Glucocorticoids treatment was administerd

to eighty two patients (90.1%) and the initial dose of prednisolone (PSL) was 0.7 ± 0.3 mg/kg/day. Cyclophosphamide (CY) was prescribed to 17 patients (18.7%). During the period of 55 ± 52 months after the onset of RRT, 18 vasculitis relapses occurred in 12 patients corresponding to an incidence rate of 0.048 episodes per person-year (95% CI: 0.029–0.076). Organ systems affected by relapses included lungs Seliciclib in vitro (n = 10), ears (n = 2), and eyes (n = 1). The duration from the onset of RRT to relapse was 49 ± 44 months and maximal duration was 156 months. At the relapse, 5 patients were not receiving immunosuppressive therapy and PSL (7.7 ± 3.4 mg/day) was prescribed for the remaining patients. Survival rates for 1, 3 and 5 years after RRT were 82.3%, 75.4% and 65.3%, respectively. The causes of deaths were infection (59.5%), cardiovascular event (24.3%), gastrointestinal bleeding (8.1%), malignancy (5.4%) and interstitial pneumonia (2.7%).

By Cox’s multivariate analysis, patient year (HR1.09, 95%CI:1.05–1.13) and pulmonary involvement (HR 3.95, 95%CI 1.77–8.83) were significant positive risks and the use of CY (HR 0.10, 95%CI 0.014–0.78) was a significant negative risk for mortality. Conclusion: Relapse could occur even after a long https://www.selleckchem.com/products/LBH-589.html period from the onset of RRT. Infection was the most frequent cause of death and pulmonary involvement was related with mortality. It is important to clarify the optimal duration of maintenance therapy after RRT. PRATT RAYMOND D, LIN VIVIAN, GUSS CARRIE, GUPTA AJAY Rockwell Medical Introduction: Triferic (Ferric Pyrophosphate Citrate) is a novel iron salt that is soluble in dialysate and crosses the dialyzer membrane. Triferic, delivered via hemodialysate donates iron rapidly and directly to apo-transferrin, bypassing the reticuloendothelial system. Methods: In two, single blind, randomized placebo controlled clinical (CRUISE) Mephenoxalone trials, iron replete HD patients received either dialysate containing Triferic at 2 μM (110 μg iron/L, combined N = 299) or placebo (standard

dialysate, combined N = 300) for up to 48 weeks. Once randomized, no changes in ESA dose or administration of IV or oral iron were allowed. During the randomized treatment period, patients meeting pre-defined anemia management criteria (ESA dose change or IV iron administration for the development of iron deficiency) completed the study and were transitioned into an open label extension. Results: Dialytic transfer of Triferic with each HD was reliable and not significantly affected by dialyzer membrane type or reuse. A greater number of placebo subjects (57%) than Triferic subjects (46%) met pre-defined criteria for a change in anemia management and transitioned into the open-label study. IV iron was required by more subjects with placebo (12%) than Triferic (2%).

The strength of the mAb 20.1-induced stimulation obtained with BTN3A1-transduced CHO cells is remarkable and considerably higher than that observed with CHO Chr6 BTN3A1 cells (Fig. 1) or RAJI cells [8]. To some extent this might be due to the three- to fourfold higher BTN3A1 expression by CHO BTN3A1 compared with CHO Chr6 BTN3A1 cells,

but it cannot be excluded that Chr6 negatively affects this type of activation. Other cell type-specific features of the presenting cells may also act on reporter cell activation, e.g. BTN3A1-transduced murine L929 cells which, like CHO BTN3A1 cells, do not present PAg, induce a much weaker, although statistically significant, mAb 20.1-triggered response (data not shown). The strong mAb 20.1-induced stimulation observed with CHO BTN3A1 cells as presenters and Vγ9Vδ2 TCR53/4-CD28+ Maraviroc order T cells as reporter cells contrasts with Vavassori et al. [12], who found an excellent PAg- but not mAb 20.1-induced activation of Vγ9Vδ2 GSK-3 inhibitor TCR-transgenic mouse reporter cells. Again differences between PAg- and mAb 20.1-induced activation could be due to differences between the presenting cells (human

