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1.
Pediatr Allergy Immunol ; 27 Suppl 23: 1-250, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-27288833

RESUMO

The availability of allergen molecules ('components') from several protein families has advanced our understanding of immunoglobulin E (IgE)-mediated responses and enabled 'component-resolved diagnosis' (CRD). The European Academy of Allergy and Clinical Immunology (EAACI) Molecular Allergology User's Guide (MAUG) provides comprehensive information on important allergens and describes the diagnostic options using CRD. Part A of the EAACI MAUG introduces allergen molecules, families, composition of extracts, databases, and diagnostic IgE, skin, and basophil tests. Singleplex and multiplex IgE assays with components improve both sensitivity for low-abundance allergens and analytical specificity; IgE to individual allergens can yield information on clinical risks and distinguish cross-reactivity from true primary sensitization. Part B discusses the clinical and molecular aspects of IgE-mediated allergies to foods (including nuts, seeds, legumes, fruits, vegetables, cereal grains, milk, egg, meat, fish, and shellfish), inhalants (pollen, mold spores, mites, and animal dander), and Hymenoptera venom. Diagnostic algorithms and short case histories provide useful information for the clinical workup of allergic individuals targeted for CRD. Part C covers protein families containing ubiquitous, highly cross-reactive panallergens from plant (lipid transfer proteins, polcalcins, PR-10, profilins) and animal sources (lipocalins, parvalbumins, serum albumins, tropomyosins) and explains their diagnostic and clinical utility. Part D lists 100 important allergen molecules. In conclusion, IgE-mediated reactions and allergic diseases, including allergic rhinoconjunctivitis, asthma, food reactions, and insect sting reactions, are discussed from a novel molecular perspective. The EAACI MAUG documents the rapid progression of molecular allergology from basic research to its integration into clinical practice, a quantum leap in the management of allergic patients.


Assuntos
Alérgenos/imunologia , Hipersensibilidade Imediata/diagnóstico , Imunoglobulina E/metabolismo , Biomarcadores/metabolismo , Humanos , Hipersensibilidade Imediata/imunologia , Hipersensibilidade Imediata/metabolismo , Hipersensibilidade Imediata/terapia , Testes Imunológicos/métodos , Medicina de Precisão/métodos
4.
Allergy ; 68(6): 748-56, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23621551

RESUMO

BACKGROUND: Replacement of peanut extracts by recombinant peanut components is an important step in allergy serologic testing. Criteria are needed for the unbiased inclusion of patients into a study to validate such a replacement. METHODS: Plasma samples from 64 peanut-positive children (42 reactors, 22 nonreactors in a double-blind, placebo-controlled food challenge) were used to compare IgE reactivity to six recombinant peanut allergens with reactivity to natural peanut proteins extracted at neutral or low pH. We tested the hypothesis that poor extractability of Ara h 9 and other basic allergens at neutral pH leads to under-representation of patients with such sensitization. RESULTS: IgE reactivity to the components did not fully explain IgE reactivity to peanut extract in 5 of 32 reactors with IgE to peanut extract ≤100 kUA /l. IgE reactivity to components was stronger than to the extract in 11 plasma samples, which was largely due to a low Ara h 8 reactivity of the extract. IgE reactivity to Ara h 9 was much lower than reactivity to other basic proteins, some of which bound IgE well in the RAST, but lost IgE reactivity upon immunoblotting. CONCLUSIONS: Conventional peanut extracts are deficient in significant IgE-binding components. The inclusion of patients for a validation study should be based on serology performed with improved peanut reagents to avoid a bias against these under-represented, potentially important allergens. To judge clinical relevance of an allergen, the reagent used for inclusion of patients needs to be efficient in detecting IgE to this component.


Assuntos
Alérgenos , Antígenos de Plantas , Arachis/imunologia , Imunoglobulina E/sangue , Hipersensibilidade a Amendoim/diagnóstico , Extratos Vegetais , Proteínas de Vegetais Comestíveis , Alérgenos/imunologia , Antígenos de Plantas/imunologia , Biomarcadores/sangue , Criança , Eletroforese em Gel de Poliacrilamida , Humanos , Immunoblotting , Hipersensibilidade a Amendoim/sangue , Hipersensibilidade a Amendoim/imunologia , Extratos Vegetais/imunologia , Proteínas de Vegetais Comestíveis/imunologia
5.
Allergy ; 66(10): 1261-74, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21623828

