In addition to IL-10 production, other facets of tolerance, namely, anergy and suppression (both in vitro and in vivo), were affinity dependent, with i.n. Ac1–9[4Y]-, [4A]- or [4K]-treated CD4+ T cells being the most, intermediate and least anergic/suppressive, respectively. These findings demonstrate that the generation of IL-10 Treg in vivo is driven by high signal strength. Antigen administered in a tolerogenic form has long been known to result in down-regulation of immune responses. Our previous studies demonstrated tolerance induction in WT B10.PL mice by i.n. administration of the N-terminal peptide of ICG-001 purchase myelin basic protein (MBP), Ac1–9[4K], the immunodominant

encephalitogenic epitope in H-2u mice, as measured by decreased EAE severity upon subsequent challenge 1. MBP Ac1–9[4K] forms highly unstable complexes with the MHC class II molecule H-2 Au2. Using MBP Ac1–9 peptide analogs

with an alanine or PD0325901 price tyrosine substitution at position four, displaying a hierarchy in affinity for H-2 Au (MBP Ac1–9[4K]<<[4A]<[4Y]), we previously found that protection from EAE correlated with peptide affinity for H-2 Au1. The Tg4 TCR Tg mouse was generated so as to circumvent the limitations imposed by low T-cell precursor frequency in the WT mice 3. The use of the Tg4 mouse model demonstrated that T-cell deletion was only transient and incomplete after a single dose of a high-affinity analog of the MBP epitope, Ac1–9[4Y]. Repeated administration resulted in down-regulation of the capacity of Tg4 CD4+ T cells to proliferate and a shift in cytokine secretion from IL-2, IL-4 and IFN-γ to IL-10 (but not TGF-β) production 4, 5. In addition to protection against EAE, the peptide-induced tolerant cells were shown Chloroambucil to suppress proliferation of responder Tg4 CD4+ T cells, both in vitro and in vivo6. The role of IL-10 in suppression was subsequently confirmed

by administration of blocking anti-IL-10R and anti-IL-10 antibodies 4, 6. Of note, peptide-induced IL-10-secreting CD4+ T regulatory cells (IL-10 Treg) were found to be distinct from naturally occurring Treg in that they did not express Foxp3 7. Furthermore, genetic depletion of FoxP3+ Treg from the CD4+ T-cell repertoire in the RAG-deficient Tg4 mouse gave rise to spontaneous EAE, the onset of which could be prevented by repetitive treatment with i.n. peptide, correlating with the generation of IL-10 Treg 8. In our most recent study, we have shown that repeated i.n. peptide treatment gave rise to IL-10 Treg that originated from Th1 cells 9. Thus, in view of the apparent correlation between protection from EAE and the affinity of MBP Ac1–9 analogs for H-2 Au, as well as the role of IL-10 in tolerance, it was of interest to investigate the ability of the analogs to induce IL-10 production.

24,25 Recent epidemiological studies have shown a strong association between ED and LUTS.26–28 In a large-scale, multinational survey, Rosen et al.26 reported that LUTS are independent risk factors for sexual dysfunction in older men. Demir et al. also reported that ED was diagnosed in 65.2% of patients with moderate LUTS and 81.8% of patients with severe LUTS, and metabolic syndrome may play a key role in the pathogenesis of both selleck screening library ED and LUTS.27 From the link between ED and hypercholesterolemia, as well as the link between ED and

