com) (Fig. 2). In addition to the highly conserved exonic regions, two intronic regions (introns 2 and 7) of the WASP were found to have apparent high

evolutionary conservation. PCR-sequencing, however, did not detect any variants. We previously described the termination codon mutation in the WASP gene in a Thai family affected with classic WAS [13]. This study further reported the clinical manifestations and long-term follow-up of seven unrelated patients with molecular diagnosis of classic WAS. In addition to the previously reported mutation, four different recurrent mutations were identified, including two missense mutations, an insertion and a 4-bp deletion in intron 8. One novel nonsense (c.55C > T, p.Q19X) IWR-1 chemical structure mutation was also detected. No causative mutations in the coding, promoter and conserved intronic regions could be identified in case 2. The patient had classic Apoptosis inhibitor WAS with a score of 4, and no WASP expression could be detected in his cells by immunoblot analysis (courtesy of Dr. Hubert B. Gaspar and Dr. Kimberly C. Gilmour, UK). It remains possible that the mutation could be located in the noncoding parts of the gene including regulatory regions. Our patients with classic WAS

had the age of onset ranging from 6 days to 8 months. Of these seven cases, two developed AIHA, which included the previously reported patient (case 1) with the c.1507T > A (p.X503R) mutation (Table 1). As there are no available HLA-matched donors, this patient has been given monthly IVIG and sulfamethoxazole-trimethoprim prophylaxis. learn more The missense mutations (p.R86N) at position 86, one of the common hot spot mutations found in the WASP gene, were identified in two unrelated patients. One with a WAS score of 4 carried

the c.256C > T (p.R86C) mutation. The other with a WAS score of 5 harboured the c.257G > A (p.R86H) mutation. The missense mutations at position 86 (p.R86N) have been found to be commonly associated with the XLT phenotype. However, some patients with these particular mutations can have a more severe phenotype with a score of 3–5 [10, 12, 17]. The previously reported c.1272insG (p.G424GfsX494) and IVS8 + 3 to 6del GAGT mutations in patients with classic WAS were also detected in the Thai population. The novel nonsense (c.55C > T, p.Q19X) mutation expected to result in the formation of a truncated protein lacking most of the functional domains was identified in one patient with severe WAS. He developed pneumonia with hepatosplenomegaly at 2 months of age caused by CMV. As microcephaly was observed at birth, congenital CMV infection cannot be excluded. Previous studies described CMV infection in patients with WAS both prior to and following HSCT [10, 18-20], and it resulted in a fatal outcome in the majority of cases. The treatment guideline for CMV infection in patients with WAS, however, has not been well established.

The histological changes were evaluated in a blinded fashion by Dr Bradley Weeks (Department of Veterinary Pathology, Texas A&M University, College Station, TX, USA). Results are displayed as the mean ± standard error of the mean (s.e.m.) of five to six animals per group. These experiments were repeated three times. Differences between groups for skin test, CFUs and flow

cytometric results were compared by Student’s two-tailed t-test. The real-time RT–PCR data were analysed by the GraphPad Prism (version 4·03, 2005; GraphPad, Inc., Tipifarnib San Diego, CA, USA) software package for the Mann–Whitney non-parametric test to compare BSA-treated and TNF-α treated guinea pigs. P-values of < 0·05 were considered

statistically significant. As shown in Fig. 1a, 6 weeks after vaccination BCG-vaccinated guinea pigs exhibited a strong skin test response 24 h after injection with 2 µg of PPD, while TNF-α-treated animals showed a significantly (P < 0·03) enhanced dermal response when compared to the BSA-injected group. Lymph nodes draining the site of vaccination were homogenized and plated for viable BCG. As shown in Fig. 1b, the CFUs were reduced selleck chemical significantly (P < 0·006) in the lymph nodes of TNF-α-treated guinea pigs when compared with the BSA-injected animals. No significant differences in the CFUs were seen in the spleen after TNF-α injection (Fig. 1b). The T cell proliferative ability of lymph node and spleen cells from TNF-α- and BSA-injected guinea pigs vaccinated with BCG was determined by the

