To determine if the stimuli enhanced Selleckchem Afatinib the S6 phosphorylation, PDC were stimulated with CpGA or loxoribine in the presence of IL-3 and intracellular p-S6 expression was determined with flow cytometric staining (Fig. 1b). CpGA stimulation resulted in the same fluorescence intensity as IL-3 treatment alone, while loxoribine stimulation slightly increased the p-S6 expression. CpG-A was a more effective stimulus than loxoribine to induce IFN-α secretion (Fig. 1c). While 20 ng/ml rapamycin inhibited loxoribine-induced IFN-α secretion by 64%, it inhibited CpG-A-induced IFN-α secretion by only 20%, despite almost complete suppression of mTOR-signalling. In contrast, secretion of the proinflammatory cytokines IL-6 and TNF-α was inhibited

by rapamycin with similar efficacy in both stimulation conditions (Fig. 1d). The observed inhibitory effects of rapamycin were not due to

general impairment of PDC function, because no inhibition of CXCL-10 secretion was observed (Fig. 1d) and rapamycin did not induce apoptosis, as demonstrated by the absence of active caspase-3 (data not shown). As mTOR inhibition decreased cytokine secretion by PDC, we reasoned that mTOR stimulation might increase cytokine production. Therefore we added 10 nM VO-OHpic trihydrate, a specific inhibitor of PTEN, during PDC activation. The upstream signalling pathway that activates mTOR is initiated by phosphatidylinositol 3-kinase (PI3K), which generates 3-phosphorylated inositol lipids (PIP3) [23]. PTEN is a negative regulator of PIP3K-signalling

because it dephosphorylates PIP3 [24], and therefore inhibition of PTEN can abrogate negative regulation of mTOR phosphorylation. see more Cyclooxygenase (COX) The addition of VO-OHpic trihydrate to TLR-activated PDC in a concentration that increased generation of PDC from human CD34+ progenitor cells [25] did not, however, affect p-S6 expression and cytokine production by PDC (data not shown), suggesting that PI3K-mTOR signalling is not limited by PTEN in human PDC. Together, these data show that a clinically relevant concentration of rapamycin inhibits proinflammatory cytokine production by TLR-7-activated PDC and TLR-9-activated PDC, while it suppresses IFN-α secretion in TLR-7-activated PDC but almost not in TLR-9-engaged PDC. To study the effects of mTOR inhibition on the T cell stimulatory capacity of PDC, we activated PDC with TLR ligands for 18 h and then added allogeneic CD3+ T cells. After activation in the presence or absence of rapamycin, PDC were washed carefully to remove rapamycin before T cells were added. Activation of PDC via TLR-7 in the presence of rapamycin increased their capacity to stimulate T cell proliferation, while the addition of rapamycin during TLR-9 activation did not (Fig. 2a). The increased proliferation of T cells upon mTOR inhibition in TLR-7-activated PDC was confined to enhanced expansion of the CD4 compartment (Fig. 2b), and was observed in both memory (CD45RO+) and naive (CD45RA+) T cells (Fig. 2c).

Modulation of the S1P/S1P1 receptor pathway might have some therapeutic potential in hepatic IRI-induced kidney injury. “
“Fibroblast

growth factor 23 (FGF-23) is a recently discovered regulator of phosphate and mineral metabolism. Its main PI3K inhibitor physiological function is the enhancement of renal phosphate excretion. FGF-23 levels are inversely related to renal function and in patients with chronic kidney disease (CKD) elevation in FGF-23 precedes the rise of serum phosphate. Studies have demonstrated an important role for FGF-23 in the development of secondary hyperparathyroidism through an effect on parathyroid hormone and calcitriol. In cross-sectional studies FGF-23 has been associated with surrogate

markers of cardiovascular disease such as endothelial dysfunction and arterial stiffness. FGF-23 has also been associated with both progression of CKD and mortality in dialysis patients. The discovery of FGF-23 has provided a profound new insight into bone and mineral metabolism, and it may become an important biomarker and therapeutic target in CKD. Patients with chronic kidney disease (CKD) have a significantly increased risk of cardiovascular disease (CVD) compared with age-matched individuals with normal kidney function.1 Mineral abnormalities complicating CKD such as hyperphosphatemia, calcitriol deficiency and secondary hyperparathyroidism (SHPT) are associated with increased cardiovascular (CV) and overall Target Selective Inhibitor Library screening mortality.2–4 Proposed mechanisms for this relationship click here include endothelial dysfunction, arterial stiffness, left ventricular hypertrophy (LVH) and vascular calcification.5 The term ‘Chronic Kidney Disease-Mineral Bone

