Patients were not selected on viral load (VL). Subjects included were 29 males, four females, with a median age of 38.69 (25–67), 4 median years of infection (<1–17), a median CD4+ count of 240.2 (51–336) and median VL of 101 669 (45≥500 000). For longitudinal studies, these patients were sampled prior to and 1, 4, 8–12 months post-initiation of HAART (Supporting Information Table

2). In addition, 31 chronically infected asymptomatic treatment-naive I-BET-762 cost HIV+ subjects were studied (Supporting Information Table 3). Chronic untreated patients were identified as being treatment naïve with a stable CD4+ count above 300, as measured on at least two occasions (from time of diagnosis and at 6–12 monthly intervals) prior to sampling, not requiring therapy. This group had a median age of 37.87 (26–53), eight of which were females and 23 were males, with a median CD4+ T-cell count of 672.5 (277–1439) and median VL of 17 451 (<50–18 779) and 5.5 (1–16) median years of infection. Control HIV sero-negative blood samples were purchased from the National Blood Pirfenidone Transplantation Service at St George’s Hospital Tooting, UK, and tested in parallel with samples from HIV+ subjects. For controls subjects, where information was available the intention was to match the patients as closely as possible in terms of age, and to attempt to match in terms of gender if possible. Although all recruited patients were studied, not all

subjects could be analysed for all parameters included in this study, which was linked to blood volume, yield and CD4+ T-cell count at the time of experimentation. Peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation (Lymphoprep: Axis-Shield PoC AS, Oslo, Norway). CD4+CD45RO+CD25− effector and CD4+CD45RO+CD25+ Treg-cell populations were isolated using Dynabeads T regulatory cell isolation kit (Invitrogen, Paisley, UK) as described previously 15. Purity of isolated fractions was confirmed by immunostaining Nitroxoline to be >95% for effector and Treg populations (Supporting Information Fig. 5). All assays were carried out in RPMI-1640 Glutamax 25 mM HEPES media

(Invitrogen), 10% human AB serum (Lonza, Sweden), and supplemented with 20 μg/mL Gentamycin (Sigma-Aldrich, UK) as described previously 15 by co-cultuting 2.5×103 effector cells, with at least two ratios of Treg cells. Cells were stimulated with Dynal anti-human CD3/CD28 coated magnetic beads (bead: effector cell ratio, 2:1) (Invitrogen) for 5 days. Each well received 0.5 μCi of (3 H)-thymidine (Perkin Elmer, UK) for the last 16 h of culture. As described previously 15 2×104 effector cells were cultured with varying ratios of Treg cells and stimulated with 2:1 (bead:effector cell) Dynal anti-human CD3/CD28 coated magnetic beads. After the addition of Brefeldin A (Sigma-Aldrich) cultures were maintained for 16 h before ICS for IFN-γ (PE-IFN-γ) and Interleukin-2 (APC-IL-2, both BD Pharmingen, UK) or appropriate isotype control mAbs.

Neither index of mind-mindedness related to infant temperament. We conclude that mind-mindedness is best characterized as a facet of the specific caregiver–child relationship, while also being influenced by stable cognitive–behavioral

traits in the mother. “
“This paper presents two methods that we applied to our research to record infant gaze in the context of goal-oriented actions using different eye-tracking devices: head-mounted and remote eye-tracking. For each type of eye-tracking system, we discuss their advantages and disadvantages, describe the particular experimental setups we used to study infant looking and reaching, and explain how we were able to use and synchronize these systems with other sources of data collection (video recordings and motion capture) to analyze gaze Venetoclax chemical structure and movements directed toward

