Tick-borne encephalitis virus (TBEV) is an emerging human pathogen that causes potentially fatal disease with no specific treatment. Mouse monoclonal antibodies are protective against TBEV, but little is known about the human antibody response to infection. Here, we report on the human neutralizing antibody response to TBEV in a cohort of infected and vaccinated individuals. Expanded clones of memory B cells expressed closely related anti-envelope domain III (EDIII) antibodies in both groups of volunteers. However, the most potent neutralizing antibodies, with IC50s below 1 ng/ml, were found only in individuals who recovered from natural infection. These antibodies also neutralized other tick-borne flaviviruses, including Langat, louping ill, Omsk hemorrhagic fever, Kyasanur forest disease, and Powassan viruses. Structural analysis revealed a conserved epitope near the lateral ridge of EDIII adjoining the EDI-EDIII hinge region. Prophylactic or early therapeutic antibody administration was effective at low doses in mice that were lethally infected with TBEV.

• MeSH
• Survival Analysis MeSH
• Epitopes immunology   MeSH
• Immunoglobulin G administration & dosage   immunology   MeSH
• Encephalitis, Tick-Borne immunology   prevention & control   virology   MeSH
• Cohort Studies MeSH
• Cells, Cultured MeSH
• Humans MeSH
• Antibodies, Monoclonal administration & dosage   genetics   immunology   MeSH
• Mice, Inbred BALB C MeSH
• Mice MeSH
• Antibodies, Neutralizing administration & dosage   genetics   immunology   MeSH
• Viral Envelope Proteins genetics   immunology   MeSH
• Antibodies, Viral administration & dosage   genetics   immunology   MeSH
• Amino Acid Sequence MeSH
• Sequence Homology, Amino Acid MeSH
• Encephalitis Viruses, Tick-Borne drug effects   immunology   physiology   MeSH
• Cross Reactions immunology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Names of Substances
• Epitopes MeSH
• Immunoglobulin G MeSH
• Antibodies, Monoclonal MeSH
• Antibodies, Neutralizing MeSH
• Viral Envelope Proteins MeSH
• Antibodies, Viral MeSH

INTRODUCTION: Frequently observed multiple sensitizations to several animals highlights the importance of a molecular diagnosis, distinguishing between sensitizations specific to single species and sensitizations due to cross-reactivity. OBJECTIVE: The aim of our study was to assess the usefulness of a molecular diagnosis in the description of sensitization profiles in allergy patients living in Central Europe, with a particular focus on animal-derived molecules. METHODS: The molecular diagnosis was performed using the ImmunoCAP ISAC microarray. Results of 1,255 allergy patients were subjected to statistical analysis. RESULTS: The highest sensitization rates were observed for uteroglobin Fel d 1 (31.8%) and kallikrein Can f 5 (16.4%), followed by animal lipocalins Can f 1 (13.9%), Equ c 1 (6.2%), Fel d 4 (5.3%), Can f 2 (4.2%), and Mus m 1 (4.1%). Sensitization rates to serum albumins Fel d 2, Can f 3, Equ c 3, and Bos d 6 were very low, with the highest being 3.2% to Fel d 2. Detailed subanalysis confirmed the dominant role of Fel d 1 or Can f 5 and/or Can f 1 in cat- or dog-sensitized patients, respectively. Further analysis focused on lipocalins and albumins confirmed a high rate of cosensitizations within both groups. CONCLUSION: The sensitization to animal allergen molecules is very frequent in Central Europe. The most common is sensitization to species-specific cat uteroglobin Fel d 1 and dog kallikrein Can f 5, followed by sensitizations to animal lipocalins. Our data suggest that commonly observed multiple sensitizations detected by extract approach can be explained not only by true cosensitization, but also by cross-reactivity, mainly in the frame of lipocalins. Cross-reactive serum albumins are minor sensitizers and are probably not important from this point of view.

