CD4+CD25+ cells utilize Fas–FasL interactions to inhibit activation of hapten-specific CD8+ T cells by Fas-expressing DC, immune CD8+ T cells from sensitized WT mice were cultured with hapten-presenting DC purified from sensitized WT or lpr mice in the presence of naïve WT CD4+CD25+ or CD4+CD25− cells and IFN-γ production by the immune CD8+ T cells was assessed by ELISA. To assess the possibility that CD4+CD25− or CD4+CD25+ cells produce IFN-γ during this culture, we tested IFN-γ production by immune CD8+ T cells cultured with hapten-presenting DC only. The results indicated that additional amounts of IFN-γ were not produced when CD8+ T cells were cultured with DC and CD4+CD25− T cells when compared with CD8+ T cell/DC cultures (Fig. 5A). In fact IFN-γ production was slightly decreased in CD8+ T cell/DC/CD4+CD25− T-cell cultures, although this was not a significant decrease and most likely due to competition between the T cells for access to the DC.