three-dimensional objects within a common time frame. Finally, for each method, we briefly present some results from our studies to illustrate the different levels of analyses that may be carried out using these different types of eye-tracking devices. These examples aim to highlight check details some of the novel questions that may be addressed using eye-tracking in the context of goal-directed actions. “
“Prosocial behaviors are a diverse group of actions that are integral to human social life. In this study, we examined the ability of 18- and 24-month-old infants to engage in three types of other-oriented behaviors, Sucrase specifically helping, sharing, and comforting. Infants in both age groups engaged in more prosocial behavior on trials in which an unfamiliar adult experimenter required aid (experimental conditions) than on those in which she did not (control conditions) across two of the three prosocial tasks (i.e., helping and sharing). The infants engaged in these behaviors with similar frequency; however, there was no correlation between the tasks. The implications for the construct

of prosocial behavior and the presence of a prosocial disposition are discussed. “
“The present study used event-related potentials (ERPs) to monitor infant brain activity during the initial encoding of a previously novel visual stimulus, and examined whether ERP measures of encoding predicted infants’ subsequent performance on a visual memory task (i.e., the paired-comparison task). A late slow wave component of the ERP measured at encoding predicted infants’ immediate performance in the paired-comparison task: amplitude of the late slow wave at right-central and temporal leads decreased with stimulus repetition, and greater decreases at right-anterior-temporal leads during encoding were associated with better memory performance at test. By contrast, neither the amplitude nor latency of the negative central (Nc) component predicted infants’ subsequent performance in the paired-comparison task.

These findings suggest that minocycline administration does not suppress MMPs at

mRNA and protein levels but that it suppresses gelatinase activities by upregulating TIMPs. Thus, MMP-targeted therapies should be designed after the mechanisms of candidate drugs have been considered. “
“K. Seidel, J. Vinet, W. F. A. den Dunnen, E. R. Brunt, M. Meister, A. Boncoraglio, M. P. Zijlstra, H. W. G. M. Boddeke, U. Rüb, H. H. Kampinga and S. Carra (2012) Neuropathology and Applied Neurobiology38, 39–53 The HSPB8-BAG3 chaperone complex is upregulated in astrocytes in the human brain affected by protein aggregation diseases Aims: HSPB8 is a small heat shock protein that forms a complex this website with the co-chaperone BAG3. Overexpression of the HSPB8-BAG3 complex

in cells stimulates autophagy and facilitates the clearance of mutated aggregation-prone proteins, whose accumulation is a hallmark of many neurodegenerative disorders. HSPB8-BAG3 could thus play a protective role in protein aggregation diseases and might be specifically upregulated in response to aggregate-prone protein-mediated toxicity. Here we analysed HSPB8-BAG3 expression Z-VAD-FMK purchase levels in post-mortem human brain tissue from patients suffering of the following protein conformation disorders: Alzheimer’s disease, Parkinson’s disease, Huntington’s disease and spinocerebellar ataxia type 3 (SCA3). Methods: Western blotting and immunohistochemistry techniques were used to analyse HSPB8 and BAG3 expression levels in fibroblasts from SCA3 patients and post-mortem brain tissues, respectively. Results: In all diseases investigated, we observed a strong upregulation of HSPB8 and a

moderate upregulation of BAG3 specifically in astrocytes in the cerebral areas affected by neuronal damage and degeneration. Intriguingly, no significant change in the HSPB8-BAG3 expression levels was observed within neurones, irrespective of their localization or of the presence of proteinaceous aggregates. Conclusions: We propose Thiamine-diphosphate kinase that the upregulation of HSPB8 and BAG3 may enhance the ability of astrocytes to clear aggregated proteins released from neurones and cellular debris, maintain the local tissue homeostasis and/or participate in the cytoskeletal remodelling that astrocytes undergo during astrogliosis. “
“Cockayne syndrome (CS) and xeroderma pigmentosum (XP) are caused by deficient nucleotide excision repair. CS is characterized by cachectic dwarfism, mental disability, microcephaly and progeria features. Neuropathological examination of CS patients reveals dysmyelination and basal ganglia calcification. In addition, arteriosclerosis in the brain and subdural hemorrhage have been reported in a few CS cases. Herein, we performed elastica van Gieson (EVG) staining and immunohistochemistry for collagen type IV, CD34 and aquaporin 4 to evaluate the brain vessels in autopsy cases of CS, XP group A (XP-A) and controls.

