IgE, IgG1, and IgG4 Reactivity to Dermatophagoides pteronyssinus Glycosylated Extract in Allergic Patients.

BACKGROUND: House dust mites are important allergen sources and some of these allergenic proteins may contain carbohydrate moieties, which are able to be isolated using lectins, as Concanavalin A (ConA). This study aimed to investigate allergenicity (IgE) and antigenicity (IgG1 and IgG4) of ConA-unbound and ConA-bound Dermatophagoides pteronyssinus (Dpt) crude extracts using sera of mite-allergic patients as well as inhibition capacity of antibody binding. MATERIAL AND METHODS: We obtained mannose-enriched and mannose-depleted fractions from Dpt by ConA affinity chromatography. Both ConA-bound and ConA-unbound fractions were evaluated by ELISA and Western Blotting for specific IgE, IgG1, and IgG4 reactivity with sera obtained from 95 mite-allergic patients (DP+) and 92 nonallergic (NA) subjects. Inhibition ELISA was used to assess cross-reactivity between Dpt extract and its fractions. RESULTS: Among the DP+ patients, no difference was found between ConA-unbound and ConA-bound fractions regarding the levels of specific IgE, IgG1, and IgG4. Nonallergic subjects had the same levels of specific IgG1 to both ConA-unbound and ConA-bound fractions, although for specific IgG4, values were higher for ConA-bound. A positive correlation was found among specific IgE, IgG1, and IgG4 levels when Dpt was compared to ConA-unbound and ConA-bound fractions. Recognition of crude Dpt by IgE, IgG1, and IgG4 was highly inhibited by ConA-unbound and ConA-bound fractions. Western Blotting revealed a broad spectrum of bands ranging from 14 to 116 kDa recognized by specific IgE and IgG4. However, IgG1 reached higher frequency values on high molecular weight polypeptides. CONCLUSION: ConA-unbound and ConA-bound fractions derived from D. pteronyssinus crude extract revealed important components involved in the IgE recognition in allergic patients as well as IgG1 and/or IgG4 in allergic and healthy subjects.

Acetonic Fraction of Bidens pilosa Enriched for Maturase K Is Able to Control Cerebral Parasite Burden in Mice Experimentally Infected With Toxoplasma gondii.

Toxoplasma gondii infection can cause abortions or congenital infection for a vast number of domestic animals and humans, leading to economic loss in veterinary sciences, as well as severe consequences for immunocompromised patients. Bidens pilosa Linné has been used in ethnopharmacology for treatment of diseases, as malaria, diabetes and hepatitis, in addition to its use as antioxidant, antiallergic, anti-inflammatory, and antiviral. The components of this plant have never been studied before for treatment of toxoplasmosis, and the conventional drugs currently used to treat this disease have high degree of toxicity. Thus, the aim of this study was to evaluate the effect of B. pilosa against T. gondii, by analyzing a total extract of this plant in parallel with a fraction obtained by precipitation in acetone. Also, it was assessed if the acetonic fraction could present lectinic activity, followed by its identification by mass spectrometry. It was observed with the experimental models designed that both total extract and acetonic fraction of B. pilosa were able to control T. gondii infection by in vitro and in vivo experiments, in addition to their low toxicity to host cells. Both total extract and acetonic fraction of this plant display capacity to impair replication of T. gondii tachyzoites. Interesting, the B. pilosa acetonic fraction treatment for 10 days after infection decreases significantly the number of T. gondii brain cyst in comparison with controls. The protein isolated from B. pilosa acetonic fraction was characterized as a novel lectin identified as maturase K. Taken together, these findings open new perspectives to treat patients infected by T. gondii. Future studies will be necessary to investigate the precise mechanism underlying the control of T. gondii infection to impair the replication of this parasite in the host cells after treatment with B. pilosa maturase K.

Biological and Enzymatic Characterization of Proteases from Crude Venom of the Ant Odontomachus bauri.