A375 cells versus rodent cells), but could also be due to different reporter cell origin (cell line versus artificially in vivo-matured Vγ9Vδ2 T cells [12, 14]) and, finally, different TCR specificities might also play a role. Surprisingly, CHO Chr6 BTN3A1 cells, as well as several of the Chr6-containing hybridomas (Fig. 1), lacked the capacity to activate the reporter cells in the presence of the sec-butylamine. Alkylamines and aminobisphosphonates inhibit FPPS and therefore have been proposed to act by causing accumulation of IPP in the presenting cells [5]. Thus, either inhibition of FPPS activity is not the common feature that makes both substances Vγ9Vδ2 T-cell activators or alkylamines require additional cellular compound(s), which are missing in CHO Chr6 BTN3A1 cells, to exert FPPS inhibition. Independent evidence that, in addition to BTN3A1, human Chr6 is needed to convert rodent

cells into mediators of PAg-dependent Vγ9Vδ2 T cell activation was obtained using total human PBMCs as reporter cells. In this experiment we did not include mAb 20.1, since binding to BTN3A1 expressed by the γδ T cells would have complicated interpretation. Forskolin datasheet Figure 2 summarizes data comparing zoledronate-pulsed CHO Chr6 and CHO Chr6 BTN3A1 cells for induction of CD69 expression by the Vγ9Vδ2 T-cell population contained in total PBMCs. Most importantly, only the Vδ2+ T cells (essentially identical to Vγ9Vδ2 T-cell population) were activated. Furthermore, activation was better with CHO Chr6 BTN3A1 than with CHO Chr6 cells, while CHO (not shown) and CHO BTN3A1 cells failed to induce a PAg response. This experiment provides independent experimental proof that, in addition to BTN3A1, other gene(s) on Chr6 are indeed mandatory for PAg action.

Two ml 0·5% bovine serum albumin (Sigma, St Louis, MO, USA) in IsoFlow (Beckman

Coulter, Lane Cove, NSW, Australia) was then added and the tubes centrifuged at 300 g for 5 min. After decanting supernatant, Fc receptors were blocked with 10 μl human immunoglobulin (Intragam, CSL, Parkville, Australia) Alpelisib for 10 min at room temperature. Five μl of appropriately diluted anti-CD8 FITC (BD), anti-CD3 PerCP-Cy5·5 (BD), CD28 PE-Cy7 (BD) and PE-conjugated granzyme B (BD), 10 μl undiluted perforin (BD) or isotype control monoclonal antibody was added for 15 min in the dark at room temperature. Cells were analysed within 1 h. Samples were analysed by live gating using FL3 staining versus side-scatter (SSC). A minimum of 10 000 CD3-positive, low-SSC selleck screening library events were acquired in list-mode format for analysis. Control staining of cells with anti-mouse immunoglobulin (Ig)G1-PE/IgG-PC5 was performed on each sample and background readings of < 2% were obtained as described previously

[8]. Significant co-stimulatory molecule expression on T cells other than CD28 requires T cell stimulation similar to that required for intracellular cytokine production [14]. For CD154, CD152, CD137 and CD134, 1-ml aliquots of blood (diluted 1:2 with RPMI-1640 medium) were placed into 10 ml sterile conical polyvinyl chloride (PVC) tubes (Johns Professional Products, Sydney, Australia). Phorbol myristate (25 ng/ml) and ionomycin (1 mg/ml) were added to stimulate the T cells.

