Our results show that HIV-specific

CD8+ T cells contribute significantly to IL-10 production in the peripheral blood and that this subset modulates monocyte activation. Constitutive IL-10 gene transcription was reported to be upregulated in multiple cell subsets among PBMCs in chronically HIV-infected individuals but there is uncertainty as to whether this is universally reflected in increased spontaneous or antigen-driven cytokine production [7]. We therefore analysed the Veliparib research buy fractions of IL-10-producing cells among circulating CD4+ T cells, CD8+ T cells, CD19+ B cells and CD14+ monocytes ex vivo, after stimulation with either 0.05% DMSO or HIV-1 gag peptides, in three subject

groups: patients who were antiretroviral (ART) naïve (n = 31, median viral load – 17 964 copies/mL) or fully suppressed on ART for >1 year (n = 20) and HIV-uninfected healthy controls (n = 5). Study participants’ characteristics are described in Table 1. The gating strategy used to identify IL-10-producing cells is shown in Supporting Information Fig. 1. Constitutive IL-10 release (0.05% DMSO control) was detected in all cell subsets analysed; the proportion of IL-10-producing cells was highest among CD19+ B cells and CD14+ find more monocytes in all three groups but there were no significant differences among the groups for each cell subset analysed, suggesting that constitutive IL-10 expression was not increased at the protein level in this patient cohort

(Supporting Information Fig. 2). By contrast, we observed significant IL-10 secretion in response to HIV-1 gag stimulation, predominantly in CD8+ T cells. These IL-10+ CD8+ T cells were rare but reproducibly detected in ART-naïve viraemic individuals, at a median frequency of 0.01% (range 0–0.13%, tenfold greater than the 0.05% DMSO control). Frequencies among ART-treated and uninfected Urease subjects were <0.001 and 0%, respectively (p < 0.01, Fig. 1A and B). Although the proportion of IL-10+ cells was lower among CD8+ T cells than monocytes, CD8+ T cells were the major contributors to IL-10 production in response to HIV-1 gag, due to the higher absolute numbers of CD8+ T cells in the peripheral blood (Fig. 1C). Phenotypic analysis of HIV-specific IL-10+ CD8+ T cells revealed that the majority were CD25- and FoxP3-negative and a substantial minority expressed CXCR3, a ligand for inflammatory chemokines that promotes migration to sites of inflammation and differentiation towards an effector phenotype [15] (Fig. 1D). We also investigated the expression of the gut-homing integrin alpha-4/beta-7 on HIV-specific IL-10+ CD8+ T cells, since IL-10 expression is upregulated in gut-associated lymphoid tissue (GALT) during acute HIV-1 infection [16].

Methods: Three hundred and twenty cases of biopsy-proven DPLN with ≥10% crescents (cDPLN) were included in this study. Another

180 DPLN patients without crescents were enrolled as a control group. Their clinicopathological data and long-term outcome were compared. Results: There were 280 females and 40 males with an average age of 31.8 ± 11.3 years followed for a median period of 7 years. Compared with the control HM781-36B mw group, cDPLN patients had a significant lower rate of clinical remission (CR+PR) (90.3% vs 96.5%, p = 0.036) for longer period (10.1 ± 7.9 vs 8.9 ± 7.6 months, p = 0.154), much higher rate of treatment failure (9.7% vs 3.5%, p = 0.036) and relapse (41.5% vs 37.8%, p = 0.511). The 5-, 10-and 15-year cumulative renal survival rates of cDPLN and the control group were 87% vs 90.8%, 73.3% vs 81.6% and 58.7 vs 81.6%, respectively. At the time of biopsy, higher percentage of crescents (HR 1.030, P = 0. 001), fibro-cellular crescents (HR 1.025, P = 0. 002), glomerular sclerosis (HR 1.033, P = 0. 022), impaired renal function (HR 1.519, P < 0.001), decreased eGFR (HR3.567, P = 0.003), higher levels of NAG enzyme (HR 1.009, P = 0. 014), urinary C3 (HR 1.046, P = 0. 024), serositis history (HR 2.814, P = 0. 013), failure to achieve clinical remission (HR 0.144,

