BACKGROUND: The house dust mite (HDM) allergen Der p 18 belongs to the glycoside hydrolase family 18 chitinases. The relevance of Der p 18 for house dust mite allergic patients has only been partly investigated. OBJECTIVE: To perform a detailed characterization of Der p 18 on a molecular, structural and immunological level. METHODS: Der p 18 was expressed in E. coli, purified to homogeneity, tested for chitin-binding activity and its secondary structure was analyzed by circular dichroism. Der p 18-specific IgG antibodies were produced in rabbits to localize the allergen in mites using immunogold electron microscopy and to search for cross-reactive allergens in other allergen sources (i.e. mites, crustacea, mollusca and insects). IgE reactivity of rDer p 18 was tested with sera from clinically well characterized HDM-allergic patients (n = 98) and its allergenic activity was analyzed in basophil activation experiments. RESULTS: Recombinant Der p 18 was expressed and purified as a folded, biologically active protein. It shows weak chitin-binding activity and partial cross-reactivity with Der f 18 from D. farinae but not with proteins from the other tested allergen sources. The allergen was mainly localized in the peritrophic matrix of the HDM gut and to a lower extent in fecal pellets. Der p 18 reacted with IgE from 10% of mite allergic patients from Austria and showed allergenic activity when tested for basophil activation in Der p 18-sensitized patients. CONCLUSION: Der p 18 is a rather genus-specific minor allergen with weak chitin-binding activity but exhibits allergenic activity and therefore should be included in diagnostic test panels for HDM allergy.

• MeSH
• antigeny roztočů domácího prachu chemie   genetika   imunologie   MeSH
• antisérum chemie   MeSH
• bazofily cytologie   účinky léků   imunologie   MeSH
• chitin chemie   imunologie   MeSH
• Escherichia coli genetika   metabolismus   MeSH
• exprese genu MeSH
• interakční proteinové domény a motivy MeSH
• klonování DNA MeSH
• konformace proteinů, alfa-helix MeSH
• konformace proteinů, beta-řetězec MeSH
• králíci MeSH
• lidé MeSH
• proteiny členovců chemie   genetika   imunologie   MeSH
• protilátky krev   chemie   izolace a purifikace   MeSH
• Pyroglyphidae chemie   ultrastruktura   MeSH
• rekombinantní proteiny chemie   genetika   imunologie   MeSH
• respirační alergie chemicky indukované   imunologie   patofyziologie   MeSH
• sbalování proteinů MeSH
• sekvence aminokyselin MeSH
• sekvenční homologie aminokyselin MeSH
• sekvenční seřazení MeSH
• vazba proteinů MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigeny roztočů domácího prachu MeSH
• antisérum MeSH
• chitin MeSH
• Der p 18 allergen, Dermatophagoides pteronyssinus MeSH Prohlížeč
• proteiny členovců MeSH
• protilátky MeSH
• rekombinantní proteiny MeSH

Cryptochromes are a ubiquitous group of blue-light absorbing flavoproteins that in the mammalian retina have an important role in the circadian clock. In birds, cryptochrome 1a (Cry1a), localized in the UV/violet-sensitive S1 cone photoreceptors, is proposed to be the retinal receptor molecule of the light-dependent magnetic compass. The retinal localization of mammalian Cry1, homologue to avian Cry1a, is unknown, and it is open whether mammalian Cry1 is also involved in magnetic field sensing. To constrain the possible role of retinal Cry1, we immunohistochemically analysed 90 mammalian species across 48 families in 16 orders, using an antiserum against the Cry1 C-terminus that in birds labels only the photo-activated conformation. In the Carnivora families Canidae, Mustelidae and Ursidae, and in some Primates, Cry1 was consistently labeled in the outer segment of the shortwave-sensitive S1 cones. This finding would be compatible with a magnetoreceptive function of Cry1 in these taxa. In all other taxa, Cry1 was not detected by the antiserum that likely also in mammals labels the photo-activated conformation, although Western blots showed Cry1 in mouse retinal cell nuclei. We speculate that in the mouse and the other negative-tested mammals Cry1 is involved in circadian functions as a non-light-responsive protein.

• MeSH
• antisérum chemie   MeSH
• Canidae fyziologie   MeSH
• čípky retiny - opsiny genetika   MeSH
• čípky retiny fyziologie   účinky záření   ultrastruktura   MeSH
• cirkadiánní rytmus fyziologie   účinky záření   MeSH
• exprese genu MeSH
• fylogeneze * MeSH
• Hominidae fyziologie   MeSH
• imunohistochemie MeSH
• konformace proteinů MeSH
• kryptochromy chemie   genetika   MeSH
• magnetické pole MeSH
• medvědovití fyziologie   MeSH
• Mustelidae fyziologie   MeSH
• proteinové domény MeSH
• ptáci fyziologie   MeSH
• savci klasifikace   fyziologie   MeSH
• světlo MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antisérum MeSH
• čípky retiny - opsiny MeSH
• kryptochromy 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
• antisérum chemie   imunologie   MeSH
• ELISA metody   MeSH
• genistein analýza   chemie   imunologie   MeSH
• hapteny analýza   chemie   imunologie   MeSH
• imunoglobulin G chemie   imunologie   MeSH
• isoflavony analýza   imunologie   MeSH
• králíci MeSH
• Medicago sativa chemie   MeSH
• peroxidasa chemie   MeSH
• radioimunoanalýza metody   MeSH
• senzitivita a specificita MeSH
• sérový albumin hovězí chemie   imunologie   MeSH
• skot MeSH
• specificita protilátek imunologie   MeSH
• zkřížené reakce imunologie   MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antisérum MeSH
• biochanin A MeSH Prohlížeč
• genistein MeSH
• hapteny MeSH
• imunoglobulin G MeSH
• isoflavony MeSH
• peroxidasa MeSH
• sérový albumin hovězí MeSH

Seven monoclonal anti-idiotypic antibodies (anti-ID MAbs, Ab2) were generated against virus-neutralizing and hemagglutination-inhibiting monoclonal antibody (Ab1) specific for tick-borne encephalitis (TBE) virus. Six of these anti-ID MAbs inhibited the binding of Ab1 to the virus antigen, thus classifying these anti-ID antibodies as Ab2 beta or AB2 gamma. Inhibition tests with heterologous anti-TBE sera revealed that these anti-ID MAbs were not recognized by anti-TBE antibodies and therefore they do not carry an internal image of TBE virus antigen. Hence, the anti-ID MAbs may be classified as Ab2 gamma type. None of the anti-ID MAbs induced production of antiviral antibodies nor protective immunity in syngeneic Balb/c mice. Using these anti-ID MAbs four nonoverlapping idiotopes were identified on Ab1 variable region.

• MeSH
• antisérum chemie   MeSH
• ELISA MeSH
• mapování epitopu MeSH
• monoklonální protilátky biosyntéza   chemie   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• neutralizační testy MeSH
• protilátky anti-idiotypické biosyntéza   chemie   MeSH
• protilátky virové biosyntéza   chemie   MeSH
• viry klíšťové encefalitidy imunologie   MeSH
• zkřížené reakce MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antisérum MeSH
• monoklonální protilátky MeSH
• protilátky anti-idiotypické MeSH
• protilátky virové MeSH