[7] In tissue sections, entomophthoromycotina are easily differentiated from other fungi by their characteristic hyphal morphology. The hyphae are broad, ribbon-like, aseptate or sparsely septate, with right- or wide-angle branching.[47] The histological inflammatory reaction shows

predominance of lymphocytes, Selleck SRT1720 plasma cells, epitheloid cells, multinucleate giant cells and histiocytes.[1] In addition, entomophthoromycosis is characterised by an important histological finding in the form of eosinophilic hyaline material around hyphae in haematoxylin and eosin (H&E) stained sections (Splendore–Hoeppli phenomenon).[2, 44] The characteristic histological and histochemical feature of basidiobolomycosis are illustrated in Fig. 2.[25] However, Splendore–Hoeppli phenomenon is not pathognomonic of entomophthoromycosis, as it is also seen in other infections e.g. sporotrichosis and schistosomiasis.[37] Typically, there is no evidence of angioinvasion, necrosis or tissue infarction.[21] Diagnosis of the disease remains difficult and may initially be missed. The fungus may be rare in tissue sections and when present is often fragmented.[2] Additionally, focal hyphae may appear

in only part of the specimen.[1] Moreover, fungal elements stain poorly with H&E and are not well demonstrated with fungus-specific tissue stains as periodic acid Schiff.[2] Examination of the fluorescent dye (Blankophor) wet-mount preparation under fluorescent microscopy increases the sensitivity of diagnosis.[18] Isolating the fungus by culture and molecular confirmation have epidemiological significance and also help in definitive diagnosis MLN2238 nmr and determining the susceptibility to antifungal agents.[18] Cultures should be inoculated

soon after tissue procurement, since the organisms do not survive at 4°C.[39] Entomophthoromycotina produce characteristic colonies on standard mycologic media e.g. SDA, potato dextrose agar or corn meal agar.[48] The colonies are dense, waxy, deeply furrowed and folded with a rapid growth at 37°C. The propulsion of conidia is characteristic of the genus. Conidia are forcibly ejected and stick to the Petri dish lid, thus clouding the view into culture with time.[2, 46] Although culture remains the ‘gold standard’ for disease diagnosis and species identification[2]; yet, recovery of fungi in the culture could Grape seed extract also be problematic. Countless results of negative cultures have been reported throughout the literature.[2, 49] This may be due to the aggressive processing of the specimen that occurs before plating, where fungal hyphae are often damaged and become non-viable.[2] Because of these difficulties in culture techniques and because successful management relies on early diagnosis; it has been agreed that microscopic identification of characteristic fungi should be considered significant even if the offending fungus couldn’t be recovered in culture.

[34] The misclassification of ectopy may also explain the discrepancy of findings across studies due to the lack of standardized criteria in

addition to variations in age and parity of participants. One of the most important methodological limitations find more of cross-sectional data is the imprecision of the timing of cervical ectopy in relation to HIV acquisition, which can introduce bias. Hence, studies have often been unable to assess the appearance of the cervix at the time of HIV acquisition.[12, 26] If cervical ectopy facilitates HIV acquisition and transmission, then it is important to identify other factors that influence the development of ectopy. Prior studies have noted an association between hormonal forms of contraception, primarily oral contraceptive pills, and the injectable depot medroxyprogesterone acetate, with increased ectopy[12]; this effect is likely mediated by the influence of estrogen on columnar epithelium.[5, 9, 35] Additionally, C. trachomatis has been shown to preferentially infect columnar cells, and hence, ectopy may increase exposure of susceptible cells to infection.[4]

C. trachomatis increases the susceptibility to acquiring HIV infection in women.[36] The interrelationships between cervical ectopy, hormonal contraception, C. trachomatis, and HIV are important selleck products to discern in young women, given that cervical ectopy, hormonal contraception use, and C. trachomatis are highly prevalent in this population. Additional mechanisms by which the cervical mucosa can be disrupted include Papanicolaou smears, trauma during sexual intercourse, as well as certain intravaginal practices by women in certain social settings. Because human studies cannot ethically

be designed to demonstrate HIV acquisition with or without Anidulafungin (LY303366) cervical ectopy, animal studies or ex vivo studies (i.e., explants, tissues samples) may provide the data to arrive at this causal association. Future studies would need to be mindful of additional confounding factors that could affect HIV acquisition, including STIs, ulcerative lesions, phase of menstrual cycle, inflammation, bacterial vaginosis, exudate, and alcohol use (see Table 2). It is difficult to reconcile the divergent results of observational studies assessing the impact of cervical ectopy on the increased risk of HIV acquisition. Ectopy is difficult to measure, and even when present, it is difficult to interpret. A recent review study did not find any evidence for the routine treatment of cervical ectopy.[37] Given that cervical ectopy is a common feature of the immature cervix, this may contribute to the disproportionate risk of HIV infection faced among young sexually active women in resource-limited settings, particularly in the hyperendemic regions of sub-Saharan Africa.