P < 0.001) and relapse (HR 11.634, P = 0. 020), were the independent risk factors for worse renal survival of cDPLN patients. Multivariate find more Cox analysis showed the percentage of glomerular sclerosis was the most important risk factor of ESRD. Conclusion: cDPLN had worse treatment response and lower probability of renal survival than those without crescents. Ten clinicopathological features including a higher percentage of crescents, fibro-cellular crescents, glomerular sclerosis, impaired renal function, higher NAG enzyme, urinary C3, history of serositis, failure of achieving clinical remission and relapse were independent predictors of an unfavorable renal outcome. IKEUCHI HIDEKAZU, HIROMURA KEIJU, TSHILELA

KADIOMBO A, KAYAKABE KEN, SAKURAI NORIYUKI, SAKAIRI TORU, KANEKO YORIAKI, MAESHIMA AKITO, NOJIMA YOSHIHISA Department of Medicine and Clinical Science, Gunma University much Graduate School of Medicine Introduction: In this study we sought to identify predictive factors for renal insufficiency in patients with lupus nephritis (LN). Methods: We retrospectively analyzed 155 biopsy proven LN patients (21 male, 134 female) at our department between 1976 and 2012. Renal histology was classified by ISN/RPS 2003 classification. A renal endpoint was defined as doubling of serum creatinine (S-Cr) or end-stage renal disease. Results: The mean age at renal biopsy was 36.5 ± 13.2 years.

1C. Crosses indicate the death of individual mice at the marked time point. Data were obtained from three separate experiments. “
“Male patients with female-stem-cell donors have better prognosis compared to female-to-male combinations due to Y-encoded minor histocompatibility antigens recognized by female-alloimmune-effector lymphocytes in the context of a graft-versus-leukemia (GvL) effect. We provide data

in a dog-model that the minor histocompatibility antigen UTY might be a promising target to further improve GvL-immune reactions after allogeneic-stem-cell transplantations. Female-canine-UTY-specific T cells (CTLs) were stimulated in vitro using autologous-DCs loaded with three Selleckchem GPCR Compound Library HLA-A2-restricted-UTY-derived peptides (3-fold-expansion), and specific T cell responses were determined in 3/6 female dogs. CTLs specifically recognized/lysed autologous-female-peptide-loaded DCs, but not naïve-autologous-female DCs and monocytes. They mainly recognized bone-marrow (BM) and to a lower extent DCs, monocytes, PBMCs and B-cells from DLA-identical-male littermates

and peptide-loaded T2-cells in an MHC-I-restricted manner. A UTY-/male-specific reactivity was also obtained in vivo after stimulation of a female dog with DLA-identical-male PBMCs. In summary, we demonstrated natural UTY processing and presentation in dogs. We showed that female-dog CTLs were specifically stimulated by HLA-A2-restricted-UTY peptides, thereby enabling recognition of find more DLA-identical-male cells, mainly BM cells. These observations suggest UTY as a promising candidate-antigen to improve GvL-reactions

in the course of immunotherapy. Allogeneic-stem-cell 2-hydroxyphytanoyl-CoA lyase transplantation (alloSCT) represents the only curative therapy for many patients with haematological-malignancies including leukemia. The therapeutic-effect is mediated by donor-derived immune-effector cells infused with donor-lymphocyte transfusion (DLT) after transplantation. This approach is successful in treating relapsed myeloid-malignancies [1]. The favourable graft-versus-leukemia (GvL) effect of donor-lymphocytes is mainly mediated by allo-reactive T cells recognizing antigens (Ags) on hematopoietic-cells including the malignant leukemic-cells of the patients [2, 3]. These T cells can also be reactive towards healthy-tissues and cause graft-versus-host-disease (GvHD) [4, 5]. Own clinical observations demonstrated that in haploidentical-transplantations female-donors (especially mothers) show a higher GvL-effect against male-recipients (particularly sons) compared to all other haploidentical donor-recipient combinations [6, 7] (H. J. Kolb, unpublished data). These reactions might be due to the existence of male-associated antigens [8]. The Y-chromosome coded minor histocompatibility antigen (mHA) UTY (ubiquitously-transcribed-tetratrico-peptide-repeat-gene, Y-linked) could be a new immunotherapeutically useful potential candidate-target structure [8, 9].