LUTS, it is possible to derive a relationship between OAB and hypercholesterolemia, although there has been a controversial study that reported that only obstructive LUTS is associated with ED.28 As mentioned previously, several studies have been conducted to investigate the relationship between OAB and hypercholesterolemia in animal models.9–11 Son et al.10 reported detrusor overactivity

in hypercholesterolemic rats. In this study, Sprague–Dawley rats were fed a daily 1% cholesterol diet for 8 weeks to induce hypercholesterolemia, and a 2-week treatment of 3 mg/mL NG-nitro-L-arginine methyl ester was added to induce intimal changes AZD1208 purchase that would make rats vulnerable to atherosclerosis, and this method is the same method used to create vasculogenic ED rat models. As a result, the cholesterol group had shorter voiding intervals (377.6 ± 205.4 versus 121.8 ± 79.6 s, P Liothyronine Sodium < 0.01) and a smaller

functional bladder volume (1.4 ± 0.7 versus 0.7 ± 0.3 mL, P < 0.05) on cystometrography compared to the control group. Rahman et al.9 also reported similar results around the same time. To induce hypercholesterolemia, they fed Sprague–Dawley rats a diet consisting of 2% cholesterol and 10% lard for 6 months, and then performed awake cystometry. Twelve of 15 hyperlipidaemic rats had bladder overactivity, with multiple episodes of bladder contractions with or without voiding, beginning soon after infusion and occurring throughout bladder filling, while only one of nine controls showed bladder overactivity. These observations were further corroborated in another rat model by Huang et al.11, who also used a 2% cholesterol and 10% lard diet to induce hypercholesterolemia and observed that the micturition interval was significantly shorter and mean volume per void was significantly less in high-fat diet rats than in control rats. In addition, there are several studies suggesting a link between DO and metabolic syndrome. A link between detrusor overactivity and metabolic syndrome was also reported. A study that employed a fructose-fed rat (FFR) model, which is often employed to study metabolic syndrome, reported that unstable bladder contractions suggestive of DO occurred in 62.5% of male FFRs, compared with none in controls.

Forty-seven patients with anti-GBM disease were enrolled in this study. Forty-eight healthy individuals were used as normal controls. The levels of serum BAFF and APRIL were assessed using commercially available enzyme linked immunosorbent assay kits. The association between the levels of serum BAFF and APRIL, and the clinical and pathological parameters were further evaluated. The serum levels STI571 nmr of BAFF and APRIL in patients with anti-GBM disease were significantly

higher than that in normal controls (12.3 ± 14.1 ng/mL vs. 0.9 ± 0.3 ng/mL, P < 0.001; 19.1 ± 22.9 ng/mL vs. 1.6 ± 4.6 ng/mL, P < 0.001), respectively. The levels of serum APRIL were correlated with the titres of anti-GBM antibodies (r = 0.347, P = 0.041), and the levels of serum BAFF were associated with the percentage of glomeruli with crescents (r = 0.482, P = 0.015) in patients with anti-GBM disease. The levels of serum BAFF and APRIL were raised in patients with anti-GBM disease and might

be associated with disease activity and kidney damage. “
“Angiotensin-(1–7) (Ang-(1–7)) opposes angiotensin-II-induced cell growth, matrix accumulation and fibrosis in cardiac tissue. However, the role of Ang-(1–7) in the pathogenesis of renal fibrosis is uncertain. This study observed the effects of Ang-(1–7), on its own or in combination with losartan, an angiotensin-receptor blocker, on five-sixths RG7204 ic50 nephrectomized rats. Male Sprague–Dawley rats underwent five-sixths nephrectomy, Ribociclib mouse and then were either untreated, treated with Ang-(1–7), treated with losartan, or treated with a combination therapy of Ang-(1–7) and

losartan. After 8 weeks, renal function was assessed by measuring systolic blood pressure, serum creatinine and proteinuria. The effect of nephrectomy on the renin–angiotensin system was examined by measuring plasma levels of Ang-II and Ang-(1–7). The extent of glomerulosclerosis and tubulointerstitial fibrosis was assessed by periodic acid-Schiff staining and Masson-trichrome staining. The expression of plasminogen activator inhibitor-1, fibronectin and angiopoietins-Tie-2 was investigated by immunohistochemistry and western blot. In the groups of treated rats, serum creatinine, proteinuria and markers of glomerulosclerosis, such as fibronectin and plasminogen activator inhibitor-1, were ameliorated compared with the untreated, nephrectomized rats. Plasma Ang-(1–7) levels were elevated in all treatment groups, but the plasma Ang-II levels were reduced in the Ang-(1–7)-treated group and the combination therapy group. The ratio of Ang-1/Ang-2 was increased in the combination therapy group compared with two other treatment groups. Ang-(1–7) ameliorated the renal injury of nephrectomized rats. The combination of Ang-(1–7) treatment alongside losartan exerted a superior effect to that of Ang-(1–7) alone on regression of glomerulosclerosis.