Our study quantified the intracellular CTLA-4 expression of Tregs in peripheral blood and found Autophagy Compound Library cost the expression of CTLA-4 was lower in HIV-infected SPs than in asymptomatic HIV-infected patients and AIDS patients, and that the level of CTLA-4 expression was inversely correlated with CD4+ T cell counts, but not correlated with viral load. It is reported that the intensity of CTLA-4 expression correlates with the suppressive capacity of cloned human CD4+CD25+ T cell populations and that the function of CTLA-4 is intimately

related to its expression (21, 22). Our results indicate that lower expression of CTLA-4 in HIV-infected SPs may limit the function of Tregs, which may contribute to the maintenance of functional immune

status in this population. These results agree with the findings described by Nilsson et al. who found that Tregs in lymphoid tissues express less CTLA-4 in non-progressors than in regular progressors (13). However, because expression of CTLA-4 is induced by T cell stimulation, further research might explore whether the lower expression level of CTLA-4 within Tregs can be attributed to the slower progression of HIV-infected SPs. This study uniquely shows the complex dynamics of the proportion and absolute number of Tregs in peripheral blood of HIV-infected SPs, which may have important clinical impacts for the prediction of the clinical progress of HIV infection. The LY294002 price authors thank Kumi Smith, Tristan Bice, and Naomi Juniper for their editing assistance. The study was supported by the Ministry of Health Science and Technology Special Mega Grant on Major Infectious

Disease (2008ZX1001-001), the Fund of the National Natural Science Foundation of China (30600532), the 973 Program for the Development of National Significant Elementary Research (2006CB504206), and a grant of the Key Laboratory of Liaoning Province (2008S242). HSP90 “
“Pandemic influenza H1N1 virus (A[H1N1]pdm09) emerged in 2009. To determine the phylogeography of A(H1N1)pdm09 in a single population, 70 strains of the virus were isolated from university students or trainee doctors at Tobetsu, Hokkaido, Japan, between September and December 2009. The nucleotide sequences of the HA1 region of the HA genes and described phylogenetic relationships of the strains circulating among them were analyzed. It was found that the 70 isolates could be phylogenetically separated into three groups and that two epidemics were caused by different groups of the virus. The three groups were also distinguishable from each other by three amino acid changes: A197T, S203T and Q293H. The substitution of S203T, which is located in the antigenic site, suggests antigenic drift of the virus. In March 2009, the first outbreak caused by swine-origin influenza virus A/H1N1 occurred in Mexico City.

All experiments were approved by the VAMC-Institutional Animal Care and Use Committee. Bone marrow (BM)-derived DCs (BMDCs) were generated from the femurs, tibias and pelvic bones of euthanized mice. The bones were cleaned with sterile Kim Wipes, both ends of each bone were cut, and the bone marrow was flushed out. Contaminating erythrocytes were lysed using ACK lysis buffer for 5 min at room temperature. Cells (1 × 106/mL/well) were cultured in 24-well plates using RPMI 1640 basal medium supplemented with 10% foetal bovine serum, 1% penicillin/streptomycin solution (Hyclone, Thermo Fisher Scientific, Waltham, MA, USA),