Hymenoptera venoms constitute an interesting source of natural toxins that may lead to the development of novel therapeutic agents. The present study investigated the enzymatic and biological characteristics of the crude venom of the ant Odontomachus bauri. Its crude venom presents several protein bands, with higher staining for six proteins with gelatinolytic activity (17, 20, 26, 29, 43 and 48 kDa). The crude venom showed high proteolytic activity on azocasein at optimal pH 8.0 and 37 °C. In the presence of protease inhibitors as aprotinin, leupeptin and EDTA, the azocaseinolytic activity was reduced by 45%, 29% and 9%, respectively, suggesting that the enzymes present in the crude venom belong to the three classes of proteases, with the serine proteases in greater intensity. The crude venom degraded the fibrinogen α-chain faster than the ß-chain, while the fibrinogen γ-chain remained unchanged. In biological assays, O. bauri venom showed hemolytic and coagulant activity in vitro, and defibrinating activity in vivo. In addition, the venom showed antimicrobial activity against Staphylococcus aureus and Escherichia coli as well as antiparasitic activity on Toxoplasma gondii infection in vitro. In that sense, this study sheds perspectives for pharmacological applications of O. bauri venom enzymes.

Evaluation of colostrum as an alternative biological sample for the diagnosis of human congenital toxoplasmosis.

BACKGROUND: Toxoplasmosis is a zoonosis caused by Toxoplasma gondii, an intracellular protozoan parasite able to infect a wide range of hosts, including humans. Congenital infection can cause severe damage to the fetus. Thus, it is important to detect antibodies against the parasite to confirm clinical manifestations. Considering that all immunoglobulin isotypes may be present in biological samples from newborns and their mothers, this study aimed to evaluate the ability to diagnose recent toxoplasmosis by using colostrum, as an alternative noninvasive way to obtain biological samples, as well as to determine correlation rates between antibodies from serum samples to detect IgG, IgM and IgA isotypes against T. gondii. METHODS: A total of 289 puerperal women from Clinical Hospital of Federal University of Uberlândia (mean age: 24.8 years, range: 14 - 43 years) took part in this study. Serum and colostrum samples from these patients were analyzed using ELISA and immunoblotting assays for soluble antigens from T. gondii. RESULTS: ELISA immunoassays with serum samples showed reactivity in 47.0, 6.9 and 2.8 % of samples to anti-T. gondii IgG, IgM and IgA, respectively, in comparison with colostrum samples, which showed reactivity in 46.0, 7.9 and 2.8 % of samples to the same isotypes. Also, significant correlation rates of anti-T. gondii antibody levels between serum and colostrum samples were observed. Interestingly, reactivity to IgM and/or IgA in colostrum and/or serum confirmed clinical manifestations of congenital toxoplasmosis in three newborns. Immunoblotting assays showed that it is possible to detect IgG, IgM and IgA antibodies against various antigens of T. gondii in serum and colostrum samples. IgG antibodies in serum and colostrum samples recognized more antigenic fractions than IgM and IgA antibodies. Serum IgG detected more antigenic fractions than IgG antibodies present in the colostrum of the same patient. In contrast, specific IgA present in colostrum recognized a higher number of antigens than IgA present in serum samples of the same patient. CONCLUSIONS: Overall, the results show that it is important to investigate the occurrence of congenital toxoplasmosis, even at puerperal period. Furthermore, this study demonstrates that T. gondii-specific IgG, IgM and IgA antibodies in serum and colostrum samples from puerperal women may be detected with a significant correlation, suggesting that colostrum may also be used as an alternative biological sample to efficiently diagnose recent human toxoplasmosis.

Pesquisa de anticorpos anti-Neospora caninum em bovinos leiteiros, cães e trabalhadores rurais da região Sudoeste do Estado de Mato Grosso / Inquiry of antibodies anti-Neospora caninum in dairy cattle, dogs and rural workers of the south-west region of Mato Grosso State

Considerando a importância da neosporose interferindo na produtividade animal, avaliou-se a frequência de anticorpos anti-Neospora caninum em amostras de soros de bovinos leiteiros da Região Sudoeste do Estado de Mato Grosso, Brasil, complementando-se com amostras sorológicas colhidas de cães e de humanos que conviviam nas mesmas propriedades rurais amostradas. Um total de 1.036 amostras de soros foram analisadas, sendo 932 de fêmeas bovinas leiteiras, 37 de cães e 67 de humanos, provenientes de 24 propriedades e examinados por meio da reação de imunofluorescência indireta (RIFI). As amostras de soros humanos reagentes foram testadas novamente por Western-blotting para confirmação dos resultados. Anticorpos anti-N. caninum foram encontrados em 499 bovinos (53,5%), em pelo menos um animal positivo por propriedade, em 25 caninos (67,6%) e em sete humanos (10,5%). Não houve diferença significativa no número de bovinos positivos por faixa etária. Os resultados obtidos indicam uma ampla disseminação de N. caninum na região estudada.