To further test the functional attributes of Fab specific for the two-domain RTL1000, we utilized an Fab specific for RTL1000 that was also cross-reactive with a similar Selleckchem PLX4032 antigenic determinant on RTL342m (α1β1 domains of DR2 linked to mMOG-35-55 peptide). DR2 Tg

mice were immunized with mMOG-35-55 peptide/CFA/pertussis toxin (Ptx) to induce EAE and were treated with pre-formed complexes of 2E4 Fab:RTL342m, the control D2 Fab:RTL342m (specific for RTL2010 that comprised DR4–GAD-555-567 described in Fig. 8C) or TRIS buffer (Fig. 5). As shown in Fig. 5, mice receiving RTL342m plus TRIS buffer were effectively treated, whereas a 2:1 ratio of 2E4 Fab:RTL342m almost completely neutralized the RTL therapeutic effect on EAE. In contrast, a 1:1 ratio of Fab:RTL342m had less neutralizing activity as assessed by daily EAE scores (Fig. 5A) and by the entire experimental effect on EAE for each group as assessed by the cumulative disease index (CDI) (Fig.

5B). Importantly, D2 Fab (also used at a 2:1 ratio) did not neutralize the therapeutic effect of RTL342m on EAE, indicating specificity of the 2E4 Fab for the two-domain RTL342m. In a recent phase I safety study in DR2+ MS subjects 34 to be treated with Fulvestrant clinical trial RTL1000 or placebo, we observed detectable baseline plasma levels of two-domain RTL-like structures in 4 of 13 donors (31%). This observation suggested the natural occurrence of two-domain Thymidylate synthase structures that could be derived from four-domain intermediates possibly shed from MHC-II expressing APC upon immunization. Using the power of our conformationally sensitive Fabs, we evaluated the appearance and persistence of naturally occurring two-domain MHC-II structures in human MS subjects. Fab 1B11 is specific for the two-domain HLA-DR conformation. It was found to bind to all HLA-DR-derived RTLs (with no peptide specificity), but not to other human and murine allele-derived RTLs or four-domain HLA-DR molecules (Fig. 6A). Serum or plasma samples were diluted 1:10 and adsorbed onto plastic wells pre-coated with the TU39 mAb (that detects all forms of MHC), washed and reacted with 1B11 Fab specific

for HLA-DR-derived RTLs, followed by the addition of enzyme-labeled anti-Fab and substrate for ELISA detection. As shown in Fig. 6B, the 1B11 Fab detected RTL-like material in serum or plasma from the healthy control pool as well as all six MS subjects tested at baseline, with detected levels of protein ranging from 13 to 1100 ng/mL. These results indicate for the first time the existence of soluble serum MHC-II structures with a distinct RTL-like conformation that differs from the classical membrane-bound MHC conformation. Increased signal for two-domain MHC-II was also observed in subject ♯42 after 30 min of infusion of 200 mg RTL1000 and in subject ♯44 after 2 h of infusion of 100 mg RTL1000, consistent with increased levels of injected RTL1000.

The cells were then washed in cold PBS solution and 1 mL of freshly prepared eBioscience Fix/Perm Buffer was added to each sample before incubating at 4°C for 40 min in the dark. After a second wash, 2% (2 μL) normal rat serum was added and the cells were incubated again at 4°C for 15 min. Anti-human Foxp3-PE was added and incubated at 4°C for 30 min in the dark. In another tube, anti-Foxp3-FITC (eBioscience) and anti-CTLA-4-PE (BD Biosciences) were added at the same time as anti-Foxp3-PE Ab. The appropriate isotype-matched control Abs were used to define positivity. The cells were washed twice with PBS

and fixed in 1% polyformaldehyde. Cells MG-132 research buy were analyzed on a FACSAria (BD Biosciences, San Jose, CA, USA) with FACSDiva software. T-lymphocytes were identified by gating on CD3+ T cells and side scatter, and Tregs were identified as CD25-positive and