50 μm 2-mercaptoethanol (SIGMA, St Louis, MO, USA), 10 mm HEPES (Hyclone) and 20 ng/mL murine GM-CSF (R&D Systems, Minneapolis, MN, USA). The culture medium was changed completely every 2–3 days PI3K inhibitor with Y-27632 price fresh medium containing GM-CSF. The subset of DCs thus generated is referred to as myeloid-derived DCs (12). The cells were cultured and tested for the expression of DC markers at days 7, 10 and 14. Dendritic cell phenotyping targeted loosely adherent cells, collected by gentle pipetting, for the expression of DC markers. Day 14 was determined to be the most optimal time for maximal generation of DCs, because >95% cells expressed the DC differentiation markers. Cell viability was also determined by

the trypan blue exclusion test. In all the batches tested, the viability was >95%. The cells harvested from the BM or spleens were considered immature DCs. The immature DCs were exposed to various antigens for 18 h, whereupon the conditioned media (CM) and cells were harvested. Lipopolysaccharide (LPS) (SIGMA) was

dissolved as per the manufacturer’s instructions and used as a positive control at a concentration of 1 μg/mL. Peripheral blood mononuclear cells (PBMCs) were isolated from buffy coats of heparinized blood from anonymized healthy volunteer donor pools, using centrifugation on Ficoll–Hypaque gradients (SIGMA). Monocytes were isolated from PBMCs by positive selection using CD14+ learn more beads (Miltenyi Biotech, Boston, MA, USA). The CD14+ cells were cultured in RPMI 1640 with 10% FBS and 1% penicillin/streptomycin solution containing hGMCSF and hIL-4 (R&D systems), 50 and 14 ng/mL, respectively, for 5 days, until the cells were expressing >90% CD11c, CD11b and <5% CD14+. An increase in the appearance of other DC markers, such as CD86 and HLA-DR, was noted. Before specific antibody labelling, DCs were incubated with normal mouse and human IgG to block Fc receptors. Cells were then incubated with 200 μg/mL of antibody solution for 30 min in the dark at 4°C. The labelling buffer consisted of PBS with 1% FBS (21). The cells were washed and fixed with 1% paraformaldehyde and analysed using a BD Aria II cytometer using FACSDiva 6.1.1 software (Becton Dickinson, San Jose, CA, USA).

8%. However, the pooled incidence of AKI requiring RRT remained largely unaffected (pooled crude incidence, 0.86%). The increase of the pooled AKI incidence may reflect that AKIN and RIFLE criteria were the most sensitive diagnostic criteria for AKI among our studies. Besides, the study included patients undergoing noncardiac surgery[46] had the lowest Enzalutamide crude incidence

of AKI, among all the seven studies using AKIN and RIFLE criteria. These findings pointed out the impact of surgery type and diagnostic definition of AKI when considering the incidence of AKI. Importantly, since RIFLE and AKIN criteria have become the mainstays of diagnostic definition for AKI, caution should be exercised when it comes to interpret the past studies not applying these criteria for diagnosis. The strength of our meta-analysis and systemic review include the comprehensive search, the large sample size, the inclusion of latest studies with high methodological quality, multiple subgroup analyses, and low statistical heterogeneity with regards to the outcome of postoperative AKI requiring RRT. Our study also provided a review of the incidence of postoperative AKI and postoperative AKI requiring RRT in the context of the specific type of surgery and specific definition of AKI (Table 1). There were

several limitations of our study. As with all the observational studies, the causal relationship was hard to establish and there might be unknown confounders left unadjusted even after meticulous MTMR9 search for confounders. GSK2118436 Besides, the variation in types of surgery, the heterogeneity of the definition of postoperative

AKI, and the lack of the complete report of preoperative statin therapy were also problems. Different types of surgery pose different risk on postoperative AKI. In cardiac surgery, duration of CPB may be an important risk factor for AKI,[56] but this information was not provided in most studies. In other major surgeries other than cardiac surgery, the pathophysiology of renal insult is not as clear. The intensity of surgery-related insult to the kidney in different types of surgery may vary, and this effect was unable to be adjusted for. The level of emergency of the operation might also influence the risk of AKI, but this information was also unavailable for our meta-analysis. Although a dose dependent renoprotective effect was demonstrated in two studies,[43, 57] the majority of studies did not report the specific type, dosage, and duration of preoperative statin therapy. In studies reporting the detail of preoperative statin therapy, the specific type, dosage, and duration of statin therapy were often not uniform among studies. In chronic statin users, early re-institution of statin therapy after operation might be beneficial, but only one study[38] reported outcome relevant to this kind of statin exposure.