RESUMO

Here, we discuss various questions related to IgE epitopes: What are the technical possibilities and pitfalls, what is currently known, how can we put this information into hypothetical frameworks and the unavoidable question: how useful is this information for patient care or allergenicity prediction? We discuss the information obtained by (i) 3D structures of allergen-antibody complexes; (ii) analysis of allergen analogues; (iii) mimics without obvious structural similarity; (iv) mAbs competing with IgE; (v) repertoire analysis of cloned IgEs, and other developments. Based on limited data, four suggestions are presented in the literature: (i) IgE might be more cross-reactive than IgG; (ii) IgE might be more often directed to immunologically 'uninviting' surfaces; (iii) IgE epitopes may tend to cluster and (iv) IgE paratopes might have a higher intrinsic flexibility. While these are not proven facts, they still can generate hypotheses for future research. The hypothesis is put forward that the IgE repertoire of switched B-cells is less influenced by positive selection, because positive selection might not be able to rescue IgE-switched B cells. While this might be of interest for the discussion about mechanisms leading to allergen-sensitization, we need to be modest in answering the 'clinical relevance' question. Current evidence indicates the IgE-epitope repertoire is too big to make specific IgE epitopes a realistic target for diagnosis, treatment or allergenicity prediction. In-depth analysis of a few selected IgE epitope-peptides or mimitopes derived from allergen-sequences and from random peptide libraries, respectively, might well prove rewarding in relation to diagnosis and prognosis of allergy, particularly food allergy.


Assuntos
Epitopos/química , Epitopos/imunologia , Imunoglobulina E/química , Imunoglobulina E/imunologia , Alérgenos/imunologia , Animais , Mapeamento de Epitopos/métodos , Humanos , Hipersensibilidade/imunologia , Ligação Proteica
6.
Allergy ; 66(4): 517-22, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21083569

RESUMO

BACKGROUND: The modified Th2 response, defined as an IgG4 response in the absence of IgE, is suggested to protect against the development of allergic sensitization. However, studies suggesting this protective effect all had a cross-sectional design, making it impossible to study the development of both responses. AIM OF THE STUDY: We aimed to study the dynamics in IgG4 antibodies in relation to allergic sensitization in an occupational cohort of starting laboratory animal workers. Moreover, we studied the relation between exposure, antibody responses, atopy and self reported allergic symptoms. METHODS: A total of 110 starting animal workers were followed for 2 years. IgG4 antibodies against rats and mice were assessed. Workers were tested for allergic sensitization and exposure to animal allergens was estimated. Symptom status was assessed using questionnaires. RESULTS: Rat and mouse specific IgG4 antibodies were present before the development of allergy and did not significantly change over time. Allergic sensitization was related to exposure and atopic status but high levels of IgG4 showed no protective effect. In contrary, workers that developed mouse specific sensitization during follow up had higher levels of mouse specific IgG4. Symptoms were related to allergic sensitization and IgG4 levels did not influence that relationship. CONCLUSIONS: IgG4 antibodies are present before IgE antibodies develop and IgG4 levels are stable over time. In our occupational cohort, the modified Th2 response had no protective effect on development of sensitization or allergic symptoms.


Assuntos
Hipersensibilidade/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Doenças Profissionais/imunologia , Adolescente , Adulto , Animais , Animais de Laboratório/imunologia , Feminino , Humanos , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Masculino , Pessoal de Laboratório Médico , Camundongos , Ratos , Testes Cutâneos , Adulto Jovem
7.
Br J Anaesth ; 106(5): 665-74, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21414980

