These nitrate TCR and CD8 molecules at the sites of MDSC-T cell contact, thereby disrupting TCR complexes and preventing T-cell activation [20]. Other mechanisms

of MDSC-mediated suppression include l-arginine depletion from the find more environment and the sequestration of cystine leading to a reduced availability of cysteine for T-cell activation [18]. Up to now it is unclear which aspects of CTL activation and differentiation are affected by the distinct MDSC subsets. Here, we demonstrate that splenic MDSCs not only inhibit several features of early CD8+ T-cell activation (proliferation; IL-2 secretion/responsiveness; CD44, CD162, and granzyme B expression; CD62L downregulation) check details — whereby MO- and PMN-MDSCs differ in their capacity to do so and differentially depend on IFN-γ, STAT-1, interferon regulatory factor 1 (IRF-1), and NO — but at the same time stimulate other activation events, such as IFN-γ production, CD69 expression, and Fas expression. Hence, MDSC-CD8+ T-cell interactions are more intricate than anticipated and include both inhibitory and stimulatory events. Previous studies suggested that MDSCs require IFN-γ to become T-cell suppressive [11]. To gain further insight in the dependence of MO- and PMN-MDSCs on IFN-γ and IFN-γ-activated transcription factors

for their activation, EG7-OVA (where OVA is ovalbumin) tumors were grown in WT, IFN-γR−/−, STAT-1−/− (inducing “first wave” IFN-γ-dependent

genes), and IRF-1−/− (inducing “second wave” IFN-γ-dependent genes [21]) mice. In all strains, splenic CD11b+CD115+Ly6G−Ly6Chigh MO-MDSCs and CD11b+CD115− Ly6G+Ly6Cint PMN-MDSCs were expanded in the course of tumor growth and MDSC subsets were purified from the spleen when tumors reached an approximate diameter of 15 mm (Supporting Information Fig. 1). Upon coculture with OVA-stimulated TCR transgenic OT-1 splenocytes, WT MO-MDSCs suppressed T-cell proliferation in a dose-dependent manner, while IFN-γR−/− and STAT-1−/− MO-MDSCs almost completely lost their suppressive capacity (Fig. 1A and Supporting Information Fig. 2A for CFSE dilution). However, MO-MDSCs from IFN-γ–/– tumor-bearers were as suppressive as 4��8C their WT counterparts (data not shown). These data illustrate that (i) there is an absolute requirement for IFN-γ/STAT-1 to activate the suppressive potential of splenic MO-MDSCs in vitro, and (ii) this does not rely on autocrine IFN-γ signaling. Interestingly, when treating MO-MDSCs with recombinant IFN-γ, only 72% of the population phosphorylates STAT-1, illustrating MO-MDSC heterogeneity and suggesting that only the IFN-γ-responsive part of this population mediates suppression (Supporting Information Fig. 3). Remarkably, IRF-1−/− MO-MDSCs retained a partial antiproliferative capacity (Fig.

Further experiments involving studies in rhesus macaques will be required

to find optimal adjuvant formulations able to specifically shape protective immune selleck compound responses to a given pathogen. In conclusion, the findings reported here contribute to our knowledge about rhesus macaque B-cell responses and support the relevance of using non-human primates for modelling TLR-administration to people. These data will hopefully inform future vaccine design and development of adjuvant strategies. This work was supported by grants from Vetenskapsradet, the Swedish International Development Agency (Sida), the International AIDS Vaccine Initiative (IAVI), the Swedish Governmental Agency for Innovation Systems (Vinnova) and the Swedish Society of Medicine. We are grateful for the assistance of the veterinarians Drs Mats Spångberg and Helene Fredlund, and to the personnel at the Astrid Fagraeus Laboratory

