The role of nuclear factors as “Find-Me”/alarmin signals and immunostimulation in defective efferocytosis and related disorders

Efferocytosis as an apoptotic cell (AC) clearance mechanism facilitates the removal of dangerous and damaged cells, an important process in regulating normal homeostasis. Failure to correctly execute apoptosis and efferocytosis is associated with atherosclerosis, as well as chronic inflammatory and autoimmune disorders such as systemic lupus erythematosus (SLE). Effective and timely efferocytosis involves various molecules that act as “Find-Me” signals or as alarmins to quickly allow identification by phagocytic cells. In recent years, most of these molecules have been investigated, but less attention has been paid to the nuclear molecules associated with efferocytosis of ACs and necrotic cells (NCs). These molecules have several functions including acting as alarmin signals for faster recognition of ACs, facilitating the cleanup of ACs and for maintaining self-tolerance. The same group of molecules is also implicated in several inflammatory and autoimmune diseases. Previous studies have shown that these molecules also serve as targets for pharmacological agents such as necrostatins, recombinant Fcnb, anti-histone, neutralizing antibodies, calbiochem, aminophylline, activated protein C, CD24IgG recombinant fission protein, and recombinant thrombomodulin. Thus, greater understanding of these molecules/pathways will enable developments in the treatment and/or prevention of various disorders, especially autoimmune diseases. Here, we review current knowledge about the mechanisms by which nucleic acids, histones, nucleosomes and monosodium urate microcrystals (MSU) can act as alarmins/“Find-Me” signals, how they might be stimulated in defective efferocytosis and their function and importance as biomarkers for prognosis and treatment of atherosclerosis, inflammatory disorders and autoimmune diseases.     

The role of Toll-like receptors and C-type lectins for vaccination against Candida albicans

Recent progress has provided important novel insights in the processes driving the adaptive immune responses. Central to these developments is the discovery of pattern recognition receptors like TLRs and CLRs that not only induce innate immune responses, but also modulate cellular and humoral adaptive immunity. As vaccination is one of the great achievements in medicine and probably the most powerful tool to protect human and animals against infectious disease, further vaccine development and optimization of current strategies can improve health status of large groups of people. Development of a vaccine against Candida spp. should induce both cellular and humoral immune responses. While the TLRs are strong inducers of inflammatory responses, it seems that the CLRs have the potential to modulate these responses by enhancement or inhibition of cytokine production. Understanding the natural host defense mechanisms against pathogens like C. albicans therefore helps to identify the proper targets for inducing a strong adjuvant effect, in order to stimulate an effective adaptive immune response and protection.     

Modifications of surface carbohydrates on bovine spermatozoa mediated by oviductal fluid: a flow cytometric study using lectins

The objective of the present study was to characterize and quantify changes in exposed saccharide residues of bovine sperm during capacitation in oviductal fluid (ODF) using flow cytometry (FC). Bovine sperm were incubated with 0% or 50% non-luteal ODF for 30 min or 3.5 h. After incubation, sperm were labelled with 11 fluorescein isothiocyanate-labelled lectins and evaluated for lectin binding with FC. Furthermore, inhibiting sugars were used to determine specificity of lectin binding to oligosaccharides on the sperm surface. After 30 min incubation, there was a 91% decrease in fluorescence intensity of labelled sperm incubated in WGA, a 76% decline for Con A, 75% decline for BS-I and a 36% decline for DBA. These differences remained approximately the same over the 3.5-h incubation. Interestingly, although there was no reduction in UEA-I binding at 30 min, a significant reduction (23%) was observed at 3.5 h. Con A fluorescence was mostly inhibited with either alpha-d-glucose or alpha-d-mannose (86% and 90% respectively). BS-I fluorescence was reduced after prior incubation of the control samples with N-acetyl-galactosamine and galactose by 74% and 80% respectively. After prior incubation with N-acetyl-galactosamine DBA fluorescence reduced by 18% in the control samples. With UEA-I no fluorescence reduction was observed after prior incubation with l-fucose. We have demonstrated that capacitation of bovine sperm in ODF is accompanied by a quantitative reduction in individual lectin binding sites. These modifications may be crucial to the subsequent signalling events involved with sperm-zona binding, zona penetration or interaction with the oolema.     

Interest of glycolipids in drug delivery: from physicochemical properties to drug targeting

Importance of the field: The need for new products derived from natural sources for the replacement of the commonly used non-ionic surfactants containing ethylene oxide units with degradable carbohydrate headgroups has become an important area of research. Glycolipids offer a wide range of applications in the pharmaceutical and cosmetic fields and can compete with the most commonly used surfactants. Involved in molecular recognition mechanisms at the surface of cells, glycolipids are also used for drug targeting.

Areas covered in this review: The structure and pharmaceutical applications of the main glycolipid categories are summarized. The review focuses on marketed glycolipids, biosurfactants and compounds developed at laboratory scale for applications such as self-assembly or drug targeting.

What the reader will gain: This article aims to provide an overview of the different sugar-based surfactant classes and their potential uses.

Take home message: Beside their use as surfactants or absorption enhancers in basic formulations, glycolipids can build gels, niosomes, hexosomes and cubosomes, whose structure is directly related to lyotropic properties. These systems allow solubilization and entrapment of drugs. In innovative delivery systems, glycolipids are also used for drug targeting because their sugar moieties can be specifically recognized by carbohydrate-binding proteins exposed at the surface of cells.     

Exogenous gonadotropin administration affects the glycocalyx of rat endometrial epithelial cells during the period of implantation

Purpose Exogenous gonadotropins which cause superovulation are known to effect endometrial morphology, including the glycocalyx of surface epithelial cells. Certain of the carbohydrates in the glycocalyx of surface epithelial cells may be involved in the attachment and implantation of the blastocyst.Methods The effect of exogenous gonadotropins on specific carbohydrates in the glycocalyx of the rat endometrium around the time of implantation was investigated. Lectin-avidin-biotin-ferritin cytochemistry was used to ascertain which carbohydrates were affected. The lectins soybean agglutinin, fucose binding protein and wheat germ agglutinin were used.Results Statistically significantly less lectin was associated with the apical membrane of surface epithelial cells of animals following hyper stimulation than in noninjected pregnant animals.Conclusions The reduction in the carbohydrates contributes to a reduced receptivity of the endometrium for the blastocyst.     

Non-lectin component in a fermented extract from Viscum album L. grown on pines induces proliferation of lymphocytes from healthy and allergic individuals in vitro

Mistletoe preparations have been shown to express immunomodulatory properties. In order to evaluate the stimulatory potency of different mistletoe extracts, peripheral blood mononuclear cells (PBMC) from healthy and allergic/atopic individuals were exposed to aqueous or fermented extracts derived from Viscum album L. grown on apple trees (Mali-extracts) or on pines (Pini-extracts). None of them had received any mistletoe treatment. Iscador Pini was the only extract which strongly induced proliferation of PBMC in contrast to the other five preparations. On testing these extracts by Western blotting with anti-mistletoe lectin-1 (ML-1) antibody positive sera from mistletoe-treated patients, it became evident that Iscador Pini was almost devoid of lectins.The stimulatory potency of Iscador Pini for PBMC from three different groups was examined: PBMC from 35 normal controls (Group I), 23 patients with drug-induced adverse effects (Group II) and 16 individuals with allergic manifestations (Group III). Cells were exposed in 7-day cultures to the extract at concentrations between 1 and 10,000 g/ml. PBMC from 63% of Group III individuals showed strong stimulation (SI varying from 6 to 97) in contrast to only 9% from Group I and 22% from Group II individuals. Anti-ML-1 antibodies were detected in 5% and anti-IP antibodies in 11% of subjects in the three groups. They were either of the IgA or IgM type but not of the IgG type.Our findings strongly imply that a non-lectin associated antigen from Iscador Pini is able to activate PBMC from healthy and allergic/atopic individuals, thereby demonstrating sensitization to probably highly conserved plant antigens.Abbreviations AM Abnobaviscum Mali - AP Abnobaviscum Pini - HM Helixor Mali - HP Helixor Pini - IM Iscador Mali - IP Iscador Pini - LS Lactobacillus solution - ML-1/-2/-3 mistletoe lectin-1/-2/-3 - PBMC peripheral blood mononuclear cells - SI stimulation index     

Analysis of tumour necrosis factora-specific,lactose-specific and mistletoe lectin-specific binding sites in human lung carcinomas by labelled ligands

Summary Receptor sites can be visualized by labelled ligands as an alternative to receptor-specific antibodies, as substantiated for two different receptor classes. Recombinant tumour necrosis factor (TNF) was biotinylated via amino-groups and the resultant probe was applied to formalin-fixed, paraffin-embedded tissue sections of 94 primary bronchial carcinomas and to normal peripheral lung parenchyma. In addition, monoclonal antibodies specific for neuron-specific enolase (NSE) and TNF itself were used. The biotinylated -galacto-side-specific mistletoe lectin, which exhibits dose-dependent immunomodulatory and toxic potency, and two probes that specifically detect certain types of sugar receptors were employed to illustrate further the feasibility of using ligands for receptor localisation. The tumours comprised 62 small cell lung carcinomas, 10 epidermoid carcinomas, 11 adenocarcinomas and 11 large cell anaplastic carcinomas. Expression of TNF-binding sites was found in 39 of the small cell lung carcinomas and in 13 of the non-small cell lung carcinomas. Binding capacity for the TNF-specific antibody was seen in similar proportions of small cell lung carcinomas and of non-small cell lung carcinomas. None of the normal lung parenchymas revealed significant staining. Binding capacities to mistletoe lectin were seen in all normal lung parenchymas and in nearly all cases of adenocarcinoma (10/11). A correlation between the expression of NSE and the binding capacities to TNF was detected. Endogenous lectins, specific for lactose or-GalNAc, were displayed in nearly one half of the small cell lung carcinoma cases (44% or 45% respectively) and in about 25% of the non-small cell lung carcinoma cases.     

Comparison of Ulex europaeus I lectin binding and factor VIII-related antigen as markers of vascular endothelium in follicular carcinoma of the thyroid

Factor VIII-related antigen (F VIII-RAg), a well established marker for endothelial cells, and the lectin Ulex europaeus agglutinin I (UEA-1), a newly described marker, were used in immunoperoxidase techniques to demonstrate endothelial cells in 30 follicular neoplasms of the thyroid and so to assist in detection of vascular invasion. Both endothelial cell markers made visible a greater number of instances of vascular invasion than were detected on routine elastic stains, but UEA-1 lectin gave more reliable staining of endothelium lining large capsular veins and of blood vessels within the tumour than did F VIII-RAg. In vessels completely occluded by tumour, examples of endothelial staining by UEA-1 in the absence of F VIII-RAg staining were found, but some such vessels appeared to lack surviving endothelial cells and in these no staining occurred. It is concluded that UEA-1 lectin is a more reliable endothelial marker in this setting than F VIII-RAg and may assist the demonstration of vascular invasion in these tumours.