Our patient was demonstrated to have a combined mutation to both CFH and MCP. Combined mutations have been reported in approximately 3% of https://www.selleckchem.com/products/BKM-120.html patients.[3] CFH blocks the formation of

C3 convertase and accelerates its breakdown. CFH can also bind to negatively charged molecules within the kidney to regulate the activation of complement on the cell surface. The surface of glomerular endothelium shows high levels of MCP expression where it provides additional cofactor activity for CFI. Wild-type MCP should have been present in the donor kidney and the donor did not undergo MCP genotyping. It is of interest the recipient of the partner kidney also developed ABMR/TMA to a less severe degree, unfortunately neither the donor or the second recipient was tested for complement mutations. Post-transplant focus is usually on the risk of recurrent aHUS. The risk depends on the genetic abnormality involved and is higher in patients with CFI and CFH mutations and may be up to 50–100% in these groups compared with 15–20% in the group with MCP mutations.[4-6] It has been shown that 50% of patients with confirmed aHUS have recurrent disease in the

graft after transplant, and of these 90% progress to graft failure.[4, 6] Although there is increasing interest PI3K inhibitor in the role of complement in the development and propagation of acute antibody-mediated renal allograft rejection

via terminal complement activation[1] very little is known about the incidence of AMR in patients with aHUS, who would theoretically be at increased risk. Interesting to note, in the study by Le Quintrec,[2] Vasopressin Receptor that 60% of patients with recurrent aHUS had rejection. Same group demonstrated that 30% of patients with de novo TMA post transplant had a mutation in CFH or CFI.[7] Very little study has been done on the impact of complement dysregulation on the development of anti HLA antibodies however the strength of the HLA antibody formation was striking in this case. Of interest is the case report by Noone et al.[8] of a patient with ESKD secondary to spina bifida whose first graft was lost due to acute rejection and who was subsequently highly sensitized. The patient received a second transplant following a desensitization protocol with a graft to which she had 3 low titre DSA. She developed early oliguric renal failure, severe TMA that was unresponsive to standard therapy and significant increases in antibodies to the mismatched class I and II antigens. She was treated with 2 doses of eculizumab with good effect with rapid normalization of her platelets and creatinine. Subsequent renal biopsy demonstrated ABMR. Complement factor H related protein 3/1 deficiency was subsequently demonstrated.

TDP-43-immunoreactive inclusions affected more of the cortical profile in longer duration cases; their distribution varied with disease subtype, but was unrelated to Braak tangle score. Different TDP-43-immunoreactive

inclusions were not spatially correlated. Conclusions: Laminar distribution of pathological features in 10 sporadic cases of FTLD-TDP is heterogeneous and may be accounted for, in part, by disease subtype and disease duration. In addition, the feedforward and feedback cortico-cortical connections may be compromised in FTLD-TDP. “
“Angiocentric glioma (AG) is an epileptogenic benign cerebral tumor primarily affecting children and young adults, and characterized histopathologically Romidepsin by an angiocentric pattern of growth of monomorphous bipolar cells with features of ependymal

