0) for 40 min, followed by a blocking step for 1 h (Roti-Immuno Block, dilution 1:10; Roth, Karlsruhe, Germany). Primary anti-human FUBP1 antibody was incubated overnight at 4°C and subsequently labelled with a secondary antibody for 1 h at room temperature (RT) (dilution 1:500; Alexa Fluor568, donkey anti-goat IgG, Invitrogen, Darmstadt, Germany). Next, the primary antibodies for the double staining (as listed above) were added and incubated for 1 h at RT and specimens

were then labelled with an additional secondary antibody for 1 h at RT (dilution 1:500; Alexa Fluor488, goat anti-mouse IgG, Alexa Fluor488, goat anti-rabbit IgG, Invitrogen). Nuclear staining was performed with DAPI (dilution 1:1000, Invitrogen). The images were analysed and recorded on a Nikon Eclipse 80i fluorescence microscope (Nikon,

Düsseldorf, Germany). Digital images were then adjusted AZD2281 mw Ixazomib with NIS elements imaging software (Nikon). Thirty samples were analysed by fluorescence in situ hybridization (FISH) to assess for 1p and 19q deletions. The two-colour FISH assay was performed on 3-μm-thick sections using a mixed 1p36/1q25 dual colour probe and 19p13/19q13 dual colour probe set (ZytoLight SPEC, Cat. No. Z-2075 and Z-2076, Zyto-Vision, Bremerhaven, Germany). The Histology Accessory FISH Kit (Dako) was used for slide pretreatment, probe hybridization and post-hybridization processing. Nuclei were counterstained with DAPI/Antifade-Solution (Zyto-Vision). Fluorescent signals were others analysed using an Olympus BX50 fluorescent microscope with the appropriate filters (Olympus, Hamburg, Germany). Samples displaying sufficient FISH efficiency (80% fluorescent nuclei) were evaluated. Signals were scored in at least 100 non-overlapping, intact nuclei. Deletions of 1p or 19q were defined by samples with over 50% of the tumour nuclei containing only one signal. The FUBP1 gene (ENSG00000162613; ENST00000370767) was analysed for mutations in 15 tumour samples using the primers shown in Table 1. All exons listed were amplified using GoTaq polymerase (Promega, Mannheim, Germany). PCR products were treated with

ExoSAP (ExoSAP-IT, GE Healthcare, Pittsburgh, PA, USA) and sequenced in both directions using an ABI PRISM 3100 Genetic Analyser (Applied Biosystems, Foster City, CA, USA). Additionally, the number of FUBP1 mutated gliomas was increased by samples deriving from a previously studied cohort [4]. A semiquantitative score was used for the analysis of FUBP1 protein expression. The immunohistochemical staining intensity value (no staining = 0, low = 1, moderate = 2, strong = 3) was multiplied with the assigned value for the proportion of positive tumour or vascular cells (0–1% = 0, 1–10% = 1, 10–25% = 2, 25–50% = 3, > 50% = 4). MIB-1-positive cells were analysed as a percentage of all cells within the tumour and then plotted against FUBP1 expression scores followed by the Wilcoxon rank sum test.

Employees and housewives comprised selleck chemicals llc around 70% in our study group. Patients with a yearly income below NT$400 000 (US$12 000) comprised 71%. (The annual average GNP of Taiwan in 2003 was around US$ 12 000.) (Table 1) Forty-four percent of the patients indicated that they felt stress pressure from their life. The onset of first symptom occurred at the age of 37.6 years (ranging from 18 to 81 years).

The duration of urgency/frequency was 62 months (ranging from 9 to 396 months). The duration of pain was 46 months (ranging from 9 to 492 months). The average daily voiding frequency was 16 times (ranging from 9 to 50 times), including 3.7 times (ranging from 1 to 18 times) during sleeping time. While 94% had frequency and complaint, 80% suffered from pain, 53% had nocturia, 10% associated with incontinence. Forty-seven

percent of the IC patients in the study complained that their symptoms were persistent in nature. Eighty-three percent of the pain with a full urinary bladder was the prominent pain characteristic followed by 74% of pain relief after voiding. Forty-five percent reported pain when their urinary AZD2281 research buy bladders were not full. Forty percent had burning pain during voiding. Fifty-four percent of the IC patients in the study complained the type of pain was a full sensation, followed by 32% of soreness, 22% of sharp, 21% of stabbing, 11% of spasms, 8% of dull, and 4% of throbbing. Fifty-two percent of patients pointed to the pain at the lower abdomen area, 23% at suprapubic, 22% at vagina,