When we try and reach the best coverage of the immunological repertoire, we actually aim to sequence as many immunoglobulin sequences as possible, out of the whole repertoire. That is, we aim to maximize the ratio between sequenced immunoglobulins (SI) to the total number of immunoglobulins (TI) in

the organism. We aim to reach an SI : TI ratio of 1. When this SI : TI ratio has been reached, an account for the entire repertoire can be obtained. Dorsomorphin solubility dmso Smaller model organisms, therefore, provide a better starting point from which to reach this ratio. Smaller organisms contain significantly fewer cells in total and, obviously, fewer immune cells. Much smaller organisms (e.g. the round worm) are sufficient for some aspects of immunology (see refs 31,32) but not for studying the lymphocyte repertoire. Zebrafish, Danio rario, are an ideal model system for studying the adaptive immune system for two reasons: first, they have the earliest recognizable adaptive immune system whose features match the essential human elements, and second, the zebrafish RG7420 immune system has only ∼ 300 000 antibody-producing B cells, making it three orders of magnitude simpler than mouse and five

orders of magnitude simpler than human. Recent works study the zebrafish B-cell repertoire via high throughput analysis.33 An important issue in the immune receptor diversity analysis is clone identification, e.g. classification of the obtained reads into clusters, under the assumption that relatively close sequences originate from the same clonally expanded cell. V(D)J segment identification is usually carried out by performing local alignment to germline sequences [available on the International ImMunoGeneTics (IMGT) database34]. However, D segment classification is more complex because of the short length of the sequence, as opposed to V and J genes. Furthermore, nucleotide deletions and P/N additions occur frequently during somatic recombination processes at the V–D and D–J junctions. Much immunological interest is focused on the complementarity determining region 3 (CDR3) of the chains,14,18–20,22,25,33 Farnesyltransferase the most

variable locus of the three CDR regions, and especially the β chain of the TCR.14,18–20,22,25 A recent study focused only on a small portion of the TCR-β repertoire by capturing only sequences generated by a specific gene recombination.22 Read length is a critical parameter in this case, as the entire V(D)J region is ∼ 300 nucleotides in length, including its V and J segments. This has been solved by either using the 454 method (with longer reads), the Douek approach (see above) or special methods of read assembly as in refs 14,25. Once sequences are available, different perspectives portray the repertoire: the size of the repertoire; similarities between repertoires; V(D)J segment use; nucleotide insertions and deletions; CDR lengths; and amino acid distributions along the CDRs.

[2] In some areas of the sheep placenta, called placentomes, there is aggressive interdigitation between trophoblast villi on the fetal side (cotyledon) and the

uterus on the maternal side (caruncle), and at points the epithelia form a common syncytium allowing for more efficiency of gas and nutrient exchange. Pigs have a similar but more diffuse placental structure than sheep with less aggressive interdigitation.[2, 17] The human/primate uterus is a single muscular organ different structurally from the two-horned uterus of rodents (for mice see Margaret J Cook’s CT99021 concentration book at www.jax.org), pigs,[18] rabbits,[16] or sheep.[19] While the electro-mechanics of the human/primate uterus may be fundamentally different from that seen in other species,[20, 21] the uteri of rodents,[22] rabbits[23] sheep,[24] and pigs[18] respond to oxytocin, suggesting a common expression

of the receptor, and most have been used to study the mechanisms underlying uterine contractility in vitro. In addition to hormones such as estrogen (discussed elsewhere), progesterone is a key hormone of pregnancy that appears to be differentially regulated in humans and animals.[25] The particulars of the responsiveness to this hormone and its interaction with estrogen in successful pregnancy remain selleck chemicals a topic of intense investigation. In humans, the corpus luteum is the major site of progesterone expression with help from chorionic

gonadotropin released by the early conceptus.[26] Blockade of progesterone during this time causes pregnancy loss.[26] Major production of progesterone switches to the placenta by 5–6 weeks’ gestation. Maternal serum levels of progesterone raise post-conceptionally and continue to elevate beyond parturition.[25, 27] However, progesterone has been given with variable success to treat women with recurrent miscarriage[28] and antiprogesterone given late in pregnancy can cause cervical ripening and delivery in some women[29] suggesting a complex biology. Human fetal membranes can produce[30] Aurora Kinase and metabolize progesterone,[31] and locally produced progesterone metabolites may be important in uterine quiescence and activation.[32] The human uterus can produce an inhibitory progesterone receptor which increases before parturition.[33] Finally, progesterone receptor regulation at multiple levels in the cytoplasm and the nucleus may regulate functional progesterone activity leading to parturition.[34] Progesterone’s regulation during pregnancy in related non-human primates is similar to human pregnancy in several respects including dependence on early production of progesterone by the corpus luteum[35] that early pregnancy can be interrupted by antiprogestins[36] and that there is not systemic withdrawal before parturition.