differentiation (WHO grade I). We report an unusual cerebral glial tumor in a 66-year-old woman with generalized tonic-clonic seizure; the patient also had a 6-year history of headache. On MRI, the tumor appeared as a large T2-hyperintense lesion involving the right insular gyri-anterior temporal lobe, with post-contrast enhancement in the BTK inhibitor insula region. Histopathologically, the tumor involving the insular cortex-subcortical white matter was composed of GFAP-positive glial cells showing two different morphologies: one type had monomorphous bipolar cytoplasm and was angiocentric with circumferential alignment to the blood vessels, with dot-like structures positive for epithelial membrane antigen and a Ki-67 labeling index of <1%, and the other was apparently astrocytic, being diffusely and more widely distributed in the parenchyma, showing mitoses and a Ki-67 labeling index of >5%. In the anterior temporal lobe, a diffuse increase in the number of astrocytic cells was evident in part of the cortex and subcortical white matter. On the basis of these findings, we considered whether the present

ifenprodil tumor may represent an unusual example of AG with infiltrating astrocytic cells showing primary anaplastic features (AG with anaplastic features), or anaplastic astrocytoma showing primary vascular-associated ependymal differentiation (anaplastic astrocytoma with angiocentric ependymal differentiation). At present, the latter appears to be the more appropriate interpretation. “
“Malignant peripheral nerve sheath tumor (MPNST) is an uncommon type of sarcoma that arises from peripheral nerve sheaths and rarely involves the spinal roots. The origin of this tumor is thought to be Schwann cells or pluripotent cells of the neural crest. The subgroup of tumors in which malignant Schwann cells coexist with malignant rhabdomyoblasts is termed malignant triton tumor (MTT). MPNSTs can show different degrees of malignancy, but overall spinal MTTs are high-grade lesions.

Results of the apoptosis percentage are referred to this basal value. In our study, neither FPR2/ALX agonists nor CysLT1 antagonists exerted any effect on the inhibition of neutrophil survival induced by IL-8 (100 nM) at the concentrations tested (0·1 nM–1 μM) (Fig. 4). Caspase inhibitor I was used as a control of apoptosis inhibition, resulting in a complete blockade of caspase 3/7 activity. Similar results were observed using annexin V staining as a marker BTK inhibitor in vitro of apoptotic cells and propidium iodide as a control of the number

of necrotic cells (Figs 5 and 5). 15-epi-LXA4 (100 nM) could not reverse the percentage of neutrophil apoptosis arrest induced by IL-8 stimulation (21% and 23% of apoptotic cells in IL-8 alone and IL-8 plus 15-epi-LXA4, respectively). As expected, the CXCR2 antagonist SCH527123 reversed IL-8-induced apoptosis

arrest and returned the apoptotic cell index to the basal conditions (Fig. 6). Of interest, compound 43 (100 nM) by itself increased neutrophil survival in the absence of IL-8, confirming the recent published data regarding the inflammatory actions associated with this small molecule FPR2/ALX agonist [28, 32]. All the other reference compounds tested showed no effect on neutrophil survival by themselves (Fig. 6). Overall, these results indicate that 15-epi-LXA4 is inactive in reversing the survival signal induced by proinflammatory Rucaparib purchase chemokines such as IL-8 in human neutrophils, and compound 43 by itself induces proinflammatory signals in neutrophils. LXs and 15-epi-LXs are arachidonic acid-derived metabolites suggested to play an important

role as novel anti-inflammatory and pro-resolution agents. LX stable analogues display potent bioactivity in vivo in several murine model systems of acute inflammation [25] and block airway hyper-responsiveness and allergic inflammation in ovalbumin and cockroach allergen-induced airway inflammation models [26]. In addition, transgenic over-expressing mice of human FPR2/ALX receptor show shorter resolution times and doses required in response to lipoxin stable Tideglusib analogues [16], and are protected from acid-induced acute lung injury [33] and allergen-induced pulmonary inflammation [34]. FPR2 knock-down cell lines no longer signal in response to LXA4 and deficiency of FPR2 in mice decreases the ability of lipoxin A4 and annexin peptide to reduce inflammation in vivo [14, 15]. Nevertheless, all the in-vivo data supporting the role of FPR2/ALX mediating the anti-inflammatory actions of LXs has been generated in mice and differences in FPR2/ALX signalling between species cannot be discarded. Moreover, no FPR2/ALX knock-out or transgenic mice studies have been addressed to study in particular the relevance of the LX–FPR2/ALX axis in neutrophil migration in vivo. In humans, differences in FPR2/ALX expression have been observed in acute and chronic inflammatory responses.