at the Swedish Institute for Infectious Disease Control. The authors have no financial conflicts of interest. “
“α-Fetoprotein (AFP) is a tumour-associated antigen in hepatocellular carcinoma (HCC). The biological properties of AFP have been identified in its regulatory effects on immune responses of T cells and B cells. However, AFP effects on natural killer (NK) cells are still unclear. In this study, we examined the immunoregulation of AFP on NK activity. The cytolytic activity against K562 cells and Huh7 cells Anti-infection Compound Library of NK cells co-cultured

with AFP-treated dendritic cells (DCs) (AFP-DCs) was lower than that with albumin-treated DCs (Alb-DCs). Direct addition of AFP to NK cells did not alter the cytolytic activity of NK cells. Adding AFP inhibited the interleukin (IL)-12 production of DCs after stimulation with lipopolysaccharide (LPS) [Toll-like receptor (TLR)-4 ligand], PtdIns(3,4)P2 or Poly(I:C) (TLR-3 ligand), but not IL-18 production. The mRNAs of IL-12p35 and IL-12p40 were significantly inhibited in AFP-DCs compared with Alb-DCs, but those of TLR-4 or TLR-3 were not. Transwell experiments revealed that soluble factors derived from DCs played roles in inhibition of the ability of activating NK cells by AFP-DCs. Adding the neutralizing antibody of IL-12 to NK cells co-cultured with Alb-DCs resulted in a decrease of cytolytic activity to the levels of NK cells co-cultured with AFP-DCs. Adding IL-12 to NK cells co-cultured with AFP-DCs resulted in an increase of cytolytic activity to the levels of NK cells co-cultured with Alb-DCs. These demonstrated that the impairment of IL-12 production from AFP-DCs resulted in inhibition of the ability of the activation of NK cells by DCs, and thus suggests a role of AFP in HCC development. Hepatocellular carcinoma (HCC) is one of the leading causes of cancer deaths worldwide.

The neuroprotection provided by the proactive transplantation of human NSCs in the rat model of HD appears to be contributed by brain-derived neurotrophic factor (BDNF) secreted by the transplanted human NSCs. Previous studies have also demonstrated that BDNF could block neuronal injury under pathological conditions in animal models of HD.[78, 79] These findings suggest that proactively transplanted human NSCs were well integrated in the striatum

and supported the survival of host striatal neurons against neuronal injury. To develop an effective stem cell-based cell therapy for HD, it is desirable VX-809 in vitro to use genetic animal models, but earlier studies have used chemical (QA or 3-NP)-induced animal models and only a small number of studies have used transgenic HD animals. In YAC HD transgenic mice, bone marrow MSCs genetically modified to express BDNF were transplanted in striatum and induced behavioral improvement.[80] In another study in R6/2 HD transgenic mice, transplantation of adipose tissue-derived stem cells (ADSCs) improved motor function and increased the survival of striatal neurons.[81] Human striatal selleck chemicals llc neural stem cell line cells were treated with a hedgehog agonist to generate DARPP-32 cells and transplanted in R6/2 HD

transgenic mouse brain. The results were disappointing that the outcome was the same as a vehicle control injection.[82] This study is only one using human NSCs for cell therapy in HD genetic animal model. Human NSCs derived from ESCs could Olopatadine provide a viable cellular source for cell therapy in HD, since they can be expanded indefinitely and differentiate into any cell type desired. Three previous studies have shown that neurons expressing striatal markers could be induced from ESCs and brain transplantation of these ESC-derived

neurons in QA-lesioned rats leads to behavioral recovery in the animals.[83-85] We have previously written a review that focuses on the stem cell-based therapy for HD and investigators who wish to learn more about the subject are referred to the review article.[86] A summary of preclinical studies of stem cell transplantation in HD animal models is shown in Table 2. Intact BBB Lesion vol GAD + cells 0.3% No change NPC migration Lesion vol NeuN + cells Lesion vol NeuN + cells Lesion vol NeuN + cells Lesion vol NPC migration No change ESC-derived NSC (human) Noggin-primed NSC migration Amyotrophic lateral sclerosis (ALS), known as Lou Gehric disease, is a relentlessly progressive, adult onset neurodegenerative disorder characterized by degeneration and loss of motor neurons in the cerebral cortex, brain stem and spinal cord, leading to muscle wasting and weakness, and eventually to death within 5 years after the onset of its clinical symptoms.