Disorder’ (CKD-MBD) has been developed to highlight the intimate relationship between abnormalities of mineral metabolism, renal bone disease and excessive tissue calcification. The recent characterization of fibroblast growth factor-23 (FGF-23) and its important role in CKD-MBD has challenged the traditional understanding of the pathophysiology of SHPT. With an increasing number of clinical studies linking FGF-23 to clinical outcomes, we review the physiology of FGF-23 and its potential role as a biomarker and therapeutic target in CKD. The link between FGF-23 and phosphate regulation was first described in the rare inherited condition of autosomal dominant hypophosphatemic rickets, and soon after in the acquired condition of tumour-induced osteomalacia.6,7 These diseases are characterized by a common phenotype – hypophosphatemia, low or inappropriately normal calcitriol levels, urinary phosphate wasting and osteomalacia.8 The postulated phosphaturic circulating factor was subsequently identified as FGF-23 and the characteristic phenotypes in patients with conditions of FGF-23 excess or deficiency provided important early clues regarding its function.

Recently, Palbociclib mouse a blinded study utilizing a highly sensitive in vitro expansion method of detecting CTL responses failed to identify HIV-specific T cell responses in the HESN partners among HIV-discordant couples from Zambia [36]. Among HESN individuals with detectible T cell responses to HIV-1 antigens, the breadth and magnitude of the HIV-specific responses has often been significantly lower than comparable responses observed in HIV-1-infected individuals [25,37], due probably to the clear differences in antigen exposure between these subjects. Work from several groups

showing that pre-existing CTL responses against HIV-1 do not ensure a sustained resistance against infection in some persistently exposed HESN subjects who later seroconvert [38–40] further dampened interest in the potential role of T cells in sterilizing immunity. Currently, the potential role of antigen-specific T cell responses to HIV-1 in natural resistance from infection remains debated, and it is

currently unknown if HIV-1-specific T cell responses represent an active mechanism of protection or merely a marker of exposure to the virus, as suggested recently [41]. The fact that 30–60% of HESN subjects lack detectable T cell responses to HIV-1 (reviewed elegantly by Piacentini et al. and Miyazawa et al. in complementary analyses of HESN studies to date [42,43]) suggests that the presence of adaptive anti-HIV T cell responses has not been a unifying 4-Aminobutyrate aminotransferase functional attribute of HESNs. Rather, the collective evidence supports the notion that non-T cell-mediated immune

CP-690550 in vitro responses may also be involved in protection from HIV-1 in a subset of HESN subjects. Similar to adaptive T cell responses, HIV-specific IgA responses have been identified in the mucosa and sera of high-risk HIV-exposed seronegative subjects from multiple HESN cohorts [5,44–48]. HIV-specific IgA responses have also been documented in the absence of infection following oral exposure to HIV-1 through unprotected oral sex [49,50] and breast feeding [51]. Although there have been cohorts where no HIV-specific IgA has been evidenced [52], most HESN cohorts with documented mucosal exposure have evidenced detectable levels of HIV-specific IgA (see Table 2) [42,43]. Various reports have shown that HIV-specific IgA can neutralize HIV in ex-vivo assays [47,53], with most neutralizing epitopes found in gp41 and gp120 [53]. HIV-specific IgA from HESN subjects has also been shown to inhibit transcytosis across epithelial barriers, suggesting a functional mechanism of action in protection against HIV-1 infection [54,55]. In addition to direct neutralization of viral particles, HIV-specific IgA responses may also trigger antibody-dependent cellular cytotoxicity (ADCC) of infected target cells in conjunction with innate immune cells bearing the IgA-specific Fc receptor, CD89 [56,57].