14% at left low abdomen, 12% at right low abdomen, 11% at left flank area, 10% at right flank area, 9% at inguinal area, and 8% at low back area (Table 2). The factors that aggravated interstitial cystitis symptoms were screened. Clomifene Among the factors, 44% of the patients indicated that stress was the factor most frequently encountered, followed by 31% of urinary tract infection. Beverages such as tea and coffee were the most frequent fluid that would aggravate IC symptoms. Oranges and pineapple were the most noted fruit that made IC symptoms worse (Table 3). The most associated diseases were recurrent urinary tract infection (28%), migraine headache (24%), neurodermatitis (21%), and hay fever/allergic rhinitis (20%). The family history of the IC patients in this study were hypertension (18%), diabetes mellitus (14%), hay fever/allergic rhinitis (11%), heart disease (18%), urinary stone (10%), migraine headache (7%), neurodermatitis (7%) (Table 4). Thirteen percent of female patients had the history of hysterectomy and 15% had tube ligation. The average of doctor visitation was 3.2 doctors (1–37) and traditional doctor 1.3 doctors (0–10) before diagnosis. Eleven percent had a history of anti-depression or anti-anxiety drug intake. Five percent had an allergic drug intake. The average number of the children from married patients discussed was 1.9 persons. Sixty percent of these children were normally delivered.

Fifty-four patients were enrolled in the study and received study treatment (from six centres in Brazil, one in Chile, two in Colombia, two in Mexico and one in Panama). Appropriate patient selection for candidaemia studies remains challenging due to issues associated with early identification of infection and a variety of concomitant risk factors; insufficient enrolment to this study meant that the target of 210 patients was not achieved. Patient disposition is shown in Fig. 1. In total, the per protocol population (all MITT

subjects who were compliant with the study protocol) comprised 22 (40.7%) patients and 32 (59.3%) patients discontinued the study prematurely; the most common reason for discontinuation was death (n = 23, 42.6%), followed by lack of efficacy (n = 4, 7.4%), other reasons (n = 3, 5.6%), AEs (n = 2, 3.7%), lost to follow-up, and no longer willing to participate (both n = 1, 1.9%). Other reasons included a legal representative

selleckchem withdrawing informed consent, voriconazole being added to treatment due to isolation of moulds and yeast in blood culture, and doctors and relatives not accepting continuation of treatment due to diagnosis of brain death. Forty-four patients were included in the MITT population and the overall median duration of therapy with IV anidulafungin was 9.5 days (range 2–25 days). Ten patients were excluded from the MITT population because they did not selleck compound have a positive baseline culture for Candida within 96 h before study entry. Patient demographics and baseline characteristics of the MITT population are included in Table 1. All patients enrolled in this study were in the ICU. At study entry, 72.7% Histone demethylase (33/44) of patients in the MITT had been in the ICU for ≥4 days; among these patients, the overall median duration of ICU stay was 16.0 (95% CI: 8.0, 29.0) days. Within the MITT population, 14 patients were able to step-down to oral voriconazole. These patients had a shorter median duration of treatment with IV anidulafungin

(6 days), compared with that of patients who did not step-down to oral therapy (14 days). Patients who stepped-down to oral voriconazole had lower APACHE II scores and lower incidences of solid tumours and prior abdominal surgery compared with patients who remained on IV anidulafungin (Table 1). Global, clinical and microbiological response rates for the MITT population are summarised in Table 2. The primary endpoint of global response rate at EOT for the MITT population was 59.1% (95% CI: 44.6, 73.6), when 13 patients with missing responses were counted as failures. Patients with an indeterminate or missing response could not be assessed for clinical or global response at the EOT because they either received less than three doses of anidulafungin or they died of a cause other than candidaemia before the planned EOT. At day 30, the all-cause mortality rate in the MITT population was 43.