4d). These results demonstrated that

heat-killed MoLac-1 induced IFN-γ production by NK cells via IL-12 secretion from macrophages and activated NK cells in vitro. Oral administration of LAB has been reported to augment NK activity in mouse and clinical studies (Ogawa et al., 2006; Takeda et al., 2006; Koizumi et al., 2008). Oral administration of heat-killed MoLac-1 increased the population of NK cells in the spleen, but did not affect the expression of early activation marker CD69 on NK cells learn more (Fig. 6). Takagi et al. (2001) suggested that the enhancement of NK activity in splenocytes from LAB-fed mice was caused by the increased proportion of NK cells but not by the increased cytotoxicity of individual NK cells. Thus, oral administration of MoLac-1 might enhance

NK activity by increasing the population of NK cells, but further investigation such as functional assay of NK cells is needed for evaluating the possible effect on the activity of NK cells. NK cells and IFN-γ produced by NK cells are crucial to the early natural defenses against IFV infection (Stein-Streilein & Guffee, 1986; Monteiro et al., 1998). As in vitro studies demonstrated that heat-killed MoLac-1 cells induced the production of IFN-γ by NK cells, Talazoparib the in vitro immunomodulating effects of MoLac-1 might be associated with the alleviation of IFV infection; however, involvement of NK cells in the anti-infective effects of MoLac-1 is not clear and further investigation is needed. Substantial interest has been aroused in the application of nonviable microorganisms in food or food supplements. At first, the use of nonviable microorganisms could solve the problem concerning SPTLC1 the stability of active constituents in handling and preservation, and could prolong the shelf life of the products. Furthermore, nonviable microorganisms could eliminate the risks of microbial translocation, invasion, and toxin production (Taverniti & Guglielmetti, 2011). Concerns have been raised for safety aspects in the application of live bacteria in food or food supplements

(Wassenaar & Klein, 2008). In summary, we demonstrated that heat-killed MoLac-1 would have the potential to modulate innate immunity and might be useful for alleviation of symptoms of IFV infection. This strain was found to induce dose dependently IL-12p40 production by human PBMCs (data not shown). Further studies are anticipated to assess the usefulness of heat-killed MoLac-1 in clinical experiments. “
“OTHER ARTICLES PUBLISHED IN THIS MINI-REVIEW SERIES ON Th17 CELLS Function and regulation of human T helper 17 cells in health and disease. Clin Exp Immunol 2009; doi:10.1111/j.1365-2249.2009.04037.x Are T helper 17 cells really pathogenic in autoimmunity? Clin Exp Immunol 2009; doi:10.1111/j.1365-2249.2009.04039.x CD4+ T helper cells: functional plasticity and differential sensitivity to regulatory T cell-mediated regulation.

Optical coherence tomography (OCT) may help to detect increased macular volume that seems to occur frequently under FTY treatment; selleck however, macular oedema is a rare condition [109, 110]. Two deaths were reported due to herpes virus infections: a primary VZV

infection and a herpes-simplex encephalitis [9]. A PML case is being discussed [111], but thus far has not been fully elucidated. In the post-marketing setting, mainly cardiac events have been reported thus far and have led to extended cardiovascular safety monitoring [112]. Recently, the marketing authorization holder published two fatal cases of haemophagocytic syndrome (HPS) associated with a 9- and 15-month treatment period with FTY. HPS is triggered typically by (viral) infections such as Epstein–Barr virus, as in the cases described. It results in a severe disturbance of the immune system and multi-organ involvement including fever, lymphadenopathy, organomegaly, cytopenia, liver failure and various neurological symptoms. Early diagnosis and treatment of both the triggering condition and the overwhelming immune response via immunosuppressive means are crucial to reduce mortality of HPS. The described Crizotinib cost safety set-up implies several putative biomarkers, although not evaluated formally thus far in terms of prediction of response or determination of SADR development. However,