• Keywords
• Animal allergen, Animal allergy, Molecular diagnostics, Sensitization profile, Specific IgE,
• MeSH
• Allergens immunology   MeSH
• Hypersensitivity immunology   MeSH
• Child MeSH
• Adult MeSH
• Species Specificity MeSH
• Cats MeSH
• Infant MeSH
• Middle Aged MeSH
• Humans MeSH
• Lipocalins immunology   MeSH
• Adolescent MeSH
• Young Adult MeSH
• Mice MeSH
• Child, Preschool MeSH
• Dogs MeSH
• Aged MeSH
• Serum Albumin immunology   MeSH
• Cross Reactions immunology   MeSH
• Animals MeSH
• Check Tag
• Child MeSH
• Adult MeSH
• Cats MeSH
• Infant MeSH
• Middle Aged MeSH
• Humans MeSH
• Adolescent MeSH
• Young Adult MeSH
• Male MeSH
• Mice MeSH
• Child, Preschool MeSH
• Dogs MeSH
• Aged MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Europe MeSH
• Names of Substances
• Allergens MeSH
• Lipocalins MeSH
• Serum Albumin MeSH

Birch and other related trees of the families Betulaceae and Fagaceae (alder, hazel, oak, hornbeam, chestnut, and beech) constitute the birch homologous group. This grouping is primarily based on the extensive IgE cross-reactivity of allergen homologs to the major birch allergen Bet v 1. Birch pollen is the most dominant tree pollen in Northern and Central Europe and is a major cause of allergic rhinitis and, possibly, asthma symptoms. Over the last few decades, levels of birch pollen have risen and the period of exposure has increased due to climate changes. Subsequently, the prevalence of birch pollen sensitization has also increased. The cross-reactivity and sequential pollen seasons within the birch homologous group create a prolonged symptomatic allergy period beyond birch pollen alone. Furthermore, many plant food allergens contain homologs to Bet v 1, meaning that the majority of patients with birch pollen allergy suffer from secondary pollen food syndrome (PFS). As a result, the negative impact on health-related quality of life (HRQoL) in patients allergic to birch pollen is significant. The purpose of this manuscript was to narratively review topics of interest such as taxonomy, cross-reactivity, prevalence, clinical relevance, PFS, and HRQoL with regard to birch pollen allergy from a European perspective.

• Keywords
• alder, allergic rhinitis, birch, cross-reactivity, hazel,
• MeSH
• Allergens immunology   MeSH
• Antigens, Plant immunology   MeSH
• Betula immunology   MeSH
• Immunization MeSH
• Immunoglobulin E immunology   MeSH
• Humans MeSH
• Public Health Surveillance MeSH
• Prevalence MeSH
• Pollen immunology   MeSH
• Seasons MeSH
• Rhinitis, Allergic, Seasonal diagnosis   epidemiology   immunology   MeSH
• Symptom Assessment MeSH
• Cross Reactions immunology   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH
• Geographicals
• Europe epidemiology   MeSH
• Names of Substances
• Allergens MeSH
• Antigens, Plant MeSH
• Immunoglobulin E MeSH

• MeSH
• Allergens chemistry   immunology   MeSH
• Antigens, Plant chemistry   immunology   MeSH
• Gibberellins chemistry   immunology   MeSH
• Mass Spectrometry MeSH
• Immunoglobulin E immunology   MeSH
• Humans MeSH
• Food Hypersensitivity diagnosis   immunology   MeSH
• Pollen chemistry   immunology   MeSH
• Plant Proteins chemistry   immunology   MeSH
• Amino Acid Sequence MeSH
• Cross Reactions immunology   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Letter MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Allergens MeSH
• Antigens, Plant MeSH
• Gibberellins MeSH
• Immunoglobulin E MeSH
• Plant Proteins MeSH