[3H]-thymidine uptake assay after culturing the cells for 4 days in the presence of ConA or PPD. As depicted in Fig. 2, both lymph node and spleen cells proliferated well to ConA (Fig. 2a), although the response was much higher in the lymph node cells. There was no significant difference in the Olopatadine T cell response between TNF-α- and BSA-injected guinea pigs. Similarly, lymph node and spleen cells proliferated well after PPD stimulation (Fig. 2b), and the response was similar in both cell types. However, T cell proliferation was enhanced significantly (P < 0·04) in the lymph node cells of TNF-α-injected guinea pigs compared to the BSA controls (Fig. 2b). The effect of TNF-α injection on the proportions of immune cells in the lymph nodes and spleen was carried out by flow cytometry after staining the cells with the mAbs against guinea pig MHC class II, pan (CD3+) T, CD4 and CD8 T cell phenotypic markers. TNF-α injection resulted in a significant increase in the proportion of CD3+ T cells (P < 0·03) in the lymph nodes (Fig. 3a). There was no significant treatment effect on the proportions of MHC class II, CD4 or CD8+ cells in the lymph nodes (Fig. 3a) or spleen (Fig. 3b) of guinea pigs. Lymph node and spleen cells were cultured with PPD and peritoneal cells were stimulated with PPD or live M. tuberculosis for 24 h.

B10.2) (all: Santa Cruz), rabbit polyclonal α-plexA1 or -A4 (Abcam), mouse α-VSV-G (Sigma), mouse α-MV H protein (K83, produced in our laboratory) and mouse α-NP-1 (clone AD5-17F6, Miltenyi). For double stainings with mouse monoclonal antibodies, α-NP-1 was directly conjugated according to the manufacturer’s protocol (Zenon, Molecular Probes/Invitrogen). After final washing steps in PBS, fluorochrome G (Southern Biotech, Eching, Germany) was used as the mounting medium and cells

were analyzed by confocal laser scanning microscopy (Laser Scan Microscope, LSM510 Meta, Software version 3.0; Axiovert 200 microscope, objective: 100×; NA=1.4 Plan Apochromat). T cells were nucleofected with 2 μg plasmid encoding for DN-plexA1 (kindly provided by L. Tamagnone, Milano) 54 following the manufacturer’s protocol (Amaxa). For silencing of plexA1, human T cells were transfected with a two-day interval according ABT 263 to the manufacturer’s protocol (DharmaFECT, Thermal Scientific) with 100 nM siRNA targeting KU-60019 concentration plexA1 (Santa Cruz) or, for control, a scrambled siRNA (Sigma). Before cells were recruited into

the respective experiments, aliquots were harvested for nucleic acid extraction (Qiagen, RNAeasy Kit) and subsequent RT-PCR analyses. Forward 5′-ctgctggtcatcgtggctgtgct and reverse 5′-gggcccttctccatctgctgcttga primers were used for specific amplification of plexA1. Signals obtained Cell Penetrating Peptide after electrophoresis were digitalized and quantified using the AIDA software program (Raytest, Straubenhardt, Germany). Supernatants of DC or DC/T-cell co-cultures were harvested at the time intervals indicated and immunoprecipitated using 2 μg/mL rabbit polyclonal anti-SEMA3A antibody (H300, Santa Cruz). Immune complexes were washed in PBS containing 0.5 M LiCl and 1% v/v Triton X100, and analyzed by Western blot using an anti-SEMA3A mAb (R&D Systems) followed by an anti-mouse HRP-conjugated antibody (Dianova, Hamburg, Germany). Signals obtained after ECL development

were digitalized and quantified (recombinant SEMA3A-Fc was included for reference) using the AIDA software program. For conjugate analyses, DC were labelled with 1 μM R18 dye for 20 min and T cells with 1 μM CSFE (both: Invitrogen) for 5 min (each in RPMI-5% FBS at 37°C). DC and T cells (exposed to SEMA3A/6A or human IgG (150 ng/mL each) for 15 min at 37°C) were co-cultured directly in an FACS tube for the time intervals indicated, fixed with PFA (final concentration of 2% w/v in PBS), washed once with FACS buffer (low-speed centrifugation (400 rpm)) and subsequently analyzed by flow cytometry. The double-positive population representing conjugates was determined, and percentages were calculated using one sample t-test with hypothetical value set as 100 for the IgG-treated controls. Under-agarose assays were performed as described elsewhere 41. Briefly, 2.