In short, 100 ng of total RNA was transcribed into cDNA using T7-(N)6 primers. Anti-sense RNA is generated by in vitro transcription of the cDNA, which is used as a template to generate 2′-deoxyuridine, 5′-triphosphate (dUTP)-containing sense cDNA fragments. Following the removal of RNA fragments, the sense cDNA fragments were terminal-labelled with biotin. The labelled samples were hybridized to the Human Gene 1·0 gene ST array (Affymetrix). The arrays were washed and stained with phycoerythrin-conjugated streptavidin (SAPE) using

the Affymetrix Fluidics Station® 450, and the arrays were scanned in the Affymetrix GeneArray® 3000 scanner to generate fluorescent images, as described in the Affymetrix GeneChip® protocol. Group sensitization rates were compared using a χ2 test and logistic regression analyses. The challenge outcome, Cisplatin supplier whether positive or negative, was used as

the dependent variable and skin status, sex and age as the independent variables. Results were expressed as odds ratios (OR) with 95% confidence intervals (CI). The strength of reactions in the three groups was analysed with both the visual scores and ultrasound Selleck EPZ-6438 data. The Mann–Whitney two-sample rank sum test was used to compare the sum clinical scores in the groups. Using the ultrasound data, linear regression analyses of the elicitation responses were calculated for each individual and the means of the slopes and the intercepts, used as parameters for strength of the elicitation reaction, were compared

for each group using a Mann–Whitney U-test, as recommended for serial measurements. P-value < 0·05 was considered to be statistically significant. All data analysis was performed using spss version 12 (SPSS, Inc., Chicago, IL, USA). The degree of infiltration of positively stained cells was scored semi-quantitatively using a five-point scale: 0 = none, 1 = few positive, 2 = some positive, 3 = many positive and 4 = highly positive. To detect significant differences between the group mean values, the Mann–Whitney U-test was applied. Significance was determined with a P-value < 0·05. All data analysis was performed using spss version 12 (SPSS, Inc.). Analysis Celecoxib the of microarray data followed the overall strategy described previously [12]. Briefly, a single log2 scale expression measure for each probe set was attained from the low-level data files (CEL files), using the robust multi-array analysis procedure with quantile normalization [13] implemented in the Affymetrix library for the r statistical environment. Principle component analysis (PCA) was carried out with the prcomp r function. The 2·5% of the probe sets with the most extreme positive or negative loading values for the first two principal components were extracted and used for analysis of over-represented GO terms using Fisher’s exact test for proportions with Bonferroni’s correction for multiple testing.

The present study investigated the neural differentiation of BMSCs, the lesion volume and axonal regrowth of injured spinal cord after transplantation. Seven days after spinal cord injury, 3 × 105 BMSCs or PBS (control) was delivered into the injury epicenter of the spinal cord. At 8 weeks after spinal cord injury, transplantation of BMSCs reduced the volume of cavity and increased spared white matter as compared to the control. BMSCs did not express the cell marker of neurons, astrocytes and oligodendrocytes

in injured spinal cord. Transmission electron microscopic examination displayed an increase in the number of axons in BMSC rats. The effect of BMSCs on growth of neuronal process was further click here investigated by using a coculture

system. The length and the number of neurites from spinal neurons significantly increased when they Enzalutamide cocultured with BMSCs. PCR and immunochemical analysis showed that BMSCs expressed brain-derived neurotrophic factor (BDNF) and glia cell line-derived neurotrophic factor (GDNF). These findings demonstrate that transplantation of BMSCs reduces lesion volume and promotes axonal regrowth of injured spinal cord. “
“We analyzed the incidence and extent of Lewy-related α-synucleinopathy (LBAS) in the olfactory mucosa, as well as the central and peripheral nervous systems of consecutive autopsy cases from a general geriatric hospital. The brain and olfactory mucosa were immunohistochemically examined using antibodies raised against phosphorylated α-synuclein. Thirty-nine out of 105 patients (37.1%) showed LBAS in the central or peripheral nervous systems. Seven patients presented LBAS (Lewy neurites) in the olfactory lamina propria