5-Bromo-2'-deoxyuridine (BrdU) and 2'-deoxy-5-ethynyluridine (EdU) are widely used as markers of replicated DNA. While BrdU is detected using antibodies, the click reaction typically with fluorescent azido-dyes is used for EdU localisation. We have performed an analysis of ten samples of antibodies against BrdU with respect to their reactivity with EdU. Except for one sample all the others evinced reactivity with EdU. A high level of EdU persists in nuclear DNA even after the reaction of EdU with fluorescent azido-dyes if the common concentration of dye is used. Although a ten-time increase of azido-dye concentration resulted in a decrease of the signal provided by anti-BrdU antibodies, it also resulted in a substantial increase of the non-specific signal. We have shown that this unwanted reactivity is effectively suppressed by non-fluorescent azido molecules. In this respect, we have tested two protocols of the simultaneous localisation of incorporated BrdU and EdU. They differ in the mechanism of the revelation of incorporated BrdU for the reaction with antibodies. The first one was based on the use of hydrochloric acid, the second one on the incubation of samples with copper(I) ions. The use of hydrochloric acid resulted in a significant increase of the non-specific signal. In the case of the second method, no such effect was observed.

• MeSH
• Antibody Affinity MeSH
• Staining and Labeling MeSH
• Biological Transport MeSH
• Biotinylation MeSH
• Bromodeoxyuridine chemistry   immunology   metabolism   MeSH
• Deoxyuridine analogs & derivatives   chemistry   immunology   metabolism   MeSH
• DNA chemistry   metabolism   MeSH
• Fluorescent Dyes MeSH
• Microscopy, Fluorescence * MeSH
• HeLa Cells MeSH
• Humans MeSH
• Antibodies chemistry   immunology   MeSH
• Cross Reactions immunology   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• 5-ethynyl-2'-deoxyuridine MeSH Browser
• Bromodeoxyuridine MeSH
• Deoxyuridine MeSH
• DNA MeSH
• Fluorescent Dyes MeSH
• Antibodies MeSH

Development of antibodies with broad specificity recognition for sulfonamide drugs was found to be surprisingly difficult when conventional immunochemical strategies were applied to hapten design. To improve the cross-reactivity pattern of antibodies for the family of sulfonamide drugs, a novel strategy based on the single-ring (fragment-derived) hapten moieties with different spacer substituent lengths was employed for the preparation of immunogens, coating conjugates, and enzyme competitors. The rabbit antibodies raised against a common (one-ring) p-aminobenzenesulfonamide hapten moiety (attached to a carrier protein through the N-1 position) in combination with a homologous hapten-peroxidase tracer allowed the detection of 15 sulfonamide species at the maximum residue limit level using direct ELISA. The two-ring 6-(4-aminobenzensulfonylamino)hexanoic hapten mimics, previously reported in the literature as a weak generic antigen, generated surprisingly superior immune responses in rabbits. The antibodies raised against this two-ring hapten were capable of detecting at least 19 and 17 sulfonamides in a direct ELISA system at the regulatory level with sensitivities corresponding to 20 and 50% binding inhibition, respectively. A negligible cross-reaction with N4 metabolites makes it possible to measure responses of parent sulfonamides in the presence of their metabolized forms. In skimmed milk, the highest limit of detection (LOD) for sulfacetamide defined as 20% inhibition was 65.2 microg x L(-1) (IC20 value), whereas the additional 18 sulfonamides tested exhibited LODs in the range of 0.2-36.8 microg x L(-1). This sensitivity allows simple multisulfonamide tests to be established for use in the laboratory or on site.

• MeSH
• Enzyme-Linked Immunosorbent Assay methods   standards   MeSH
• Haptens immunology   MeSH
• Binding, Competitive MeSH
• Rabbits MeSH
• Milk MeSH
• Antibodies MeSH
• Antibody Specificity immunology   MeSH
• Sulfonamides analysis   immunology   MeSH
• Cross Reactions immunology   MeSH
• Animals MeSH
• Check Tag
• Rabbits MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Haptens MeSH
• Antibodies MeSH
• Sulfonamides MeSH