Foxp3-positive cells found among p38 MAPK cancer CD4+ T cells within the lymphocyte gate. The absolute number of Treg cells was determined by multiplying the proportion of CD4+CD25+Foxp3+ with the total CD4+ T cell count. CTLA-4 expression within the Tregs was identified as the proportion of CTLA-4 positive cells within the CD4+, CD25+ and Foxp3+ cells. Whole blood samples were incubated with the monoclonal antibody combinations anti-HLA-FITC/anti-CD38-PE/anti-CD8-APC/anti-CD4-APC-Cy7 for 30 min at room temperature. After lysis of red blood cells by FACS lysis buffer (BD Biosciences), cells were washed twice, fixed with 1% polyformaldehyde and analyzed via FACSAria. Lymphocytes were identified by gating on forward scatter and side scatter,

then CD4+ or CD8+ T cells were gated. The percentage of HLA-DR and CD38 expression on CD4+ and CD8+ T cells was determined. PBMC depleted of CD25+ T cells was obtained with MACS CD25 MicroBeads (Miltenyi Biotec, Auburn, CA, USA). Briefly, fresh PBMC were washed twice in PBS-containing 0.5% BSA, resuspended in 80 μL of PBS containing 0.5% BSA and 20 μL of MACS CD25 MicroBeads per 107 total PBMC, and incubated for 25 min at 4–8°C. PBMC were washed twice in PBS-containing 0.5% BSA and applied Dichloromethane dehalogenase to a magnetic column on a MidiMACS separation unit (Miltenyi Biotec). CD25- T cell fractions were collected. PBMC and PBMC depleted of CD25+ T cells were stimulated with one of three treatments: phorbol 12-myristate 13-acetate (20 ng/mL; Sigma-Aldrich, St Louis, MO, USA) and ionomycin (1 μg/mL, Sigma-Aldrich), HIV Gag peptide mix (5 μg/mL; Lianmei, Xian, China), or RPMI 1640. Cells were supplemented with 15% FCS and incubated for 18 hr. Golgiplug (BD BioSciences) was added at a final concentration of 1 μL/106 cells for the last 6 hr of incubation. Cells were washed in PBS and were stained with CD8-APC and CD3-PerCP (BD BioSciences). Following permeabilization in permeabilizing solution (eBioScience), cells were stained with IFN-γ-FITC (BD BioSciences).

Although this region acts partly as an E1A enhancer in wild-type Ad5, the enhancer function is not necessary because a sufficient amount of E1A proteins are supplied in 293 cells. The loxP-insertion site at 191 nt of SgrAI in AdLC8cluc, which is the most popular helper Deforolimus virus, is extremely close to the cis-acting packaging domain AI described above. The virus

titers of helper viruses containing loxP at 143 nt in the AflIII cleavage site or at 192 nt in the BsrGI site have been studied (24, 26) (Fig. 1a). These previous reports suggested that the titer of the virus carrying loxP at 192 nt was slightly higher than, or not different to, that of the virus carrying loxP at 143 nt. However, both groups examined

only one pair of the viruses and so far no detailed examinations have since been performed. In this report, we constructed six pairs of AdV containing upstream loxP at 143 nt or 191 nt; we then compared the resulting virus titers and examined the influence of the loxP insertion FK228 cost upstream of the packaging domain AI. We observed that the viral titers of the AdV containing loxP at 143 nt was not lower and sometimes much higher than those of the AdV containing loxP at 191 nt. In a competition analysis, where two different viral genomes compete to be packaged into a viral shell, the insertion of loxP at both 143 nt and 191 nt reduced the packaging efficiency, compared with that of the competing AdV which did not contain loxP. These results suggested that the upstream insertion of loxP influences viral packaging. The human embryonic kidney cell line, 293, was cultured in DMEM supplemented Adenosine with 10% FCS. HeLa cells, derived from human cervical cancer, were also cultured in 10% FCS-DMEM. After infection with AdV, the cells were maintained in 5% FCS-DMEM. To examine the influence of loxP insertion near the packaging