We would therefore assume that migration of activated CD8+ T cells to the GT is in part random and affected by their overall frequencies in blood, and in part driven by the expression of yet to be identified homing markers. In either case, we would assume that activated CD8+ T cells receive signals from the microenvironment that favor buy NVP-BEZ235 their retention once they reach the GT, leading to an enrichment

of these cells at the mucosal surface, which is the port of entry for many pathogens. The functionality of genital CD8+ T cells remains to be investigated in more depth. Our data thus far show that T cells from the GT produce IFN-γ but not IL-2 as has also been reported for genital T cells in SIV-infected non-human primates 34. In our study, Gag-specific CD8+ T cells from the GT expressed high levels of

granzyme B, perforin and XL765 molecular weight Ki-67, which suggests that they are highly activated cells able to immediately commence target cell lysis and proliferation. Other authors have demonstrated atypical T cells within mucosal surfaces 22 and we speculate that the high levels of lytic enzymes seen in memory-type CD8+ T cells from the GT could be a result of a specific microenvironment. In summary, data presented here show that i.m. immunization with a replication defective AdC vector in mice induces a robust transgene product-specific CD8+ T-cell response within the GT that can be enhanced by a booster immunization given i.m. The response is sustained and can still be detected 1 year after immunization. Vaccine-induced genital CD8+ T cells are functional; they carry lytic enzymes

and release cytokines upon antigenic stimulation. Taken together, the results shown should allow for guarded optimism that potent vaccines administered i.m. may induce a genital barrier to HIV-1 infection in women. In fact, systemic regimens would be preferable over mucosal ones in humans due to the logistical factors and the lack of interference by flora or menstrual cycle, which may profoundly affect mucosal vaccine efficacy. Female 6- to 8-wk-old BALB/c mice were obtained from Ace Animals (Boyertown, PA). Female 6- to 8-wk-old Thy1.1 mice were obtained from The Jackson Laboratory (Bar Harbor, ME). Resminostat Animals were housed at the Animal Facility of The Wistar Institute (Philadelphia, PA) and all experiments were performed according to the institutionally approved protocols. Purified E1-deleted Ad vectors expressing Gag of HIV-1 clade B, derived from simian serotypes C6 (AdC6) or C68 (AdC68), were produced and quality controlled as described previously 8, 35. Groups of 5–20 BALB/c mice were immunized by i.m. or mucosal routes with AdC vectors diluted to 1010 viral particles in sterile saline to a total volume of 10 μL (i.n. and i.vag.) or 100 μL (i.m.). Mice were immunized i.m. by injection into the lower leg muscle, whereas mucosal immunization was given with an automatic pipette.

These results showed that mbIL-21-CD137L-K562 cells induced the generation of high-purity human NK cells from peripheral blood mononuclear cells. Besides CD56 and CD16, the NK cell surface has many other receptors, such as the activating receptors NKG2D, NKp30, NKp44, NKp46, NKp80, CD226 and 2B4, and the inhibitory receptors KIR2DL1, KIR2DL2 and KIR3DL1. The concerted action of these receptors determines NK cell lytic activity [2]. Therefore, we

analysed expression of the receptors on the expanded NK cell surface Mitomycin C mouse via flow cytometry. The results showed that other than the down-regulation of activating receptor NKp80, the expression of all other detected activating and inhibitory receptors were increased with the expansion (Fig. 3). In short, the data showed that expression of NK cell receptors were maintained, most MG-132 mw of which were up-regulated during expansion. Because balanced expression of NK cell receptors determines NK cell lytic activity, and both activating and inhibitory receptors (except for NKp80) were up-regulated in expanded NK cells, we evaluated the effectiveness

of NK cell-mediated killing via cytotoxicity assay. The results showed that NK cell killing activity increased with expansion and reached the highest point at 3–5 weeks, then began to decrease after 6 weeks, although still significantly higher than unexpanded (resting) NK cells (Fig. 4a). These results showed that expanded NK cells were activated and functioned properly. The goal of ex-vivo expansion was to produce large numbers Nintedanib (BIBF 1120) of functional NK cells. As the expanded NK cells were functional, the next objective was to evaluate NK cell proliferation by counting the total cell numbers after trypan blue staining. The results showed that NK cells

were increased significantly after expansion (Fig. 4b). Taken together, our results provide strong evidence showing that mbIL-21 could promote sustained NK cell proliferation and produce highly cytotoxic NK cells. Because mbIL-21-CD137L-K562 induced large-scale and sustained proliferation of functional NK cells from peripheral blood mononuclear cells, we wanted to investigate the mechanisms involved. By screening the phosphorylation status of STAT-1–6 via Western blot, we found that only STAT-3 was phosphorylated continually in primary NK cells (unpublished data), which led us to hypothesize that STAT-3 activation is required for human NK cell maintenance and expansion. To test this hypothesis, we first examined the effect of IL-21 on STAT-3 phosphorylation in human NK cells.

