Any level of elevated buy MK-8669 MCT may be a falsely elevated, even very high MCT: three samples with very high IgM RF values were reduced by 17 to 39% following HBT treatment. The MCT levels became normal in all three (41·8 to 2·6 µg/l; 160 to 5·2 µg/l; 200 to 4·1 µg/l) with 94%, 97% and 98% reduction, respectively. These patients had diagnoses of rheumatoid arthritis in the first two cases and non-Hodgkin lymphoma in the latter, respectively; none had any clinical history of mast cell increase or activation. Another sample with a raised RF (in

a patient with rheumatoid arthritis) had a 47% reduction in MCT (13·9 to 7·3 µg/l). Overall, there was no clear correlation between the measured IgM RF levels and the degree of reduction in MCT. This is due probably to variability in binding of mouse IgG Fc or AZD9291 supplier to the variability in the relative total amounts of IgG RF and IgA RF in individual sera (which are not measured in the IgM RF assay). HAMA interference can

also occur in the absence of RF but appears uncommon: one sample (systemic mastocytosis) with significantly raised tryptase level (319 µg/l) had almost undetectable levels of RF but raised levels of IgG HAMA (A450 0·115). Following blocking treatment, the tryptase result remained elevated (246 µg/l) but reduced by more than 17%, but the IgG HAMA dropped to normal levels (A450 0·087). Nine of 13 samples with a >17%

reduction in tryptase after HBT absorption had positive HAMA (A450 > 0·095) and eight of these became negative for HAMA after HBT treatment (one sample insufficient for HBT treatment) (Table 1). Heterophile antibodies can also lead potentially to false negative results, but we found little evidence for this in our cohort. In one RF-negative sample there was an apparent increase in MCT level >17% after HBT treatment (18·8 to 22·2 µg/l). In two RF-positive samples GNA12 analysed, there was an apparent increase in MCT following HBT treatment (43·3 to 49·2 and 128 to 143 µg/l), 14% and 12%, respectively. Both samples showed a decrease in RF level (314 to 102 and 129 to 82). HAMA was not detected in the first of these samples and there was insufficient material to measure HAMA in the second sample. We needed to ensure that the apparent presence of IgM RF was not itself caused by HAMA. Of the 14 samples with raised IgM RF, 13 had sufficient serum remaining to allow the analysis of HAMA. Of these, three were negative for IgG HAMA with the remaining samples having very low levels (A450 values between 0·095 and 0·197), and the blocking experiments revealed no samples that appeared to have false positive RF levels due to HAMA (Table 1).

Understanding the role of primary cilia in the kidney continues to provide clues concerning the pathogenesis of cystic kidney disease as well as epithelial homeostasis and regeneration. The near ubiquitous presence of primary cilia on epithelial cells in the kidney means that their involvement should be considered in a wide range of renal diseases and injuries. We

thank the Rotary Club of Wodonga and the Australian Chapter of the PKD foundation for supporting our studies of polycystic kidney disease. The micrographs in Figures 2 and 3 of this manuscript were obtained using instruments maintained by Monash MicroImaging. The Monash Institute of Medical Research is supported by the Victorian Government’s Operational Infrastructure Support Program. “
“Lupus selleck compound nephritis (LN) is a common and important manifestation of systemic lupus erythematosus (SLE). Evidence suggests higher rates of lupus renal involvement in Asian populations, and maybe more severe nephritis, compared with other racial or ethnic groups. The management of LN has evolved considerably over the past three decades, based on observations from clinical studies

that investigated different immunosuppressive agents including corticosteroids, cyclophosphamide, azathioprine, mycophenolic acid, calcineurin inhibitors and novel biologic therapies. This is accompanied by improvements in both the short-term treatment response RAD001 rate and long-term renal function preservation. Treatment guidelines for LN have recently been issued by rheumatology and nephrology communities in U.S.A. and Europe. In view of the racial difference in disease manifestation and response to therapy, Non-specific serine/threonine protein kinase and the substantial disease burden in Asia, a panel of 15 nephrologists and rheumatologists from different Asian regions with extensive experience in