mucosa. One out of the seven cases also showed a Lewy neurite in a bundle of axons in the cribriform plate, but α-synuclein deposits were not detected in the olfactory receptor neurons. In particular, high incidence of α-synuclein immunopositive LBAS in the olfactory mucosa was present in the individuals with selleck kinase inhibitor clinically as well as neuropathologically confirmed Parkinson’s disease and dementia with Lewy bodies (6/8 cases, 75%). However, this pathologic alteration was rare in the cases with incidental or subclinical Lewy body diseases (LBD) (one out of 31 cases, 3.2%). In the olfactory bulb, the LBAS was usually present in the glomeruli and granular cells of most symptomatic and asymptomatic cases with LBD. Our studies further confirmed importance of the olfactory entry zone in propagation of LBAS in the human aging nervous system. “
“J. Duran-Vilaregut, J. del Valle, G. Manich, A. Camins, M. Pallàs, J. Vilaplana and C.

In addition to CD8+ IELs, the gut also hosts γδTCR T cells, NKT cells, and classical CD4+ T cells with αβTCR. The exact immune function of all these cells is unknown. The general tendency of these lymphocytes is to generate a tolerogenic immune response to antigens encountered in the gut lumen (20, 21). Other cellular types also participate in mounting an immune response. The most important for promoting oral tolerance are dendritic cells in the lamina propria, which infiltrate the area between

the latero-basal sides of the enterocytes and reach into the intestinal lumen with their projections, taking up antigens which are afterwards processed and presented into the mesenteric lymph nodes (22). Another important cell check details is the so-called M cell, placed as a hood over the luminal region of the PP. These M cells are in contact with MK-8669 purchase the gut content at their upper pole, allowing them to capture antigens and pass them over to the

immune milieu of the PP, where they are processed by other dendritic cells and then presented to lymphocytes in the local lymph nodes (23). It has been proved that a large proportion of intestinal dendritic cells express an enzyme called retinal dehydrogenase, (responsible for vitamin A metabolism), which produces a shift toward a tolerogenic phenotype in the case of the T helper cells that interact with these dendritic cells (24, 25). All these particularities of the enteric immune system result in generation, at the intestinal level, of Th regulatory cells, also known as iTreg, Tr1, Th3 and Th2 (26). Although intestinal T regulatory cells Casein kinase 1 are classical CD4+CD25+FoxP3+ regulatory cells, they appear in

the intestine, and not in the thymus (27). Tr1 (CD4+ IL-10+ FoxP3-) are regulatory cells which exert their function especially through the synthesis of IL-10, while Th3 (CD4+ TGF-β+ FoxP3+) rely on the release of TGF-β for the down regulation of immune responses. These regulatory subpopulations present numerous interconnections in vivo, probably leading to the existence of intermediate cellular types (28). All these characteristics make the gut a predominantly tolerogenic immune environment. The oral administration of any peptide can have three consequences: the secretion of anti-peptide IgA; a systemic immune response with the appearance of serum antibodies and cell-mediated immunity; or a state of anergy, local and/or general tolerance, which prevents an unwanted immune response when re-encountering an innocuous antigen. The first two situations are encountered in the case of pathogens with invasive potential, while the third possibility applies to commensal bacteria and dietary antigens, which do not cause local injuries or systemic immune responses (29).

First, historical

concepts related to selleck kinase inhibitor the detection of stretch by the vessel wall are reviewed, including the wall tension hypothesis, and the implications of the proposal that the arteriolar network responds to Pp changes as a system of series-coupled myogenic effectors. Next, the role of the myogenic response in the local regulation of blood flow and/or Pc is examined. Finally, the interaction of myogenic constriction and dilation with other local control mechanisms, including metabolic, neural and shear-dependent mechanisms, is discussed. Throughout the review, an attempt is made to integrate historical and current literature with an emphasis on the physiological role, rather than the underlying signaling mechanisms, of this important component of vascular control. “
“Please cite this paper as: Weiss M, Li P, Roberts MS. Estimation of sinusoidal flow heterogeneity in normal and diseased rat livers from tracer dilution data using a fractal model.

Microcirculation 19: 723–728, 2012. Objectives:  Up to now, vascular indicator-dilution curves have been analyzed by numerical integration or by fitting empirical functions to the data. Here, we apply a recently developed mechanistic model with the goal to quantitatively Fulvestrant clinical trial describe flow distribution in the sinusoidal network of normal rat livers and those with high-fat emulsion-induced NASH. Methods:  Single-pass outflow concentration data of sucrose were obtained from in situ perfused rat livers after impulse injection. The model fitted to the data consists of a continuous mixture of inverse Gaussian densities assuming a normal distribution of regional flow. It accounts for the fractal flow heterogeneity in the organ and has three adjustable parameters with a clear physiological interpretation. Results:  The model fitted the data well and revealed that the intrahepatic flow dispersion of 49.6 % in the control group increased significantly to 87.2 % in the NASH group (p < 0.01).