evaluation of lymphocyte counts may serve not only as a necessary safety measurement, but also as a therapy adherence marker. Subclinical impairment of VZV and Epstein–Barr-virus reactivity have been found recently [113]. Teriflunomide (Aubagio®) is the active metabolite of leflunomide, a disease-modifying anti-rheumatic drug (DMARD). It is an inhibitor

of the dihydroorotate dehydrogenase and interacts with de-novo pyrimidine synthesis [114]. Although the pivotal trial included 8·6% of SPMS patients [115, 116], it has been approved by the FDA and EMA for RRMS. Specific contraindications for teriflunomide Verteporfin supplier include severe hepatic or renal disorders and hypoproteinaemia (due to high plasma protein-binding) [117]. As experimental data hint at teratogenic potential, FDA prescription guidelines emphasize the restriction of teriflunomide during pregnancy [118]. It may be hypothesized that teriflunomide treatment may be especially beneficial with co-existing neuroimmunological and rheumatic disorders. Due to the long half-life of the drug and pronounced enterohepatic recirculation, teriflunomide might be an option in patients having difficulties with adherence to treatment schedules, but may be used more cautiously in patients with an impending wish for children. Oral teriflunomide is administered once daily, 7 or 14 mg (FDA approval), or 14 mg (EMA approval) [116].

05), while antagonistic cytokines like IFN-γ were increased in acute phase of KD (P < 0.05) and reduced after therapy with IVIG (P < 0.05). These results suggest that aberrantly decreased levels of NKG2D expression on NK cells and CD8+T cells might be one of the factors

led to disturbed immunological function in patients with KD. Selleckchem BIBW2992 Cytokines milieu could be important factors causing reduced expression of NKG2D. Kawasaki disease (KD) is an acute systemic vasculitis that affects infants and children. At present, the pathogenesis of KD remains to be further investigated. However, there is a large body of evidence that immunological disturbances play a key role in the pathogenesis of KD. A great many studies have found that the levels of many proinflammatory cytokines such as tumour necrosis factor (TNF)-α and interleukin (IL)-6 are elevated in acute KD, but the mechanisms resulting in aberrant immune function or overexpression of proinflammatory cytokines are not completely clear [1-3]. NKG2D is a C-type lectin-like type II transmembrane glycoprotein. It expressed on immunocompetent cells, such as natural-killer (NK) cells, CD8+ cytotoxic lymphocytes (CD8+T), NKT cells and γδT cells and participates in the regulation of innate and adaptive immune response through enhancing their killing activity. It has been

demonstrated that NKG2D expression is induced Selleck Ensartinib on NK cells and CD8+T cells by their activation [4-6]. Accumulated evidences suggest that peripheral CD8+T cells may be functionally suppressed in acute phase of KD. Previous studies have shown a reduction in the total number of CD8+T cells in the peripheral blood of KD patients [7]. However, the expression of NKG2D on NK cells and CD8+T cells in the acute phase of KD is still required to be investigated. In this study, flow cytometry (FCM) was used to detect the expression of NKG2A/NKG2D on CD8+T cells and CD3−CD56+ NK cells in patients with KD, both in the acute phase and after IVIG therapy. The cytokines regulating expression of NKG2D such as IL-1β, IL-6, TNF-α, IL-7, IL-12, IL-15, interferon (IFN)-γ Fludarabine and transforming growth factor

(TGF)-β were also evaluated in this study. Aberrantly, decreased levels of NKG2D expression were found in acute phase of KD patients, suggesting that downregulation expression of NKG2D might be one of the factors led to disturbed immunological function in KD. Forty-six children with KD admitted to the Shenzhen Children Hospital between June 2011 and April 2012 were included in the study. The patients comprised 26 males and 20 females (mean age: 26.33 ± 23.82 months; age range: 2 months–5 years). The diagnosis was carried out according to the clinical criteria of the Kawasaki Disease Research Committee of Japan. Blood samples were obtained before treatment with 2 g/kg/day intravenous immunoglobulin (IVIG, mean duration of illness, 6.3 days; range, 3–12 days) and after IVIG treatment (mean duration of illness, 12.0 days; range, 8–20 days).