We also found that memory B cells from our patients expressed higher levels of CD5 compared to healthy controls. These cells are known to produce low-affinity polyreactive antibodies (natural antibodies), which recognize autoantigens or conserved structures on self-antigens such as polysaccharide residues [21]. They have a reduced capacity to enter the cell cycle and have a longer lifespan. Although the precise role of these cells in autoimmunity is still obscure, the numbers of peripheral CD5+ B cells were found to be increased Pictilisib chemical structure in several autoimmune diseases, such as rheumatoid arthritis, primary Sjögren’s syndrome, autoimmune thyroid disease and multiple sclerosis [22]. Therefore, it seems that these cells

might play a role in the pathogenesis of autoimmune diseases [23]. The finding of low C4 levels, along with low functional C1INH in HAE, remains the most important immunological finding in this disease. C4 is important for the immune complex solubilization and removal [24]. Therefore, inherited deficiencies of C1q and C4 are associated with the chronic activation of the classical complement pathway and the development of autoimmune disease

such as lupus-like disease early in life [25]. Activation of the classical complement arm through immune complexes causes the production of C3 convertase, and the cleavage of C3 by C3 convertase leads to the production of C3b being an essential product for the immune complex removal. In addition, deficiencies of C4 render mice

Selleck AZD0530 unable to clear apoptotic cells/debris [26]. Mevorach et al. demonstrated that apoptotic materials are immunogenic and accelerate the production of autoantibody in mice not prone to autoimmunity [27]. Apoptotic material, especially when associated with microbial products in the form of immune complexes (ICs), might activate autoreactive B lymphocytes and induce serum autoantibodies [28]. One can speculate that the persistence of ICs could possibly activate B cell receptors and up-regulate the expression of TLR-9, allowing HAE patients to overproduce autoantibodies. Another possible explanation for the over-activation of B cells in HAE could be through increased signalling of the human complement receptor type 2 (CR2) on B cells. second CR2 (CD21) plays a pivotal role in the activation and proliferation of B cells and is a prerequisite for T-dependent immune responses. Engagement of CR2 with the B cell receptor lowers the threshold required for B cell activation by an antigen, enhances cell activation, reduces inhibitory signals and prevents apoptosis [29–32]. Only seven of our 61 (11·4%) patients had a defined immunoregulatory disorder. This incidence of immunoregulatory disorders is similar to the 12% found by Brickman et al. and 11·5% that was found by Farkas et al. [11,13]. It is not yet clear if this finding represents increased incidence compared to that in the general population.

chabaudi AS (34). Similarly, P. berghei, check details which has a homologous gene family, bir (35), has been shown to sequester via specific interaction with placental chondroitin sulphate A (36), the best described receptor for P. falciparum in the human placenta (27). Severe anaemia in pregnancy is an important contributor to maternal morbidity and mortality (37,38), and in malaria, endemic settings account for 7% to 18% of malaria-associated LBW (39).

Significant anaemia is observed in both B6 (20,21) and A/J mice, but ultimately is more severe in the latter, likely contributing to the lethality of the infection (40). Although anaemia may contribute to compromise of pregnancy in A/J mice, it is noteworthy that infected pregnant IFN-γ−/− B6 mice develop severe anaemia, but abort later than their IFN-γ+/+ counterparts, suggesting that anaemia may play a minor role in HSP inhibitor malaria-induced murine pregnancy loss (21). High rates of abortion have been associated with malaria infection in non-immune pregnant women during the first or second trimester (41). Pregnant malaria-naïve rhesus monkeys infected with P. coatneyi have increased rates of abortion and intrauterine growth retardation associated with significant malaria-associated placental pathology (42). Mid-gestational and pregnancy-associated recrudescent P. berghei infection in BALB/c mice results in reduced gestation time (36), reduced litter size (43) and reduced birth

weight (36,43). Consistent with these observations, both B6 and A/J mice experience poor pregnancy outcomes as a result of P. chabaudi AS infection. As evidenced by a higher rate of embryo resorption at experiment day 9, A/J mice experience accelerated pregnancy loss relative to B6 mice (20). Interestingly, the presence of haemorrhaging in embryos is more frequent and occurs earlier in B6 mice, suggesting that the precipitating mechanisms that drive embryo loss in these two mouse strains are complex Methane monooxygenase and multifactorial. Increased systemic inflammatory cytokines like TNF and IFN-γ have been observed in malaria during

pregnancy (6). Levels of TNF in particular have been associated with maternal anaemia and LBW (6,9) and this cytokine is sufficient to drive mid-gestational pregnancy loss in P. chabaudi AS-infected B6 mice (21). In this study, systemic levels of TNF and IL-1β were significantly elevated only in infected pregnant A/J mice, as early as experiment day 9, at which time resorption rates are increased. Thus, while pregnancy-protective anti-inflammatory responses may prevail early during infection in this strain (15), including elevated IL-10 production at experiment day 9, the tendency for this strain to subsequently produce inflammatory cytokines (18) is intact in pregnant mice. Interestingly, however, whereas antibody ablation of TNF successfully restored mid-gestational pregnancy in B6 mice (21), the same treatment was unsuccessful in A/J mice.