RESUMO

BACKGROUND: The significance of IgE antibodies to neuromuscular blocking agent (NMBA)-induced anaphylactic reactions during anaesthesia is unclear. We investigated the relevance of IgE to rocuronium using an in vitro technique. METHODS: Serum samples from 61 patients with anaphylactic reactions during anaesthesia were investigated. On the basis of clinical history, allergy to NMBA was considered likely in 48 patients, further assessed using intradermal skin tests for several commonly used NMBAs, including rocuronium, vecuronium, and succinylcholine. To determine the presence of rocuronium IgE in human serum, a rocuronium-human serum albumin (rocHSA) conjugate was coupled to a solid phase and a radioallergosorbent test performed. The biological effects of patient serum NMBA-IgE on histamine release were investigated using in vitro sensitized basophils from healthy blood donors. RESULTS: IgE to rocuronium was found in 23 of 48 serum samples (48%) with NMBA allergy, although only two of these were able to sensitize basophils to release histamine in response to rocHSA. IgE-responsiveness in the basophil test was only observed with conjugated rocHSA and not with unconjugated rocuronium or the other NMBAs evaluated. However, unconjugated rocuronium inhibited the histamine release induced by rocHSA. Correlation between skin-test reactivity to rocuronium and IgE to rocHSA was low (P>0.1). In contrast, striking correlation between IgE to rocuronium and skin-test reactivity to succinylcholine was found (P<0.001). CONCLUSIONS: Our results indicate that NMBA-related anaphylaxis requires not only IgE NMBA reactivity, but also altered cellular reactivity in the patient. The latter may be demonstrable by testing basophils from the patient, a skin test with (steroidal) NMBA, or both.


Assuntos
Anafilaxia/induzido quimicamente , Androstanóis/imunologia , Imunoglobulina E/sangue , Complicações Intraoperatórias/induzido quimicamente , Fármacos Neuromusculares não Despolarizantes/imunologia , Adulto , Idoso , Anafilaxia/imunologia , Androstanóis/efeitos adversos , Anestesia Geral , Especificidade de Anticorpos , Teste de Degranulação de Basófilos/métodos , Feminino , Liberação de Histamina/efeitos dos fármacos , Humanos , Complicações Intraoperatórias/imunologia , Masculino , Pessoa de Meia-Idade , Fármacos Neuromusculares não Despolarizantes/efeitos adversos , Teste de Radioalergoadsorção/métodos , Rocurônio , Testes Cutâneos/métodos
8.
Allergy ; 65(10): 1227-33, 2010 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-20121760

RESUMO

BACKGROUND: Occupational allergy forms an attractive model to study the development of allergic responses, as in some occupations it has a high incidence and develops quickly. In a cohort of starting laboratory animal workers, we previously found 20% sensitization to animal allergens within 2 years. METHODS: We compared cellular responses of incident laboratory animal workers who developed rat-specific sensitization (cases, n = 18) during 2 years of follow-up to control animal workers matched for atopic status but without sensitization after follow-up (controls, n = 18). Practically, this is a case-control study, nested within the cohort. Rat-specific IgE antibodies were measured in sera, and allergen-specific and nonspecific cytokine responses were measured in whole blood and in isolated peripheral blood mononuclear cells. RESULTS: Self-reported allergic symptoms were related to the presence of rat-specific IgE (P ≤ 0.01). Cases developed a rat allergen-specific interleukin (IL)-4 response during sensitization, while controls did not show an increased IL-4 response (at visit D: 33 vs 5 IL-4 producing cells/10(6) cells, P < 0.001). The IL-4 response was related to the levels of rat-specific IgE in cases (visit D: rho = 0.706, P < 0.001). By contrast, allergen-specific IL-10 and interferon γ (IFNγ) responses as well as nonspecific cytokine responses were comparable between cases and controls. CONCLUSION: This study is the first to show the development of an allergen-specific IL-4 response in adult human subjects during allergen-specific sensitization. This IL-4 response was quantitatively associated with the development of the specific IgE antibodies. Allergen-specific or nonspecific IL-10 and IFNγ responses showed no protective effect on the development of allergic sensitization.


Assuntos
Técnicos em Manejo de Animais , Citocinas/imunologia , Hipersensibilidade/etiologia , Interleucina-4/imunologia , Doenças Profissionais/imunologia , Ratos/imunologia , Animais , Estudos de Casos e Controles , Citocinas/sangue , Humanos , Hipersensibilidade/imunologia , Imunoglobulina E/sangue , Interleucina-4/sangue , Pessoal de Laboratório Médico , Doenças Profissionais/etiologia , Exposição Ocupacional/efeitos adversos
9.
Clin Exp Allergy ; 39(4): 469-77, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19222496