Group IV was designated as a combination group for inhalation and epidural

anesthesia. Group V was a combination group of inhalation and spinal anesthesia. Group III and group V showed significant increases in the number of rolling and sticking leucocytes and in RBC volume (peripheral stasis) when compared with group I. Blood flow and velocity significantly Erismodegib research buy increased without peripheral stasis in groups II and IV when compared with group I. Although there was no statistically significant difference in the numbers of rolling, sticking, and transmigrating leucocytes or in functional capillary perfusion, group IV had better flow hemodynamics in the peripheral microcirculation when compared with group I. The inhalation and epidural anesthesia see more combination was determined to be the ideal anesthesia technique for improved peripheral microcirculation. Spinal anesthesia, either separately or in combination with inhalation anesthesia, has adverse effects on microcirculation. © 2009 Wiley-Liss, Inc. Microsurgery, 2010. “
“The objective of this study was to compare the free muscle-musculocutaneous flaps and free perforator skin flaps used for soft tissue reconstruction of the lower extremities. Fifty-three patients whose skin and soft

tissue of the lower extremities had been reconstructed were divided into two groups: a perforator flap group, reconstructed using anterolateral thigh (ALT) free flap (23 cases), and a muscle-musculocutaneous flap group, in whom latissimus dorsi and rectus abdominus muscle-musculocutaneous free flaps were used (30 cases). Postoperative complications, long-term results, and donor site morbidities were studied in the two groups. Complete flap survival was 78.3% with four total and one

partial flap loss in the ALT group and 90.0% with one total and two partial failure in the muscle-musculocutaneous second flap group. Muscle-musculocutaneous flaps were the flaps of choice in Gustillo grade IIIB-C injuries and for reconstruction of more proximal localizations. ALT was preferred in relatively younger patients and was typically used for coverage of the distally localized defects. Flap complication rate was significantly higher in the ALT group, but the overall complication rate was similar between the groups. ALT perforator flap is a precious option for lower extremity soft tissue reconstruction with minimal donor site morbidity. Nevertheless, the beginners should be attentive to an increased rate of flap complications with the ALT flap and free axial muscle-musculocutaneous flaps would still be the tissue of choice for coverage of leg defects for a surgeon before gaining enough experience with perforator flap dissection. © 2009 Wiley-Liss, Inc. Microsurgery 2010.

We screened relevant studies according to predefined inclusion and exclusion criteria, evaluated the quality of the included studies, and performed meta-analyses

by using the Cochrane Collaboration’s Revman 5.1 software. Results:  We identified nine trials including 3098 patients. Meta-analysis showed statins can significantly decrease the serum C-reactive protein (CRP) (SMD, −0.54; 95% confidence interval (CI), −1.04 to −0.05; P = 0.03) and high sensitivity CRP (hs-CRP) level (SMD, −0.72; 95% CI, −1.14 to −0.31; P = 0.0007) of dialysis patients compared with that of the control group. However, statins did not differ significantly from the control group in increasing the serum Alb level (SMD, −0.13; 95% CI, −0.42 to 0.15; P = 0.37). Conclusions:  Statins can improve the chronic inflammation status reflected by the decreasing of serum CRP and hs-CRP levels, whereas PD-332991 there is no conclusive evidence that it can improve the nutrition status. However, this result needs to be further confirmed in more high-quality randomized clinical trials. “
“Cerebral white matter

hyperintensities (WMHs), comprised of periventricular hyperintensity (PVH) and deep and subcortical white matter hyperintensity (DSWMH), have been presumed to be predictors for future stroke, cognitive impairment and dementia in the general population. However, no longitudinal GABA Receptor studies have been performed to determine the clinical significance of WMHs in haemodialysis (HD) patients. In the present study, we investigated the influence CYC202 in vitro of WMHs as a predictor of future cardiovascular disease in HD patients. Cranial magnetic resonance imaging was performed on 179 HD patients with no past history of stroke

from April 2006 to October 2009, and the prevalence of WMHs was investigated. The patients were followed prospectively until March 2012 or death or renal transplantation. The influence of WMHs on cardiovascular events was investigated using the Kaplan–Meier method and Cox proportional hazards analysis. The patients with advanced PVH and DSWMH had a significantly higher incidence of cardiovascular morbidity than those without advanced PVH and DSWMH by Kaplan–Meier analysis. By multivariate Cox proportional hazards analysis, the presence of advanced PVH and DSWMH increased the risk of cardiovascular events, independent of other cardiovascular risk factors. In addition, the present study revealed that of the subtypes of WMHs, PVH was a stronger predictor of cardiovascular events compared to DSWMH. The present study indicates that the presence of WMHs is a novel predictor of cardiovascular events in HD patients, and that PVH is more closely associated with incident cardiovascular disease.