In conclusion, patient-centered and quality of life outcome measures are an important part of evaluating the usefulness of FFR of lower extremity wounds. Without procedure-specific assessments currently available, these outcomes can be easily measured using standardized questionnaires such as

the SF-12 or SF-36. We have shown that microsurgical flap reconstruction is a valuable reconstructive option in high-risk patients and offers a HRQoL comparable with that of the general population. In addition, successful ambulation in patients who have undergone FFR improves HRQoL, whereas quality of life is decreased significantly when failure to ambulate occurs. “
“Literature on the reconstruction of the proximal femur in skeletally immature patients with the use of an epiphyseal transplant is scarce and with variable results depending on the indication. We report Ulixertinib datasheet successful outcomes using selleck screening library a modified vascularized fibular epiphyseal transplant in a 4-year-old boy with an oncologic lesion. We discuss the advantages of supplementing the standard graft with a vascularized fibular periosteal tissue. The vascularized fibular epiphyseal transplant (VFET) is an effective option in the reconstruction of the epiphysis in skeletally immature patients, owing to the

advantage of restoring both the joint function and the growth potential in a single surgical operation.1 Multiple reported cases demonstrate the effectiveness of this complex technique in upper extremity reconstruction.1,2 However, literature is scarce regarding its use for the reconstruction of the proximal femur and hip joint.3-5 Through this article, we report the use of a VFET in the reconstruction of a proximal femur in a 4-year-old boy after an intra-articular wide excision of an epithelioid hemangioendotelioma. We also discuss PRKACG the advantages of designing the flap as a composite

vascularized epiphyseo-osteo-periosteal flap.6 © 2012 Wiley Periodicals, Inc. Microsurgery, 2012. “
“Two cases are reported of flap loss following microsurgical perforator flap breast reconstruction in patients diagnosed with a factor V Leiden mutation. Factor V Leiden is the most common inherited cause of hypercoagulability, leading to an increased risk of thrombotic events. The first patient underwent a deep inferior epigastric artery perforator flap and then had recurrent arterial thrombosis both intraoperatively and postoperatively. This patient was subsequently diagnosed with a factor V Leiden mutation. The second patient had a known factor V Leiden mutation and underwent a superior gluteal artery perforator flap, which developed thrombosis and flap loss 2 days later. Preoperative assessment of a personal or family history of unexplained venous or arterial thrombosis should prompt suspicion of a factor V Leiden mutation.

The number of clotting episodes and the premature termination of HD were studied as the primary end points. We observed intradialytic hypotension and hypocalcaemia MK-2206 datasheet as secondary outcomes Data was analyzed using SPSS Version 15. Results: The baseline Characteristics (Table 1) were comparable between groups except for the higher number of femoral access in the CD with saline group. The number of clotting episodes were

significantly lower in the CD with saline group (p = 0.02, figure 1), and the use of CD alone was not superior in this regard when compared to SF (p = 0.27). There was no symptomatic hypocalcemia in either group. The mean fall in serum calcium level was 0.3 mg/dl in the CD groups when compared to 0.1 mg in the SF group (p = 0.36). Other complications including intradialytic hypotension (p = 0.09) were comparable between the groups. Conclusion: Citrate-containing standard bicarbonate dialysate in combination with intermittent saline flushes was better at preventing circuit clotting in heparin free HD in ICU. The use of citrate did not increase the occurrence of symptomatic hypocalcaemia or intradialytic hypotension during ICU dialysis. PRATT RAYMOND D, LIN VIVIAN, GUSS CARRIE, GUPTA AJAY Rockwell Medical Introduction: Triferic added to bicarbonate concentrate crosses the dialyzer membrane and binds to apotransferrin during hemodialysis. Methods: In this double-blind RCT, 103 iron replete, CKD-HD patients were randomized

