The

wells of 96-well Nunc-immuno plates PLX3397 price (Nunc, Roskilde, Denmark) were coated with serial two-fold dilutions of antigen at 4°C overnight and then treated with 5% skim milk at 37°C for 1 hr to block nonspecific reactions. After washing five times, antigen was detected by anti-N MAb 13-27 and anti-G Mab 15-13, 13-13 and 15-10 (20). Following an additional five washes, horseradish peroxidase-conjugated anti-mouse IgG (Cappel, West Chester, PA, USA) was added to each well and incubated as above. After a final wash, the reaction was visualized with o-phenylenediamine (Sigma fast o-phenylenediamine dihydrochloride tablet sets, Sigma, St Louis, MO, USA) and stopped with H2SO4. The resulting OD490 was measured on a Model 559 Microplate Reader (Bio-Rad, Hercules, CA, USA). Monolayer cultures of NA cells were infected with each virus at a MOI of 0.01. After adsorption of virus for 1 hr, the cells were washed twice with Hank’s solution. Then fresh medium was added and the cells were incubated at 37°C. The culture media and cells were harvested at 1, 3 and 5 dpi. The virus in each sample was titrated in NA cells by focus assay as described above. For staining of viral foci, an IFA was performed using MAb 13-27 and FITC-conjugated rabbit IgG

to mouse IgG (Cappel). NA cells grown in 24-well culture plates were incubated with each virus PD0325901 in vitro at 150 FFU per well for various time periods (15, 30 and 60 min). Following washing of the cells, medium-0.5% methylcellulose mix was added and the cells were incubated at 37°C. After 2 days, cells were fixed by 4% paraformaldehyde and then permeabilized with methanol, followed by IFA as described above. Efficiency of internalization of each strain is indicated as relative focus number considering focus number at 60 min as 1. Cell-to-cell spread of each strain was examined by using NA cells grown on 24-well culture plates (Greiner Bio-one, Frickenhausen, Germany). The monolayer cells were infected with each strain at 50 FFU per well and incubated for 1 hr at 37°C. After removal of the inoculums, the cells were washed with Hanks’ solution.

A medium-0.5% methylcellulose mix was added to each well and the cells Olopatadine incubated at 37°C. After 48 and 72 hpi, the cells were fixed with 4% paraformaldehyde and then permeabilized with methanol. For staining of viral foci, an IFA was performed as described above. Photographs were taken with the Axiovert 200 or BZ-8000 digital microscope. Focus area was measured by Image J software (public software, http://rsbweb.nih.gov/ij/). Student’s t-test was applied for statistical analysis and P < 0.05 was considered to be statistically significant. We examined and compared the distribution of RC-HL strain- and R(G 242/255/268) strain-infected cells in the mouse brains by immunostaining for viral N protein. RC-HL strain-infected cells were found only in the hippocampi of the infected mouse brains (Figs 2a-2 and 2b-4 to 6).

In this study, we determined the fate and function of Lgr5-expressing cells Nutlin3a during thymic development. We show that TECs transiently express Lgr5 during fetal development and specifically marks a subset of TECs at E10.5 and E11.5. However, presence of Lgr5 is not essential for proper thymic development. Finally, lineage tracing confirmed that fetal Lgr5+ TECs do not generate detectable progeny in vivo. The presence of Lgr5 transcripts has been reported at E13.5 of thymic development in mice with a TEC-specific overexpression of β-catenin

[31]. We first set out to determine the temporal regulation of these Lgr5 transcripts in the fetal thymus. Fetal thymi of different ages were evaluated for presence of Lgr5 transcripts. Low levels of Lgr5 message were detected in the fetal thymus at E13.5 and E14.5 by RT-PCR. With increasing gestational age, the levels of Lgr5 transcripts gradually decreased and were undetectable from E19.5 onwards (Fig. 1A). To determine whether the observed Lgr5 transcripts lead to Lgr5 protein expression and to identify the cells expressing Lgr5, individual fetal thymi from Lgr5-EGFP-IRES-CreERT2 reporter mice in which EGFP marks

cells expressing Lgr5, were collected and single cell suspensions were made. First, the hematopoietic (CD45+) fraction was analyzed for the presence of Lgr5+ cells; however, at early and later embryonic age no considerable amount could be detected (Fig. 1B). Next, the epithelial fraction (CD45−EpCAM+) was analyzed by flow cytometry for EGFP expression Selleck MAPK inhibitor (Fig. 1C and D). In agreement with our transcript analysis, we found that the percentage of EGFP+ TECs was highest at E13.5 with a range from 2.17 to 7.37% (3.95 ± 1.51%). At later embryonic ages, Lgr5+ TECs