Brefeldin A (10 mg/ml) was added to prevent shedding of Protirelin the co-stimulatory molecules from the T cell surface, as reported previously [15]. The tubes were incubated in a humidified 5%CO2/95% air atmosphere at 37°C. At 16 h 100 ml 20 mM ethylenediamine tetraacetic acid/phosphate-buffered saline (EDTA/PBS) was added to the culture tubes, which were vortexed vigorously for 20 s to remove adherent cells. Three hundred microlitre aliquots of cells were treated with 2 ml FACSLyse for 10 min. Cells were centrifuged, supernatant discarded and 500 ml FACSPerm added for 10 min. Two ml 0·5% bovine serum albumin (BSA) (Sigma) in IsoFlow (Beckman Coulter) was then added and the tubes centrifuged at 300 g for 5 min. After decanting supernatant, Fc receptors were blocked with 10 ml human immunoglobulin (Intragam, CSL, Parkville, Victoria, Australia) for 10 min at room temperature. Five μl of appropriately diluted CD8 FITC [allophycocyanin (APC)-Cy7], CD3 PerCP-Cy5·5 (BD), CD28 PE-Cy7 (BD), CD45 V450 (BD) and PE-conjugated monoclonal antibodies to CD40L, CD152, CD137, CD134 or isotype control (BD) were added for 15 min in the dark at room temperature. Cells were washed and events acquired and analysed as described above.

However, prolonged-culture with IgE failed to alter the defective degranulation response in αβFFFγ2 cells (Fig. 4D). Moreover, wortmannin completely

prevented the degranulation response in αβYYYγ2 cells, but not in αβFFFγ2 cells (Fig. 4E). Since activation of Grb2-associated binder 2 (Gab2) is crucial for PI3K-signaling in mast cells 27–29, we examined tyrosine phosphorylation of Gab2 by using immunoblotting with an antibody that specifically recognizes Gab2 (Tyr452). BMMC were cultured with 0.5 μg/mL of anti-TNP IgE (IgE-3) or anti-DNP IgE (SPE-7) for 4 or 48 h. Low-dose of TNP-BSA or DNP-BSA triggered a low level of tyrosine phosphorylation of Gab2 in BMMC cultured with each IgE for 4 h, and adenosine significantly increased this phosphorylation level (Fig. 5A). In addition, prolonged-cultures of BMMC with each IgE further increased the amplified phosphorylation

Protein Tyrosine Kinase inhibitor level of Gab2. We further examined whether adenosine itself triggers tyrosine phosphorylation of Gab2 in BMMC. As shown in Fig. 5B and C, adenosine loading induced tyrosine phosphorylation of Gab2 in BMMC cultured with 0.5 μg/mL of IgE. Under the culture conditions, SPE-7 was more helpful IgE clone for the adenosine-induced Gab2 phosphorylation than IgE-3. Figure 5D shows that monovalent hapten DNP-lysine did not abolish adenosine-induced Buparlisib supplier Gab2 phosphorylation in BMMC cultured with SPE-7 for 48 h. The finding excludes the possibility that the effect of prolonged-culture with SPE-7 on Gab2 phosphorylation was due to FcεRI cross-linking. We next examined the roles of FcRβ-ITAM in the amplification of Gab2 tyrosine phosphorylation by adenosine (Fig. 6A). Upon antigen stimulation, αβYYYγ2 and αβYFYγ2 mast cells showed tyrosine phosphorylation of Gab2, whereas αβFFFγ2 and αβFYFγ2 mast cells failed to cause tyrosine phosphorylation of Gab2. The phosphorylation level in αβYYYγ2 and αβYFYγ2 cells was increased by adenosine loading. The Gab2 phosphorylation level in αβFYFγ2 cells was also somewhat amplified. In contrast, amplification of Gab2 tyrosine phosphorylation in αβFFFγ2 mast cells was thoroughly undetectable. After prolonged culture of αβFFFγ2

cells with IgE, adenosine-induced phosphorylation of Gab2 became detectable, but the level of phosphorylation was much lower than that in αβYYYγ2 cells (Fig. 6B). Collectively, Branched chain aminotransferase these results clearly indicate that FcRβ-ITAM plays an essential role in Gab2 tyrosine phosphorylation in mast cells. To clarify the molecular mechanisms of FcRβ-ITAM-dependent Gab2 phosphorylation following adenosine stimulation, we employed Fyn−/− BMMC and Lyn−/− BMMC to examine the role of Src family kinase which is thought to act upstream of Gab2. Fig. 7A and B clearly showed an indispensable role of Lyn kinase in tyrosine phosphorylation of Gab2 induced by adenosine. We further examined tyrosine phosphorylation of a signaling complex that contains Lyn in αβYYYγ2 and αβFFFγ2 mast cells following adenosine loading. Fig.