Considering the importance of neosporosis in the animal health and production, the frequency of antibodies to Neospora caninum was evaluated in dairy cattle of the Southwestern region of Mato Grosso State, Brazil, in addition to serum samples obtained from dogs and humans living in the farms. A total of 1036 serum samples were analyzed, from which 932 were from dairy bovine females, 37 from dogs and 67 from humans, from 24 farms and examined by the indirect fluorescent antibody test (IFAT). Reactive human serum samples were retested by Western-blotting to confirm the results. Antibodies to N. caninum were found in 499 cattle sera (53.5 %), with at least one positive in each farm, 25 dog sera (67.6 %) and seven human sera (10.5 %). There was no significant difference in the number of positive cattle sera according to age group. The results indicate a wide dissemination of N. caninum in the studied region.

Diversity of allergen exposure: implications for the efficacy of environmental control.

UNLABELLED: The prevalence of allergic diseases such as asthma, rhinitis, allergic conjunctivitis and atopic dermatitis has increased in the last decades. The relationship between allergen exposure, atopic sensitization and development of allergic diseases is widely described in the literature. AIM: To evaluate measures for reducing allergen exposure as part of the treatment of allergic diseases. METHODS: An analysis was made of previous studies on allergen exposure done with a similar methodology in the central region of Brazil; the study included homes, hotels, cinemas, cars, taxis, buses and scholar transportation. RESULTS: High levels of Der p 1 and Der f 1 mite allergens were found in a large proportion of the sample in most of the environments included in those studies; there were higher levels of pet allergens in cars and school transportation vehicles. CONCLUSION: The diversity of allergen exposure demonstrates the need for education about allergic diseases for patients and their families, as well as measures of reducing allergens in homes. This should be part of a global strategy of the management of allergic diseases, given that individuals live in society, not only in their houses.

Diversidade da exposição alergênica: implicações na obtenção da eficácia do controle ambiental: [revisão] / Diversity of allergen exposure: implications for the efficacy of environmental control: [review]

As doenças alérgicas, como a asma, rinite, conjuntivite alérgica e a dermatite atópica têm apresentado um aumento na sua prevalência nas últimas décadas. A relação entre exposição alergênica, sensibilização atópica e desenvolvimento de doenças alérgicas são amplamente descrita na literatura. OBJETIVO: Discutir a dificuldade no controle ambiental da exposição alergênica como parte do tratamento das doenças alérgicas. MÉTODOS: Analisar trabalhos de exposição alergênica realizados com metodologia similar na região central do Brasil, incluindo casas, hotéis, cinemas, carros, táxis, ônibus e transporte escolar. RESULTADOS: Níveis elevados dos alérgenos do grupo 1 de Dermatophagoides pteronyssinus (Der p 1) e de D. farinae (Der f 1), capazes de causar sensibilização e exacerbação de sintomas foram encontrados na maioria dos ambientes estudados em uma larga proporção das amostras, enquanto os alérgenos de animais domésticos atingiram maiores níveis em carros e veículos de transporte escolar. CONCLUSÃO: A diversidade da exposição alergênica mostra a necessidade de uma compreensão da doença alérgica pelos pacientes e familiares, e que as medidas de controle do ambiente doméstico fazem parte de uma estratégia global do tratamento das doenças alérgicas, uma vez que os indivíduos vivem em uma sociedade e não isoladas no interior de seus domicílios.