could still be detected; however, the number decreased with age; 0.02–0.64% (0.36 ± 0.19%) for E14.5, 0.05–0.242% (0.12 ± 0.10%) for E16.5 and 0.00–0.04% Mannose-binding protein-associated serine protease (0.05 ± 0.03%) at E19.5. In order to confirm that the Lgr5+ cells are indeed located within the thymus and to determine their in situ localization, fetal thymi of Lgr5-reporter embryos were analyzed by immuno-histochemistry. E10.5 complete embryos were sectioned and analyzed for the presence of Lgr5+ cells in the thymic anlage. The 3rd pharyngeal pouch at E10.5 clearly showed EGFP+ cells within the thymic primordium and these cells coexpressed the epithelial marker epithelial cell adhesion molecule (EpCAM) (Fig. 2A). At the right side of the pharyngeal region the number of EpCAM+EGFP+ cells appeared to be higher, consistent with earlier observations that there is asymmetry in developmental timing between the two sides of the embryo [32]. Next, sections of whole E11.5 embryos were analyzed. Also at E11.5, EpCAM+EGFP+ cells were clearly detectable within the thymic primordium and marked a subpopulation of fetal TECs.

Indeed, when just considering an alignment of the FHA domain region of the Pellino2 crystal structures, a sequence identity of 27.6% to the 3EGA crystal structure sequence and 25.5% to the

3EGB crystal structure sequence was observed (Fig. 1A). Modeller 9v5 21 was used to generate multiple models from both available templates of Pellino2 to examine the structure and stability of viral Opaganib Pellino modeled as an FHA domain. The best model was selected using a combination of the Modeller objective function score and a stereochemical analysis using ProCheck 22, 23 with only one outlier being identified. Subsequently, the model was minimised using MOE 2008 (http://www.chemcomp.com) in a 5 Å water sphere using the Amber99 force field to further examine its stability. Following this process, a stable 11-stranded

β-sandwich remained for viral Pellino (Fig. 1B). A topology-based comparison with Pellino2 demonstrates that the β-sandwich has the same strand orientation as that observed for the core FHA domain of Pellino2. To further assess the CHIR-99021 mw stability of our developed model, it was subjected to a 5 ns molecular dynamics simulation with a maximum root mean square deviation (RMSD) of 3.5 Å being experienced. An average structure was taken over the last 2 ns of simulation and upon examination of the secondary structure elements the 11-stranded β-sandwich had remained intact. This comparative model of viral Pellino superimposes well on the crystal structure of the Pellino2 FHA core region (Fig. 1C). This suggests that viral Pellino has the potential to form a core FHA domain without the wing appendage that is present in Pellino2. The lack of a wing appendage means that viral Pellino lacks the multiple IRAK phosphorylation sites Quisqualic acid present in Pellino2. However, viral Pellino contains most of the

highly conserved signature amino acid residues that are found in canonical FHA domains and that are required for binding to phosphorylated peptides and proteins. These five crucial residues in Pellino2 are R106, S137, R138, T187 and N188 18 and correspond to R33, S47, N48, Q85 and N86 in viral Pellino. Thus, viral Pellino contains four of the five highly conserved residues in classical FHA domains that are required for binding to phosphorylated protein-binding partners. This, in conjunction with the homology modeling described above, provides strong predictive indication that viral Pellino contains a core FHA domain. The ability of mammalian Pellinos to function as E3 ubiquitin ligases is bestowed by the presence of a C-terminal RING domain, where the eight cysteine and histidine residues are arranged in the atypical CHC2CHC2 formation. This RING domain is conserved between mammalian, nematode and Drosophila Pellinos.

The identity between NN and nurses’ strains is very selleck compound suggestive of a nosocomial acquisition from health-workers. “
“Pityriasis versicolor is a frequent mycosis and the use of systemic corticotherapy is one of its predisposing factors. This is an observational, cross-sectional, analytical and comparative study, conducted from January 2012 to January 2013 in the following outpatient clinics: Dermatology Service, Cassiano Antonio Moraes Hospital (HUCAM), Vitória, ES, Brazil;