domain AI, we constructed a new AdV, which has one loxP at 143 or 191 nt, a LacZ-expression unit, and another loxP at 466 nt, in this order (Fig. 1a). The left-end fragment of the Ad5 genome including a loxP at the AflIII site (143 nt), at which the loxP is located at approximately 150 nt after Klenow polymerase treatment, or at the SgrAI site (191 nt) was introduced into the cassette cosmid pAxcw (27); the former and latter positions were named here as 15L and 19L, respectively. The terms 15L and 19L also refer to the names of the viruses containing loxP at these sites. The resultant cosmid was termed pAx15Lcw or pAx19Lcw, respectively. The expression unit, expressing the LacZ gene under the control of the human polypeptide EF1α promoter (28), and the second loxP in this order were inserted at the SwaI site (464 nt; 454 nt in the original Ad5 genome), which is located downstream of the repeat AIIV in pAx15Lcw or pAx19Lcw; the resulting cosmid was named pAxLEFZ15L or pAxLEFZ19L, respectively.

Although vascular pedicle avulsion in breast reconstruction is an extremely rare complication, pedicle damage in free flap surgery is well documented,[11] while TD pedicle injury during axillary lymph nodes dissection is still poorly debated in literature. The most common causes of intra-operative pedicled flap failure are coupled to errors in surgical dissection, or excessive tension or torsion to the pedicle that

could give rise to flap ischemia and necrosis.[12, 13] Some new techniques for LD harvest might be effective for sparing muscle functions and achieving better aesthetic outcomes in recipient and donor sites, although increasing the chance of pedicle damage by the plastic surgeons.[14-16] In all reported five cases, the general surgeon injured Selleck BTK inhibitor the TD pedicle during axillary lymph-node dissection prior to complete breast reconstruction, damages occurring at different anatomical sites requiring different types of microsurgical repair. In two cases, an end-to-end anastomosis between the distal TDV stump and CSV was adopted as best option to flap salvage since the previously experienced shortening of the TD vein stumps after refreshing the edges could produce an unsafe primary anastomosis

limiting the flap’s arch of rotation. No doubt raised on case where a sharp, longitudinal laceration of TDV without tissue loss required a simple microsurgical repair. In case of TDV injury from previous surgery, where the scarring around TD pedicle made also CSV dissection difficult and unreliable, selleck compound surgeon was skilled enough to suddenly convert the pre-operative plan, considering the integrity of TD pedicle, from a pedicled to a free flap. In one case, the partial flap

loss probably occurred because of the shortening of arterial Erastin stumps that may have led to unsafe anastomosis under tension; moreover the strain on the vessel followed by implant positioning under the muscle may have caused arterial vasospasm, flap ischaemia and consequently occlusive clot of the vein. To salvage a LD flap from a pedicle injury, few points should be addressed. Feasibility of primary anastomosis should be always assessed, but depending on type of injury (sharp laceration, cauterization, avulsion) including or not a vessel tissue loss, as the stumps revisions may result in too short vessels contraindicating a direct under tension anastomosis. Time of injury is also important, as long lasting damage from previous surgery can severely obstruct vessels, wrapped in scar tissue not suitable for anastomosis. Finally, according to the anatomical level of injury different salvage options are available and should be preferred. For better understanding, the TD pedicle can be converted into a vascular path along a line extending from the apex of axilla to the anterior border of the muscle, where it provides two terminal branches, a horizontal and a descending.