Fresh fruit or raw kiwi fruit extracts have been used so far to investigate these effects, but the molecule(s) responsible for these health-promoting activities have not yet been identified. Kissper learn more is a kiwi fruit peptide displaying pore-forming activity in synthetic lipid bilayers, the composition of which is similar to that found in intestinal cells. The objective of this study was to investigate the kissper influence on intestinal inflammation using cultured cells and ex-vivo tissues from

healthy subjects and Crohn’s disease (CD) patients. The anti-oxidant and anti-inflammatory properties of kissper were tested on Caco-2 cells and on the colonic mucosa from 23 patients with CD, by challenging with the lipopolysaccharide from Escherichia coli (EC-LPS) and monitoring the appropriate markers by Western

blot and immunofluorescence. EC-LPS challenge determined an increase in the intracellular concentration of calcium and reactive oxygen species (ROS). The peptide kissper was highly effective in preventing the increase of LPS-induced ROS levels in both the Caco-2 cells and CD colonic mucosa. Moreover, it controls the calcium increase, p65-nuclear factor (NF)-kB induction and transglutaminase 2 (TG2) activation inflammatory response in Caco-2 cells and CD colonic mucosa. Kissper efficiently counteracts the oxidative STI571 stress and inflammatory response in valuable model systems consisting of intestinal cells and CD colonic mucosa. This study reports the first evidence supporting a possible

correlation between some beneficial effects of kiwi fruit and a specific protein molecule rather than generic nutrients. “
“Macaques provide important animal models in biomedical research into infectious and chronic inflammatory disease. Therefore, a proper understanding of the similarities and differences in immune function between macaques and humans is needed for adequate interpretation of the data and translation to the human situation. Dendritic cells are important as key regulators of innate and adaptive immune responses. Using a new whole blood assay we investigated functional Carbohydrate characteristics of blood plasmacytoid dendritic cells (pDC), myeloid dendritic cells (mDC) and monocytes in rhesus macaques by studying induction of activation markers and cytokine expression upon Toll-like receptor (TLR) stimulation. In a head-to-head comparison we observed that rhesus macaque venous blood contained relatively lower numbers of pDC than human venous blood, while mDC and monocytes were present at similar percentages. In contrast to humans, pDC in rhesus macaques expressed the interleukin (IL)-12p40 subunit in response to TLR-7/8 as well as TLR-9 stimulation. Expression of IL-12p40 was confirmed by using different monoclonal antibodies and by reverse transcription–polymerase chain reaction (RT–PCR).

The concentration (in pg/ml) was determined using a standard curve with known amounts of IL-2 added to the ELISA plate. While sustained Foxp3 Z-VAD-FMK supplier gene expression is required for the suppressive function of natural Tregs,29 its expression is also up-regulated in activated human Teffs.4–6 Thus, a challenge in the study of Tregs in humans is the difficulty in discriminating between recently activated CD25+ FoxP3+ Teffs and the

subset of resting Tregs in which FoxP3 can be expressed at similar levels. In this regard, other markers that help to discriminate Tregs from Teffs can be used in combination with FoxP3 expression for the study of freshly isolated and ex vivo activated T cells.4,30 We used unfractionated PBMC rather than purified Tregs/Teffs in order to study them within the context of a broader population of immune cells.

To study the relationship between human natural Tregs and Teffs upon polyclonal activation, total PBMC were stimulated with anti-CD3 (5, 100 or 1000 ng/ml) and the expression of FoxP3, IFN-γ and IL-2 was determined on CD4+ cells by flow cytometry at days 3, 7 and 10, as previously reported.4 This system relies on ‘presentation’ of anti-CD3 antibody to T cells by Fc receptors on antigen-presenting cells, a situation that resembles T-cell receptor (TCR) activation in response to its natural Selleck Ixazomib ligand [i.e. peptide/major histocompatibility complex (MHC) complexes] in vivo.4 In addition, as the assay is performed on total PBMC, it avoids the requirement of T-cell purification, a condition that may affect the activation state of the cells. In the absence of TCR stimulation, rTregs (defined as CD4+ FoxP3low IFN-γNeg IL-2Neg) remained fairly stable at day 3 of culture (compare Figs 1a and 1d). In contrast, as previously described,4 anti-CD3 activation of PBMC induced a dramatic increase in the percentage of FoxP3-positive cells, peaking at day 3 post-stimulation (compare Figs 1d and g, and data not shown). Furthermore, among these cells, two novel cell