Nephrology Service, HUCAM; and Leprosy Department, Maruípe Health Unit, Vitória, ES, Brazil. Patients, undergoing long-term systemic corticotherapy (or not), were assessed with respect to the presence of pityriasis versicolor. If there was mycosis, a direct mycological examination would be carried out. The spss 17.0 software was used for the statistical analysis. From the total of 100 patients, nine had pityriasis versicolor, being eight from the corticotherapy group and one from the group with no use of corticosteroids. Regarding the patients with mycosis, the prevalent age Selleckchem BIBW2992 ranged from 20 to 39 years, with six patients; six were women; seven mixed race; eight were undergoing long-term systemic corticotherapy; seven were taking low-dose systemic corticosteroids; four had leucocytosis; five had normal total cholesterol and triglycerides; and four had normal glycaemia. There was increased frequency

of pityriasis versicolor in the group undergoing systemic corticotherapy with statistical significance, corroborating the only study on the topic (1962). “
“Rhizopus is the most common genus of invasive mucormycosis, whose prognosis and outcome was not improved over the past decades. We studied the Gefitinib apoptotic-like phenotype in Rhizopus arrhizus exposed

to hydrogen peroxide (H2O2) and amphotericin B (AMB). The strain provided by Fungal Genetic Stock centre was studied about the apoptotic-like phenotype treated with different concentrations of H2O2 and AMB, and then analyzed by fluorescent microscopy (observed by Annexin-V/FITC and TUNEL staining), flow cytometry (stained with DHR123/PI), and DNA agarose gel electrophores. When R. arrhizus was treated with H2O2 and AMB, there was a loss of viability associated with different phenotype of apoptosis makers. Membrane externalization of phosphatidylserine (PS) on the cell surface, DNA fragmentation, chromatin condensation can be induced and observed obviously by Annexin-V/FITC, DAPI and TUNEL staining. DNA smear not DNA ladder was also visible in R. arrhizus. Flowcytometry of R. arrhizus cells revealed not only the increase of apoptosis cell stained with DHR123 under the nonfungicida doses but dead cells stained with PI under the fungicida concentrations.This study indicated that both H2O2 and AMB could induce the apoptotic-like phenotype in R. arrhizus. The incidence of invasive fungal disease has increased over the past two decades due to increasing numbers of immunocompromised individuals.

FACS data analysis was performed using FlowJo software. The research leading to these results has received funding from The European Community’s Seventh Framework Programme FP7 selleckchem under grant agreement no. HEALTH-F2–2008-223404, Centre for Medical Systems Biology (CMSB), and Dutch Arthritis

Foundation. The authors declare no financial or commercial conflict of interest As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors. Supporting Information Figure S1: IL-10 production by DX5+CD4+ Tcells DX5+CD4+TcellsandDX5-CD4+ T cells isolated from BALB/c mice that have received 3 injections withim mature DCs, were stimulated with anti-CD3 and anti-CD28. After 3 days IL-10 production was a ssessed by flow cytometry. Each bar represents one mouse. The experiment is performed more than 3 times. DX5+CD4+ T cells and DX5-CD4+ T cells are from the same mouse Supporting Information

Figure S2: gating strategy for figure 2The expression of the surface molecules (PDL-1, PDL-2, CD40, CD80, CD86 andMHC class II) on the modified bone marrow derived DCs was analyzed after dead cells were excluded based on FSC and SSC. The same Opaganib supplier gating strategy is applied for other surface molecules Supporting Information Figure S3: gating strategy for Figure 4 and 5CD4+T cells were isolated from D011.10 (OVA specific) mice. To identify IFN-γ production by OVA specific CD4+T cells, lymphocytes were first gated based on FSC and SSC, subsequently CD4 and KJ1-26 positive cells were gated and further analyzed for IFN-γ production. “
“The modulatory effects of solar ultraviolet radiation (UVR) on the immune system have been widely studied. As the skin is the main target of UVR, our purpose was to compare the impact of two contrasting ways to be exposed to sunlight on the

skin innate immunity. Hairless mice were UV irradiated with a single high UV dose (shUVd) simulating a harmful STK38 exposure, or with repetitive low UV doses (rlUVd) simulating short occasional daily exposures. Skin samples were taken at different times post-UV irradiation to evaluate skin histology, inflammatory cell recruitment, epidermal T cell population and the mitochondrial function of epidermal cells. The transcriptional profiles of pro-inflammatory cytokines, chemokines, antimicrobial peptides and TLRs were evaluated by RT-PCR and ELISA in tissue homogenates. Finally, a lymphangiography was performed to assess modification in the lymphatic vessel system. A shUVd produces a deep inflammatory state characterized by the production of pro-inflammatory cytokines and chemokines that, in turn, induces the recruitment of neutrophils and macrophages into the irradiated area.