The prevalence of allergic diseases such as asthma, rhinitis, allergic conjunctivitis and atopic dermatitis has increased in the last decades. The relationship between allergen exposure, atopic sensitization and development of allergic diseases is widely described in the literature. AIM: To evaluate measures for reducing allergen exposure as part of the treatment of allergic diseases. METHODS: An analysis was made of previous studies on allergen exposure done with a similar methodology in the central region of Brazil; the study included homes, hotels, cinemas, cars, taxis, buses and scholar transportation. RESULTS: High levels of Der p 1 and Der f 1 mite allergens were found in a large proportion of the sample in most of the environments included in those studies; there were higher levels of pet allergens in cars and school transportation vehicles. CONCLUSION: The diversity of allergen exposure demonstrates the need for education about allergic diseases for patients and their families, as well as measures of reducing allergens in homes. This should be part of a global strategy of the management of allergic diseases, given that individuals live in society, not only in their houses.

An opposite role is exerted by the acarian Myocoptes musculinus in the outcome of Toxoplasma gondii infection according to the route of the protozoa inoculation.

Infection with Toxoplasma gondii leads to a Th1 immune response. Alternatively, the acarian Myocoptes musculinus induces a disease in BALB/c mice that involves Th2 immune mechanisms. In this study, we investigated whether infestation by M. musculinus induces Th2 immune response in C57BL/6 mice and if this response influences the T. gondii-induced Th1 response when mice are inoculated by intraperitoneal or oral route. The animals were infected with M. musculinus and one month later with T. gondii ME-49 strain and the survival and immune response were monitored. The co-infected animals displayed higher mortality rate and the spleen cells showed a decreased IFN-gamma and elevated IL-4 and IL-5 production. These changes were associated with severe pneumonia and wasting condition. On the other hand, when mice were orally infected with 100 T. gondii cysts, co-infection prolonged the survival rates and ameliorated intestinal lesions in association with a significant drop in IFN-gamma levels in sera. These results indicate the interference of Th2 response induced by M. musculinus in a T. gondii-induced Th1 response. Altogether, these data demonstrate the profound interactions between the immune response induced against unrelated organisms T. gondii and M. musculinus, and suggest that this type of interactions may impact clinical disease.

Alergia, exposição alergênica e condições socioeconômicas / Allergy, allergen exposure and socio-economical conditions

Nos últimos trinta anos tem ocorrido um aumento expressivo na prevalência das doenças alérgicas. Esse rápido aumento não pode ser justificado exclusivamente por fatores genéticos, sendo associada etiologia multifatorial às doenças alérgicas, onde a exposição à poluição ambiental (ozônio, exaustão de diesel, fumaça de cigarros, entre outras) e fatores relacionados ao estilo de vida das famílias têm importante influência na expressão dessas doenças, em especial a asma.

Strongyloides ratti antigenic components recognized by IgE antibodies in immunoblotting as an additional tool for improving the immunodiagnosis in human strongyloidiasis.

IgE antibody response in human strongyloidiasis was evaluated by enzyme-linked immunosorbent assay (ELISA) and immunoblotting (IB) using Strongyloides ratti saline extract as heterologous antigen. A total of 50 serum samples of patients who were shedding S. stercoralis larvae in feces (group I, copropositive), 38 of patients with other intestinal parasites (group II), and 38 of subjects with negative results in three parasitologic assays (group III, copronegative) were analyzed. Levels of IgE anti-Strongyloides expressed in ELISA Index (EI) were significantly higher in patients of group I (1.32) than in group II (0.51) and group III (0.81), with positivity rates of 54%, 0%, and 10.5%, respectively. Fifteen S. ratti antigenic components were recognized in IB-IgE by sera of group I, with frequency ranging from 8% to 46%. In group II, only two antigenic bands (101, 81 kDa) were detected in a frequency of 10% and no reactivity was found in group III. Sera with EI values > 1.5 recognized five from 13 specific antigenic bands (70, 63, 61, 44, 7 kDa). It can be concluded that these five antigenic components recognized by IB-IgE using S. ratti antigen might be employed as an additional tool for improving the immunodiagnosis in human strongyloidiasis.