RESUMO

Despite its well-known association with IgE-mediated allergy, IgG4 antibodies still have several poorly understood characteristics. IgG4 is a very dynamic antibody: the antibody is involved in a continuous process of half-molecules (i.e. a heavy and attached light-chain) exchange. This process, also referred to as 'Fab-arm exchange', results usually in asymmetric antibodies with two different antigen-combining sites. While these antibodies are hetero- bivalent, they will behave as monovalent antibodies in most situations. Another aspect of IgG4, still poorly understood, is its tendency to mimic IgG rheumatoid factor (RF) activity by interacting with IgG on a solid support. In contrast to conventional RF, which binds via its variable domains, the activity of IgG4 is located in its constant domains. This is potentially a source of false positives in IgG4 antibody assay results. Because regulation of IgG4 production is dependent on help by T-helper type 2 (Th2) cells, the IgG4 response is largely restricted to non-microbial antigens. This Th2-dependency associates the IgG4 and IgE responses. Another typical feature in the immune regulation of IgG4 is its tendency to appear only after prolonged immunization. In the context of IgE-mediated allergy, the appearance of IgG4 antibodies is usually associated with a decrease in symptoms. This is likely to be due, at least in part, to an allergen-blocking effect at the mast cell level and/or at the level of the antigen-presenting cell (preventing IgE-facilitated activation of T cells). In addition, the favourable association reflects the enhanced production of IL-10 and other anti-inflammatory cytokines, which drive the production of IgG4. While in general, IgG4 is being associated with non-activating characteristics, in some situations IgG4 antibodies have an association with pathology. Two striking examples are pemphigoid diseases and sclerosing diseases such as autoimmune pancreatitis. The mechanistic basis for the association of IgG4 with these diseases is still enigmatic. However, the association with sclerosing diseases may reflect an excessive production of anti-inflammatory cytokines triggering an overwhelming expansion of IgG4-producing plasma cells. The bottom line for allergy diagnosis: IgG4 by itself is unlikely to be a cause of allergic symptoms. In general, the presence of allergen-specific IgG4 indicates that anti-inflammatory, tolerance-inducing mechanisms have been activated. The existence of the IgG4 subclass, its up-regulation by anti-inflammatory factors and its own anti-inflammatory characteristics may help the immune system to dampen inappropriate inflammatory reactions.


Assuntos
Hipersensibilidade/imunologia , Imunoglobulina E/imunologia , Imunoglobulina G/imunologia , Células Th2/imunologia , Alérgenos/imunologia , Animais , Humanos , Imunoglobulina E/metabolismo , Imunoglobulina G/biossíntese , Pancreatite/imunologia , Pancreatite/metabolismo , Pênfigo/imunologia , Pênfigo/metabolismo , Células Th2/metabolismo
10.
Allergy ; 64(8): 1202-8, 2009 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-19245426

RESUMO

BACKGROUND: Recall bias may provide discrepant relationships of pet exposure with sensitization and asthma development. We studied prospectively effects of pets at home on development of sensitization, asthma and respiratory symptoms from birth up to age 8 years. METHODS: Event history analysis was performed on annually registered data of 2951 children, participating in the PIAMA birth cohort study. RESULTS: Children with a cat or dog at home at 3 months of age had a significantly lower prevalence of sensitization to inhalant allergens at age 8, but not of asthma. A cat decreased the risk of house dust mite sensitization at age 8 [odds ratio (OR) = 0.68, 95% confidence interval (CI) 0.49-0.95], a dog of pollen sensitization (OR = 0.49, 95% CI: 0.29-0.83). A cat or dog at home did not significantly affect asthma incidence in each subsequent year. From 2 years of age onwards, the incidence of wheeze (OR = 1.52, 95% CI: 1.12-2.05) and a dry cough at night (OR = 1.28, 95% CI: 1.05-1.57) was higher in children with a dog, whereas removal of a dog increased the risk of developing asthma symptoms. Comparing analyses using prospectively and retrospectively collected data on diagnosed asthma showed important recall bias. CONCLUSIONS: Our prospective study shows a protective effect of early presence of pets at home on sensitization to inhalant allergens, but no prevention of asthma development. Furthermore, children with pets had more frequent transient or intermittent asthma symptoms. Parental report of asthma by recall may provide spurious results of these associations.


Assuntos
Asma/epidemiologia , Gatos/imunologia , Cães/imunologia , Alérgenos/imunologia , Animais , Asma/imunologia , Criança , Poeira/imunologia , Feminino , Humanos , Hipersensibilidade/epidemiologia , Hipersensibilidade/imunologia , Imunização , Incidência , Masculino , Ácaros/imunologia , Países Baixos/epidemiologia , Pólen/imunologia , Estudos Prospectivos , Fatores de Risco
11.
Curr Opin Immunol ; 10(6): 634-9, 1998 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-9914221

RESUMO

Immediate hypersensitivity to indoor or outdoor allergens is strongly associated with asthma or hay fever, respectively. Recent progress has defined the sequences, tertiary structures and enzymatic functions of many of the proteins involved; furthermore, the immune responses to these proteins have been examined; however, the mechanisms responsible for the dichotomous response remain elusive. The resolution of such mechanisms may explain the large increase in the prevalence of eosinophil-rich inflammation of the lower respiratory tract.