populations were distinguished based on the expression levels of FoxP3 and the effector cytokines IFN-γ and IL-2. These cells were PLEK2 identified as CD4+ FoxP3HI IFN-γNeg IL-2Neg and CD4+ FoxP3Low IFN-γPos IL-2Pos (Fig. 1g,h), representing activated Tregs and Teffs, respectively.4,6 From these experiments, the highest expression of FoxP3 was observed at day 3 using 100 ng/ml of anti-CD3 (Fig. 1g and data not shown); this concentration was used in the subsequent assays. In addition, aTeffs were further defined as IFN-γPos, which include both FoxP3Neg and FoxP3Low cells. In order to address the mechanism of CD4+ FoxP3HI cell generation, we determined the expression of Ki-67, a marker of cell proliferation.31 At day 3 post-TCR stimulation, 20% of CD4+ FoxP3HI cells were Ki-67 positive (Fig. 1i), supporting the conclusion that this cell population is expanded through proliferation.

13 Although incompletely documented, non-human primates appear to possess subpopulations of dendritic cells (DCs) and B cells that are similar

to those present in humans.14,15 Non-human primates are therefore valuable for studies aimed at investigating immune responses induced by human pathogens and vaccine components aimed for human use.16,17 Several reports indicate that TLR ligands show potency as vaccine adjuvants when tested in rhesus macaques18–20 or in human clinical trials.21–23 Subsets of human DCs and B cells express distinct repertoires of TLRs and they respond to TLR stimulation accordingly.2,24,25 Unlike rodents, rhesus macaques express a similar repertoire of TLRs on immune cells such as DCs and B cells as humans.26 Some differences between the human and rhesus macaque immune systems have been reported.17 An improved understanding about similarities selleck chemicals and disparities between human and non-human primate immune functions is therefore important and would provide valuable information for translating non-human

primate studies for the design of clinical trials aimed at testing new vaccine and treatment strategies. In this study, we performed a side-by side comparison of the phenotypes of human and rhesus DCs and B cells and we examined their responsiveness to well-defined ligands targeting TLR3, 7/8, and 9. We further asked if IFN-α comparably enhanced B-cell functions such as proliferation and differentiation into antibody-producing cells as Dabrafenib molecular weight observed in culture systems of human cells. We found similar responses in human and rhesus primary cell cultures to TLR ligand stimulation in terms of B-cell proliferation and induction of IFN-α production by pDCs. In both species, B-cell proliferation to the TLR7/8 ligand (-L) and CpG class C showed a significant increase in the presence of IFN-α. Some phenotypic differences between human and rhesus B cells were observed as the cells differentiated

why into antibody-producing cells, although in both species TLR stimulation promoted maturation of B cells into IgM-producing cells and this effect was enhanced in the presence of IFN-α. Untreated and healthy rhesus macaques of Chinese origin, 5–6 years old, were housed in the Astrid Fagraeus laboratory at the Swedish Institute for Infectious Disease Control. Housing and care procedures were in compliance with the provisions and general guidelines of the Swedish Animal Welfare Agency. All procedures were approved by the Local Ethical Committee on Animal Experiments. The animals were housed in pairs in 4-m3 cages and enriched daily. All blood samplings were performed under sedation with ketamine at 10 mg/kg (100 mg/ml Ketaminol; Intervet, Sollentuna, Sweden). All animals were confirmed negative for simian immunodeficiency virus, simian T-cell lymphotropic virus, and simian retrovirus type D.

These data suggest that mediators synthesized by the pathogen during infection regulate both protective as well as detrimental responses

to the host. Thus, discovery and characterization of Mtb-secreted proteins could be an approach to identify novel therapeutic and diagnosis targets as well as biomarkers of disease. Lectins are classically defined as a family of proteins with the ability to specifically bind carbohydrate moieties. A number of pathogens have been demonstrated to express Fluorouracil supplier such molecules, which are involved in recognition and invasion processes 17, 18. For example, Pseudomonas aeruginosa produces several membrane-associated lectins that promote attachment to epithelial cells and contribute to its virulence 19. In addition, bacterial lectins could be released into the extracellular milieu and play an important role during infection as demonstrated by experiments using Bordetella18. These data suggest that both membrane-expressed and secreted lectins participate in host–microbial interactions. In the case of Mtb, the heparin-binding hemagglutinin adhesin (HBHA) is one of the most studied cell surface-expressed lectins

and it has been shown to be critical for bacterial dissemination in vivo20. Moreover, the existence of at least 11 hypothetical lectins from Mtb21 suggests that these molecules may be an important component of the host–mycobacteria interplay. Consistent with this, click here Non-specific serine/threonine protein kinase active TB (ATB) patients have been found to display increased levels