Strongyloides ratti antigenic components recognized by IgE antibodies in immunoblotting as an additional tool for improving the immunodiagnosis in human strongyloidiasis

IgE antibody response in human strongyloidiasis was evaluated by enzyme-linked immunosorbent assay (ELISA) and immunoblotting (IB) using Strongyloides ratti saline extract as heterologous antigen. A total of 50 serum samples of patients who were shedding S. stercoralis larvae in feces (group I, copropositive), 38 of patients with other intestinal parasites (group II), and 38 of subjects with negative results in three parasitologic assays (group III, copronegative) were analyzed. Levels of IgE anti-Strongyloides expressed in ELISA Index (EI) were significantly higher in patients of group I (1.32) than in group II (0.51) and group III (0.81), with positivity rates of 54 percent, 0 percent, and 10.5 percent, respectively. Fifteen S. ratti antigenic components were recognized in IB-IgE by sera of group I, with frequency ranging from 8 percent to 46 percent. In group II, only two antigenic bands (101, 81 kDa) were detected in a frequency of 10 percent and no reactivity was found in group III. Sera with EI values > 1.5 recognized five from 13 specific antigenic bands (70, 63, 61, 44, 7 kDa). It can be concluded that these five antigenic components recognized by IB-IgE using S. ratti antigen might be employed as an additional tool for improving the immunodiagnosis in human strongyloidiasis.

Heterologous antigen extract in ELISA for the detection of human IgE anti-Strongyloides stercoralis

Extrato contendo larvas de Strongyloides ratti foi usado na padronização de um ELISA para detecção de IgE gênero-específica na estrongiloidíase humana. Foram analisadas 40 amostras de soro de pacientes monoinfectados que estavam eliminando larvas de S. stercoralis nas fezes (Grupo I), 40 de pacientes com outros parasitos intestinais (Grupo II), e 40 indivíduos copronegativos (Grupo III). Níveis de IgE gênero-específica (índice ELISA: EI) foram significativamente maiores no Grupo I (EI = 1,43) do que no II (EI = 0,70) e III (EI = 0,71), mostrando positividade de 55 por cento, 2,5 por cento e 0 por cento, respectivamente. Similarmente, soros dos pacientes copropositivos (Grupo I) apresentaram níveis significativamente maiores de IgE total (866 IU/mL) quando comparados com os soros dos Grupo II (302 IU/mL) e III (143 IU/mL). Uma significativa correlação positiva foi encontrada entre os níveis de IgE específica a Strongyloides sp. e IgE total nos soros de pacientes com estrongiloidíase. Em conclusão, extrato heterólogo de S. ratti mostrou ser uma ferramenta útil para detecção de IgE gênero-específica por ELISA, desta forma contribuindo para melhor caracterização do perfil da resposta imune na estrongiloidíase humana.

Heterologous antigen extract in ELISA for the detection of human IgE anti-Strongyloides stercoralis.

Strongyloides ratti larval extract was used for the standardization of ELISA to detect genus-specific IgE in human strongyloidiasis. Forty serum samples from monoinfected patients shedding S. stercoralis larvae (Group I), 40 from patients with other intestinal parasites (Group II), and 40 from copronegative healthy subjects (Group III) were analyzed. Genus-specific IgE levels (ELISA Index: EI) were significantly higher in the group I (EI = 1.43) than groups II (EI = 0.70) and III (EI = 0.71), showing positivity rates of 55%, 2.5% and 0%, respectively. Similarly, sera from copropositive patients had significantly higher levels of total IgE (866 IU/mL) as compared to those from group II (302 IU/mL) and III (143 IU/mL). A significant positive correlation was found between levels of Strongyloides specific-IgE and total IgE in sera from patients with strongyloidiasis. In conclusion, S. ratti heterologous extract showed to be a useful tool for detecting genus-specific IgE by ELISA, contributing for a better characterization of the immune response profile in human strongyloidiasis.

[IgA, IgE and IgG subclasses to Candida albicans in serum and vaginal fluid from patients with vulvovaginal candidiasis]. / IgA, IgE e subclasses de IgG anti-Candida albicans no soro e lavado vaginal de pacientes com candidíase vulvovaginal.