5, 3, 24 and 72 h after exposure. The cerebellum and hippocampus were subjected to Western analysis for VEGF, iNOS, eNOS, nNOS and AQP4 expression; ELISA analysis for cytokine and chemokine levels; and immunohistochemistry for GFAP/AQP4, RECA-1/RITC and TUNEL. Aminoguanidine (AG) was administered to determine the effects of iNOS after smoke inhalation. Both the cerebellum and hippocampus showed a significant Y-27632 supplier increase in VEGF, iNOS,

eNOS, nNOS and AQP4 expression with corresponding increases in inflammatory cytokines and chemokines and increased AQP4 expression and RITC permeability after smoke exposure. AG was able to decrease the expression of iNOS, followed by VEGF, eNOS, nNOS, RITC and AQP4 after Raf targets smoke exposure. There was also a significant increase in TUNEL+ cells in the cerebellum and hippocampus which were not significantly reduced by AG. Beam walk test revealed immediate deficits after smoke inhalation which was attenuated with AG. The findings suggest that iNOS plays a major role in the central nervous system inflammatory pathophysiology after smoke inhalation exposure with concomitant increase in proinflammatory molecules, vascular permeability and oedema, for which the

cerebellum appears to be more vulnerable to smoke exposure than the hippocampus. “
“J.-F. Ma, Y. Acyl CoA dehydrogenase Huang, S.-D. Chen and G. Halliday (2010) Neuropathology and Applied Neurobiology36, 312–319 Immunohistochemical evidence for macroautophagy in neurones and endothelial cells in Alzheimer’s disease Aim: To determine the pathological structures associated with macroautophagy in Alzheimer’s disease (AD) and any relationship to disease progression. Methods: Immunohistochemistry using antibodies to beclin-1, Atg5 and Atg12, early macroautophagy markers and LC3, the mammalian homologue of the later macroautophagy marker Atg8, were localized in formalin-fixed, paraffin-embedded medial temporal lobe sections of AD cases at variable neuritic disease stages.

Double immunofluorescence labelling was used to co-localize these macroautophagy markers with Aβ and phospho-tau (AT8) and correlations performed using Spearman rank tests. Results: Atg12 immunoreactivity in AD was either dispersed in the soma and dendrites or concentrated in tau-immunoreactive dystrophic neurites and some neurofibrillary tangles. Fewer Atg12-immunopositive neurones were observed with longer disease durations. Atg12-immunoreactive endothelial cells were found spatially associated with Aβ-positive plaques, with more Atg12-immunoreactive capillary endothelial cells with higher neuritic disease stage. These findings were confirmed by the other autophagy markers beclin-1, Atg5 and LC3.

IgM+ B cells in the CD3−CD19−MHC II+ population in the infected mice were mostly IgD−B220− and were distinct from those in uninfected mice (Fig. 2b). The morphology of each population was examined (Fig. 2c). CD11chi DCs and MHC II+CD11c−CD3−CD19−IgM+ cells from the infected mice were homogeneous in size and staining patterns. However, MHC II+CD11c−CD3−CD19−IgM− cells

were heterogeneous in size and may have included multiple cell types. The proportion of these MHC II+CD11c−CD3−CD19−IgM− cells in the peripheral blood and bone marrow were also examined (Fig. 2d). These cells increased in spleen, blood and bone marrow on days 6 and 8 post-infection, suggesting that greater numbers of them were being generated in the bone marrow. Since it became clear that the