Two variants of immunoassay for the determination of biochanin A (5,7-dihydroxy 4'-methoxy isoflavone), i.e., a radioimmunoassay (RIA) and an indirect ELISA, have been developed and evaluated. Both methods employ the same rabbit antiserum to a 7-O-carboxymethyl-5-hydroxy-4'-methoxyisoflavone-bovine serum albumin (BSA) conjugate. A 125I-labeled hapten-tyrosine methyl ester (TME) conjugate was used as a radioligand for the RIA. The indirect ELISA uses immunogen-coated microtitration plates and a peroxidase-labeled antirabbit Ig antibody. Both methods are specific for biochanin A with a comparable sensitivity (3.1 pg/tube for RIA; 5.3 pg/well for ELISA); however, their sensitivity to individual cross-reactants differs. The main cross-reactants are sissotrin (the cross-reactivity 15.7% for RIA; 120% for ELISA), 5-hydroxy, 4',7-dimethoxy isoflavone (51.5% for RIA; 46.5% for ELISA), prunetin (4.5% for RIA; 5.0% for ELISA), genistein (0.8% for RIA; 2.8% for ELISA) and formononetin (0.4% for RIA; 0.3% for ELISA). These methods were used for the analysis of biochanin A in alfalfa and in several nonleguminous plants.

• MeSH
• Immune Sera chemistry   immunology   MeSH
• Enzyme-Linked Immunosorbent Assay methods   MeSH
• Genistein analysis   chemistry   immunology   MeSH
• Haptens analysis   chemistry   immunology   MeSH
• Immunoglobulin G chemistry   immunology   MeSH
• Isoflavones analysis   immunology   MeSH
• Rabbits MeSH
• Medicago sativa chemistry   MeSH
• Peroxidase chemistry   MeSH
• Radioimmunoassay methods   MeSH
• Sensitivity and Specificity MeSH
• Serum Albumin, Bovine chemistry   immunology   MeSH
• Cattle MeSH
• Antibody Specificity immunology   MeSH
• Cross Reactions immunology   MeSH
• Animals MeSH
• Check Tag
• Rabbits MeSH
• Cattle MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Immune Sera MeSH
• biochanin A MeSH Browser
• Genistein MeSH
• Haptens MeSH
• Immunoglobulin G MeSH
• Isoflavones MeSH
• Peroxidase MeSH
• Serum Albumin, Bovine MeSH

High sensitivity radioimmunoassay of 3beta, 7alpha-dihydroxy-5-androsten-17-one (7alpha-OH-DHEA) has been developed and evaluated. The method is based on polyclonal rabbit antisera raised against 19-O-(carboxymethyl)oxime bovine serum albumin conjugate and bridge- and position homologous [(125)I]iodotyrosine methyl ester as a tracer. Sensitivity of the assay amounted to 3.12 fmol (0.95 pg)/tube, precision as a mean intra- and interassay coefficient of variation was 7.1 and 10.6%, respectively, and the average recovery of the analyte added to steroid-free serum was 110%. Out of the steroids occurring in human serum which may interfere with the assay, the only important cross-reactants were dehydroepiandrosterone and 3beta, 7beta-dihydroxy-5-androsten-17-one (7beta-OH-DHEA) with cross-reactivities of 1.95 and 1.16%, respectively. The levels of free (unconjugated) 7alpha-OH-DHEA have been determined in 29 sera from healthy volunteers (23 females and 6 males), and from 48 patients (43 females and 5 males) in which dehydroepiandrosterone and its sulfate (DHEA/S) had been measured for various endocrinopathies. The levels in healthy subjects ranged from 0.21 to 6.57 (mean 2.33+/-1.50) nM, while those of the patients from 0 to 5. 99 (mean 1.46+/-1.52) nM. The levels of 7alpha-OH-DHEA in patients significantly correlated with those of DHEA and its sulfate.

• MeSH
• Dehydroepiandrosterone analogs & derivatives   blood   MeSH
• Hydroxysteroids analysis   blood   MeSH
• Rabbits MeSH
• Rats MeSH
• Humans MeSH
• Radioimmunoassay methods   MeSH
• Regression Analysis MeSH
• Sensitivity and Specificity MeSH
• Chromatography, High Pressure Liquid MeSH
• Cross Reactions immunology   MeSH
• Animals MeSH
• Check Tag
• Rabbits MeSH
• Rats MeSH
• Humans MeSH
• Male MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• 7-hydroxydehydroepiandrosterone MeSH Browser
• Dehydroepiandrosterone MeSH
• Hydroxysteroids MeSH