40; P = 0.05). On the other hand, the mitochondrial antioxidant enzyme glutathione reductase decreased with severe agonal state (P = 0.003), while the

activity of glutathione-S-transferase declined with increased storage time (P = 0.005) and severe agonal state (P = 0.02). Conclusion: Our data highlight the influence of pre- and post mortem factors on preservation of mitochondrial function with implications for studies on brain pathology employing stored human BMS-777607 molecular weight samples. “
“Hypothermia has been shown to have neuroprotective effects in various models of neurological damage. However, its effects on pediatric status epilepticus (SE) are relatively unknown. In order to understand the effects of hypothermia on pediatric SE, we conducted experiments to determine the neuroprotective effects of mild hypothermic pretreatment in a model of pediatric SE. Juvenile (21-day-old) rats were subjected to mild hypothermic or normothermic conditions prior Paclitaxel price to intraperitoneal injections of pilocarpine. We

analyzed the seizure response of these animals via electroencephalogram and conducted ex-vivo analysis for apoptotic cells in the hippocampus via a TUNEL assay. We found that mild hypothermia increased both seizure latency and time to SE onset. It also reduced the overall average spike frequency and spike area compared to normothermia controls. Furthermore, the number of apoptotic cells was reduced in the hippocampus. In conclusion, these data indicate that mild hypothermia reduces both seizure activity and neurotoxicity in a pilocarpine model of pediatric these SE. This expands previous findings examining the neuroprotective effect of hypothermia by showing neuroprotection in a pediatric model of SE. We believe these findings will help researchers find better preventative treatments for

pediatric SE in the future. “
“R. H. Xia, N. Yosef and E. E. Ubogu (2010) Neuropathology and Applied Neurobiology36, 388–398 Selective expression and cellular localization of pro-inflammatory chemokine ligand/receptor pairs in the sciatic nerves of a severe murine experimental autoimmune neuritis model of Guillain–Barré syndrome Aims: To determine if specific pro-inflammatory chemokine ligand/receptor pairs expressed in the peripheral nerves of Guillain–Barré syndrome patients are expressed in a severe murine experimental autoimmune neuritis (sm-EAN) model and to determine their cellular localization as a prerequisite to designing potentially therapeutic interventions in vivo. Methods: Sm-EAN was induced in 8–12-week-old female SJL/J mice using bovine peripheral nerve myelin emulsified in complete Freund adjuvant with pertussis toxin and recombinant mouse interleukin-12 acting as co-adjuvants, with appropriate controls. Mice were evaluated for neuromuscular weakness and weighed daily. Dorsal caudal tail and sciatic nerve motor electrophysiological studies were performed at expected maximal severity.

This dual role was also seen in our results on HPC expansion: when used

alone, IL-32 led to twice the number of HPCs, whereas in combination with SCF, IL-32 significantly reduced cell expansion induced by SCF. Apart from its in vitro effects, IL-32 also increased the number of HPCs in vivo in a model of chemotherapy-induced BM suppression, thereby alleviating BM regeneration. The fact that, as with IL-1β 50, one injection of IL-32 sufficed, speaks in favor of the activation of secondary mechanisms. Interestingly, a rodent form of IL-32 has not yet been identified 44; the human homolog can, however, activate murine macrophages to secrete TNF-α 46. TNF-α has a detrimental effect on HPC renewal 51. Therefore, other bystander effects, in combination with the expansion potential of IL-32, are most likely responsible for a sustained stem cell renewal in click here a well-established mouse model 24. In conclusion, the combination of unbiased microarray analyses of IL-1β-stimulated ECs with a hypothesis-driven filtering by gene annotation allowed the targeted identification of cytokines with previously unknown hematopoietic growth factor

potential. The most outstanding discovery was that IL-32 induced the expansion of functional HPCs in vitro and in vivo, thus attenuating chemotherapy-related BM cytotoxicity; on the other hand, IL-32 reduced an SCF-dependent cell expansion. Future in vitro and in vivo studies will help to further define the role of IL-32 within hematopoiesis. Cord blood specimens www.selleckchem.com/products/BIBW2992.html were collected from full-term deliveries,