Thus, in the absence of SAP or CD84, CD4+ T cells are unable to form stable conjugates with B cells and cannot deliver help to B cells.47,51 In addition, this prevents the CD4+ T cells from receiving signals from B cells that regulate the formation or maintenance of Tfh cells. While

it is thought that Tfh cell development is a multi-step process with initial activation on DCs, followed by secondary signals provided by B cells, several recent findings have challenged this view. Many reports have demonstrated that Tfh cell numbers are decreased in the absence of B cells or when T–B cell interactions are disrupted.5,9,16,35,36 However, we recently showed that in the absence of antigen presentation by B cells, Tfh cell development (as indicated by surface phenotype and GC localization) could at least partially be PD-0332991 purchase rescued in the presence of abundant antigen, which prolonged presentation by DCs.9 Consistent with this, a recent study found that Akt inhibitor Tfh cells also developed in B cell-deficient mice in response

to chronic viral infection.52 This suggests that the requirement for B cells results not from a unique signal that B cells provide, but because Tfh cells need prolonged antigen stimulation and B cells often quickly become the only cells capable of presenting antigen to the T cells.9 A requirement for prolonged antigen presentation is consistent with data indicating a crucial role of TCR signalling in Tfh cell development. For example, many of the features of Tfh cells, such as up-regulation of CXCR5 and PD-1 and down-regulation of CD127, are observed in T cells following TCR stimulation.3,6,53,54 Moreover, it has been shown that high-affinity GBA3 T cells are preferentially selected to become Tfh cells.55 The restriction of antigen presentation to the B cells presumably occurs ordinarily because, first, the B cell receptor allows for efficient uptake of antigen and secondly, as the T cells move

into the B cell follicle and then the GC, these are the antigen-presenting cells (APCs) which the T cells encounter. Furthermore, several new papers support the idea that early activation on DCs is able to drive differentiation of Tfh cells. They demonstrated that CD4+ T cells with a Tfh cell phenotype – high CXCR5, PD1, IL-21 and Bcl-6 expression – could be identified early on in the response (e.g. day 3)21–23 in the interfollicular zone or outer follicle.21,22 This early appearance of Tfh-like cells was independent of B cells;21,23 however, the continued maintenance of these cells was disrupted in the absence of B cells.21–23 This suggests that a role of the second round of signalling, usually provided by B cells, may be to maintain a Tfh cell phenotype or the survival of Tfh cells rather than to drive unique differentiation events. Generation of the different Th lineages is associated with the action of particular cytokines.

We then employed H5N1 infection as

a model to study the antiviral activity of α-defensin-induced MxA. The viral plaque assay in Fig. 4A shows that, similar to IFN-α-pretreated HGECs, α-defensin-1, -2, and -3-pretreated cells significantly inhibited H5N1 replication, suggesting a functional MxA protein. On the other hand, β-defensin-1, -2, -3, and LL-37-pretreated HGECs poorly inhibited viral replication. These findings Lenvatinib clinical trial were confirmed by microscopically observed cytopathic effects (data not shown). To confirm the antiviral activity of MxA against H5N1, we transfected HGECs with MxA-targeted siRNA, treated the cells with α-defensin-1 overnight, and then infected them with H5N1 virus. MxA-targeted siRNA greatly reduced levels of MxA mRNA expression by 95%, (Fig. 4B) and effectively abolished inhibition of viral replication by 93% in H5N1-infected HGECs (Fig. 4C). These findings were supported by microscopically observed cytopathic effects (Fig. 4D). α-defensins are known as major proteins secreted by PMNs [[32]]. In the physiological condition of healthy gingiva, PMNs and their products are present in the tissue and the crevicular fluid in the gingival sulcus [[33, 34]]. In vitro culture of PMNs (5 × 106 cells/mL)

for 6 h led to secretion of α-defensins in supernatants (which ranged from 90 479 to 98 714 pg/mL). To investigate the role of the PMN-derived α-defensins selleck screening library in MxA expression, we cultured HGECs with 6 h PMN supernatants. Under this condition, expression of MxA at both mRNA and protein levels in HGEC was observed after 6 h and 24 h treatment, G protein-coupled receptor kinase respectively (Fig 5A and B). The MxA-inducing activity was diminished when neutralizing antibody against