Flow cytometric analysis was performed and positive events, i.e. antigen-specific T cells, were identified as a percentage of CD3+ CD8+ T cells. At least 50 000 events were obtained in the CD3+ CD8+ CD4− CD13−

CD19− population. The following antibodies (Abs) obtained from Beckman Coulter were used: anti-CD3-phycoerythrin-Texas red (clone Selleckchem GSI-IX CHT1) and anti-CD8α-FITC (clone T8) for positive gating, and anti-CD4-PCy5 (clone 13B8.2), anti-CD13-Pcy5 (clone SJ1D1) and anti-CD19-Pcy5 (clone J4.119) for negative gating. Positive tetramer staining was compared with staining with the iTag negative control tetramer. This gating strategy has been found to reliably identify ‘low-frequency’ events, for example melanoma-specific and Melan-A/melanoma antigen recognized by T-cell-1 (MART-1)-1 reactive CD8+ T cells, if the negative control tetramer reagent (loaded with an irrelevant peptide) is used to set the negative gate.22 Flow cytometry analysis was performed using an FC500 flow cytometer from Beckman Coulter (Krefeld, Germany). Eighty-eight overlapping peptides from TB10.4 were tested for binding

to five HLA-A molecules (A*0101, A*0201, A*0301, A*1101 and A*2402) and three HLA-B molecules (B*0702, B*0801 find more and B*1501). Binding to each allele is reported as a percentage relative to a positive control peptide for the respective MHC class I allele. With a cut-off of 20% binding as compared with the positive control peptide, we identified the following numbers of positive binding epitopes: two of 88 for A*0101, 17 of 88 for A*0201, two of 88 for A*0301, three of 88 for A*1101, 10 of 88 for A*2402, seven of

88 for B*0702, zero of 88 for B*0801 and 12 of 88 for B*1501 (Fig. 1, Table 1). The alleles HLA-A*0201 and HLA-A*2402 were among the most frequent MHC class I–peptide binders; they bound 20% and 11% of the candidate peptides, respectively. Also, HLA- B*1501 was among the top MHC class I-binding alleles; it bound to 14% of the TB10.4 peptide library. The prediction program syfpeithi (http://www.syfpeithi.de) picked up most TB10.4 epitopes for HLA-A*0201, A*2402 and A*1101; 17 of 17, old five of seven and two of three binding epitopes showed a syfpeithi score ≥ 10. For other MHC class I alleles, the program showed a lower success rate; for example, for B*0701 and B*1510, one of seven and five of 12 binding epitopes showed a syfpeithi score ≥ 10. Thirty-three of 88 candidate peptides bound at least to one MHC class I allele; the epitopes could be found throughout the whole amino acid sequence but with some clustering at the N- and C-termini (Fig. 2). Screening of TB10.4 peptides for binding to the eight most frequent Caucasian alleles revealed extensive cross-binding of the identical or closely related peptides to different MHC class I molecules.

Nevertheless, our analysis is focused on hypothesis-generation,

hence it is speculative in its attempt to integrate disparate observed molecular events to elucidate PGD pathogenesis. Also, this study has several limitations to its methodology, which must be addressed in the future. The antigen microarray used only screened a small fraction of all the proteins constituting the lung proteome, perhaps as few as click here 1%. Furthermore, this analysis gives no information about time-sequence causality of suggested processes involved. Prospective follow-up studies are needed to confirm our findings, as well as to elucidate how the reactive proteins as well as their down-stream components behave functionally over time in respect to the pathogenesis of PGD. The authors wish to thank Dr Noam Shental for advice on statistical design and analysis and Yoni Boxman for support and advice on scientific issues.

The work of P.H.H. was supported by a grant from the Lundbeck Foundation. The work of E.D. was partially supported by a grant from the Leir Charitable Foundation. The authors have no financial conflicts of interest. Additional Supporting Information may be found in the online version of this article: Figure S1. Concordance see more between IgG and IgM reactivity changes. Figure S2. Distributions of autoreactivities including both bronchiolitis obliterans syndrome and primary graft dysfunction status. Please note: Wiley-Blackwell are not responsible for the content or functionality of any supporting materials supplied

by the authors. Any queries (other than missing material) should be directed to the corresponding Isoconazole author for the article. Figure S1. Concordance between IgG and IgM reactivity changes. Figure S2. Distributions of autoreactivities including both BOS and PGD status. “
“Our previous study demonstrated that T helper (Th) cells from patients with rheumatoid arthritis (RA) display an altered expression profile of Notch receptors and enhanced activation of Notch signalling. The aim of this study was to investigate the role of distinct Notch receptors and ligands in the activation and differentiation of collagen II (CII)-reactive Th cells upon antigen-specific restimulation. Spleen mononuclear cells (SMNCs) from CII-immunized DBA/1J mice were restimulated by culturing with CII. CII-specific proliferation and differentiation of T cells were determined by tritiated thymidine (3[H]-TdR) incorporation and flow cytometric analysis, respectively. The mRNA expression of Notch receptors and Hes1 was assessed by real-time polymerase chain reaction (PCR).