of anti-HBHA Ab during active disease 22, 23, suggesting that mycobacterial lectins may elicit specific immune responses. We have utilized a previously generated non-redundant lectin data bank 24 in order to identify lectins from Mtb, a major human pathogen. In the present study, we have demonstrated a secreted 13 kDa ricin-like lectin from Mtb (sMTL-13). sMTL-13 was detected in pleural biopsies from ATB patients and led to an increased IFN-γ production by PBMC from patients during active disease. Importantly, ATB patients display high titers of serum IgG against sMTL-13, a response found to be rapidly decreased following successful treatment. These data report a secreted Mtb lectin with antigenic activity in human TB and suggest it may be useful as a biomarker of disease therapy. We have previously generated a non-redundant lectin database for searching lectin domains from Arabidopsis thaliana genome 24. To further evaluate the presence of such domains in an important human pathogen, Mtb, we have adapted this database and identified a single hypothetical lectin encoded by the Rv1419 gene. Figure 1A shows the bioinformatics characterization of the Rv1419 gene. Its open reading frame (ORF) contains 474 nucleotides and the aa sequence encodes a hypothetical protein of 157 residues containing a signal peptide and a predicted molecular mass of 16.8 kDa.

Intravenous (i.v.) Ig (IVIG) also provides an important adjunctive treatment to control airway inflammation, reducing oral steroid requirements in severe bronchial asthma [4–7]. Protein Tyrosine Kinase inhibitor The efficacy of IVIG is due largely to IgG, which is a major portion of IVIG. Several roles of IgG in IVIG therapy in autoimmunity have been proposed [8–10], and the functions of IgG in IVIG therapy in allergic diseases are also envisaged to inhibit inflammatory reaction. Although these reports suggest that i.v.-administered

IgG have functions to protect against allergies and asthma, the precise target and mechanisms in allergic airway inflammation have not yet been revealed. In a murine experimental model, intranasal instillation of antigen-specific IgG reportedly FK228 clinical trial reduce eosinophilic inflammation and goblet cell hyperplasia induced by antigen challenge, suggesting that topical IgG reportedly counteracts allergic pulmonary inflammation that is dependent upon Fc and interferon (IFN)-γ[11]. However, clinical use of these therapies in bronchial asthma is currently limited because of the lack of evidence.

Clarifying the role of FcRs leads potentially to the development of a new strategy to manage asthmatic airway disorders. The role of antigen-presenting cells (APCs), including dendritic cells (DCs), in the pathogenesis of asthma has been clarified. When allergens are encountered in the airways, DCs in the airway epithelium capture allergens

and migrate to the draining lymph nodes, where they reside in a mature, antigen-priming mode [12]. There, antigen-specific T cells are induced to differentiate into Th effector cells or regulatory cells by these DCs. Thus, DCs are important in the initiation of T cell differentiation and activation and contribute indirectly to the development of Anacetrapib airway inflammation. Targeting the inhibitory Fc receptor on DCs can potentially inhibit induction of the Th2 cytokine response. We hypothesized that i.v. IgG administration (IVIgG) inhibits allergic inflammation through inhibitory FcRs on immune cells to induce a Th2 response. Among several types of FcRs, FcγRIIb is a unique inhibitory FcR which regulates immune cell function [13]. To verify the inhibitory effects of IVIgG and FcγRIIb in bronchial asthma, we pursued the mechanisms of IVIgG using murine models of allergic airway inflammation induced by ovalbumin (OVA) sensitization and aerosol challenge. As IVIgG, we analyse the effects of both mouse IgG and xenogenic (rabbit) IgG to analyse the functions on FcRs.

, 1997; Wu et al., 2005; Xu et al., 2005; Yamashita et al., 2008). The amino acid sequences of the NS3 helicase domain of JEV exhibited 65%, 44% and 23% homology to those of DEN, YFV and HCV, respectively (Yamashita et al., 2008). The crystal structures of the NS3 helicases of DEN (Xu et al., 2005) and YFV (Wu et al., 2005) are similar to that of JEV, but slightly different from HCV (Yao et al., 1997). Yamashita et al. (2008) emphasized that the distance between domains 1 and 2 of HCV helicase is longer than