Furthermore, CD38− chronic lymphatic

leukemia cells show impaired chemotactic responses to CXCL12 in vitro, and, consequently, are thought to home less efficiently to lymphoid tissues 33, 34. The in vivo analyses of CD38-deficient mice have confirmed the impaired chemotactic migration of DCs and granulocytes towards chemotactic signals. CD38 activity also controls lymphocyte proliferation and apoptosis 23, which indirectly have an impact on leukocyte trafficking. CD38 can also regulate leukocyte traffic by interactions that are not dependent on its enzymatic activity 3, 23. On the cell surface, CD38 is normally expressed as a dimer, and is concentrated in lipid rafts. It can laterally interact with integrin α4 and CXCR4, classical adhesion and chemokine receptors, respectively, and this supramolecular complex may fine-tune leukocyte migration. Moreover, CD31, another classical adhesion molecule that is particularly important for leukocyte transmigration, is www.selleckchem.com/ALK.html a non-substrate ligand for CD38; ligation of CD38 by CD31, triggers signaling cascades in lymphocytes, and may also directly bind leukocytes to endothelial cells. CD157 triggers the same catalytic reactions as CD38, therefore also generating ADPR, cADPR and NAADP 23, 26; however, CD157 is attached to the cell membrane via a GPI-linkage, whereas CD38 is a transmembrane CYC202 supplier protein. CD157

is expressed both on endothelial cells and myeloid leukocytes and it interacts with integrins on the cell surface of monocytes. Via this integrin interaction, the

ligated CD157 triggers MycoClean Mycoplasma Removal Kit signals that enhance the polarization of monocytes, and enhance their chemotaxis towards fMLP and transmigration through the endothelial monolayer 35. NAD+ can also post-translationally modify surface proteins 23, 26, 36. In this reaction, which is catalyzed by ectoenzymes belonging to the ADP-ribosyltransferase (ART) family, one or more ADP-riboses are covalently attached to specific amino acid residues. In terms of leukocyte trafficking, L-selectin and the purinergic P2X7 receptor on the leukocyte surface are two important targets of ARTs. In mice, ART2-modified L-selectin is rapidly shed from the cell surface, with potential consequences for leukocyte extravasation, and ADP-ribosylated P2X7 triggers signals, which ultimately lead to T-cell apoptosis 37, 38. Thus, extracellular NAD+ also functions as a classical danger signal, as well as regulating leukocyte traffic. Enzymes regulating extracellular ATP metabolism are intimately connected to leukocyte trafficking. The balance between ATP and its dephosphorylated products ADP, AMP and adenosine determines whether the microenvironment is pro-inflammatory (ATP), pro-thrombotic (ADP) or anti-inflammatory (adenosine). ATP and ADP mediate their effects by binding to the purino-receptor of the P2X and P2Y families, whereas adenosine binds to the G-protein coupled A1, A2a, A2b or A3 receptors 26, 39.

While chest CT and conventional chest X-ray are generally used to assess bronchiectasis, these techniques fail

to detect a large proportion of bronchial pathologies. To date, there are no studies that demonstrate effective preventive or therapeutic measures against bronchiectasis in PAD patients. One of the major underlying reasons for the lack of studies is the difficulty to agree on a consensus protocol to reliably create quantitative data on bronchial pathology in a multi-centre setting. The international Chest CT in Antibody Deficiency Group (http://www.Chest-CT-Group.eu) aims to establish and validate a score for bronchiectasis and other structural lung disease for documenting the natural course of lung disease in PAD patients and potential effects in interventional selleck chemicals llc studies. Preliminary data of the group show a steady increase of the prevalence of bronchiectasis with age from approximately 40% in patients aged less than 20 years to almost 80% in patients above 60 years in a large multi-national cohort of CVID patients. Assessing the prevalence and course of airway disease is only a prerequisite for improving the health of the patients. Which intervention is the most promising to improve efficacy over the present management? The Selleck Ibrutinib role of antibiotic therapy has not been assessed

thoroughly to date, and present practices range from no therapy to preventive antibiotic maintenance therapy. Different antibiotics may have differing effects which are not purely anti-bacterial, such as improvement of sputum rheology properties or anti-inflammatory effects, as shown for azithromycin in patients with cystic fibrosis [11]. Hypertonic saline, which proved effective in improving sputum