lupus nephritis – the Steering Group for the Asian Lupus Nephritis Network (ALNN) – met and discussed the management of lupus nephritis in Asian patients. The group has also reviewed and deliberated on the recently published recommendations from other parts of the world. This manuscript summarizes the discussions by the group and presents consensus views on the clinical management and treatment of adult Asian patients with LN, taking into account both the available evidence and expert opinion in areas where evidence remains to be sought. Systemic lupus erythematosus (SLE) is a potentially severe autoimmune disease that demonstrates variations in incidence, prevalence, disease activity and prognosis according to race and ethnicity.[1-3] Renal involvement affects over 60% of patients with SLE, and is a major contributor to morbidity and mortality.[4, 5] A systematic review of SLE in Asia has shown higher rates of renal involvement in Asian patients (21–65% at diagnosis and 40–82% at follow-up) compared with Caucasians.

Such maternal immunological imprinting and in-utero exposure of the fetus resulting in adverse pregnancy outcomes are best exemplified in pregnancies with autoimmune conditions such as APS, SLE, myasthenia gravis and primary Sjögren’s syndrome. Risks for fetus and neonate Patients with APS often have anti-phospholipid autoantibodies that are reactive against phospholipid proteins, such as β2-glycoprotein, cardiolipin, tissue plasminogen activator, thrombin, protein C and platelet antigens. The pathogenicity of anti-phospholipid autoantibodies is often associated with IgG classes and they target proteins that are involved in thrombosis, platelet and complement pathway

activation, monocyte and endothelial cell functions MK-1775 mouse [75]. These autoantibodies can be either agonistic or antagonistic in nature. They contribute to the pathologies of APS by promoting thrombotic events, impairing endothelial Gemcitabine cost cell function and provoking overt inflammatory responses in the maternal circulation and placental tissues. This may lead to vasoconstriction, impaired endothelial function and placental dysfunction that restrict blood supply to the placenta and result in placental ischaemia

and/or hypertensive disorders. Such a cascade of events can lead to a range of poor pregnancy outcomes such as RSA, IUGR, pre-eclampsia or stillbirth. Mild to moderate thrombocytopenia is common in APS, and this can worsen in pregnancy [9]. The causes of APS-associated thrombocytopenia are poorly understood: unlike immune thrombocytopenia (ITP), specific antibodies against the major platelet adhesion receptors (GPIIb-IIIa or GPIb-V-IX) are uncommon. Pregnant

women with SLE carry not only a risk of maternal and fetal morbidity, but also risks of long-term disability to the newborn. The immunopathologies of SLE pregnancy display several features of those seen in APS. Thus, it is not surprising that SLE pregnancy shares many of the adverse risks and poor outcomes of APS, such as maternal morbidity, IUGR, pre-eclampsia, stillbirth or preterm birth [9]. In addition, the autoimmune conditions of SLE and APS are often exacerbated during pregnancy and contribute further to the disease burden and DOK2 dysfunction of the maternal circulation and renal system. The deposition of anti-nuclear proteins, anti-dsDNA, anti-basement membrane autoantibodies and autoreactive antibodies in kidney glomeruli can cause nephritis that results in further damage to the already compromised kidney function. This, in turn, exacerbates the hallmark signs of pre-eclampsia, such as hypertension and proteinuria. In addition, neonates of mothers with SLE or primary Sjögren’s syndrome are at risk of developing neonatal lupus syndrome and congenital heart block [9, 10]. These neonatal conditions often occur in mothers who are seropositive for anti-Ro/SSA and/or anti-La/SSB autoantibodies.