CD3−CD19−MHC II+ population contained B cells, these IgM+ cells were excluded from further study, and we thereafter focused on CHIR99021 MHC II+CD11c−CD3−CD19−IgM− cells. The phenotypes of each MHC II+CD3−CD19−IgM− subset were examined next (Fig. 3a). MHC II+CD3−CD19−IgM−CD11chi cells are conventional DCs. Most of this population expressed CD11b, F4/80 and the costimulatory molecules CD80 and CD86. During P. yoelii infection, the proportion of cells expressing F4/80 was reduced, whereas that of cells expressing Ly6C was increased. Additionally, expression of CD40, CD80 and CD86 was increased. Selleck Mitomycin C MHC II+CD11cintCD3−CD19−IgM− cells, most of which expressed Ly6C, CD11b, CD80 and CD86, were a minor population in uninfected mice. This population may have contained several distinct subsets, including pDCs that express B220 and PDCA-1. Some cells in this group expressed NK1.1, suggesting that this group included NK DCs or interferon-producing killer DCs [23]. After 8 days post-infection, MHC II+CD11cintCD3−CD19−IgM− cells that expressed B220 and PDCA-1 had almost disappeared. Expression of their costimulatory molecules was upregulated. MHC II+CD11c−CD3−CD19−IgM−

cells, which may have contained several different cell types including those expressing B220, Ly6G, Ly6C, NK1.1, CD11b, and F4/80 were a minor population in uninfected mice, as were IgD+ B cells. Eight days post-infection, the number of these cells increased, whereas those expressing B220, 5-Fluoracil datasheet Ly6G, IgD, NK1.1, and F4/80 had almost disappeared. Thus, this population of MHC II+CD11c−CD3−CD19−IgM− cells in infected mice was distinct from those in uninfected mice and lacked expression of many cell type specific markers. Approximately 41% of this population expressed Ly6C and most appeared to express PDCA-1 to a moderate degree. To examine whether MHC II+CD11c−CD3−CD19−IgM− cells increase during P. yoelii infection in the absence of B and T cells, we infected Rag-2−/− mice with P. yoelii (Fig. 3b). After infection with P. yoelii, splenocytes from Rag-2−/− mice exhibited striking differences from those of wild-type mice. Infected Rag-2−/− mice (5.6 ± 0.8 × 107; parasitemia, 37.4 ± 21.9%) had more spleen cells than uninfected Rag-2−/− mice (1.1 ± 0.4 × 107).

The impact of TCR repertoire diversity on Treg-cell function is controversial. Regarding the prevention of autoimmune disease, previous studies on the effective suppression of EAE through Treg cells with

limited TCR repertoires came to divergent conclusions 47, 48. A recent study by Adeegbe et al. found that limited TCR diversity of transferred Treg cells was a risk factor for autoimmune disease in IL-2Rbeta−/− mice 49. Intriguingly, non-obese diabetic mice were recently shown to select a low diversity Treg-cell TCR repertoire 50. Understanding the parameters that govern Treg-cell homeostasis will be critical for the design of future Treg-cell-based intervention strategies. Sufficient availability of organ-specific antigen must be considered in translational attempts to manipulate organ-specific autoimmunity https://www.selleckchem.com/JNK.html with engineered Treg cells of known self-peptide specificity. Otherwise, exogenous therapeutic Treg cells may be lost quickly after transfer. Previous studies suggested that organ-specific self-antigen preferentially drives the survival and/or expansion of organ-specific Treg-cell clones 11, 13, 21, 22. Our results also support the view that the antigen specificity of Treg cells changes by anatomical location, although

MK-1775 cost TCR sequences of recovered Treg cells from pLNs and mLNs were largely overlapping. This may be the result of two possible scenarios. Either Treg cells recirculate less than naïve T cells or differences are due to selective local survival. Importantly, our study infers that Treg-cell diversity is connected to diversity and availability of specific self- and foreign-antigen and thus the amount of DCs presenting it on MHC class II. In accord, it was recently shown that DC ablation Liothyronine Sodium reduced Treg-cell frequencies 51, 52, whereas an increase of DC numbers by FLT3L treatment led to expansion of peripheral naturally occurring Treg cells 52,

53. However, in the latter report, it was concluded that Treg-cell proliferation was mainly IL-2 dependent. In our study, we also recognized IL-2 as a master regulator that controls the absolute size of the Treg-cell pool. We propose that an optimal and maximally broad organ-specific Treg-cell TCR repertoire is continuously shaped by inter- and intraclonal competition for diverse antigen. Within a peripheral Treg-cell niche, sufficient population diversity seems to be crucial for proper Treg-cell function. Hence, in future studies, HT-sequencing analysis of Treg-cell diversity may be suitable to predict the relative risk of T-cell-mediated diseases. C57BL/6-Foxp3eGFP (here: WT) 54, C57BL/6-Foxp3.LuciDTR-4 36, and C57BL/6-Tg(TcraTcrb)425Cbn/J (here: OT-II/TCR-Tg) 55 mice have been described. The Thy1.