that in most flavivirus NS3 helicases. This leads to the conclusion that MLN8237 chemical structure the HCV helicase has a larger ATP-binding pocket than other flaviviruses, and that the folding of domain 3 of the HCV helicase is unique, whereas the folding of JEV is very similar to those of other flaviviruses, including DEN and YFV (Yamashita et al., 2008). Superposition of JEV, DEN, YFV and HCV helicases further clarified that the HCV helicase has a unique conformation in the NTPase-binding region and domain 3 in comparison with JEV, DEN and YFV helicases (Yamashita et al., 2008). In particular, the conformation of motifs I and II of HCV helicase was different from Doxorubicin nmr that of JEV, DEN and YFV helicases. The distance between motifs I and II

of Cα of HCV and the other flaviviruses was 6.7 and 3.5 Å, respectively (Yamashita et al., 2008). There was also a 4.7 Å difference in the distance of Nz of Lys200 in the motif I between JEV and HCV, suggesting that HCV helicase has a wider ATP-binding pocket than other flaviviruses (Yamashita et al., 2008). In contrast to the structure of motifs I and II, that of motif VI was well conserved among the flavivirus helicases, including Rucaparib purchase HCV. Although a subtle difference is observed, the ATP-binding residues in JEV, DEN, YFV, and HCV helicases are well conserved, suggesting that flavivirus

helicases possess similar mechanisms of ATP hydrolysis, which reflects the lack of specificity of compounds 1 and 2. The virtual screening performed allowed the noncompetitive mode of action of 3 and 4 to be confirmed, as they were not identified as hits for the ATP-binding site. Although the antiviral activity of the identified hits needs to be confirmed in experimental studies, the reliability of the computational results obtained is enhanced by several factors. As mentioned, the refined crystal structure of the catalytic domain of JEV NS3 helicase/NTPase was utilized to construct the pharmacophore model. Moreover, the residues constituting the ATP-binding site were identified in the mutational analysis. Finally, the application of consensus screening procedure improved the hit ranking list. The consensus scoring procedure has been demonstrated to improve virtual screening results significantly (Feher, 2006). It was reported that consensus scoring usually substantially enhances virtual screening performance, contributing to better enrichments.

Our experiments showed clearly that this is possible. Red cells were able to inhibit the IC-mediated stimulation of macrophages. Conversely, IC-loaded red cells were able to stimulate TNF-α production Selleck Ganetespib by macrophages in the absence of free ICs. Although the pro-inflammatory [12] and anti-inflammatory potential [8] of erythrocytes have been recognized separately, in this

study we highlight how the two can occur simultaneously, and explore their relationship to the CR1 level. We hypothesized that the ability of erythrocytes to serve as inhibitors of IC-mediated production of TNF-α by macrophages varies with the level of CR1 expression. For this purpose we selected donors on the basis of their red cell CR1 expression as low, medium Selleck Palbociclib or high expressors. Because the IC binding capacity is the critical factor in determining the buffering capacity of the red cell, we also measured this parameter. Surprisingly, the IC binding capacity did not show a good relationship with the inhibitory capacity of red cells. We observed that medium and high expressor red cells were able to inhibit IC-mediated macrophage stimulation equally effectively, despite their having clearly

different IC binding capacities. Conversely, low expressors inhibited less effectively than medium expressors, despite the two groups having a similar IC binding capacity. One possible explanation for these results is that both medium and high expressor red cells mafosfamide were capable of binding most of the free opsonized ICs available, despite having different IC binding capacities. Closer examination of Fig. 1b shows that medium expressors had a slightly higher IC binding capacity than low expressors. Therefore, it is likely that our assay for IC binding capacity lacked the sensitivity to detect differences in CR1-mediated IC binding at the lower end of the spectrum. Although the data did not show a straightforward relationship between the IC binding capacity and the inhibitory ability of the red cells,

the CR1 level showed a better relationship with the medium and high expressors, being more effective inhibitors than the low expressors. Lastly, we demonstrated that IC-loaded red cells are effective stimulators of TNF-α from macrophages. This is in agreement with a previous observation that IC-loaded red cells induce production of IL-1 when they interact with macrophages [12], although the mechanism was not clearly recognized at that time. Surprisingly, there was no difference in the stimulatory capacity in relation to the CR1 level of expression. One possible explanation is that even the lowest level of CR1 when saturated with ICs is able to maximally stimulate macrophages by cross-linking their Fcγ receptors. Our findings have several important clinical implications. A number of infectious and autoimmune disorders such as malaria, SLE, hepatitis B and HIV are characterized by the production of ICs [16–18,25–28].