to either Triferic dialysate (2 μM or 110 μg iron/L) or placebo, provided as premixed liquid bicarbonate. Dabrafenib price Two strata were prospectively defined by baseline ESA dose (Epoetin equivalent units): 1 (<13,000 U/week) and 2 (≥13,000 U/week). ESA prescriptions were managed by a centralized Anemia Management Center to facilitate consistent adherence to the protocol-specified hemoglobin target range of 95 to 115 g/L. IV iron administration was protocol defined. Results: The

primary end-point was the change in prescribed ESA dose from baseline to end of treatment (EoT). Triferic required 35% less prescribed ESA compared to placebo (p = 0.045) in the primary analysis. The subgroup analysis examined the effect of Triferic in patients with relative ESA resistance (baseline ESA doses ≥13,000 U/week) compared to those who were normo-responsive to ESA. (Table) Triferic reduced ESA utilization in both subgroups, Cyclin-dependent kinase 3 compared to placebo controls. Subgroup size was not large enough for statistical significance. However the results were similar in each subgroup i.e. a reduction in prescribed ESA favoring Triferic. The effect was numerically larger in the hypo-responsive group. Reticulocyte Hgb was better maintained with Triferic than in Placebo. The adverse and serious adverse events in the Triferic group were typical for CKD-HD patients and similar in type, frequency and severity to placebo. There were no anaphylaxis events in this study and no death was attributed to Triferic.

Cells were fixed and stained with anti-IL-17A-PE, according to the manufacturer’s protocol (♯555028 BD Biosciences) and analyzed on the FACS calibur. Forty and sixty-four hours post stimulation, 1 μCi of [3H]-thymidine (ICN Biochemicals) was added to each well containing 50 000 of unseparated splenocytes and lymph node cells; for CD4+ and CD8+ cells 25 000 cells find more were used, followed by additional 8 h incubation. Plates were harvested with the TOMTEC cell harvester and [3H]-thymidine

incorporation was measured usina a TRILUX Microbeta counter (PerkinElmer Life Science). Data were obtained from triplicate samples for each treatment. Flat-bottom Immulon 2HB plates (Fisher Scientific) were coated overnight with 3 μg/mL of capture anti-mouse IL-17 antibody (R&D Systems, Minneapolis, MN) in 1× PBS. Plates were blocked with 2% BSA and 5% sucrose in 1× PBS at room temperature for 1 h. Recombinant mouse IL-17 (standard curve) and the supernatant from

the in vitro stimulation were diluted 1:2, then added in duplicate to the ELISA plates and incubated for 2 h at room temperature. Plates R788 datasheet were washed and incubated with biotinylated anti-mouse IL-17 (R&D Systems) for 1 h at 37°C, followed by additional washes and incubation with neutravidin–alkaline phosphatase for 30 min at room temperature. Plates were then developed with the AP substrate, para-nitrophenyl phosphate (Pierce), in 0.2% diethanolamine substrate buffer (Pierce) and were read at 405 nm in a SpectraMax spectrophotometer (Molecular Devices). Similar procedures were used for IFN-γ, IL-2 and IL-4 ELISAs, according to the manufacturer’s protocol. lck-DPP2 kd and littermate controls were immunized