α-defensins was added to the culture, whereas neutralizing antibodies against type I IFN (IFN-α and IFN-β) had no effect (Fig. 5B). These data suggest that PMN-derived α-defensins were responsible for the observed MxA expression. The immunostaining results to detect epithelial MxA were obtained using the oral, but not the sulcus, side of periodontal tissue (Fig. 2) because the epithelium at the sulcus side, especially for the junctional epithelium, is generally lost or torn during the surgical procedure. Fig. 6A depicts anatomic landmarks of the gingival sulcus. In this study, we were able to obtain two specimens of gingival sulcus area from healthy periodontal tissue. We then investigated localization of MxA protein in the healthy sulcus and also in relation to α-defensin. Fig. 6C shows that MxA protein was consistently expressed throughout epithelial cells of periodontal tissues. MxA staining was especially intense in the junctional epithelium (Fig. 6C). α-defensins were identified in small round cells with PMN morphology, most of which were found in the connective tissue layer (Fig. 6E). Migratory PMNs in junctional epi-thelium were also observed and highlighted in Fig. 6D.

We hope for continuous EFIS-EJI support for future meetings, which is indispensable as it provides travel grants for a significant number of young immunologists who attend the conference. The next conference is planned for September 2012 and the details will be posted on http://www.img.cas.cz/tatra/ approximately one year in advance of the meeting. Perhaps we will see you there. Radek Špíšek Department of Immunology, Charles University, 2nd Faculty of Medicine, University Hospital Motol, Prague, Czech Republic e-mail: [email protected]

“
“The behavior of self-reactive T cells 3-deazaneplanocin A mw in the peripheral immune system has often been studied by following the fate of adoptively transferred antigen-specific T cells in antigen expressing mice. In most cases, after a period of expansion, such cells undergo a slow clonal deletion, accompanied by the onset of anergy and/or suppression in the remaining cells. Here, we demonstrate that at initial frequencies approaching those found in normal repertoires, it is possible to completely avoid deletion Apoptosis inhibitor and still maintain peripheral tolerance. At starting numbers of <1000 T cells, stimulation by chronic self-antigens resulted in a period of robust clonal expansion, followed by a steady plateau phase extending

beyond 4 months. Despite their Bay 11-7085 stable persistence, the self-reactive T cells did not convert to a Foxp3+ fate. However, they displayed a considerable block in their ability to make IL-2, consistent with the onset of anergy — in a precursor frequency or deletion independent fashion. In an adaptive immune repertoire, the frequency of T cells that are specific for any given pathogen is thought to be very low. Although the precise numbers are difficult to estimate, in the mouse, it is thought to range in frequency from 1/1000 to 1/100,000 [1, 2] and numerically as low as 20 per mouse [3, 4]. The robust clonal expansion and differentiation that follows antigen recognition in vivo, is therefore geared to expanding

these rare precursors to large numbers of potent effector cells, in a short amount of time. However, the same process can be lethal if the target epitope is derived from a self-antigen. Therefore, the vertebrate has evolved several processes to curtail self-reactive T cells. After central tolerance deletes a large proportion of these, very few escape to the periphery. This makes it even more difficult to isolate and follow their behavior in unmanipulated animals (until an autoimmune process activates and expands them). Instead, we and others have routinely used adoptive transfer model systems that infuse a traceable population of self-reactive T cells into mice and follow their fate.