Reaction products were diluted five times upon addition of 10 mmol l−1 Tris-HCl (pH 8.3) buffer. Adapters were produced by mixing equimolar amounts of complementary oligonucleotides (Eurogentec, Seraing, Belgium): (5′-CTCGTAGACTGCGTACC-3′ and 5′-CGGGTACGCAGTC-3′ for HpyCH4IV; 5′-GACGATGAGTCCTGAC-3′ and 5′-TAGTCAGGACTCAT-3′ for MseI) and heating the mixture to 95 °C followed by slow cooling to ambient temperature. One microlitre of the diluted restriction-ligation mixture was used for amplification in a reaction volume of 25 μl containing

1 μmol l−1 AZD2281 clinical trial M HpyCH4IV primer with one selective residue (underlined) (5′-Flu-GTAGACTGCGTACCCGTT-3′), 1 μmol l−1 MseI primer with four selective residues (underlined) (5′-GATGAGTCCTGACTAATGAA-3′), 0.2 mmol l−1 of each deoxynucleoside triphosphate, Ixazomib nmr and 1 U of Taq DNA polymerase (Roche Diagnostics) in 1× reaction buffer containing 1.5 mmol l−1 MgCl2. Amplification was performed as follows. After an initial denaturation step at 94 °C for 4 min in the first 20 cycles, a touchdown procedure was applied: 15 s denaturation at 94 °C; 15 s annealing at 66 °C with the temperature for each successive cycle lowered by 0.5 °C and 1 min

of extension at 72 °C. Cycling was then continued for further 30 cycles with an annealing temperature of 56 °C. After completion of the cycles, incubation at 72 °C for 10 min was included before the reactions were cooled to room temperature. Products were diluted 1 : 10 with distilled water. To 1 μl of diluted product, 0.25 μl of ET400-R size marker (GE Healthcare, others Diegem, Belgium) and 8.75 μl of distilled water were added. Following 1-min denaturation step at 94 °C, the samples were

quickly cooled to room temperature and analysed on a MegaBACE 500 automated DNA analysis platform equipped with a 48-capillary array according to the instructions of the manufacturer (GE Healthcare). AFLP data were imported into BioNumerics v. 6.0 Software (Applied Maths, Sint-Martens-Latem, Belgium) and analysed by UPGMA clustering using the Pearson correlation coefficient. The analysis was restricted to DNA fragments in the range from 60 to 300 bp. Clinical isolates were identified to the species level based on the similarity of their AFLP profile to those of the type strains. To assign specific AFLP genotypes, the obtained profiles were also inspected visually and scored for presence/absence of clearly recognisable DNA fragments. Two profiles differing by at least one clearly recognisable DNA band were considered to represent different genotypes.

Patients with

PD0332991 mouse HCV infection with and without fibrosis were similar apart from the level of HCV-RNA (Table 1). The group of co-infected patients varied in gender distribution and age compared with HCV-infected patients and healthy controls (P < 0.05) (Table 1). The CD4+ count was as expected significantly lower in patients with HIV co-infection (P < 0.05). The distribution of HCV genotypes was comparable in the three hepatitis groups, and significant associations between genotype, ALT, HCV-RNA and fibrosis were not found (data not shown). According to our definition of fibrosis and cirrhosis, 12 of the 25 HCV-infected patients with a liver stiffness above 8 kPa had a fibroscan defined as cirrhosis. However, no difference in any aspects was found between HCV-infected LY2835219 price patients with fibrosis and cirrhosis (data not shown). To evaluate chronic immune activation, the frequency of activated T cells (CD38+ HLA-DR+) within the CD4+ as well as the CD8+ compartment were determined. The median frequency of both CD4+- and CD8+-activated T cells were elevated in HIV/HCV co-infected patients (2.2%; 1.4–2.6 and 7.0%; 4.1–9.2, respectively), compared with HCV-infected patients