clearance in cystic fibrosis patients, may also be beneficial in PAD patients. Other measures, such as dornase alpha, nasal irrigation and physiotherapy, could also be effective, but have not yet been assessed formally. Most challenging, however, would be an effort to develop an Ig replacement strategy also which is more physiological than the present practice. Is it feasible to replace serum IgA and IgM together with IgG systemically? In antibody-deficient patients, systemic replacement with serum IgA could lead potentially to the delivery of secretory IgA in the airway lumen, which is a natural process in healthy people. Indeed, these patients do not lack the expression of polymeric immunoglobulin receptor (pIgR), which is involved in the transepithelial transport of polymeric IgA and IgM (J-chain-positive IgA and IgM) on mucosal surfaces. However, this approach might not be as effective as desired for PAD patients, as serum IgA is mainly monomeric. It may eventually be more effective to apply Ig directly to the luminal site of the airways. Again, a number of challenges have to be met and are summarized in Table 1.

Real-time PCR is a practical, rapid, non-invasive screening test for excluding IFI in paediatric leukaemia. The high NPV makes real-time PCR a promising tool to use this prior to initiating EAFT in antibiotic-resistant febrile neutropenic patients; this would avoid toxicity, cost and hospitalisation for EAFT (ClinicalTrials.gov identifier:NCT00624143). “
“The

aim of this study was to evaluate the incidence of candidaemia, consumption of fluconazole and susceptibility of blood Candida isolates at a tertiary MK2206 hospital. From January 1999 to September 2006, all candidaemic episodes were identified and available strains were evaluated for the susceptibilities of antifungal agents. Annual selleck chemicals llc defined daily doses of antifungal agents were collected. There had been 909 Candida isolates detected from the bloodstream of 843 patients during the study period. Among them, 740 isolates were available

for the susceptibilities of antifungal agents. The incidence density of candidaemia was 28 episodes per 10 000 patient-days. Species distribution of 909 isolates did not vary annually, but varied greatly in the units of the hospital. Candida parapsilosis was the more prominent (30.1%) isolate in the paediatric units, where C. tropicalis and C. glabrata were less common (12.3% and 1.4% respectively). Resistance rates for itraconazole, fluconazole and voriconazole were 6.9%, 3.8% and 3.8% respectively. There were 25 (3.4%) isolates resistant to amphotericin-B. Although fluconazole usage increased over time (r2 = 0.45; P = 0.07), fluconazole resistance did not increase accordingly (P = 0.33). In our institution in which the incidence of candidaemia was high, fluconazole resistance among blood Candida isolates remained rare. “
“The aim of this study was to investigate the relationship between fungal exposure prior to hospitalisation and ensuing onset

of invasive mould infections (IMI) in patients at risk. Patients admitted to the Cyclin-dependent kinase 3 Department of Haematology, Oncology and Transplant Surgery of the Medical University Innsbruck received a questionnaire regarding fungal exposure prior to hospital stay. Questions inquired heavy fungal exposures up to 5 days before hospitalisation. A total of 234 patients were enrolled in this study. Multiple fungus exposures were associated with the onset of community-acquired IMI in patients with haematological malignancies. In univariate analysis, haematological malignancies (P = 0.013) and allergy to dust, pollen or moulds (P = 0.015) were significantly associated with fungal infections. In multivariate analysis, logistic regression showed that haematological patients (P = 0.015) and patients with allergy (P = 0.015) were significantly more frequently infected with fungi. Hospital-independent fungal sources highlight risk-factors for IMI in severe immunocompromised patients and the rate of community-acquired IMI does increase.

Results: GSAP immunoreactivity exhibited Proteasome inhibitor distinct morphological features, such as fine granular cytoplasmic deposits, dense nodular and patchy deposits, beads and string-like deposits, and diffuse dot-like deposits. In both AD and control brains, a fairly small subset of cerebral cortical and hippocampal neurones expressed fine

granular cytoplasmic deposits, while diffuse dot-like deposits were more frequently found in the neuropil and neuronal processes, particularly enriched in the hippocampal CA2 and CA3 regions. Among GSAP-immunoreactive deposits, dense nodular and patchy deposits, located in the neuropil and closely associated with PS1 expression and Aβ deposition, indicated the most distinguishing features of AD pathology. Conclusions: Aberrant regulation of GSAP expression plays a key role in acceleration of γ-cleavage BMN 673 purchase of APP-CTF and accumulation of Aβ in AD brains. “
“There is little immunohistochemical information about the early