Taken together, the available data suggest that AGS might be treated with reverse transcriptase inhibitors (RTIs: compounds that can potentially disrupt the replication cycle of both exogenous retroviruses and endogenous retro-elements).

Indeed, considering this possibility, Stetson et al. [26] dosed the Trex1-null mouse with the nucleoside analogue RTI azidothymidine (AZT) – but without obvious effect on the lethal phenotype. However, Doitsh et al. [43] showed, in the context of HIV-1 infection of CD4+ T cells, that AZT inhibits DNA elongation but not early DNA synthesis, indicating that it might be necessary to block reverse transcription at an earlier stage in order Sorafenib in vivo to avoid accumulation of immunostimulatory DNA. Taking this insight into account, Beck-Engeser et al. [44] have rescued the lethal Trex1-null murine phenotype by treatment with a combination of RTIs. On the assumption of no ‘off-target’ mechanism, this truly remarkable experiment indicates that the accumulation of cytosolic DNA in Trex1-null cells can be ameliorated by inhibiting endogenous retro-element cycling.

Importantly, we are aware of these results having been recapitulated in ITF2357 mouse an independent laboratory. RTIs are prescribed worldwide to children and adults (with HIV-1 infection), so that their pharmacodynamic, safety and toxicity profiles are already well characterized. There is no reason to predict that patients with AGS will demonstrate a distinct safety/toxicity profile when treated with these drugs, and so we are actively considering a trial of RTIs in AGS patients. One thing to note here is that any regimen employed will need to incorporate drugs capable of crossing the blood–brain barrier, an issue of no relevance in the Trex1-null mouse which does not demonstrate a neurological phenotype. The production of autoantibodies

against nucleic acids has been variably documented in AGS. Of note, Trex1-deficient mice [26] develop organ-targeted autoantibodies against cytosolic cardiac proteins, probably related to the lethal inflammatory myocarditis seen in these animals. Furthermore, a possible role of autoantibodies in AGS pathogenesis is indicated by substantial rescue of Cyclic nucleotide phosphodiesterase the Trex1-null mouse after crossing onto a B cell-deficient background [27]. Notably, these double knock-out mice demonstrate sustained increased levels of interferon, suggesting that interferon alone is not sufficient, on its own, to drive disease. The implication of lymphocytes and autoantibody production in AGS pathogenesis suggests possible therapeutic strategies, including the use of already licensed agents to deplete B cells. Other compounds of possible interest might include the use of medications, alone or as adjuvants, directed toward the probable presence of autoreactive T cells, such as mycophenolate mofetil. That such agents are established and often already approved for use in children – albeit for other indications – may facilitate clinical trial design and development.

Leprosy is a chronic infectious disease caused by Mycobacterium leprae (ML) affecting the peripheral nerves and skin. The major cause of disabilities observed in leprosy is the result of immunological reactions. These reactional episodes are classified as either reversal reaction (RR) or erythema nodosum leprosum.[1] buy Inhibitor Library It is well recognized that cell-mediated immunity is required for an effective response to ML infection.[2] Several studies have established that the production of T helper type 1 cytokines like interferon-γ (IFN-γ) by antigen-specific CD4+ T cells is critical in triggering a protective

immune response against ML.[3] These cells, found in the centre of tuberculoid granuloma, commonly present a memory phenotype.[4] Indeed, ML-specific CD8+ cytotoxic T cells have also been identified in tuberculoid leprosy lesions and appear to benefit their host via granulysin-mediated bacillus killing.[5-7] Reversal reaction, the major cause of the nerve function

impairments resulting in disability and deformity, is characterized by the appearance of new leprosy lesions and the inflammation of existing ones. The immunopathology underlying RR consists of an increased cell-mediated immune response accompanied by CD4+ T cells and macrophage activation in addition to increased expression of pro-inflammatory mediators such as IFN-γ,tumour necrosis factor, interleukins 6, 2 and 12p40, and matrix