CpG ODN is a ligand for TLR9 and therefore was not expected to signal through TRIF because TLR9 signals exclusively via the MyD88-dependent pathway. As discussed above, TLR4, for which LPS is a ligand, utilizes either MyD88 or TRIF as adaptor molecules. In this case it appeared that the observed effects were diminished by deletion of either MyD88 or TRIF, but that the effect of MyD88 deletion was more marked. From this data it was concluded that signalling through both the MyD88-dependent and the MyD88-independent/TRIF-dependent pathways could initiate

changes in lineage commitment in developing haematopoietic cells in vitro, which was dependent on the inducing ligand. The data from experiments involving influenza viruses demonstrated that, although they have been shown to activate MyD88-dependent signalling in B lymphocytes, https://www.selleckchem.com/products/ensartinib-x-396.html in this instance their effects were INCB024360 ic50 mediated by a mechanism that was not dependent on either MyD88 or TRIF. As the above evidence demonstrated that the effects of LPS on BMDC generation were dependent, in part, on both MyD88 and TRIF, and LPS has been shown to be a ligand for TLR4,16 which interacts with both adaptors, it was important to directly confirm the role of TLR4 in modulating the effects of LPS on BMDC production in vitro. To assess this, bone marrow cells from C57Bl/6 (TLR4+/+) and TLR4−/− mice were cultured in the presence of GM-CSF, with or without Poly I, Poly I:C, LPS or CpG, for 6 days. The

production of BMDCs was assessed by monitoring the surface expression of CD11c and MHCII. The results (Fig. 4) confirm a requirement for TLR4 in signalling initiated by LPS. Stimulation of TLR4+/+ bone marrow cultures with Poly I, Poly I:C, LPS or CpG resulted in a striking reduction in the production of CD11c+/MHCII+ BMDC, similar to that observed in BALB/c bone marrow cultures. A similar reduction in BMDC production was observed in cultures of TLR4-deficient bone marrow containing Poly I, Poly I:C or CpG. By contrast, TLR4-deficient bone marrow cultures containing LPS displayed a level of BMDC production comparable to that observed in unstimulated

cultures. This evidence supports the previous findings that MyD88 and TRIF are involved in signalling downstream from LPS and confirms a role for TLR4 in regulating changes in BMDC production. Type A influenza viruses have been shown to be strong inducers PJ34 HCl of type 1 IFNs,17 which are a major component of the antiviral response, inducing an antiviral state in uninfected cells18. It therefore seemed possible that the effects seen in our experiments in response to influenza A viruses could be mediated by type 1 IFNs. To investigate this we first examined the effects of influenza viruses on the generation of BMDCs in cultures of bone marrow cells from IFNAR−/− and IFNAR+/+ mice in the presence of GM-CSF. Cultures containing IFNAR+/+ bone marrow cells displayed reduced CD11c+/MHCII+ BMDC production in response to the addition of Jap, X31 or PR8 virus (Fig.

They are not only involved in tissue development and homeostasis but also perform various immune regulatory functions 1–4. They are efficient effector cells of the innate immune system as they have the capacity to respond to parasite, viral or bacterial infections. In addition, eosinophils have an important role in bridging innate and adaptive immunity. In particular, activated eosinophils are crucial in promoting TH2 responses by secreting TH-cell polarizing cytokines such as IL-4 and IL-13, and this release of IL-4 also promotes rapid differentiation of B cells into IgM plasma cells 5, 6. Thus, in T-cell-dependent immune responses eosinophils are required for the early protective IgM response

7, 8. In contrast, the generation of an antigen-specific IgG response seems not to be affected as in eosinophil-deficient ΔdblGATA-1 Ceritinib molecular weight mice B-cell maturation in germinal centers and their differentiation into memory B cells and plasma cells were shown to be normal 9. Eosinophils, however, are crucial for the long-term survival of plasma cells in the BM as in their absence the plasma cells quickly die by apoptosis. Thus, as a major source of the plasma cell survival factors APRIL and IL-6, eosinophils Sunitinib in vitro have an essential function in the