OBJECTIVE: To determine a target recognized by anti-Bh autoantibody, found in the serum of a patient with the unusual coexistence of systemic sclerosis (SSc) and psoriatic arthritis (PsA). METHODS: Antigens recognized by the anti-Bh serum were characterized by indirect immunofluorescence on HeLa cells, by conventional immunoblotting using nuclear extract or partially purified preparation of heterogenous nuclear RNP (hnRNP) proteins, and by 2-dimensional immunoblotting. For the analysis of cross-reactivity and immunofluorescence patterns, autoantibodies were affinity-purified by blot elution and then retested. RESULTS: Comparison of the reactivity of the anti-Bh antibody with the monoclonal antibody 4F4 against both the hnRNP C proteins, together with the determination of biochemical properties of the autoantigens, led to the identification of C1 and C2 core proteins as the targets for the anti-Bh autoantibody. CONCLUSION: Several essential components of the spliceosome are targeted by autoantibodies that are present in the sera of patients with systemic rheumatic diseases. We also found that the hnRNP core proteins C1 and C2 are recognized by the autoantibody present in the serum of a patient with SSc and PsA. C1 and C2 hnRNP proteins should be added to the several intracellular autoantigens recently shown to be cleaved by interleukin-1beta-converting enzyme-like enzymes during apoptosis.

• MeSH
• Autoantigens analysis   MeSH
• Autoantibodies immunology   MeSH
• Chromatography, Affinity MeSH
• Fluorescent Antibody Technique, Indirect MeSH
• HeLa Cells immunology   metabolism   MeSH
• Heterogeneous-Nuclear Ribonucleoprotein Group C MeSH
• Heterogeneous-Nuclear Ribonucleoproteins MeSH
• Humans MeSH
• Antibodies, Monoclonal MeSH
• Arthritis, Psoriatic complications   immunology   MeSH
• Ribonucleoproteins immunology   MeSH
• RNA, Heterogeneous Nuclear immunology   MeSH
• Aged MeSH
• Scleroderma, Systemic complications   immunology   MeSH
• Cross Reactions immunology   MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Aged MeSH
• Publication type
• Journal Article MeSH
• Case Reports MeSH
• Research Support, Non-U.S. Gov't MeSH
• Comparative Study MeSH
• Names of Substances
• Autoantigens MeSH
• Autoantibodies MeSH
• C1 HNRNP MeSH Browser
• Heterogeneous-Nuclear Ribonucleoprotein Group C MeSH
• Heterogeneous-Nuclear Ribonucleoproteins MeSH
• HNRNPC protein, human MeSH Browser
• Antibodies, Monoclonal MeSH
• Ribonucleoproteins MeSH
• RNA, Heterogeneous Nuclear MeSH

Spirochetal microorganisms were isolated from female Ixodes ricinus in Slovakia. Morphological, immunochemical and molecular biological analysis showed that the microorganism shared several common antigens with Borrelia species while other genetic traits were distinct and not related to Borrelia burgdorferi sensu lato. Lyme disease patient's serum contained antibodies reacting with antigens of this microorganism. On the one hand the cross-reacting antigens represent a risk of false positive results in laboratory diagnostics, while on the other hand they have a certain potential for vaccine development against Lyme disease.

• MeSH
• Antigens, Bacterial immunology   MeSH
• Bacterial Proteins analysis   MeSH
• DNA, Bacterial analysis   MeSH
• Electrophoresis, Polyacrylamide Gel MeSH
• Microscopy, Electron MeSH
• Ixodes microbiology   MeSH
• Humans MeSH
• Lyme Disease immunology   MeSH
• Polymerase Chain Reaction MeSH
• Antibodies, Bacterial immunology   MeSH
• Spirochaetales genetics   immunology   isolation & purification   ultrastructure   MeSH
• Blotting, Western MeSH
• Cross Reactions immunology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Antigens, Bacterial MeSH
• Bacterial Proteins MeSH
• DNA, Bacterial MeSH
• Antibodies, Bacterial MeSH