57 by 21 days. Vessel diameters did not change whereas complexity Selleck BI 6727 and density did, signaling remodeling. Conclusions:  This new automated analysis identified design parameters for tissue engraftment and could be used in other models of graft vessel biology to track proliferation and pruning of complex vessel beds. “
“Please cite this paper as:

Guo, Itoh, Toriumi, Yamada, Tomita, Hoshino and Suzuki (2011). Capillary Remodeling and Collateral Growth Without Angiogenesis After Unilateral Common Carotid Artery Occlusion in Mice. Microcirculation 18(3), 221–227. Objective:  To clarify the mechanisms of blood flow restoration after major artery occlusion, we presented first dynamic changes in cortical vessel morphology observed through a cranial window in mice after unilateral common carotid artery (CCA) occlusion. Methods:  The density and diameter of capillaries, as well as diameters of pial arteries, were measured by confocal laser-scanning microscopy and fluorescent microscopy, respectively. Possible angiogenesis was evaluated find more by detecting any outgrowth of endothelial cells from pre-existing vessels or intussusception

in Tie2-GFP mice. Results:  Immediately after unilateral CCA occlusion, cerebral blood flow (CBF) index, the reciprocal of mean transit time, reduced significantly and returned to the previous level after 14 days. Repeated observation of the cortical vessels did not reveal any angiogenesis, whereas the cortical capillary diameter increased by 74% after 14 days. The anterior cerebral artery (ACA) and collateral vessels connecting ACA and middle cerebral artery also dilated significantly. The capillary dilatation to the size of arteriole in the settings of collateral growth and CBF restoration suggested capillary remodeling. Conclusions:  Our results indicate that capillary remodeling, pial artery dilatation and collateral growth without angiogenesis are sufficient mechanisms to restore normal cerebral blood flow

after unilateral CCA occlusion. “
“This chapter contains sections titled: Mouse Embryo Manipulations for Live Imaging Imaging Vascular Development and Microcirculation Using Confocal Microscopy of Vital Fluorescent Markers Live Imaging of Mammalian Embryonic Development and Circulation with OCT Summary References “
“Please these cite this paper as: Doyle and Haas (2010). The Angiogenic Response to Skeletal Muscle Overload is not Dependent on Mast Cell Activation. Microcirculation17(7), 548–556. Objective:  To determine if mast cell activation in skeletal muscle contributes to overload-induced angiogenesis. Methods:  Extensor digitorum longus muscle was overloaded through extirpation of the synergist muscle tibialis anterior. Muscles were removed after 1, 2, 4, 7 or 14 days, and mast cell density and degranulation were quantified by histology. The mast cell stabilizer, cromolyn, was administered acutely or chronically to test if mast cell degranulation contributes to overload-induced angiogenesis.

School-age children were recruited from Welamosa primary school. Stools were microscopically examined for soil-transmitted helminth eggs and two groups of ten children, either geohelminth-infected or geohelminth-uninfected were included for immunological studies. Within the geohelminth-infected selleck chemicals llc children, four had A. lumbricoides, four had hookworms, one had both A. lumbricoides and hookworm and one had both T. trichiura and hookworm infections. Plasmodium spp. infections were absent as determined both by microscopy and by quantitative PCR analysis of donor blood.

The median age (11 years) and gender ratio was identical in geohelminth-infected and geohelminth-uninfected groups of children. To determine the immunological reactivity of geohelminth-infected versus geohelminth-uninfected children, we analyzed Ag-specific T-cell responses to BCG vaccine, P. falciparum pRBC or uRBC. BrdU incorporation by CD4+CD25+

cells was assessed to measure effector T-cell proliferation. T-cell proliferation to BCG and pRBC was lower in helminth-infected children (Fig. 1A) compared to uninfected children (geomeans 8.7 versus 13.5% and 8.6 versus 15.9%; p-values of 0.021 and 0.005, respectively), whereas proliferation in medium only or in response to uRBC did not differ between the groups (data not shown). As the observed helminth-dependent differences in immune responses could be the result of helminth-induced Treg, CD25hiFOXP3+ Autophagy inhibitor price T-cell numbers and costimulatory molecules were compared in helminth-infected and helminth-uninfected individuals. Similar proportions of CD4+ T cells from the two groups expressed CD25 (20 versus 25%; p=0.85), and there were similar populations of CD25hi T expressing