long-term maintenance of humoral immunity 9. Eosinophils produce and store a wide range of cytokines whose release is dependent on the nature of the activating stimulus 10–12. Eosinophils respond by piecemeal degranulation leading to a highly controlled secretion of specific mediators 13. Full activation of eosinophils may induce de-granulation and thus a rapid release of tissue-destructive cationic proteins. Activated eosinophils may also respond by the ejection of extracellular traps consisting of mitochondrial DNA and granule-derived mediators 14. Human eosinophils

have been shown to express constitutively not only the TH2-related cytokines IL-4, IL-13 and IL-10 but also IL-12 and IFN-γ, below which are characteristic of TH1 responses 11, 15. Upon immunization, IL-4 production by eosinophils is up-regulated, although similar effects were seen when animals were injected with aluminum potassium sulfate (alum) alone, an adjuvant commonly used in antigen priming 7, 8. To further investigate the activation of eosinophils and their expression of cytokines, BALB/c mice were immunized with the T-cell-dependent antigen 2-phenyl-oxazolone (phOx) precipitated in alum or emulsified with CFA. Eosinophil activation was monitored in the primary response and also after secondary challenge with soluble antigen. We found that only in the presence of antigen was a stable activation of eosinophils and continuous expression of plasma cell survival factors achieved. By contrast, injection of adjuvant alone only transiently enhanced cytokine production. Together, these data suggest that in immune responses, eosinophils are primed to become effector cells that prevent plasma cells from undergoing apoptosis.

Collectively, our findings

support the concept that the use of Cox inhibitors can counteract the goal of vaccines, by inhibiting the generation of plasma cells which produce antibodies, important for fighting infections. Human B lymphocytes www.selleckchem.com/products/acalabrutinib.html isolated from peripheral blood mononuclear cells (PBMC) were cultured in RPMI-1640 (GIBCO/Invitrogen, North Andover, MA) supplemented with 10% fetal bovine serum, 2 mm l-glutamine, 5 × 105 m 2-mercaptoethanol, 10 mm HEPES and 50 μg/ml gentamicin. CpG oligodeoxynucleotide (ODN) 2395 5′-TCGTCGTTTTCGGCGCGCGCCG-3′ was purchased from the Coley Pharmaceutical Group (Wellesley, MA) and used to stimulate B cells at a concentration of 1 μg/ml. Stimulation of BCR was performed using a rabbit anti-human F(ab′)2 anti-IgM antibody fragment (Jackson ImmunoResearch Laboratories, West Grove, PA) at 2 μg/ml. Arachidonic acid (Nu-Chek Prep, Elysian, MN) dissolved in ethanol was supplemented in culture at a concentration of 10 μm. Mitomycin

C (Sigma-Aldrich, St Louis, MO) was added to cell cultures to prevent cell division, acting as a control for carboxyfluorescein Selleck Rapamycin succinimidyl ester (CFSE) analysis. SC-58125 and NS-398, (Cayman Chemical, Ann Arbor, MI) small molecule Cox-2 selective inhibitors, were dissolved in dimethyl sulphoxide (DMSO), and used at concentrations of 5, 10 and 20 μm. Cox-2 inhibitors were added on days 0, 3 and 5 of culture unless otherwise stated. Units of peripheral blood were obtained from healthy donors [not taking any non-steroidal anti-inflammatory

drugs (NSAIDs) or other medications] under ethical permission provided by the Research Subjects Review Board at the University of Rochester. B cells were isolated as described previously.11,12 Briefly, PBMC were isolated using Ficoll–Paque (Amersham Biosciences, Piscataway, NJ) gradient centrifugation. The B cells were labelled with CD19 Dynabeads (Invitrogen) and CD19 Dynabead-cell rosettes were disrupted using CD19 Detachabead (Invitrogen). Cells obtained by this method of isolation were > 98% CD19+. B cells were purified from Cox-2-deficient mice (B6.129P2-Ptgs2tm1Unc) and wild-type control splenocytes (Taconic Farms Inc., Hudson, NY) using a CD19 magnetic antibody cell sorter (MACS) separation protocol (Miltenyi CHIR-99021 cost Biotec, Auburn, CA). Purified CD19+ B cells were cultured with lipopolysaccharide (LPS; 10 μg/ml) for 72 hr. Positively isolated CD19+ human B cells (5 × 105 cells/ml) were cultured in 96-well round-bottom plates for 7 days in the presence of CpG ODN 2395, anti-IgM and arachidonic acid (10 μm). Vehicle control or Cox-2 selective inhibitors, SC-58125 or NS-398, were added at onset of culture and on days 3 and 5. Levels of IgM and IgG in the supernatants were assessed by enzyme-linked immunosorbent assay (ELISA; Bethyl Laboratories, Montgomery, TX) on day 7 as described previously.