Assuntos
Alérgenos/imunologia , Hipersensibilidade Respiratória/imunologia , Animais , Asma/imunologia , Modelos Animais de Doenças , Humanos , Imunoglobulina E/imunologia , Exposição por Inalação
12.
Lancet ; 365(9454): 142-9, 2005.
Artigo em Inglês | MEDLINE | ID: mdl-15639296

RESUMO

BACKGROUND: Increasing evidence suggests that patterns of T-cell immunity to inhalant allergens in genetically diverse human populations are more heterogeneous than previously assumed, and that covert differences in expression patterns might underlie variations in airway disease phenotypes. We tested this proposition in a community sample of children. METHODS: We analysed data from 172 individuals who had been recruited antenatally to a longitudinal birth cohort study. Of the 194 birth cohort participants, data from the 147 probands (age range 8.6-13.5 years) who consented to blood collection were included along with data from 25 consenting siblings (mean age 11 years [range 7.4-17.4]). We ascertained clinical phenotypes related to asthma and allergy. We measured T-cell responses to allergens and mitogens, together with blood eosinophils and IgE/IgG antibodies, and assessed associations between these indices and clinical phenotypes. FINDINGS: Atopy was associated with allergen-specific T-helper (Th)2 responses dominated by interleukin 4, interleukin 5, interleukin 9, interleukin 13, whereas interleukin 10, tumour necrosis factor alpha, and interferon gamma responses were common to both atopics and non-atopics. The wheal size from skin prick with allergen was positively associated with in-vitro interleukin 5 and interferon gamma responses, and negatively associated with interleukin 10. Asthma, especially in atopics, was strongly associated with eosinophilia/interleukin 5, and bronchial hyper-responsiveness (BHR) was associated with eosinophilia plus polyclonal interferon gamma production. BHR in non-atopics was associated with elevated allergen-specific and polyclonal interleukin 10 production. INTERPRETATION: Parallel immunological and clinical profiling of children identified distinctive immune response patterns related to asthma and wheeze compared with BHR, in atopics non-atopics. Immunological hyper-responsiveness, including within the Th1 cytokine compartment, is identified as a hallmark of BHR. RELEVANCE TO PRACTICE: These findings highlight the heterogeneity of immune response patterns in asthmatic children, including those with seemingly homogeneous Th2-driven atopic asthma. Further elucidation of the covert relationships between wheezing phenotypes and underlying immunophenotypes in this age group will potentially lead to more effective treatments for what is an unexpectedly heterogeneous collection of disease subtypes.


Assuntos
Asma/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Adolescente , Animais , Asma/fisiopatologia , Hiper-Reatividade Brônquica , Criança , Eosinofilia , Humanos , Hipersensibilidade Imediata/imunologia , Interleucinas/metabolismo , Fenótipo , Pyroglyphidae/imunologia , Sons Respiratórios , Testes Cutâneos
14.
Mol Immunol ; 25(4): 345-54, 1988 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-3398859

RESUMO

The mechanism of activation of the classical pathway of human complement by house-dust extracts and its relevance to atopic disease was studied. Our results confirm that for most sera of adults, house-dust extract is, on a weight basis, a more potent C-activator than aggregated human IgG or lipopolysaccharide (endotoxin). Naturally occurring IgM antibodies directed against ubiquitous polysaccharides appeared to be the dominant factor in the C1 activation by house-dust extracts in human sera. Large variations were found between sera with respect to the concns of these IgM antibodies as measured by C1 activation or fixation of haemolytic complement. The IgM antibody titre was, however, not associated with atopic disease. Consequently, we do not support the hypothesis put forward by Berrens et al. (1978) (Allergol. Immunopath. 6, 45-54) that there might be a relation between atopy and enhanced reactivity of serum complement with allergenic extracts. More than 90% of the C-activating potential of allergen extracts like house dust was found in the fractions with high mol. wt material (mol. wt greater than 100 K). Therefore, these antigens are easily separated from the known IgE-binding major allergens of house-dust mite and cat dander.