In contrast to previously used empirical functions, the present model exhibits a power-law tail (∼t−2.4), which is a signature of fractal microvascular networks. Conclusions:  The approach offers the Histone demethylase possibility to determine hepatic blood flow heterogeneity in perfused livers and to evaluate the functional implications. “
“Please cite this paper as: Neitzke, Harder, and Plagemann (2011). Intrauterine Growth Restriction and Developmental Programming of the Metabolic Syndrome: A Critical Appraisal. Microcirculation 18(4), 304–311. According to the “small baby syndrome hypothesis,” low birthweight and intrauterine growth restriction (IUGR) occurring in westernized countries mainly through altered placental flow, have been linked to increased metabolic syndrome risk in later life. Independency and causal mechanisms of this phenomenological association are a matter of controversy.

PAR-1, PAR-2 and PAR-3 were amplified with 35 cycles (94 °C for 30 s, 55 °C for 30 s, 72 °C for 60 s). PAR-4 was amplified with 35 cycles (94 °C for 30 s, 55 °C for 30 s, 72 °C for 30 s). Beta-actin (β-actin) was used as positive control using the following primer sequences: Selumetinib in vitro β-actin (sense) 5′-CCAAGGCCAACCGCGAGAAGATG-3′ and β-actin (antisense) 5′-AGGGTACATGGTGGTGCCGCCAG-3′; yielding a expected PCR product of 587 bp. Beta-actin was amplified

with 35 cycles (94 °C for 60 s, 60 °C for 90 s, 72 °C for 60 s). Negative control was performed for each reaction and included the omission of the reverse transcriptase or the omission of cDNA in the PCR mix. PCR products were resolved on a 1.5% agarose gel for visualization. Flow cytometry analysis was performed of the freshly isolated naïve CD14+ monocytes and the CD14+ monocytes cultured for 24 h with experimental conditions. Briefly, the freshly isolated naïve CD14+ monocyte cell pellet was washed in PBS containing 1% BSA and 0.1% Na-azide and subsequently used for incubation with fluorochrome-labelled antibodies. The CD14+ monocytes cultured with experimental

conditions for 24 h were placed on ice for 1 h. Subsequently, medium with CD14+ monocytes was transferred to 1.5-ml tubes and centrifuged at 900 g for 5 min at room temperature. Supernatants were harvested; the remaining CD14+ cell pellet was washed in PBS containing 1% BSA and 0.1% Na-azide, and centrifuged at 900 g for 5 min at room temperature. After centrifuging, Sodium butyrate freshly isolated naïve CD14+ monocytes as well as cultured CD14+ monocytes Cobimetinib were incubated with APC-conjugated monoclonal mouse anti-human CD14 antibody, PE-conjugated monoclonal mouse anti-human PAR-1 (ATAP2) antibody, FITC-conjugated monoclonal mouse anti-human PAR-2 (SAM11) antibody, PE-conjugated monoclonal mouse anti-human PAR-3 (8E8) antibody, FITC-conjugated polyclonal rabbit anti-human PAR-4 (APR-034-F)

antibody, PE-conjugated monoclonal mouse anti-human TF (HTF-1) antibody, and APC-, PE- and FITC-conjugated isotype control antibodies for 30 min at 4 °C in the dark. After a final washing and centrifuging step, cells were fixated in 2% paraformaldehyde. All cells were analysed using the FACS Calibur (BD Biosciences) and FlowJo software (Tree Star Inc., Ashland, OR, USA). For cytokine assays, naïve PBMCs and naïve CD14+ monocytes recuperated for 24 h and subsequently cultured according to the experimental conditions for 24 h were used. Supernatants were harvested, transferred to 1.5 ml tubes, centrifuged at 900 g for 5 min at room temperature and cryopreserved at −80 °C. Cytokine production (IL1-β, IL-6, IL-8, IL-10 and TNF-α) was determined in triplicate. Standard and positive control recovery for each ELISA assay was between 90–110%.