without fibrosis (1.5%; 1.1–1.9, P = 0.03 and 3.4%; 2.1–8.7, P = 0.03), and healthy controls (1.3%; 1.1–1.7, P = 0.01 and 3.5%; 2.5–4.1, P < 0.001) (Fig. 2). There were no differences in activated CD4+ and CD8+ T cells between the two groups of mono-infected

patients and the healthy controls (Fig. 2). CD4+ Tregs, CD8+ Tregs and Th17 cells were determined to evaluate the composition of pro- and anti-inflammatory Glutathione peroxidase lymphocyte subsets. Patients with HCV infection with fibrosis (5.0%; 4.5–5.6) as well as without fibrosis (5.6%; 4.2–6.4) had significantly higher frequencies of CD4+ Tregs compared to healthy controls (4.4%; 3.4–4.7, P = 0.03 and P < 0.001, respectively) (Fig. 3A). Furthermore, the HIV/HCV co-infected patients appeared with even higher frequencies of CD4+ Tregs (6.5%; 6.0–7.0) compared with HCV-infected patients without fibrosis (P = 0.01) and to healthy controls (P < 0.001). To further describe the composition of CD4+ Tregs, three CD4+ Tregs subpopulations were determined based on co-expression of CD45RA and Foxp3 (Fig. 1). HCV-infected patients with fibrosis and HCV infected without fibrosis as well as HIV/HCV co-infected patients had significantly lower frequencies of resting Tregs compared with healthy controls (P < 0.001, P = 0.001 and P = 0.005, respectively) (Fig. 4A). No difference was observed between the three groups of patients. In contrast, the frequency of activated Tregs was higher in both HCV-infected patients and HIV/HCV co-infected patients compared with healthy controls, although, significant difference was only observed when comparing HCV-infected patients without fibrosis and healthy controls (P = 0.022) (Fig. 4B).

Thus, it is likely that the antiviral activity R428 manufacturer of the CL-46 NCRD significantly exceeds that of SP-D. We also confirm the substantially greater mannan-binding activity of CL-43. We attempted to determine the structural

differences that could account for increased antiviral activity of these proteins. The ridges around the primary carbohydrate binding site show considerable divergence among collectins, perhaps in response to a need to recognize different pathogens. One obvious difference between all serum collectins and SP-A or SP-D is the presence of a hydrophobic residue at position 343. We have shown that the R343V or R343I mutants of hSP-D-NCRD have greatly increased antiviral Selumetinib ic50 activity compared to the wild-type hSP-D-NCRD [28]; hence, this is one important difference accounting for the increased antiviral activity of bovine serum collectin NCRD. Another difference relates to the presence of small amino acid insertions immediately N-terminal to residue 325. As CL-43 had particularly strong mannan-binding and antiviral activity, for this paper we produced and tested addition of the RAK sequence to the R343V (or R343I) mutant of hSP-D-NCRD. Although the combined mutations greatly increased mannan-binding activity, antiviral activity was decreased when compared to R343V (or R343I). This finding indicates that the mechanisms of binding to mannan

and to IAV, while similar, are not identical and involve a complex interplay between residues on the two ridges that flank the primary carbohydrate binding site. High mannose oligosaccharides on the IAV hemagglutinin are important for recognition and neutralization by SP-D [6]. Important P-type ATPase differences in the detailed structure of oligomannose sugar chains on IAV and mannan, or in the macromolecular patterns of sugars of mannose-rich sugars on IAV and mannan, may account for the differences in recognition of these ligands by specific NCRD. It is