cells (5.4 versus 4.7%; p=0.57) as well as of CD25hiFOXP3 co-expressing T cells (0.7 versus 0.8%; p=0.68; Fig. 1B) in the CD4+ population. In a subset of the donors the expression check details of the activation markers CTLA-4 and GITR was assessed. Within these small sub-groups (four infected and seven uninfected), no significant differences were observed in expression of these two markers on either CD4+FOXP3+ or CD4+CD25hiFOXP3+ T cells (data not shown). To examine the functional capacity of Treg, CD4+CD25hi T cells were depleted from PBMC by magnetic beads. Following CD4+CD25hi T-cell depletion, CD4+CD25hi T-cell populations decreased from 1.74 to 0.67% and in parallel the CD4+CD25hiFOXP3+ population diminished from 0.90 to 0.33% (p<0.001 for both, Fig. 2A) in total CD4+ T cells. In three donors with very low numbers of CD4+CD25hi T cells, depletion failed and they were excluded from further analysis. Proliferation in response to different stimuli was measured in CD4+CD25hi T-cell-depleted and mock-depleted populations.

Hybridization to Affymetrix Human Gene 1·0 ST arrays AZD6244 cell line (764 885 probe sets, representing 28 869 annotated genes), staining, washing and scanning (Scanner 3000) procedures were performed as described by Affymetrix and performed by the Erasmus MC Center for Biomics. Probe set summarization, array QC and annotations

of the probe sets were performed using Affymetrix ‘Gene Expression Consolle’ (Affymetrix). All the different QC metrics analysed met the standards required by Affymetrix and showed an overall comparability of the signal distribution obtained from the different arrays. Principal component analysis was used to assess the underlying structure of the data set and define correlation relationships among samples (Partek Inc., St Louis, MO USA). Probe sets expressed differentially among conditions were identified using the class comparison tool implemented in BRB ArrayTools (National Cancer Institute, Bethesda, MD, USA). Briefly, we identified genes that were expressed differentially among the two classes using a random-variance t-test. The random-variance t-test is an improvement over the standard separate t-test as it permits

sharing information among genes about within-class variation without assuming that all genes Forskolin have the same variance. Genes were considered statistically significant if their P-value was less than 0·0001. A stringent significance threshold was used to limit the number of false positive findings. A ‘per gene’ estimate of the false discovery rates among genes passing the test was also computed. The false discovery rate associated with a row of the table is an estimate of the proportion of the genes with univariate P-values less than or equal to the one in that row that represent false positives. The Benjamini–Hochberg method for false discovery rate control was used for this estimation [32,33]. Genes passing the test threshold were clustered and displayed as a heatmap using Spotfire (Spotfire Inc., Somerville, MA, USA). The change in gene expression of a number of genes (IDO, IL-6, IL-8, CXCL10) as measured by microarray was confirmed

by real-time reverse transcription–polymerase Ergoloid chain reaction (RT–PCR). In brief, ASC were precultured under control, MLR (in transwell culture systems) or cytokine conditions and trypsinized at day 7. Total RNA was isolated and cDNA synthesized as described previously [34]. Quantitative gene expression was determined using TaqMan Universal PCR Master Mix and assays-on-demand for IDO (Hs 00158027.m1), IL-6 (Hs 00174131.m1), IL-8 (Hs00174114.m1) and CXCL10 (Hs 00171042.m1) (all Applied Biosystems, Foster City, CA, USA) on a StepOnePlus (Applied Biosystems). Data were analysed using paired t-test or Wilcoxon’s signed-rank test depending on the distribution of the data as tested with the Kolmogorov–Smirnov test for normality.

The wild-type strain, click here A. sobria 288, grown in 3  mL NB (0.5), was collected by centrifugation and the cells suspended in 0.3  mL distilled water. The suspension was

heated in boiling water for 10  min. The suspension was centrifuged to separate the precipitates from the supernatant. The supernatant was used as the source of the DNA template in PCR amplification and each set of oligonucleotides was used as a primer. The length of the DNA fragment amplified by the first and second sets was the 2251  bp and 1569  bp band, respectively. Subsequently, the amplified DNA in the reaction mixture was purified by treatment with phenol. The nucleotide sequence of each DNA was then determined by the dideoxy chain termination method. The protein