They found that a considerable proportion of myofibroblasts co-express the EC marker CD31 and the (myo) fibroblast markers α-SMA and FSP1 in all three models. They also used an endothelial lineage-traceable transgenic mouse line (Tie2-Cre; R26R-stop-EYFP) NVP-BKM120 chemical structure to demonstrate that yellow fluorescence protein expression was present in a substantial proportion of activated fibroblasts, suggesting the existence of endothelial origin myofibroblasts. Further, they analysed kidneys 6 months after a single injection of STZ in CD1 mice. Double staining demonstrated that around 40% of all fibroblast-specific protein-1-positive and 50% of the α-SMA-positive cells in STZ kidneys were also CD31 positive. In kidneys of 22-week-old

COL4A3 knockout (homozygous null) mice, a model for Alport disease, co-expression of CD31 was observed in 45% of all α-SMA-positive fibroblasts and 60% of all FSP1-positive fibroblasts, suggesting that these fibroblasts are likely of endothelial origin and that EndoMT may contribute substantially to the accumulation of fibroblasts in the development and progression of renal fibrosis. Endothelial-mesenchymal transition is a specialized form of EMT.24 Compared with EMT, relatively little is known at this stage about EndoMT. For further understanding of EndoMT, we will briefly review EMT. During EMT, tubular cells

lose epithelial cell phenotype and acquire mesenchymal characteristics. Sotrastaurin Yang and Liu described four key steps at the cellular level essential for the complete process of EMT: (i) loss of epithelial adhesive properties; (ii) de novo expression of α-SMA and actin reorganization; (iii) disruption of the tubular basement membrane; and (iv) enhanced migration and invasive capacity

of the transformed cells.25 Of note, the phenotype of cells undergoing transition may contain both epithelial and mesenchymal (myofibroblast) properties.13 The phenotypic conversion of epithelial cells into fibroblasts is regulated by a complex molecular process.13 Metalloproteinases25,26 or membrane assembly inhibitors27 initiate the process by dismantling the local basement membrane with proteolytic digestion while local upregulation of epidermal growth factor (EGF), insulin-like growth factor II or fibroblast growth factor-2, or activation of TGF-β1 facilitate the process not of EMT.13 The most prominent inducers of EMT are TGF-βs 1–3.28,29 The TGF-βs may be involved sequentially28,29 dependent on the types of tissue and injury.13 EGF and TGF-β1 synergistically induce EMT in renal proximal tubular epithelial cells.30 Insulin-like growth factor II induces rapid EMT and a redistribution of β-catenin from the plasma membrane to the nucleus, as well as intracellular sequestration and degradation of E-cadherin.31 Fibroblast growth factor-2 induces MMP-2 and MMP-9 activity providing a mechanism for basement membrane disintegration and migratory access of transforming epithelium to the interstitium.

