RESUMO
Loxoscelism is a severe human envenomation caused by Loxosceles spider venom. To the best of our knowledge, no study has evaluated the presence of antibodies against Loxosceles venom in loxoscelism patients without treatment with antivenom immunotherapy. We perform a comparative analysis for the presence of antibodies capable of recognizing Loxosceles venom in a group of patients diagnosed with loxoscelism and in a group of people without loxoscelism. Methods The detection of L. laeta venom, Sicarius venom and recombinant phospholipases D from Loxosceles (PLDs) in sera from people with loxoscelism (Group 1) and from healthy people with no history of loxoscelism (Group 2) was evaluated using immuno-dot blot, indirect ELISA, and Western blot. Results We found naturally heterophilic antibodies (IgG-type) in people without contact with Loxosceles spiders or any clinical history of loxoscelism. Either serum pools or single sera from Group 1 and Group 2 analyzed by dot blot tested positive for L. laeta venom. Indirect ELISA for venom recognition showed titles of 1:320 for Group 1 sera and 1:160 for Group 2 sera. Total IgG quantification showed no difference in sera from both groups. Pooled sera and purified IgG from sera of both groups revealed venom proteins between 25 and 32 kDa and the recombinant phospholipase D isoform 1 (rLlPLD1), specifically. Moreover, heterophile antibodies cross-react with PLDs from other Loxosceles species and the venom of Sicarius spider. Conclusions People without contact with the spider venom produced heterophilic antibodies capable of generating a cross-reaction against the venom of L. laeta and Sicarius spiders. Their presence and possible interference should be considered in the development of immunoassays for Loxosceles venom detection.(AU)
Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto , Pessoa de Meia-Idade , Fosfolipase D/isolamento & purificação , Venenos de Aranha/toxicidade , Anticorpos Heterófilos/sangue , Antivenenos/uso terapêutico , Ensaio de Imunoadsorção Enzimática/métodos , Immunoblotting/métodosRESUMO
BACKGROUND: Declining immune function poses an important clinical challenge worldwide and supplementation with natural products that possessing immune enhancing properties is a promising approach for preventing or delaying immune function decline. Cocoons from yellow silkworms are a significant source of lutein, and this unexplored silk extract could be a viable alternative source for dietary lutein. This study assessed immunomodulatory activities of the silk lutein extract. Female BALB/c mice orally received lutein, either as silk or marigold extracts (10 or 20 mg/kg daily), or vehicle only (1% tween 80 in PBS pH 7.4) for 4 weeks. Natural killer (NK) cell activity, specific antibody production, lymphocyte subpopulations, mitogen-induced lymphocyte proliferation, and cytokine production were examined. RESULTS: Silk lutein extract increased NK cell activity, and the effect was dose-related whereas marigold lutein extract was ineffective. Silk lutein extract dose-dependently enhanced antibody production in pre-immunized mice but marigold lutein extract had no effect. Feeding with silk lutein extract increased the populations of CD3+ and CD4 + CD3 + cells. Silk lutein extract also stimulated concanavalin A- and lipopolysaccharide-induced proliferations of T and B lymphocytes, respectively. Moreover, silk lutein extract increased IL-2 and IFN-γ production while the effect of marigold lutein extract was undetectable. CONCLUSIONS: Together, silk lutein extract enhanced both innate and adaptive immune functions. This preparation may prove to be an effective supplement for strengthened immunity.
Assuntos
Exoesqueleto/química , Bombyx/imunologia , Fatores Imunológicos/análise , Luteína/imunologia , Seda/imunologia , Extratos de Tecidos/imunologia , Animais , Anticorpos Heterófilos/sangue , Linfócitos B/efeitos dos fármacos , Bombyx/metabolismo , Proliferação de Células/efeitos dos fármacos , Feminino , Citometria de Fluxo , Flores/imunologia , Interferon gama/análise , Interleucina-10/análise , Interleucina-2/análise , Interleucina-4/análise , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/imunologia , Luteína/isolamento & purificação , Camundongos Endogâmicos BALB C , Extratos Vegetais/imunologia , Pupa/imunologia , Pupa/metabolismo , Seda/química , Linfócitos T/efeitos dos fármacos , Tagetes/imunologia , Extratos de Tecidos/farmacologiaRESUMO
BACKGROUND: Declining immune function poses an important clinical challenge worldwide and supplementation with natural products that possessing immune enhancing properties is a promising approach for preventing or delaying immune function decline. Cocoons from yellow silkworms are a significant source of lutein, and this unexplored silk extract could be a viable alternative source for dietary lutein. This study assessed immunomodulatory activities of the silk lutein extract. Female BALB/c mice orally received lutein, either as silk or marigold extracts (10 or 20 mg/kg daily), or vehicle only (1% tween 80 in PBS pH 7.4) for 4 weeks. Natural killer (NK) cell activity, specific antibody production, lymphocyte subpopulations, mitogen-induced lymphocyte proliferation, and cytokine production were examined. RESULTS: Silk lutein extract increased NK cell activity, and the effect was dose-related whereas marigold lutein extract was ineffective. Silk lutein extract dose-dependently enhanced antibody production in pre-immunized mice but marigold lutein extract had no effect. Feeding with silk lutein extract increased the populations of CD3+ and CD4 + CD3 + cells. Silk lutein extract also stimulated concanavalin A- and lipopolysaccharide-induced proliferations of T and B lymphocytes, respectively. Moreover, silk lutein extract increased IL-2 and IFN-γ production while the effect of marigold lutein extract was undetectable. CONCLUSIONS: Together, silk lutein extract enhanced both innate and adaptive immune functions. This preparation may prove to be an effective supplement for strengthened immunity.
Assuntos
Animais , Feminino , Camundongos , Bombyx/imunologia , Extratos de Tecidos/imunologia , Luteína/imunologia , Seda/imunologia , Exoesqueleto/química , Fatores Imunológicos/análise , Pupa/imunologia , Pupa/metabolismo , Bombyx/metabolismo , Extratos de Tecidos/farmacologia , Luteína/isolamento & purificação , Anticorpos Heterófilos/sangue , Extratos Vegetais/imunologia , Linfócitos B/efeitos dos fármacos , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/imunologia , Linfócitos T/efeitos dos fármacos , Interleucina-4/análise , Interferon gama/análise , Interleucina-2/análise , Interleucina-10/análise , Tagetes/imunologia , Flores/imunologia , Seda/química , Proliferação de Células/efeitos dos fármacos , Citometria de Fluxo , Camundongos Endogâmicos BALB CRESUMO
INTRODUCTION: Interferon beta (IFN-ß) is a treatment for relapsing remitting multiple sclerosis. However, the therapeutic use of recombinant proteins induces a humoral immunologic response resulting in the induction of binding (BAb) or neutralizing (NAb) antibodies against the biological product. The presence of neutralizing antibodies has been associated with decreased IFN-ß treatment efficacy. MATERIALS AND METHODS: Two tumor cell lines (K562 and U937) were cultivated with human recombinant IFN-ß1a at different concentrations and lengths of time in order to measure the expression of intracellular ISG15, an inducible molecule in the IFN-ß1a signaling cascade. Blood was obtained from non-immunized and IFN-ß1a immunized (100,000 IU) New Zealand rabbits. The presence of BAb was evaluated by ELISA. For NAb detection, sera 1:20 dilution were added to the IFN-ß1a-stimulated cell lines, and ISG15 expression was evaluated by flow cytometry. RESULTS: K562 cells provided the better cell line for the assay, stimulated with a dose of 1,000 IU of IFN-ß1a, and a 1:100 dilution for the primary antibody and a 1:200 dilution for the secondary antibody. ISG15 expression was compared between cells alone or cultivated with IFN-ß1a. Mean fluorescence intensity (MFI) for ISG-15 expression median was 198 arbitrary units (AU) with interquartile ranges of 173-231 AU for non-stimulated cells and 430 AU with interquartile ranges of 316-611.5 AU for IFN-ß1a stimulated cells (p<0.01). Immunized rabbit sera decreased the expression of ISG-15 in K562 cells stimulated with IFN-ß1a, whereas non-immunized rabbit sera did not. CONCLUSIONS: This rabbit model demonstrates that ISG15 expression evaluated with flow cytometry can be used as a detection assay for NAb.
Assuntos
Anticorpos Heterófilos/sangue , Citocinas/biossíntese , Citometria de Fluxo/métodos , Interferon beta/imunologia , Testes de Neutralização , Ubiquitinas/biossíntese , Animais , Reações Antígeno-Anticorpo , Citocinas/genética , Ensaio de Imunoadsorção Enzimática , Humanos , Imunização , Interferon beta/farmacologia , Células K562/metabolismo , Masculino , Coelhos , Proteínas Recombinantes/farmacologia , Transdução de Sinais/efeitos dos fármacos , Células U937/metabolismo , Ubiquitinas/genética , Regulação para Cima/efeitos dos fármacosRESUMO
Antibody in human sera that induces lysis of sheep erythrocytes in hemolytic assay was investigated. The present study showed that the presence in serum of the thermostable cytolytic anti-sheep red blood cells antibodies is dependent on the Schistosoma mansoni infection, and this is more frequent in adults than in children. The thermostable characteristic of hemolysins in normal sera was not dependent on the presence of Ascaris lumbricoides, Trichuris trichiura or hookworm geo-helminths. Further, thermostable complement-activating heterophile antibodies were noticed in children in association with massive number of S. mansoni eggs. The results were obtained by using the z- and the chi-square tests. The z-test allows us to formulate a one-sided alternative, i.e., a tendency of one of the attributes. On the other hand, the chi-square test analyzes the independence between attributes by using a contingency table. Besides the obtained results being interesting in the field of schistosomiasis mansoni, they can provide a new insight into the use of statistics in medical science.
Assuntos
Anticorpos Heterófilos/imunologia , Anticorpos Antiprotozoários/imunologia , Eritrócitos/imunologia , Schistosoma mansoni/imunologia , Esquistossomose mansoni/imunologia , Adolescente , Adulto , Animais , Anticorpos Heterófilos/sangue , Criança , Fezes/parasitologia , Helmintos/imunologia , Hemólise , Temperatura Alta , Humanos , Imunoensaio/métodos , Modelos Estatísticos , Contagem de Ovos de Parasitas , Schistosoma mansoni/metabolismo , Ovinos , TemperaturaRESUMO
Foram investigados anticorpos de soro humano que provocam lise de eritrócitos de carneiro em ensaios hemolíticos. O presente estudo mostra que a presença de anticorpos citolíticos termoestáveis contra hemácias de carneiro é dependente da infecção por Schistosoma mansoni e é mais freqüente em adultos do que em crianças. A característica termo estável da hemolisina em soros normais não é dependente da presença de Ascaris lumbricoides, Trichuris trichiura ou ancilostomídeos. Além disto, anticorpos heterófilos hemolíticos termo estáveis ativadores de complemento foram demonstrados em crianças com associação de altas cargas de ovos de S. mansoni. Os resultados foram obtidos usando-se os testes z- e o qui quadrado. O teste z- nos permite formular uma alternativa "one side", isto é, a tendência de um dos atributos. Por outro lado o teste do qui quadrado analisa a independência entre atributos usando-se uma tabela de contingência. Ao lado dos interessantes resultados obtidos no campo da esquistossomose mansoni, a abordagem estatística pode apontar novos caminhos para o tratamento de dados na ciência médica.
Assuntos
Adolescente , Adulto , Animais , