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Introduction: Diagnosing and managing drug hypersensitivity is challenging because there are no clear limits between different types of drug reactions. Distinguishing between type A (predictable) and type B (hypersensitivity) reactions when a drug is introduced on the market is not easy. When many people use a drug, adverse reactions can occur, conditioned by diverse genetic profiles, viral infections or concomitant therapy. Occasionally the only tool clinicians have on which to base the diagnosis is the clinical history. Skins tests orin vitrotests sometimes have low sensitivity or are unavailable, and drug provocation tests may be dangerous or strictly forbidden in case of severe cutaneous reactions. Areas covered: This paper reviews the diagnosis and management of the two main types of immunological reactions: IgE-mediated immediate drug hypersensitivity reactions (IDHRs) and non-immediate drug hypersensitivity reactions (NIDHRs). Expert commentary: Although Europe and the United States use different diagnostic methods, patients with history of drug hypersensitivity must avoid the suspicious drug, and clinicians must assess tolerance to safe alternatives under medical surveillance. Sometimes desensitization may be required. There is a consensus about the need to perform genetic testing for specific drugs and give patients proper documentation to prevent future exposure to culprit drugs. [ABSTRACT FROM AUTHOR]

Immunoaffinity columns (IACs) are a well-established tool in the determination of regulated mycotoxins in food and feed commodities. However, they also have the potential to become attractive pre-concentration and clean-up materials for the determination of masked (also called modified) mycotoxins, which have been recognised as important contributors to the toxicological hazard deriving from fungal spoilage of goods. However, the information available in the literature concerning the cross-reactivity of DON-IACs against the major conjugates (DON-3-G, 15-AcDON and 3-AcDON) is incomplete and often contradictory. We have carried out a detailed characterisation of the cross-reactivity of the four main IACs brands against DON and its conjugates as well as an assessment of the competition among the analytes. Only one IAC enabled the simultaneous analysis of all relevant DON forms while two missed 15-AcDON and the fourth one missed DON-3-G and 3-AcDON. In the case of the multivalent IAC, the analytes modified at the C-3 position compete for the antibody binding with preference for 3-AcDON (less spatially hindered) while DON-3-G has the more-hindered access to the active sites. Taking into consideration the levels of DON conjugates existing in real samples, the cross-reactivity of one DON-IAC allows a quantitative analysis of all of these analytes. Important but rather neglected aspects such as the continuous supply of IACs with identical characteristics, and of columns which are strictly blank, are also addressed in this paper. [ABSTRACT FROM AUTHOR]

Soybean is considered one of the "big eight" foods that are believed to be responsible for 90% of all allergenic reactions. Soy allergy is of particular importance, because soybeans are widely used in processed foods and, therefore, represent a particularly insidious source of hidden allergens. Although significant advances have been made in the identification and characterization of soybean allergens, scientists are not completely certain about which proteins in soy cause allergic reactions. At least 16 allergens have been identified. Most of them, as with other plant food allergens, have a metabolic, storage, or protective function. These allergens belong to protein families which have conserved structural features in relation with their biological activity, which explains the wide immunochemical cross-recognition observed among members of the legume family. Detailed analysis of the structure-allergenicity relationships has been hampered by the complexity and heterogeneity of soybean proteins. A variety of technological approaches have been attempted to decrease soybean allergenicity. This paper provides a comprehensive review of the current body of knowledge on the identification and characterization of soybean allergens, as well as an update on current hypoallergenization techniques. [ABSTRACT FROM AUTHOR]

Systemic lupus erythematosus (SLE or lupus) is a complex disease with a multifactoral etiology, with genetic, hormonal, and environmental influences. Molecular mimicry as a result of viral infection may contribute to the development of lupus. The pattern of autoantibody development in lupus is consistent with initiation through molecular mimicry, as the initial autoantigenic epitopes that have been observed are limited and cross-reactive with viral proteins. Autoantibody specificity may then later diversify to other autoantigens through B-cell epitope spreading. Epstein-Barr virus (EBV) is an excellent candidate to be involved in molecular mimicry in lupus. EBV infection has been associated with lupus through serological and DNA studies. Infection with EBV results in the production of the viral protein Epstein-Barr virus nuclear antigen-1 (EBNA-1), antibodies against which cross-react with lupus-associated autoantigens, including Ro, Sm B/B', and Sm D1, in lupus patients. The immune response against EBV, and EBNA-1 in particular, differs among lupus patients and healthy controls, with controls maintaining a limited humoral response and failing to produce long-standing cross-reactive antibodies. We hypothesize that the humoral immune response to EBNA-1 in susceptible individuals leads to the generation of cross-reactive antibodies. Through the process of epitope spreading, these cross-reactive antibodies target additional, non-cross reactive autoepitopes, spread to additional autoantigens, and become pathogenic, leading eventually to clinical lupus. This paper reviews some of the current literature supporting roles for EBV exposure and epitope spreading in SLE. [ABSTRACT FROM AUTHOR]

Many zeranol immunoassay test kits cross-react with toxins formed by naturally occurring Fusarium spp. fungi, leading to false-positive screening results. This paper describes the evaluation and application of recently published, dry reagent time-resolved fluoroimmunoassays (TR-FIA) for zeranol and the toxin α-zearalenol. A ring test of bovine urine fortified with zeranol and/or α-zearalenol in four European Union National Reference Laboratories demonstrated that the TR-FIA tests were accurate and robust. The α-zearalenol TR-FIA satisfactorily quantified α-zearalenol in urine fortified at 10-30 ng ml -1 . The specificity-enhanced zeranol TR-FIA accurately quantified zeranol in the range 2-5 ng ml -1 and gave no false-positive results in blank urine, even in the presence of 30 ng ml -1 α-zearalenol. Zeranol TR-FIA specificity was demonstrated further by analysing incurred zeranol-free urine samples containing natural Fusarium spp. toxins. The TR-FIA yielded no false-positive results in the presence of up to 22 ng ml -1 toxins. The performance of four commercially available zeranol immunoassay test kits was more variable. Three kits produced many false-positive results. One kit produced only one potential false-positive using a protocol that was longer than that of the TR-FIA. These TR-FIAs will be valuable tools to develop inspection criteria to distinguish illegal zeranol abuse from contamination arising from in vivo metabolism of Fusarium spp. toxins. [ABSTRACT FROM AUTHOR]

CD34 is a cell surface glycoprotein expressed by hematopoietic progenitors and endothelial cells. It is widely used in the clinic for isolation of human hematopoietic stem cells. In recent years large animals are gaining increasing importance in biomedical research for the study and therapy of human diseases. Sheep has proved to be an useful experimental model for preclinical trials in transplantation procedures. Unfortunately, the lack of specie-specific monoclonal antibodies (MABS) recognizing hemopoietic progenitor cells hampers the use of this animal in experimental hematology. The aim of this paper was to determine whether commercial monoclonal antibodies specific for human CD34 molecule could cross-react with hematopoietic progenitor cells (HPC) present in sheep umbilical cord blood (UCB). Six antihuman CD34 MABS, recognizing the three different epitope classes, were tested in flow cytometry on purified mononuclear cells (MNC) isolated from cord blood of both species. None of the MABS used in this trial seemed to be able to identify HPC from sheep UCB. These data suggest that the panel of monoclonal antibodies used for cross reactivity detection has to be expanded with recently produced reagents. Further studies should be directed towards the production of ovine specific anti CD34 MABS. [ABSTRACT FROM AUTHOR]

House dust mite (HDM) is a main perennial allergen causing allergic rhinitis (AR). It has been shown that HDM cross-reacts with a variety of other allergens. Presently, allergen-specific immunotherapy (AIT) is an effective way for management of mono-sensitized HDM+ AR patients. However, management approaches to polysensitized HDM-sensitized AR patients are not standardized yet. This article reviews the data available in the literature for cross-reactivity between HDM and inhalant or food allergens, the diagnosis of cross-reactivity in HDM-sensitized AR patients, and the effect of immunotherapy on cross-reactivity in HDM-sensitized AR patients; which may help to develop effective therapeutic strategies for management of polysensitized HDM-sensitized AR patients in the future. Pan-allergen proteins such as tropomyosin, arginine kinase (AK), glutathione S-transferase (GST), and hemocyanin are responsible for cross-reactivity between HDM and other allergens. To distinguish genuine or cross-reactive sensitization, molecular- or component-resolved diagnosis is suggested to apply in HDM-sensitized AR patients. The effect of HDM immunotherapy to treat the associated cross-reactivity in HDM-sensitized AR patients is still contradictory, and might be dependent on the degree of homology between two allergens. Furthermore, targeting tropomyosin might be a promising way to treat HDM patients with allergen cross-reactivity AIT: allergen-specific immunotherapy; AK: arginine kinase; AR: allergic rhinitis; CRD: component-resolved diagnostics; Der f: Dermatophagoides farina; Der p: Dermatophagoides pteronyssinus; EAACI: European Academy of Allergy and Clinical Immunology; GST: glutathione S-transferase; GWAS: genome-wide association study; HDM: house dust mite; IgE: immunoglobulin E; RAST: radioallergosorbent test; sIgE: specific IgE; SIT: specific immunotherapy; SCIT: subcutaneous immunotherapy; SLIT: sublingual immunotherapy; SPT: skin prick test [ABSTRACT FROM AUTHOR]

Bioinformatic amino acid sequence searches are used, in part, to assess the potential allergenic risk of newly expressed proteins in genetically engineered crops. Previous work has demonstrated that the searches required by government regulatory agencies falsely implicate many proteins from rarely allergenic crops as an allergenic risk. However, many proteins are found in crops at concentrations that may be insufficient to cause allergy. Here we used a recently developed set of high-abundance non-allergenic proteins to determine the false-positive rates for several algorithms required by regulatory bodies, and also for an alternative 1:1 FASTA approach previously found to be equally sensitive to the official sliding-window method, but far more selective. The current investigation confirms these earlier findings while addressing dietary exposure. [ABSTRACT FROM AUTHOR]

Introduction: Beta-lactams (BL) are the main cause of allergic drug reactions mediated by specific immunological mechanisms. Reactions can be IgE or effector T-cell mediated. The new antigenic determinants are recognized by the immunological system in the context of the common beta-lactam structure or the specific differences in the side chains of the antibiotics of this family plus the protein carrier. Areas covered: We have reviewed the recent clinical literature concerning new clinical entities, the progress in diagnosis including the difficulties for in in vivo and or in vitro testing as well as the new algorithms proposed for delabelling subjects classified as allergic to beta-lactams, and recommendations for desensitization procedures. Expert opinion: The knowledge gained over the last years on beta-lactam hypersensitivity has enabled a better understanding and management of cases with allergic reactions to beta-lactams. [ABSTRACT FROM AUTHOR]

Fruits are an important source of vitamins and antioxidants that can effectively delay aging and contribute to health and well-being of the human kind. However, they are growing to be one of the primary elicitors of food allergies around the world. Fruit allergens can induce an IgE-mediated (Immunoglobulin E) reaction, presenting with a symptom like localized oral allergy syndrome (OAS). Numerous studies showed that varying environmental and cultivation conditions can influence the fruit allergen content during flowering and ripening stages. Further, the variety, harvesting maturity, and storage conditions can also significantly influence the allergenicity potential. For example, unripe apples and tomatoes have lower levels of allergens compared to ripened fruits. Researchers have also reported that modified atmosphere packaging (MAP) can help reduce Mal d 3 content present in apples during storage. Post-harvest processing like peeling is also considered a good method to help reduce the overall allergenicity in few fruits whose peel might contain majority of the allergens. This review will discuss the overall influence of both pre-harvest and post-harvest factors on the fruit allergens. We will also discuss the progress regarding the cause, symptoms and diagnostic methods of fruit based allergies. [ABSTRACT FROM AUTHOR]

Introduction: Proton pump inhibitors (PPIs) are one of the most prescribed drug classes. PPIs are remarkably safe, with minimal side effects, most of which are related to drug’s pharmacokinetic interaction profiles. However, hypersensitivity reactions can occur and anaphylactic reactions to PPIs have been described. Areas covered: A literature search in PubMed was performed to review the evaluation and management of anaphylaxis to PPIs. Clinicians should have a high level of suspicion and a drug allergy workup should be carried out by allergists, in a proper facility. Skin tests with PPIs are the only accurate methods to identify the culprit drug; they are also quite specific to solve cross-reactivity problems among drugs of this group. Expert commentary: A label of hypersensitivity to the whole group is generally not correct and tolerance to PPI with negative skin tests should be established with a negative oral challenge test. When an alternative drug among the whole group cannot be found, anti-H2 can be prescribed or a PPI desensitization protocol can be applied. [ABSTRACT FROM AUTHOR]

Salivarian trypanosomes evade the host immune system by continually swapping their protective variant surface glycoprotein (VSG) coat. Given that VSGs from various trypanosome stocks exhibited cross-reactivity (Camargo et al., Vet. Parasitol. 207, 17-33, 2015), we analyzed here which components are the antigenic determinants for this cross-reaction. Soluble forms of VSGs were purified from four Venezuelan animal trypanosome isolates: TeAp-N/D1, TeAp-ElFrio01, TeAp-Mantecal01, and TeGu-Terecay323. By using the VSG soluble form from TeAp-N/D1, we found that neither the inositol-1,2-cyclic phosphate moiety of the cross-reacting determinant nor the carbohydrate chains were exclusively responsible for its cross-reactivity. Then, all four purified glycoproteins were digested with papain and the resulting peptides were separated by high-performance liquid chromatography. Dot blot evaluation of the fractions using sera from trypanosome-infected animals yielded peptides that possessed cross-reaction activity, demonstrating for the first time that proteinaceous epitopes are also responsible for the cross-reactivity of trypanosome VSGs. [ABSTRACT FROM AUTHOR]

Protein components in food can trigger immune-mediated response in susceptible individuals. International law requires risk assessment to be undertaken by competent individuals to minimize food safety risk to consumers. Historically, allergen control legislation has been food-focused and on the requirement for on pack labeling, and the need for formal food recalls in the event of misleading or inappropriate labeling. In order to develop a mechanism for decision makers when assessing allergenic risk from plant-derived materials, the aim of this research was to consider a more holistic risk assessment method whereby rather than just using the food-based approach, an additive element in terms of considering the families of proteins is included. This approach reflects the need for food professionals to fully understand the role of proteins in triggering an allergic response to plant material and the health risk to individuals who show cross-reactivity to such proteins. [ABSTRACT FROM PUBLISHER]

Shellfish are diverse, serve as main constituents of seafood, and are extensively consumed globally because of their nutritional values. Consequently, increase in reports of IgE-mediated seafood allergy is particularly food associated to shellfish. Seafood-associated shellfish consists of crustaceans (decapods, stomatopods, barnacles, and euphausiids) and molluskans (gastropods, bivalves, and cephalopods) and its products can start from mild local symptoms and lead to severe systemic anaphylactic reactions through ingestion, inhalation, or contact like most other food allergens. Globally, the most commonly causative shellfish are shrimps, crabs, lobsters, clams, oysters, and mussels. The prevalence of shellfish allergy is estimated to be 0.5–2.5% of the general population but higher in coastal Asian countries where shellfish constitute a large proportion of the diet. Diversity in allergens such as tropomyosin, arginine kinase, myosin light chain, and sarcoplasmic binding protein are from crustaceans whereas tropomyosin, paramyosin, troponin, actine, amylase, and hemoyanin are reported from molluskans shellfish. Tropomyosin is the major allergen and is responsible for cross-reactivity between shellfish and other invertebrates, within crustaceans, within molluskans, between crustaceans vs. molluskans as well as between shellfish and fish. Allergenicity diagnosis requires clinical history, in vivo skin prick testing, in vitro quantification of IgE, immunoCAP, and confirmation by oral challenge testing unless the reactions borne by it are life-threatening. This comprehensive review provides the update and new findings in the area of shellfish allergy including demographic, diversity of allergens, allergenicity, their cross-reactivity, and innovative molecular genetics approaches in diagnosing and managing this life-threatening as well as life-long disease. [ABSTRACT FROM PUBLISHER]

The pre-erythrocytic stage of infection by malaria parasites represents a key target for vaccines that aim to eradicate malaria. Two important broad immune evasion strategies that can interfere with vaccine efficacy include the induction of dendritic cell (DC) dysfunction and regulatory T cells (Tregs) by blood-stage malaria parasites, leading to inefficient priming of T cells targeting liver-stage infections. The parasite also uses ‘surgical strike’ strategies, whereby polymorphism in pre-erythrocytic antigens can interfere with host immunity. Specifically, we review how even single amino acid changes in T cell epitopes can lead to loss of binding to major histocompatibility complex (MHC), lack of cross-reactivity, or antagonism and immune interference, where simultaneous or sequential stimulation with related variants of the same T cell epitope can cause T cell anergy or the conversion of effector to immunosuppressive T cell phenotypes. [ABSTRACT FROM PUBLISHER]

Hemiscorpius lepturus ( H. lepturus), one of the most venomous scorpions in tropical and sub-tropical areas, belongs to the Hemiscorpiidae family. Studies of antibodies in sera against the protein component of the venom from this organism can be of great use for the development of engineered variants of proteins for eventual use in the diagnosis/treatment of, and prevention of reactions to, stings. In the present in vitro study, the proteins of H. lepturus venom, which could specifically activate the production of immunoglobulin G (IgG) in victims accidently exposed to the venom from this scorpion, were evaluated and their cross-reactivity with venoms from two other important scorpion species including Androctonus crassicauda and Mesobuthus eupeus assessed. H. lepturus venom was analyzed with respect to its protein composition and its antigenic properties against antibodies found in sera collected from victims exposed to the venom of this scorpion within a previous 2-month period. The cross-reactivity of the H. lepturus venom with those from A. crassicauda and M. eupeus was assessed using ELISA and immunoblotting. Electrophoretic analysis of the venom of H. lepturus revealed several protein bands with weights of 8-116 KDa. The most frequent IgG-reactive bands in the test sera had weights of 34, 50, and 116 kDa. A weak cross-reactivity H. lepturus of venom with venoms from A. crassicauda and M. eupeus was detected. The results of immunoblotting and ELISA experiments revealed that H. lepturus venom activated the host immune response, leading to the production of a high titer of antibodies. Clearly, a determination of the major immunogenic components of H. lepturus venom could be valuable for future studies and ultimately of great importance for the potential production of recombinant or hypo-venom variants of these proteins. [ABSTRACT FROM AUTHOR]

Streptococcus pneumoniae is a major cause of childhood morbidity and mortality worldwide. A heptavalent polysaccharide-protein conjugate vaccine (PCV) has proven highly effective in preventing pneumococcal disease in industrialized countries. Two higher-valent pneumococcal conjugate vaccines are now widely available, even in the poorest countries. These differ from each other in the number of serotypes and carrier proteins used for their conjugation. Some have assumed that the only meaningful clinical difference between PCV formulations is a function of the number of serotypes each contains. A careful review of recent clinical data with these and several unlicensed PCV formulations points to important similarities but also that some key properties of each vaccine likely differ from one another. [ABSTRACT FROM AUTHOR]

Rheumatoid factors (RFs) are antibodies recognizing other antibodies usually by binding to the Fc part, while heterophilic antibodies (HAbs) are antibodies reacting with immunoglobulins (Igs) from other species. In particular, RFs have been found to cause false positive results in sandwich immunoassays. In this work, we analyzed RF-positive and RF-negative sera for content of cytokines and for heterophilic reactions by enzyme-linked immunosorbent assay and bead-based sandwich immunoassays. All sera, including those with RFs, contained insignificant amounts of cytokines and chemokines, but RF-positive sera showed large false positive values for several cytokines when analyzed by fluorescent bead-based multiplex immunoassays. This non-specific binding could be minimized by reagents designed to block HAbs, i.e. by selected animal IgGs. Furthermore, sera positive for RFs reacted with several animal IgGs, when these were immobilized on beads or coated on the polystyrene surface in enzyme-linked immunosorbent assays. This reaction could be inhibited by human IgG and by agents designed to inhibit heterophilic reactions (i.e. mixtures of IgGs from different species). In conclusion, RFs and HAbs represent an identical/overlapping set of antibodies, causing false positive reactions in sandwich and other immunoassays. Such assays must be conducted in the presence of appropriate blocking agents, e.g. HBR+, and must be carefully controlled. [ABSTRACT FROM AUTHOR]

West Nile virus (WNV) is an enveloped RNA virus in the family Flaviviridae and belongs to Japanese encephalitis virus serocomplex group. The WNV has a wide geographic distribution that includes Africa, Europe, Asia, America and Australia. Recently, it has re-emerged as an important pathogenic organism, illustrated by the series of WNV outbreaks in North America and in Europe. Several hundred people are sacrificed by WNV infection every year. WNV can infect many mammals, birds, reptiles and amphibians. A variety of diagnoses for WNV infection have been developed, such as virus isolation, nucleotide amplification, antigen detection and serology. Flaviviruses, including WNV, share common nucleotide sequences and antigenic epitopes. Understanding these properties that can influence cross-reactivity is important for accurate diagnosis, especially because areas with multiple flaviviruses are currently expanding. Herein, the authors outline the different diagnostic methods for detecting WNV infection as well as important considerations in using these methods. [ABSTRACT FROM AUTHOR]

Lomefloxacin has been increasingly used in veterinary medicine to treat microbial infections. To avoid using a complicated instrumental method to detect lomefloxacin residue in food, a simple and convenient indirect competitive enzyme-linked immunosorbent assay method has been developed in this study. The antibody generated from immunogen of bovine serum albumin-lomefloxacin showed high sensitivity toward lomefloxacin with an IC50 value of 0.35 ppb in PBS buffer and was suitable to be used as a screening assay to detect lomefloxacin residue in food products. The ELISA (Enzyme-Linked ImmunoSorbant Assay) developed in this study was compared with a commercial ELISA kit and significant improvement was achieved in terms of sensitivity and specificity. The antibody prepared showed excellent specificity with certain cross-reactivity with only two drugs including norfloxacin (17.5%) and fleroxacin (8.8%) among commonly used (fluoro)quinolones. The assay measured drug residue in defatted milk spiked with lomefloxacin with an inter-assay coefficient of variation <22.2% and an intra-assay coefficient of variation <18.2%. The average recovery rates at 0.1, 0.3, 0.6, 1.0, and 2.0 ppb were in the range of 120-85% for inter-assay and in the range of 113-90% for intra-assay. [ABSTRACT FROM AUTHOR]

Methyl methacrylate (MMA) is a respiratory irritant and dermal sensitizer that has been associated with occupational asthma in a small number of case reports. Those reports have raised concern that it might be a respiratory sensitizer. To better understand that possibility, we reviewed the in silico, in chemico, in vitro, and in vivo toxicology literature, and also epidemiologic and occupational medicine reports related to the respiratory effects of MMA. Numerous in silico and in chemico studies indicate that MMA is unlikely to be a respiratory sensitizer. The few in vitro studies suggest that MMA has generally weak effects. In vivo studies have documented contact skin sensitization, nonspecific cytotoxicity, and weakly positive responses on local lymph node assay; guinea pig and mouse inhalation sensitization tests have not been performed. Cohort and cross-sectional worker studies reported irritation of eyes, nose, and upper respiratory tract associated with short-term peaks exposures, but little evidence for respiratory sensitization or asthma. Nineteen case reports described asthma, laryngitis, or hypersensitivity pneumonitis in MMA-exposed workers; however, exposures were either not well described or involved mixtures containing more reactive respiratory sensitizers and irritants. The weight of evidence, both experimental and observational, argues that MMA is not a respiratory sensitizer. [ABSTRACT FROM AUTHOR]

The major problem in agriculture is that of the fungal pathogens. In this era, where biological control is at focus, and is the centre of crop protection as well as environmental protection, the synthesis of new bio bodies is of utmost need. Fungicides available in the market, though of potential, are pathogen specific and highly pollutive. An attempt was made to raise polyclonal antibodies against Aspergillus niger. Following the particular standardised immunisation schedule, regular injections and periodic tapping were carried out. The IgG purified was used to check cross-reactivity with different crop fungi like Fusarium solani, Fusarium oxysporum f. sp. cicer (FOC), Rhizoctonia bataticola, Botryodiplodia theobromae, Curvularia sp., Alternaria porri and also with Aspergillus niger by two different methods. Liquid test media and the radial growth inhibition test performed in solid media were used to check the inhibition of fungi and cross-reactivity. The results were subjected to analysis of variance (ANOVA) by using Tukey's test at the significance level of p < 0.05. The antibodies were active against all the fungi for more than 15 days except for A. niger in which from the seventh day onwards spore germination was observed. The probable role of antibodies to detect the common antigenic molecule that may be present in all the tested fungi and their role in inhibiting these fungi is discussed. [ABSTRACT FROM AUTHOR]

Gap junction proteins are a highly homologous family of 21 connexins. Here, the authors describe a tissue-specific technical artifact complicating analysis of connexin32 protein expression in the central nervous system. The authors show that in brain, but not liver, eight commonly employed antibodies exhibit a higher affinity for a cross-reactive protein that masks the detection of connexin32. Cross-reactivity is evident in Western blot analyses when proteins are subjected to reducing/denaturing conditions but not immunoprecipitation or immunofluorescent applications. Through bioinformatic analyses, tested by sucrose gradient fractionation and immunoblotting of lysates from connexin null-mutant mice, the authors show that the cross-reactive protein is not found in the same cellular compartments as connexin32 and is likely not a member of the connexin family. These findings are presented with the intent of helping to reduce the amount of time laboratories currently expend in validating changes in connexin32 expression in the central nervous system. [ABSTRACT FROM AUTHOR]

Characterisation of potential antigen(s) in the teliospore wall of Tilletia indica, the causal agent of Karnal bunt (KB) of wheat (Triticum aestivum) is necessary to generate specific immunoprobes for the development of reliable immunodiagnostic procedures. The intact teliospores of KB were used to generate anti-teliospore antibodies for the development of microtitre ELISA. The anti-teliospore antibodies cross-reacted with Tilletia foetida, Puccinia recondita and Puccinia striiformis suggesting that these species share one or more antigenic components. These antibodies recognised two unique immunoreactive bands of molecular weight ~34 kDa and ~15 kDa present in the spore wall fraction of T. indica after western blotting. The periodate treatment alone and sequential treatment of periodate followed by trypsin lost the immunoreactivity partially and completely, respectively. This suggests that cross-reactivity among bunt and rust of wheat fungal pathogens is most likely due to peptidal/oligosaccharide epitopes overlapping the spore wall's glycoprotein. [ABSTRACT FROM AUTHOR]

Genetic and environmental factors are believed to influence development of systemic lupus erythematosus (SLE). Endogenous retroviruses (ERV) correspond to the integrated proviral form of infectious retroviruses, which are trapped within the genome due to mutations. ERV represent a key molecular link between the host genome and infectious viral particles. ERV-encoded proteins are recognized by antiviral immune responses and become targets of autoreactivity. Alternatively, ERV protein may influence cellular processes and the life cycle of infectious viruses. As examples, the HRES-1 human ERV encodes a 28-kDa nuclear autoantigen and a 24-kDa small GTP-ase, termed HRES-1/Rab4. HRES-1/p28 is a nuclear autoantigen recognized by cross-reactive antiviral antibodies, while HRES-1/Rab4 regulates surface expression of CD4 and the transferrin receptor (TFR) through endosome recycling. Expression of HRES-1/Rab4 is induced by the tat gene of HIV-1, which in turn down-regulates expression of CD4 and susceptibility to re-infection by HIV-1. CD4 and the TFR play essential roles in formation of the immunological synapse (IS) during normal T-cell activation by a cognate MHC class II peptide complex. The key intracellular transducer of T-cell activation, Lck, is brought to the IS via binding to CD4. T-cell receptorζ (TCRζ) chain binds to the TFR. Abnormal T-cell responses in SLE have been associated with reduced lck and TCRζ chain levels. HRES-1 is centrally located on chromosome 1 at q42 relative to lupus-linked microsatellite markers and polymorphic HRES-1 alleles have been linked to the development of SLE. 1q42 is one of the three most common fragile sites in the human genome, and is inducible by DNA demethylation, a known mechanism of retroviral gene activation. Molecular mimicry and immunomodulation by a ERV, such as HRES-1, may contribute to self-reactivity and abnormal T and B-cell functions in SLE. [ABSTRACT FROM AUTHOR]

Gammaridean and caprellid amphipods, crustaceans of the order Amphipoda, inhabit laver culture platforms and, hence, are occasionally found in nori (dried laver) sheets. Amphipods mixed in nori may cause allergic reactions in sensitized patients, as is the case with other crustaceans, such as shrimp and crab, members of the order Decapoda. In this study, dried samples of amphipods (unidentified) found in nori and fresh samples of gammaridean amphipod (Gammarus sp., not accurately identified) and caprellid amphipod (Caprella equilibra) were examined for allergenicity and allergens using two species of decapods (black tiger prawn and spiny lobster) as references. When analyzed by ELISA, sera from crustacean-allergic patients reacted to extracts from amphipod samples, although less potently than to the extracts from decapods. In IgE-immunoblotting, a 37-kDa protein was found to be the major allergen in amphipods. Based on the molecular mass and the cross-reactivity with decapod tropomyosin evidenced by inhibition ELISA and inhibition immunoblotting, the 37-kDa protein was identified as amphipod tropomyosin. [ABSTRACT FROM AUTHOR]

The competitive enzyme immunoassays for detection of Campylobacter jejuni, C. coli and C. fetus subsp. fetus have been developed. Rabbit and hen immunoglobulins were prepared for these purposes. The working conditions of ELISAs, such as the concentrations of immunoreactants, incubation temperatures and time, and the composition of the substrate have been established. The detection limits were in the range 5.0 104-3.2 106 cfu/ml. The application of chemiluminescent substrates did not result in any significant improvement of the assay's detectability and sensitivity. Prepared antibodies showed rather high specificity and cross-reactivity profiles, and both rabbit and hen immunoglobulins were similar. Only IgY to C. jejuni cross-reacted with seven strains of C. jejuni and two other Campylobacter spp. A limited number of naturally and artificially contaminated food samples were tested. The results obtained by means of an enzyme immunoassay were compared with those obtained from PCR or commercially available Singlepath® Campylobacter GLISA-Rapid Test. Poultry products were naturally contaminated with Campylobacters. The wild species were identified as C. jejuni and C. coli. [ABSTRACT FROM AUTHOR]

Immunology of the central nervous system (CNS) is a growing field of study. Until recently the brain was considered an 'immunologically privileged' site. It is increasingly apparent that the CNS has a significant but tightly regulated capability to mount an inflammatory and immune response. This article serves as an introduction to the special section at the start of this issue on neuroimmunology. We also focus on several immunological concepts that are particularly relevant in the context of neuroimmunology—cross-reactivity, the immunological synapse and the nature of the immune response to transplantation in the CNS. We conclude that the fundamental concepts are common to all branches of immunology. Better understanding of the basic mechanisms will blur the borders between the different areas of immunology. [ABSTRACT FROM AUTHOR]

The article focuses on a study in which, the author examined the cross-reactivities of Stachybotrys chartarum, Aspergillus niger/fumigatus, and Penicillium notatum with affinity-purified rabbit sera. Only affinity-purified antibodies and the most sensitive immunoassay technique were used for the cross-inhibition studies. The detection of antibodies to mold antigens is used as an adjunct in the diagnosis of allergies, aspergillosis, infectious states, and farmer's lung disease. Rabbit anti-Aspergillus and anti-Penicillium and sera from patients with precipitating antibodies against these fungi have been analyzed by immunoglobulin G, enzyme-linked immunosorbent assay (ELISA) inhibition and immunoblot. The protocol for optimal fungal antigen extract preparation was based on earlier procedures, with some modifications. The results of the immunodiffusion, Western blot, and ELISA testing on different dilutions of control and immune rabbit sera against Aspergillus, Penicillium, and Stachybotrys extracts are shown.

A polarization fluoroimmunoassay (PFIA) based on rabbit polyclonal antibodies was developed and optimized for the detection and quantification of linear alkylbenzene sulfonates (LAS). Hapten mimics with different lengths of the alkyl radical and positions of the phenyl ring were synthesized to prepare fluorescent conjugates (tracers) for PFIA. Structures of target compounds, LAS derivatives and tracers were confirmed by nuclear magnetic resonance and mass spectrometry. Fluorescent tracers in combination with antibodies raised against different LAS immunogens were used for the development of class-selective PFIAs based on different structural combinations of the aliphatic chain moiety. The most sensitive assay was found for the homologous antibody-tracer combination based on the 3-(p-sulfophenyl)butyric acid hapten mimic. The PFIA has a limit of detection of 0.5 mg/L, calibration curve midpoint of 9 mg/L and a dynamic range of 3 to 85 mg/L. Cross-reactivity studies demonstrated the assay is specific towards the sulfophenyl moiety of anionic surfactants while the recognition of aliphatic alkylsulfonates was not significant (cross-reactivity<5%). Total assay time for 10 samples was 7 min, thus allowing fast and reliable detection of LAS in the low mg/L range. [ABSTRACT FROM AUTHOR]

A polarization fluoroimmunoassay (PFIA) based on previously raised rabbit polyclonal antibodies was developed for detection and quantification of the sulfonylurea herbicide metsulfuron-methyl (MSM) in surface water samples. A 'restricted' hapten, consisting of only phenylsulfonamide moiety of MSM molecule was used to synthesize a fluorescein-labelled derivative of MSM (tracer). The limited assay cross-reactivity was observed towards most of sulfonylurea herbicides, with exception of primisulfuron-methyl and chlorimuron-ethyl. The optimized MSM PFIA had a dynamic range from 30 to 1000 μg ι[sup -1] with an IC[sub 50] value of 123 μg ι[sup -1] and a lower detection limit of 5 μg ι[sup -1]. Intra- and inter-assay precision over the of MSM calibration curve dynamic range were 0.9 and 7.1%, respectively. Influence of several organic solvents on MSM PFIA performance was investigated, and the assay was found to tolerate up to 10% v/v of methanol, DMSO and acetone. Application of MSM PFIA technique to the analysis of surface water samples taken from lakes and rivers in Bavaria, and spiked with MSM at concentration of 50 μg ι[sup -1], gave mean MSM recoveries of 108.6–110.8%. [ABSTRACT FROM AUTHOR]

Two fluorescence polarization immunoassays (FPIA) for detection of the insecticide DDT (DDT-specific FPIA) and the sum of DDT isomers and metabolites (DDT class-specific FPIA) were developed. Various fluorescein-labeled tracers were synthesized and studied in order to achieve a high sensitivity using different monoclonal antibodies. For DDT-specific assay the sensitivity, estimated as limit of detection, was 12 ng/mL, with a linear working range between 20 and 1000 ng/mL. For DDT class-specific assay the sensitivity was 3 ng/mL, with a linear working range between 5 and 1000 ng/mL. Regarding specificity to other chlorinated pesticides, both immunoassays showed no cross-reaction (< 0.1%) with 2,4-D, endosulfan, diuron and hexachlorocyclohexanes. Both FPIAs were successfully applied to the analysis of DDT in spiked tap water samples. The FPIAs for DDT are simple, rapid and homogeneous (without separation and washing steps). Total time of analysis is less than 1 min per sample. The developed FPIAs could be used for rapid screening of water samples for contamination with DDT and its isomers and metabolites. [ABSTRACT FROM AUTHOR]

Two different hapten designs have been suggested for production of generic antibodies. The first approach was based on the immunogen prepared by conjugating simazine mimic to carrier proteins through the Cl position of the s-triazine herbicide. For sulfonylurea herbicides a single ring hapten strategy was designed in order to produce antibodies with dominant selectivity towards arylsulfonyl or triazine moieties of metsulfuron-methyl. The best monoclonal antibody raised against simazine-derived hapten mimic (clone B10/B8/D2) exhibited in direct ELISA a broad pattern for a group of methylthio and methoxy s-triazines. Cross-reactivities based on simazine ( = 100%) were 417, 125, 25, 50, 28, 42 and 11% for simetryn, ametryn, prometryn, terbutryn, aziprotryn, atraton and atrazine, respectively. The superiour generic pattern was found for monoclonal antibody raised against s-triazine moiety of metsulfuron-methyl herbicide (clone 2C8/C8). This antibody showed in indirect ELISA the cross-reactivity values 100, 142, 95, 60, 40, 167, 83 and 21% for metsulfuronmethyl, cinosulfuron, triasulfuron, primisulfuron-methyl, thifensulfuron-methyl, chlorsulfuron and prosulfuron and tribenuron-methyl, respectively. The assays using both monoclonal and polyclonal antibodies operated within the ppt–ppb calibration ranges. [ABSTRACT FROM AUTHOR]