Examples are the miRNA cluster 99b/125a-5p/let7e, miR-187 and miR-146b, which are induced by LPS in an IL-10-dependent manner, while miR-511 is induced by dexamethasone. M. Pagani (Milan) presented miRNA profiles in 17 lymphocyte subsets and evidence for the importance of miR-125b in the regulation of genes related to T-cell differentiation (IFNG, BAY 80-6946 cost IL2RB, IL10RA, PRDM1). Concerning

vaccines and infections, the mechanism of action of MF59, an oil-in-water emulsion adjuvant, was described by E. De Gregorio (Siena). Based on the immune response of immune individuals in endemic areas, K. Matuschewski (Berlin) summarized his findings on the rational development of a whole-organism anti-malaria vaccine, while V. Barnaba (Rome) described the polyclonal CD8+ T-cell response to apoptotic self-antigens related to the chronic evolution of hepatitis C. The multi-level host responses to influenza GSK126 mouse A virus infection was studied by E. Wilk (Braunschweig) who recorded the transcriptome of the lungs from C57Bl/6J mice over a period of 60 days and presented an extensive description of the transcriptional changes occurring during the switch from innate to acquired immunity. In the B-cell section, E. Ferretti (Genova) reported that IL-31R is expressed in

follicular B lymphoma cells and that its ligand IL-31 triggers tumor cell proliferation, while J. Freitag (Jena) described the attempts and strategies to establish a retrogenic Carnitine palmitoyltransferase II mouse that expresses transgenic anti-HEL membrane IgM receptors. After the morning symposia and workshops, a keynote lecture focussed on advanced technologies in immunology. E. O’Connor (Valencia) discussed the most recent methods, including

the spectacular tool that is mass-spectrometric cytometry, which allows the simultaneous analysis of several dozen of parameters (cell phenotype and functions) in the same cell. Autoimmunity and chronic inflammation, control of humoral immunity and antigen-presenting cells were some of the topics addressed in the early afternoon. F. Aloisi (Rome) discussed how Epstein Barr virus has gained increased credibility as the main culprit of some major B-cell-related autoimmune diseases (SLE, RA, MS, among others) over recent years. D. Engel (Bonn) discussed how pathogenic Th1 cells are generated in postoperative ileus. The renaissance of transcriptional “Th1” programs was further highlighted by M. Löhning (Berlin) who showed that LCMV infection reprograms Th2 cells into a stable GATA-3+ T-bet+ “Th2+1” hybrid cell subset. Finally, L. Maggi (Florence) provided correlative evidence that “Th1+17” cells play a role in in chronic rheumatic inflammation. During a symposium on humoral immunity, J. Wienands (Göttingen) identified signal transducers that are involved in the differential activation of IgG memory versus naive IgM B cells. V. T. Chu (Berlin) showed that eosinophils play a critical role in the memory plasma cell survival niche of the bone marrow, and R.

1B). As shown in the figure, we co-precipitated pro-IL-16 and MHC class II molecules and confirmed the association between pro-IL-16 and MHC class II molecules. More importantly, the level of pro-IL-16

was increased by LPS treatment of resting B cells for 15 min, and increased selleck expression of pro-IL-16 protein was inhibited by anti-I-Ad MHC class II antibody treatment. This inhibitory effect was haplotype-specific and was not detected when we used a monoclonal antibody (10-3.6.2) specific to an unrelated haplotype (I-Ak) (data not shown). To characterize the form of IL-16 present in 38B9 resting B cells, we performed Western blot analysis using a commercial antibody specific to the C-terminal part of mouse IL-16, which can recognize both precursor and mature forms of IL-16 (Fig. 1C). Extracts prepared from 38B9 cells showed a single band at 80 kDa, representing pro-IL-16, but there was no band at 20 kDa (C-terminal mature form of IL-16) or at 60 kDa (remaining N-terminal part of pro-IL-16). In contrast, control EL4 cells, which are mouse CD8+ T cells known to express IL-16, showed only a single band at 20 kDa, indicating the presence of the mature form of IL-16. These results suggest that the precursor form of IL-16, rather than the mature form, is predominantly learn more expressed in 38B9 resting B cells. We assumed that

cleaved mature IL-16 was rapidly secreted rather than stored in the cytoplasm of B cells because we detected the expression of caspase-3, which is involved in pro-IL-16 cleavage, in 38B9 resting B cell lysates through Western blot analysis (data not shown). Collectively, we confirmed that pro-IL-16 is associated with MHC class II molecules