learn more metalloproteinases 2 and 9, resulting in an inflammatory response in the skin and peripheral nerves.[8-11] Several lines of evidence suggest that CD4+ ML-responsive T cells with a T helper type 1 phenotype may be responsible for the immune-mediated damage occurring during RR.[12] The impact of HIV infection on the profile of the cell-mediated immune in response to ML is still unknown. Preliminary reports focusing on co-infection suggested that HIV infection Exoribonuclease does not affect the clinical classification of leprosy.[13] Although CD4+ T-cell-mediated immunity is compromised in HIV infection, it is broadly accepted that HIV infection does not lead to the multibacillary lepromatous form of the disease, as was previously believed.[14, 15] In a longitudinal study conducted with a cohort of co-infected patients in Brazil, it was noted that 66·7% of the co-infected patients were paucibacillary[11]. In addition, analyses of bacillary loads in multibacillary patients demonstrated that HIV+ patients presented a lower bacillary load than HIV− patients before multidrug therapy, which suggests that co-infected patients tended to have the tuberculoid form and lower bacillary loads.[16] As highly active antiretroviral therapy (HAART) has become more readily available for the treatment of AIDS in countries where leprosy is endemic, more than 40 cases of RR associated with immune reconstitution inflammatory syndrome have been reported.

2A–F). PD-1 has been implicated in the negative regulation of T lymphocyte function during chronic viral infections Cetuximab ic50 17. Therefore, we next analyzed whether PD-1 expression was detectable on NK cells from Tx patients. Our results demonstrate a significant up-regulation of PD-1 expression on all NK cells from patients with PTLD (36±24%), as compared with those from asymptomatic pediatric Tx patients (UVL: 16±3%; LVL: 15±5%) or HC (14±6%) resembling “exhausted” T-cell phenotypes (Fig. 3A). PD-1 up-regulation

was also detected on CD56brightCD16± and CD56dimCD16+ NK-cell subsets from PTLD patients (Fig. 3B and C) as well as on the unusual CD56dimCD16− and CD56−CD16+ subsets (data not shown). In addition, a trend of PD-1 up-regulation on NK cells was noted in chronic HVL carriers (22±13%) (Fig. 3A and B). We next analyzed the ability of CD56brightCD16± NK cells to respond by IFN-γ production and of CD56dimCD16+ NK cells to up-regulate CD107a (as a measurement

of active granule (perforin) exocytosis and NK cytotoxic potential) 18 to non-specific stimulation (pro-inflammatory Type-1 promoting cytokines), or to EBV-antigen-specific stimulation with autologous lymphoblastoid cell lines (LCL). In particular, hrIL-12p70+hrIL-18 stimulation triggered strong IFN-γ responses from CD56brightCD16± NK cells from asymptomatic Tx patients (UVL: 30±14%, LVL: 33±16%; HVL: 25±15%) and HC (32±10%) (Fig. 4A). EBV-antigen-specific stimulation with LCL triggered lower levels of IFN-γ RG7422 supplier release as compared with the non-specific stimulation, but still most effective with CD56brightCD16± NK cells

Methocarbamol from HC (6±4%) and LVL (6±3%) patients (Fig. 4B). Surprisingly, although NK cells from UVL patients showed IFN-γ responses to hrIL-12p70+hrIL-18 stimulation comparable to those from HC or to asymptomatic patients that carry an EBV load (LVL and HVL) (Fig. 4A), they displayed lower IFN-γ (UVL: 3±3%) responses following EBV-antigen-specific LCL (Fig. 4B). In contrast, PTLD patients showed impaired IFN-γ production by CD56brightCD16± cells to non-specific (13±12%) as compared with UVL, LVL and HC or to EBV-specific stimulation (2±3%) as compared with LVL and HC (Fig. 4A and B) suggesting their profound functional alteration. Furthermore, while the CD107a response was not significantly modulated by hrIL-12p70+hrIL-18 cytokine treatment (Fig. 4C), it was significantly boosted by EBV-LCL stimulation resulting in CD107a+ CD56dimCD16+ NK cells from HC (4±2%) and LVL (3±3%) patients (Fig. 4D). Similar to the IFN-γ response, the CD107a response to EBV-LCL stimulation was decreased in UVL patients (1±2%) as compared with that of HC and LVL carriers (Fig. 4D). Conversely, both PTLD (1±1%) and HVL (1±1%) patients presented with significantly decreased CD107a+ CD56dimCD16+ NK cells in response to LCL trigger (Fig. 4D).