stage of Pick body formation, due to the extremely limited opportunities of studying Pick’s disease at the incipient or subclinical stage. We report a 62-year-old man without any clinical manifestations of Pick’s disease, who died of B-cell lymphoma of the brainstem. Post mortem examination revealed many Pick bodies without obvious neuronal loss mainly in the left frontal and temporal lobes. Three brains of patients with typical Pick’s disease (disease duration: 7, 11 and 16 years) were also examined. Pick bodies were immunopositive for phosphorylated tau and 3-repeat tau, and less consistently for p62 in both incipient and typical cases. In the incipient case, borderline positivity for ubiquitin was evident in only a few Pick

bodies, whereas in the typical cases many Pick bodies showed obvious positivity for ubiquitin. These findings suggest that Pick bodies are rarely ubiquitinated in the early stage of Pick body formation. “
“Department of Laboratory Medicine, National Center for Global Health and Medicine Department of Laboratory Medicine, National Hospital Organization Kanagawa Hospital Director of a hospital, National Hospital Tobramycin Organization Komoro Kogen Hospital Department of Laboratory Medicine, National Hospital Organization Yokohama Medical Center The Gallyas method is a silver impregnation technique that is essential in the field of neuropathology because of its high sensitivity for the detection of argentophilic inclusion bodies in the central nervous system. In Japan, the Gallyas method has improved and is widely used as the “modified Gallyas method”. However, this method is not popularly used in general pathology laboratories because of the need for special reagents, several staining processes, and skilled techniques. The objective of the current study was to provide a simplified Gallyas method.

Mice were vaccinated with peptide-pulsed DC on days 6, 9, 12, and 19 post tumor injection. Tumor-bearing mice were irradiated 6 days after tumor

injection and reconstituted with 104 naïve pmel-1 spleen cells together with 107 congenic spleen cells, with or without CD25 and CD122 Rapamycin concentration depletion (Fig. 4A). After multiple DC vaccinations, Pmel-1 T cells still contracted immediately when co-transferred with undepleted spleen cells after the last vaccination. However, CD25 and CD122 depletion led to a prolonged expansion and delayed contraction of pmel-1 T cells. These results suggested that the suppression of tumor-reactive T cells mediated by CD25+ and CD122+ T cells could not be overcome by multiple vaccinations alone. Tumor-growth in melanoma-bearing mice subjected to check details reconstitution with pmel-1 T cells together with CD25- and CD122-double depleted spleen cells was significantly delayed compared with mice that received pmel-1 T cells together with undepleted spleens (Fig. 4B). To further characterize pmel-1 T cells in different organs, and in tumors, treated mice were sacrificed on day 44.

Spleen, blood, and tumors were collected for the analysis of the abundance of pmel-1 T cells (Fig. 4C). The percentage of CD8+ T cells that were GFP+(pmel-1 T cells) found in the spleen and blood or in the tumors of mice reconstituted with depleted spleen cells was double that of mice reconstituted with undepleted spleen cells. The majority (around 67%) of pmel-1 T cells, and a significant fraction of non-pmel-1 T cells found in the spleen produced IFN-γ (Fig. 4D), with or without depletion. Thus, pmel-1 T cells in peripheral tissues of tumor-bearing mice were functional effector/memory T cells. However, depletion did increase the percentage of IFN-γ producing non-pmel-1 T cells, primarily due to an increased frequency of peptide-specific

T cells. A much lower percentage of pmel-1 T cells (18%) found in tumors were able to produce IFN-γ as compared with pmel-1 T cells found in spleens (62%). These results Elongation factor 2 kinase strongly suggested that functional inactivation of pmel-1 T cells occurred locally in tumor sites. Interestingly, this inactivation could be ameliorated by CD25 and CD122 depletion, which almost doubled the percentage of IFN-γ-producing pmel-1 T cells from 18 to 34%. A much more dramatic increase of IFN-γ-producing, peptide-specific non-pmel-1 T cells was found in tumors from mice reconstituted with CD25- and CD122-depleted cells (5–23%). This could result from both an increased frequency and functionality of these tumor-specific T cells in tumor sites after depletion of Treg. Because both the expansion and survival of vaccine-induced pmel-1 T cells and lymphopenia-driven proliferation of CD122+CD8+ T cells are IL-7 dependent 6, we sought to determine whether administration of excess IL-7 would minimize the competition and improve the proliferation and expansion of pmel-1 T cells.