and that it is involved in MHC class II-mediated inhibitory signalling in resting B cells. It is known that cleavage of the C-terminal portion of pro-IL-16 Thiamine-diphosphate kinase by caspase-3 yields the mature form of IL-16 [23, 24]. Mature IL-16 is secreted, and the N-terminal fragment of pro-IL-16 or full-length pro-IL-16 translocates into the nucleus where pro-IL-16 or full-length pro-IL-16 induces G0/G1 cell-cycle arrest [18, 19]. Cytoplasmic pro-IL-16 can therefore be considered as a precursor of secreted IL-16, while pro-IL-16 in the nuclear compartment acts as cell-cycle regulator. Those previous reports and our observation of an association between pro-IL-16 and MHC class II-mediated negative signalling in resting B cells prompted us to determine whether pro-IL-16 has an inhibitory effect on B cell proliferation, as shown in T cells. Consequently, we initially examined the intracellular location of pro-IL-16 in resting B cells (Fig. 2). Western blot analysis of nuclear and cytoplasmic fractions prepared from resting B cells demonstrated that pro-IL-16 was present in both the cytoplasmic and nuclear compartments (Fig. 2A).

The AcT 5diff Cap Pierce Hematology

Analyzer (Beckman Coulter®, Suarlée, Belgium) was used to perform the full blood count quantifying numbers leucocytes (lymphocytes, monocytes, eosinophils, basophils and neutrophils); the proportion of each cell type was expressed as the percentage of total leucocytes. Thirty-nine participants provided blood samples for enumeration of leucocytes (uninfected n = 11, infected n = 11 and co-infected n = 17). Cercarial E/S material (0–3 h RP) was prepared as previously described [4, 8, 25] and used as a stimulant of the WB cultures. Alternatively, aliquots of total 0–3 h RP were treated with sodium metaperiodate (smp0–3 h CT99021 manufacturer RP), or ‘mock’-treated (m0–3 h RP), to disrupt glycan residues [8, 26]. WB cultures were stimulated with total 0–3 h RP (50 μg/mL), smp0–3 h RP (25 μg/mL), m0–3 h RP (25 μg/mL), the positive control ligand zymosan (50 μg/mL; Sigma-Aldrich, Dorset, UK) or culture medium without antigen (un-stimulated control). All cultures were conducted in the presence of 5 μg/mL polymyxin B (Sigma-Aldrich) to neutralize any potential endotoxin contamination in antigen preparations. Zymosan was chosen as a nonparasite antigen FK506 control as it is a heterogeneous mixture of protein–carbohydrate complexes and

thus is more comparable to cercarial E/S material than purified bacterial antigens (e.g. LPS). Cytokine production (IL-8, TNFα and IL-10) in the WB culture supernatants (diluted between 1:2 and 1:10) was measured by specific ELISA kits (TNFα and IL-8, Invitrogen; IL-10, R&D Systems Europe Ltd, Oxford, UK) according to the manufacturer’s guidelines. Results are given for each patient as mean cytokine production from triplicate wells in response to each stimulant minus the cytokine production for the corresponding WB sample cultured in the absence of stimulant

(i.e. medium only). Statistical analyses were conducted using the software package IBM Statistics, version 19. S. mansoni infection intensity (log(x + 1)-transformed epg) was compared by gender, age group (5–20 years (‘children’) and ≥20 years(‘adults)) and infection status (infected and co-infected) tested via anova using sequential Aurora Kinase sums of squares to account for gender and age before comparison between infection statuses. Age groups were selected according to epidemiological patterns of schistosome infection in the Diokhor Tack community as a whole [22, 23]. Log(x + 1)-transformed S. haematobium ep10 mL was compared by gender and age group via anova for the co-infected group. S. mansoni and S. haematobium infection intensities were log(x + 1)-transformed to meet parametric assumptions, and the homogeneity of error variances and normality of anova residuals was confirmed using the Levene’s test and Shapiro–Wilk test, respectively.