Recently, the delineation of human memory B cells by expression of CD27 has been challenged by the characterization of CD27-negative B cells (IgD-CD27-), indicating molecular imprints

of memory B cells (somatic hypermutation and immunoglobulin class-switch) [9,10]. Plasmablasts or plasma cells can be identified readily by an increased expression of CD38 and CD27 compared to memory B cells. The most immature peripheral B cell population in humans has been characterized in detail recently by the concomitantly high expression of CD24 and CD38 [11–13]. A CD21lowCD38low B cell subset has been shown to be expanded in autoimmune diseases and immunodeficiencies [14–16]. Recently, this B cell population has been selleck monoclonal humanized antibody inhibitor characterized

as tissue homing, innate-like B cells, containing autoreactive unresponsive B cell clones [16,17]. Using these flow cytometric approaches, changes in the peripheral Tyrosine Kinase Inhibitor Library manufacturer B cell pool have been documented to take place at distinct differentiation stages according to the underlying diseases. Several autoimmune diseases are characterized by an expansion of plasmablasts/plasma cells in the peripheral blood, indicating aberrant B cell development and activation [18]. In contrast, impairment of central or peripheral B cell development takes place in several immunodeficiencies [1,14]. Of interest, B cell regeneration after stem cell Celecoxib transplantation or B cell-depleting therapies seems to follow a tightly regulated chronology of B cell reappearance [12]. However, age-dependent reference values for distinct B cell

populations are reported only rarely [19,20]. Therefore, we analysed and quantified different peripheral B cell populations in a cohort of individuals ranging from neonates to adults and tried to establish age-dependent reference values for distinct peripheral blood B cell populations, which can help in the characterization of impaired or disturbed peripheral B cell development. Between November 2007 and August 2009 221 healthy individuals aged 1 month to 50 years were enrolled in this study. The group of healthy individuals consisted of children who were referred to the out-patient clinic at the Children’s Hospital of the University of Würzburg for diagnostic blood testing. Immunological, infectious or haemato-oncological diseases were ruled out in these children. Most of the individuals underwent routine blood testing before minor surgical or diagnostic procedures. Additionally, healthy medical students as well as employees of the University Hospital Würzburg donated blood samples on a voluntary basis. The study was reviewed by the ethics committee of the University of Würzburg and was performed according to the modified declaration of Helsinki. Venous blood was collected, anti-coagulated with ethylenediamine tetraacetic acid (EDTA) and processed within 24 h.

There were 635 accepted abstracts, and a total of 145 oral presentations. In addition Doxorubicin purchase to all this immunology, the meeting had a vibrant social program (as discussed below). The registration fee of the main conference was kept affordably low, taking into account the difficult economic situation in which all of us currently live and the cuts that have hit the research community in recent years. Fortunately, the meeting received crucial support from 7 silver and 17 bronze sponsors (http://www.immunology2011.it/sponsor.asp),

7 minor sponsors, 6 pharmaceutical companies for the clinical symposia and the cooperation of 2 media operations, including the European Journal of Immunology. As a teaser, just before the opening ceremony, the opening symposium entertained the fascinating new developments in microscopy that allow cells of the immune system to be tracked in vivo, capturing the dynamics of cellular movements and interactions. While M. Gunzer (Magdeburg/Essen) observed neutrophils at work, M. Iannacone (Milano) followed lymphocytes in a viral infection. How microscopy can be used to identify and track individual molecules was discussed by M. Reth (Freiburg), who provided evidence for an oligomeric Smad inhibitor resting state of the B-cell antigen receptor and the perturbation

of this state by activation. The opening ceremony started with the two national anthems followed by a concert given by a duo over from Modena: the Butterflies. Francesca Bergamini, vocals, and Alessandra Fogliani at

the piano, performed songs in German, Italian, Spanish and English (Fig. 1). The first keynote lecture of the meeting was sponsored by EFIS and given by Prof. Klaus Rajewsky (Boston, USA). He presented his in-depth analysis of B-cell activation and the role of c-myc and IKK in the pathogenic transformation for the survival and expansion of lymphoma cells. At the end of the opening ceremony, the President of the DGfI, Prof. Dieter Kabelitz (Kiel), awarded Prof. Hans-Hartmut Peter (Freiburg) honorary membership of the DGfI for his extraordinary impact on clinical immunology and rheumatology, and his contributions to the understanding of immunodeficiencies. After the opening session, high up on the PalaRiccione terrace with its impressive view of the sea bathed in a beautifully colored sunshine, a famous brass band from Münster (the NorthWestBrass, led by Kapellmeister Roland Göhde, Fig. 2) had the opportunity to present a new poly-functional program – from J. S. Bach to Bob Dylan, passing through Gershwin, Henry Mancini, The Beatles, Abba – to more than 600 persons who were also interested in testing the speed of evaporation of 350 bottles of ice-cold Prosecco (from Travani A. et al., Arzene, Italy, a total of 262.