with 100 μg of OVA in CFA (Sigma) s.c.. Ten to fourteen days later mice were boosted with 100 μg of OVA in IFA (Sigma) s.c. Ten to fourteen days after boosting, the mice were sacrificed, and the draining lymph nodes were harvested for in vitro stimulation with OVA. Fixed human HEp-2 cells (Antibodies) were stained with mouse serum according to the manufacturer’s instructions, except the secondary 3-oxoacyl-(acyl-carrier-protein) reductase antibody was FITC-conjugated F(ab)2 goat antimouse IgG (Jackson Immunoresearch). The slides were mounted with ProLong Gold antifade reagent (Invitrogen) and digitally photographed with a Nikon E400 fluorescence microscope. We thank Dr. Albert Tai for stimulating discussions and help with the immunofluorescence experiments. We also thank Greta Fabbri for assistance with some of the qRT-PCR data. The work was supported by NIH RO1 AI043469 (BTH) and by the Esche Fund (BTH). Conflict of interest: The authors declare no financial or commercial conflict of interest. Detailed facts of importance to specialist readers are published as ”Supporting Information”. Such documents are peer-reviewed, but not copy-edited or typeset. They are made available as submitted by the authors.

MDDCs, differentiated and infected as above, were pulsed for 3 h with 3 µg/ml of a CEF peptide pool containing 23 human leucocyte antigen (HLA)-ABC-restricted T cell epitopes from human cytomegalovirus, Epstein–Barr and influenza viruses (CEF) (Anaspec Inc., Fremont, CA, USA). The negative fraction obtained from the monocyte isolation (to serve as the pool of autologous T cells) was suspended at 1 × 107 cells/ml in 5 mM CellTrace™ carboxyfluorescein succinimidyl ester (CFSE) 10 min at 37°C and 5% CO2 in 15 ml polypropylene conical tubes MK-8669 mouse in the dark. The cells were then washed, incubated for 5 min on ice, pelleted by centrifugation and suspended at 1 × 106 cells/ml in complete media. A total

of 250 000 CFSE-labelled autologous cells from the negative fraction AZD9291 and 25 000 DC from each condition were co-cultured together in the dark for 7 days at 37°C and 5% CO2 with a negative control culture containing colchicine (100 ng/ml) (Sigma-Aldrich, Milwaukee, WI, USA). Co-cultures were then transferred to 5-ml polypropylene round-bottomed tubes and stained with PE-conjugated

anti-CD8 antibodies (R&D Systems). CD8+ T cell proliferation was measured by flow cytometric analysis (CFSE dilution). Only those cultures that proliferated in response to the CEF antigen pool beyond the level of media controls were included in the analysis (six of 12). Data were analysed using paired t-tests or the Wilcoxon rank-sum test when appropriate for identification of statistically significant differences (P ≤ 0·05 was considered significant) between experimental groups using Sigma Plot 8·0 (Systat Software Inc., Chicago, IL, USA). Monocytes isolated from PBMCs of healthy donors using CD14+ magnetic bead isolation expressed high GNA12 surface levels of CD14, CD40 and MHC I and low levels of surface DC-SIGN/CD209, CD83, CD80, CD86 and MHC II (Fig. 1a), consistent with the published literature [3,61]. Immature MDDCs differentiated from monocytes using GM-CSF and IL-4 expressed low surface levels of CD14 and high levels of DC-SIGN (Fig. 2). Immature MDDC also expressed higher levels of surface CD83,

CD80, CD86, CD40, MHC-I and MHC-II (Fig. 1b). Finally, after incubation of the iMDDC with the maturation-inducing cytokine cocktail consisting of TNF-α, IL-1β, IL-6 and PGE2 for 48 h, mMDDC were observed to express high levels of CD83, CD80, CD86, CD40, CCR7 and MHC-I and MHC-II, with a low level of DC-SIGN expression and negligible CD14 expression (Fig. 1c). Therefore, monocytes, iMDDCs and mMDDCs all expressed surface molecules consistent with that reported in the literature [58]. After a 24-h incubation with HIV-1 and 48 h of culture, HIV-1 DNA was detected consistently in HIV-1-infected cultures (Fig. 2). There was no detectable HIV-1 DNA in the mock-infected cultures over the same period of time (Fig. 4). Phenotypic analysis.