after informed consent was obtained from the mothers, and HPCs were immunomagnetically isolated as previously described 52. This study was approved by the ethical review board of the Charité. Human umbilical cord ECs were harvested and cultured as described previously 3. Confluent ECs of passages two to four were stimulated with IL-1β for 4, 8 and 16 h, and cells were harvested by collagenase (0.1% in PBS). CD34+ HPCs were used post isolation. Cell pellets were dissolved in RNA lysis buffer (Qiagen, Hilden, Germany) supplemented Selleck Gefitinib with β-mercaptoethanol (10 μg/mL) and stored at −80°C. Lysed cells were mixed with 0.2 mL of chloroform for 3 min at room temperature and then centrifuged at 11 500 rpm for 15 min at 4°C. The upper aqueous phase was collected in RNAse-free Eppendorf tubes and mixed with 0.5 mL isopropanol for 10 min. Supernatants were aspirated after recentrifugation, pellets were resuspended in 75% ethanol in DEPC-H20 air-dried and the RNA quantity was measured by spectrophotometry. Samples were run through an RNeasy column (Qiagen) and precipitated with ethanol. Total RNA was analyzed by Affymetrix 133 plus 2.0 arrays (Affymetrix, Santa Clara, CA) as previously described 53. Signal intensities for probe sets were derived using Affymetrix’s Microarray Suite version 5.

Under normal conditions, NK cells circulate in blood but are largely excluded

from lymph nodes. However, by using a mouse model, Sallusto check details and colleagues clearly demonstrated that circulating NK cells were selectively recruited to lymph nodes after stimulation by some, but not all, injected matured DCs in a CXCR3-dependent manner and that these NK cells provide an early source of IFN-γ that is mandatory for subsequent Th1 polarization [18]. These authors concluded that for vaccines that rely on Th1 responses, vaccine adjuvants should be selected according to their capacity to induce the recruitment of NK cells in antigen-stimulated lymph nodes. The findings in the present study are consistent with recently published data Venetoclax nmr from our group where mature αDC1s, from healthy blood donors, efficiently recruited NK cells in a CXCL9-dependent manner [16]. In that study, αDC1s were also able to induce IFN-γ production when co-cultured with resting autologous NK cells, but only if CD40 ligation was provided. As NKT cells express a similar chemokine receptor profile as NK cells [19], CD40 ligands could possibly be provided by co-recruited CD1d-restricted NKT cells which

are known to upregulate CD40L when recognizing endogenous glycolipids presented on mature DCs [26]. The polarization of naïve CD4+ T cells towards Th1 is highly dependent on IL-12 produced by mature DCs after re-stimulation with CD40 ligands [27, 28]. IFN-γ has been described to support such IL-12-dependent Th1 polarization by increasing the ability of mature DCs both to produce IL-12 upon re-stimulation with CD40-ligands [29] and to support TCR-mediated expression of the IL-12 receptor in naïve T cells [30]. Further, to induce an efficient Th1-polarized antitumour response, a vaccine DC must also efficiently induce rare tumour-specific naïve

CD8+ T cells to become CTLs. Indeed, we also found that αDC1s had a higher production of CD8+ T cell-recruiting chemokines CCL3/MIP-1α and CCL4/MIP-1β upon CD40 ligation. Interestingly, findings in mice from Germain for and colleagues indicate that after immunization but before antigen recognition, naive CD8+ T cells in vaccine-draining lymph nodes upregulate the chemokine receptor CCR5 [20]. This upregulation permits these cells to be attracted to sites of antigen-specific DC–CD4+ T cell interaction where the chemokines CCL3/MIP-1α and CCL4/MIP-1β are produced. Importantly, interference with this actively guided recruitment reduced the ability of CD4+ T cells to promote memory CD8+ T cell generation, which indicates that these chemokines are of high importance for the immune system to optimally activate rare antigen-specific CD8+ cells.