18 A STAT-5 phosphorylation assay was used to gauge IL-7 responsiveness in CD4+ and CD8+ cells. The increase of the percentage of P-STAT-5 cells, and an example of constitutive P-STAT-5 and IL-7-induced P-STAT-5, in HD and NHP are shown in Fig. 5(a,b). In NHPs, (n = 15) 84·4 ± 10·8% and 60·6 ± 12% of CD4+ and CD8+ cells showed an increase of P-STAT-5 cells in response to IL-7 stimulation. Similar numbers were obtained in PBMCs from HDs (n = 10): 87·6 ± 7·6% and 62·3 ± 15·4% in CD4+ and Opaganib concentration CD8+

cells, respectively. CD4+ and CD8+ subsets showed comparable responses to IL-7 stimulation as measured by STAT-5 phosphorylation in NHPs and HDs. In HDs and NHPs more CD4+ cells than CD8+ cells showed STAT-5 phosphorylation (consistent with higher levels of IL-7Rα expression on CD4+ T cells) but the amplitude (measured by MFI) was not statistically different between CD4+ and CD8+ cells. The presence of regulatory cells was assessed by expression analysis of CD25 and FoxP3 in the CD4+, CD8+ and CD4+ CD8+ T-cell compartments (gating strategy see Supplementary Fig. S2). In NHPs, the

CD4+ T-cell compartment showed the following frequencies: 16·5% (median values) were CD25intermediate (CD25interm.) and 0·5% stained for CD25bright; in CD4+ CD8+ T cells: 19·6% cells were CD25interm. and 1·4% were CD25bright; in CD8+ T cells: 1% were CD25interm. and 0·07% CD25bright (Table 2). Compared with HDs, higher frequencies of CD4+ CD25interm. T cells and CD4+ CD8+ CD25interm./bright,

and CD8+ CD25bright T cells (21%) were detected in PBMCs from NHPs. Analysis RAD001 concentration of FoxP3 expression in the different CD25+/− T-cell compartments showed that the majority of CD25bright T cells in NHPs were FoxP3+ (87·5% of CD4+ and 76% of CD4+ CD8+ and CD8+ T cells), whereas ADP ribosylation factor only 10–20% of CD25interm. T cells showed FoxP3 expression (Fig. 6a). In contrast, almost no FoxP3 expression could be observed in human CD4+ CD8+ CD8+ T cells that exhibited the CD25interm. phenotype. Analysis of PBMCs from four of eight HDs showed that CD4+ CD8+ and CD8+ T cells, which displayed a CD25bright phenotype, stained also positive for FoxP3. In summary, statistically higher frequencies (P ≤ 0·05) of CD4+/− CD25interm.FoxP3+/−, CD4± CD8± CD25interm./high FoxP3+/− and CD8± CD25interm./high FoxP3+/− were detected in NHPs than in HDs. Expression of the IL-7Rα on NHP CD25bright T cells was inversely correlated with expression of FoxP3, which is similar to the situation described in humans.25 More than 90% of NHP CD4+ CD8+ CD25interm./bright FoxP3+ T-cell subsets did not express the IL-7Rα, whereas the majority of CD4+ CD8+ CD25+/− FoxP3− (33–67%) were positive for IL-7Rα (% of IL-7Rα expression is shown for CD4+ T cells in Fig. 6b). The same trend was observed in HDs. However, 9% of human CD4+ CD25bright FoxP3+ T cells expressed the IL-7Rα (Fig. 6b), this was not true for the same T-cell subset in NHPs (3·8%).