PURPOSE: To determine the levels of IgA, IgE, IgG and subclasses (IgG1, IgG4) antibodies specific to C. albicans in serum and vaginal washes from women with or without vulvovaginal candidiasis in order to evaluate the role of these antibodies in the immunopathogenesis of the disease. METHODS: Thirty women with clinical symptoms of vulvovaginal candidiasis (15 positive vaginal culture to C. albicans, 11 negative culture and 4 positive culture to non-C. albicans) and 12 asymptomatic control women were selected. Serum and vaginal wash samples were obtained for the detection of anti-C. albicans antibodies by ELISA. RESULTS: Symptomatic patients with positive culture showed significantly higher levels of specific IgA in vaginal washes and lower in serum than those with negative culture. Specific serum IgE levels were very low compared to vaginal IgE. High levels of total specific IgG were found in serum and vaginal washes in both groups, regardless the fungal presence or absence. Specific IgG1 and IgG4 levels were significantly higher only in vaginal washes from symptomatic patients with positive culture, with a slightly higher IgG1/IgG4 ratio, indicating that the IgG1 antibody response may be predominantly involved in the fungal clearance. CONCLUSION: Our results indicate a pronounced antibody response of IgA, IgG1 and IgG4 to C. albicans in vaginal washes in symptomatic patients with positive culture, suggesting a important role of these antibodies in the local immune response triggered by the presence of the fungus.

IgA, IgE e subclasses de IgG anti-Candida albicans no soro e lavado vaginal de pacientes com candidíase vulvovaginal / IgA, IgE and IgG subclasses to Candida albicans in serum and vaginal fluid from patients with vulvovaginal candidiasis

OBJETIVO: Determinar níveis de anticorpos IgA, IgE, IgG e subclasses (IgG1, IgG4) específicos a C. albicans no soro e lavado vaginal de mulheres com ou sem candidíase vulvovaginal para avaliar o papel destes anticorpos na imunopatogênese desta doença. MÉTODOS: Foram selecionadas 30 mulheres com sintomas clínicos de candidíase vulvovaginal (15 com cultura de secreção vaginal positiva para C. albicans, 11 com cultura negativa e quatro com cultura positiva para Candida não-albicans) e 12 mulheres controles assintomáticas (nove com cultura negativa). Amostras de soro e lavado vaginal foram obtidas para a detecção de anticorpos anti-C. albicans por ELISA. RESULTADOS: Pacientes sintomáticas com cultura positiva apresentaram níveis de IgA específicas significativamente maiores no lavado vaginal e menores no soro do que aquelas com cultura negativa. Níveis séricos de IgE específica foram extremamente baixos em relação ao lavado vaginal. Altos níveis de IgG total específica foram encontrados no soro e lavado vaginal em ambos os grupos, independente da presença do fungo. Níveis de IgG1 e IgG4 específicas foram significativamente maiores somente no lavado vaginal de mulheres sintomáticas e cultura positiva, com relação IgG1/IgG4 ligeiramente maior, indicando que a resposta de anticorpos IgG1 possa estar predominantemente envolvida na resolução da infecção fúngica. CONCLUSÕES: Nossos resultados indicam resposta acentuada de IgA, IgG1 e IgG4 anti-C. albicans no lavado vaginal de mulheres sintomáticas com cultura positiva, sugerindo importante papel destes anticorpos na resposta imune local estimulada pela presença do fungo

Prevalencia e avidez de anticorpos anti-virus da hepatite C em pacientes hemodialisados na area de Uberlandia, Minas Gerais, Brasil / Prevalence and avidity of hepatitis C antibodies in hemodialysis patients in the area of Uberlandia, Minas Gerais, Brazil

O virus da hepatite C(HCV), tem sido identificado como sendo o principal agente causador das hepatites nao-A, nao-B. A deteccao dos anticorpos IgG anti-HCV e realizada pelo ELISA convencional (ELISAc) e a sua avidez pode ser determinada pelo ELISA modificado (ELISAm).Foram estudados 34 pacientes hemodialisados (M:19; F:15), com media de idade de 45+- 2,3 anos, recebendo regularmente hemotransfusoes, quanto a presenca de anticorpos IgG anti-HCV (ELISAc) bem como sua avidez (ELISAm). A prevalencia para anti-HCV foi de 23,5 por cento (8/34), sendo que 62,5 por cento (5/8) eram do sexo feminino, verificando-se maior prevalencia diretamente relacionada com o tempo de hemodialise e o numero de transfusoes sanguineas. Todas as amostras soropositivas apresentaram indice de avidez superior a 80 por cento caracterizando quadro de infeccao cronica. Conclui-se que pacientes hemodialisados constituem grupo de risco potencial para a infeccao pelo HCV, sendo necessario medidas de isolamento para prevenir a disseminacao desta infeccao