Assuntos
Ativação do Complemento , Via Clássica do Complemento , Poeira , Imunoglobulina M/imunologia , Polissacarídeos/imunologia , Adolescente , Adulto , Antígenos/análise , Bactérias/isolamento & purificação , Complemento C1/imunologia , Testes de Fixação de Complemento , Humanos , Pessoa de Meia-Idade , Peso Molecular , Rinite Alérgica Perene/imunologia , Rinite Alérgica Sazonal
15.
Mol Immunol ; 38(1): 1-8, 2001 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-11483205

RESUMO

Unlike other immunoglobulin G (IgG) subclasses, IgG4 antibodies in plasma have been reported to be functionally monovalent. In a previous paper, we showed that the apparent monovalency of circulating IgG4 antibodies is caused by asymmetry of plasma IgG4-a large fraction has two antigen-binding sites resulting in bispecificity. We postulated that the generation of bispecific antibodies was caused by a post-secretion mechanism, involving the exchange of IgG4 half-molecules (i.e. one heavy and one light chain). This hypothesis was based on the observed instability of the inter-heavy chain disulfide bonds of IgG4. To investigate this instability, we constructed IgG4 mutants and analyzed the covalent interaction between the heavy chains by sodium dodecyl sulfate-poly acrylamide gel electrophoresis (SDS-PAGE) under non-reducing conditions. The mutation to serine of one of the hinge cysteines involved in the inter-heavy chain bond formation, Cys226, resulted in a more stable rather than a more labile inter-heavy chain linkage. Moreover, we confirmed that mutating the IgG4 hinge sequence Cys-Pro-Ser-Cys to the IgG1 hinge sequence Cys-Pro-Pro-Cys also markedly stabilizes the covalent interaction between the heavy-chains. These two observations suggested an explanation for the observed instability of the inter-heavy chain disulfide bonds: the formation of an alternative, intra-chain cystine. Obviously, this intra-chain cystine cannot be formed in the mutant where Cys226 is replaced by Ser, and cannot easily be formed in the mutant with the IgG1 hinge sequence (Cys-Pro-Pro-Cys) due to the restricted torsional freedom of prolines. We, therefore, postulate that the lack of a covalent heavy-chain interaction in a subpopulation of IgG4 reflects an equilibrium between inter- and intra-chain cystines. Based upon the published structure of the IgG4-related hinge-deleted IgG1 myeloma protein Mcg, we propose a model for the two forms of IgG4 and for the half-molecule exchange reaction, which might result in the formation of bispecific IgG4 antibodies.


Assuntos
Dissulfetos , Imunoglobulina G/química , Cadeias Pesadas de Imunoglobulinas/química , Animais , Humanos , Imunoglobulina G/genética , Cadeias Pesadas de Imunoglobulinas/genética , Camundongos
16.
Mol Immunol ; 23(6): 669-73, 1986 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-3092033

RESUMO

Activation of guinea-pig complement by human IgM antibodies after interaction with a particulate antigen is well established. Human IgM antibodies directed against meningococcal group-C capsular polysaccharide, however, were not able to fix guinea-pig complement in a classical complement fixation test with soluble antigen. In the same test, human complement was readily activated by these antibodies. This inability of human IgM antibodies to activate guinea-pig complement after interaction with soluble antigen was confirmed in other antigen systems. In contrast, efficient activation of both human and guinea-pig complement was found with the same antigen in a particulate form.


Assuntos
Anticorpos Antibacterianos/imunologia , Ativação do Complemento , Via Clássica do Complemento , Imunoglobulina M/imunologia , Neisseria meningitidis/imunologia , Polissacarídeos Bacterianos/imunologia , Animais , Complemento C1/metabolismo , Testes de Fixação de Complemento , Cobaias , Hemólise , Humanos , Solubilidade , Especificidade da Espécie
17.
Mol Immunol ; 28(11): 1225-32, 1991 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-1720504

RESUMO

Fourteen synthetic peptides of 15 amino acid residues length, overlapping by five residues and spanning the entire sequence of the major allergen Der p II from the house dust mite Dermatophagoides pteronyssinus were synthesized. These peptides were coupled to CNBr-activated Sepharose-4B and used as solid-phase antigens in epitope mapping studies using human IgE antisera. These antibodies bound predominantly to the peptide comprising residues 65-78, the binding of which was inhibited by native Der p II. In addition these antisera bound, to a lesser extent, to the peptide that comprised residues 1-15, which binding was not inhibited by native Der p II. Thus, we found one sequential epitope for a number of IgE sera.


Assuntos
Alérgenos/genética , Epitopos/genética , Ácaros/imunologia , Alérgenos/imunologia , Sequência de Aminoácidos , Animais , Reações Antígeno-Anticorpo , Antígenos de Dermatophagoides , Mapeamento Cromossômico , Humanos , Imunoglobulina E/imunologia , Dados de Sequência Molecular , Teste de Radioalergoadsorção
18.
J Immunol Methods ; 19(1): 87-93, 1978.
Artigo em Inglês | MEDLINE | ID: mdl-621410

RESUMO

Hapten (trinitrophenyl) was coupled to antigen (ovalbumin). The haptenated antigen was bound by anti-ovalbumin antibody and binding was quantitated with 125I-labelled anti-hapten antibodies. Thus, with a single radioactive reagent antibodies against a variety of antigens can be detected, whilst the problems inherent in a labelled antiglobulin binding test are avoided. In the ovalbumin system, the haptenated antigen binding test proved to be approximately 20 times as sensitive as the iodinated ovalbumin binding test.


Assuntos
Anticorpos/análise , Antígenos , Haptenos , Radioisótopos do Iodo , Ovalbumina , Sefarose , Soroalbumina Bovina , Trinitrobenzenos
19.
J Immunol Methods ; 161(2): 177-86, 1993 May 26.
Artigo em Inglês | MEDLINE | ID: mdl-7685040

RESUMO

Epitope mapping with synthetic peptides bound to derivatized paper discs has been investigated using the discs both as a solid-phase matrix for peptide synthesis as well as the solid phase in immunologic testing procedures without detachment of the peptides from the paper discs. Using the T bag (tea bag) method the simultaneous synthesis and subsequent immunologic testing of large numbers of peptides was demonstrated for the feline major allergen Fel d I. A total of 15,000 paper disc-bound peptides, comprising the 146 nonapeptides overlapping by eight amino acid residues on both chains, were synthesized simultaneously with 100 paper discs per T bag. Using these paper disc-bound peptides as the solid phase in radioimmunoassays the binding sites found coincided with those detected in the PEPSCAN with the commercially available epitope mapping kit and with the binding sites that had been found with Sepharose-coupled peptides. The signal to background-ratio in the paper disc-RIA was comparable to that in the PEPSCAN and the reproducibility was good. The bound antibodies could be eluted from the paper disc-bound peptides, permitting regeneration and repeated use of the paper discs for immunologic testing. This method was shown to be a useful alternative to the PEPSCAN and to have the major advantage that large numbers of antibodies could be tested with large numbers of peptides simultaneously.


Assuntos
Alérgenos/imunologia , Epitopos/análise , Glicoproteínas , Fragmentos de Peptídeos/imunologia , Animais , Anticorpos Monoclonais/imunologia , Gatos , Ensaio de Imunoadsorção Enzimática , Humanos , Imunoglobulina E/sangue , Mioglobina/imunologia , Radioimunoensaio
20.
J Immunol Methods ; 87(1): 51-7, 1986 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-3950425

RESUMO

We have developed a modification of the solid-phase immunoassay for the detection of antibodies of specified isotype. This modified assay involves interaction of the antibody in solution with a haptenated antigen, followed by binding of the antigen-antibody complex to an anti-hapten immunoadsorbent. The antibody is subsequently detected with a suitably labelled antiglobulin reagent. The advantages of this test are: (1) The procedure is more convenient, especially in test protocols where multiple antigens are used. Because the antigens are in solution, computer-controlled dispensation of the antigens is possible. (2) The efficiency of antibody binding is increased, presumably because of the higher avidity of the antigen-antibody interaction in homogeneous solution compared to that in a heterogeneous system. Therefore, less antigen per test is required, and a solid phase with a lower capacity can be used.


Assuntos
Antígenos/imunologia , Haptenos/farmacologia , Teste de Radioalergoadsorção/métodos , Radioimunoensaio/métodos , Alternaria/imunologia , Análise de Variância , Humanos , Alótipos de Imunoglobulina , Ácaros/imunologia , Trinitrobenzenos/imunologia
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