of interest that binding of mAb 246-02 and 3C3-C-20, which is reduced to RAK, is partially or fully restored for RAK+R343V, implying that the combination of the insertion and substitution restore a structural feature in hSP-D-NCRD that is recognized by these mAb. We plan, in future studies, to determine the crystal structures of these and other mutant versions of the SP-D NCRD. Although the RAK+R343V (or I) double mutants did not result in increased antiviral activity compared to single mutants, we are pursuing other strategies including substitutions for D325 in combination with the R343V substitution and have found increased activity (Hartshorn KL, Seaton B, and Crouch EC, unpublished data). Hence, we still feel the approach of altering residues on the ridges flanking both sides of the lectin site is a productive approach to developing NCRD that could be of therapeutic use in IAV.

3C). IFN-α2b and IFN-α5 effects were almost identical over the broad range of concentrations tested (Supporting Information Fig. 3). The necessary role of IFNAR was revealed by neutralizing anti-human IFNAR2 mAb (Supporting Information Fig. 4). The CD3-redirected cytolytic assay using OKT3 mAb-coated p815 target cells is commonly

used to evaluate the TCR/CD3-triggered cytotoxicity that entails release of perforin and granzymes, and surface relocation of CD107a. Furthermore, Caki-1 cells, sensitive to TRAIL- but not to FasL-induced cell death, can be used as target cells to assess TRAIL-mediated cytotoxicity 15. Figure 3 strikingly shows that IFN-α enhanced CD3-redirected cytotoxicity (Fig. 3D–E) as well as TRAIL-mediated cytolysis (Fig. 3F–G). Neutralizing anti-TRAIL and anti-FasL mAb revealed the exclusive selleck contribution of TRAIL in the lysis of Caki-1 cells (Fig. 3G). FK506 chemical structure No significant differences were found between the IFN-α2b and IFN-α5 subtypes in any of these assays (Figs. 1–3 and Supporting Information Figs. 1–4). Following CD27- and CD45RA-based phenotypic classifications of CD8+ T

cells 16, negatively selected total CD8+ T cells were sorted into naïve (CD45RAhighCD27high), memory (CD45RA−CD27+) and effector (CD45RA+CD27− and CD45RA−CD27−) cells. For comparative studies, naïve and memory CD8+ T cells were stimulated as above. Regardless of whether cells were naïve or memory, cell division was not noticeable before 72 h of culture and required CD3/CD28-triggering (Supporting Information

Fig. 5A and B). At day 4 of culture, naïve CD8+ T cells from some individuals (3/8) showed a transiently delayed proliferation in the presence of IFN-α (Supporting Information Fig. 5C). However, from day 5, the extent of division was always higher in cells receiving CD3/CD28/IFNAR-derived signals (observed in 8/8 individuals) (Fig. 4A and Supporting Information Fig. 5A and C). By Methamphetamine contrast, once division started, CD3/CD28-induced proliferation of memory cells was always delayed by IFN-α (Fig. 4A and Supporting Information Fig. 5B). Interestingly, IFN-α increased the survival of both CD3/CD28-triggered naïve and memory CD8+ T cells (Supporting Information Fig. 5D and E). IFN-α-derived type-3 signals significantly increased the expansion of human naïve CD8+ T cells whereas they reduced the fold expansion of memory CD8+ T cells (Fig. 4B). When the expression of IFN-γ, Granzyme-B and TRAIL was assessed by flow cytometry analysis, we found that IFN-α enhanced the expression of these three effector molecules both in naïve and memory CD8+ T cells (Supporting Information Fig. 6). However, the fold-change increases in protein induction attributable to IFN-α were markedly higher in naïve cells (Supporting Information Fig. 6). Figure 4C shows that regardless of whether the cells were naïve or memory, the amounts of secreted IFN-γ were higher in cells receiving IFN-α as a signal-3.