investigated in this study was shown to be a lipase and its amino acid sequence was deduced. Antiserum against the lipase was prepared by injecting the peptide GGDDNKGDTTSSLDYC-NH2, which is a keyhole limpet hemocyanin conjugate and composed of the 15 amino acid residues at the amino terminal end of the protein under investigation, into rabbits. Preparation of the antiserum was entrusted to the Peptide Institute (Mino, Osaka, Japan). A portion of overnight preculture of A. sobria 288 (asp−, amp−) (1  mL) was inoculated into 100  mL  NB (0.5). Bacteria were grown at 37°C with shaking at 140  r.p.m. At 6  hrs, 12  hrs, and 24  hrs, 20  mL of culture liquid was removed and the cells separated from the culture supernatant by centrifugation. https://www.selleckchem.com/products/nivolumab.html BCKDHB Proteins in the

culture supernatant of A. sobria 288 (asp−, amp−) were precipitated by treatment with TCA as follows: TCA solution was added to 1.0  mL of culture supernatant to reach a concentration of 10%. The mixture was left for 30  min at room temperature and the insoluble materials yielded collected by centrifugation. After rinsing with ethanol, the precipitates were suspended in  100 μL Tris-HCl buffer (pH 7.4). The cells recovered by centrifugation were suspended in 2  mL of 10  mM Tris-HCl buffer (pH 7.5). The cell suspension was divided equally into two tubes (1  mL/tube). A periplasmic fraction of the cells was prepared by treatment with polymyxin B (22). Polymyxin B solution (1  mL) was added to a tube containing cell suspension. The mixture was incubated at 4°C for 15  min. The concentration of polymyxin B in the mixture was 6500 U/mL. After incubation, the mixture was centrifuged (12,000 g for 15  min). The supernatant obtained was used as the periplasmic fraction. An outer membrane fraction of the cells was prepared by treatment with sodium lauryl sarcosinate by the method of Filip et al. (23). Briefly, the cells of another tube were broken by sonication, and the insoluble materials precipitated by centrifugation at 10,000 g for 20  min. To solubilize the cytoplasmic membranes selectively, the precipitates were suspended with sodium lauryl sarcosinate solution.

This observation is consistent with our results showing a better MΦ activation in the presence of NK cells in response to LASV, reaching selleck the levels observed after MOPV infection, regarding the expression of CD40, CD80, and CD86. LASV induced a limited activation in isolated MΦs with moderate levels of type I IFN mRNA [9]. However, this modest basal activation may initiate a positive loop of activation between MΦs and NK cells, leading finally to a robust NK-cell activation. It would be interesting to determine if this mutual activation of MΦs and NK cells occurs in LASV-infected patients or NHP. Indeed, as MΦ activation seems to be crucial to control

Arenavirus infection, such a mechanism could play an important role in the control of LF in survivors. Type I IFNs are well-known mediators of antiviral Selleckchem Sirolimus responses and are crucial for the activation of NK cells [14]. Our results suggest that, in addition

to cell contact, low levels of type I IFN are sufficient to mediate NK-cell activation, without triggering IFN-γ production or killing infected cells. Finally, we show here for the first time that, in our in vitro model, the pathogenicity of Arenaviruses does not seem to affect NK-cell activation. Further studies are required, to determine the role of NK cells in viral replication and T-cell responses in vivo in an animal model. Unlike NK/DC cross-talk, the interactions between NK cells and MΦs have not been studied in detail although the activation of NK cells in response to MΦs infected with many pathogens or stimulated by exogenous stimuli has already been reported [28, MYO10 29]. We show here that MΦs are involved in NK-cell activation, whereas DCs are not. This approach confirms the important role of MΦs in mediating NK-cell activation and, more generally, provides new insights and hypotheses into the immune mechanism operating during LF. The VeroE6 and K562 cells were grown in DMEM supplemented with 1% penicillin-streptomycin and 5% and 10% FCS respectively (all from Invitrogen). Mopeia

(AN21366 strain [2]) and Lassa (AV strain [30]) viruses were grown in VeroE6 cells at 37°C, with 5% CO2. Viral supernatants were harvested and used as the virus stock and the absence of mycoplasma was confirmed. LASV and MOPV titers were determined as described previously [6, 8]. Inactivated LASV and MOPV were obtained after 2-h heating at 60°C and at least two freeze/thaw cycles. Virus-free supernatants of VeroE6 cells were used for mock experiments. All experiments with LASV were carried out in biosafety level 4 facilities (Laboratoire P4 Jean Mérieux-Inserm, Lyon). Monocytes and peripheral lymphocytes were isolated from the blood of consenting healthy donors provided by the Etablissement Français du Sang (Lyon, France), as previously described [6].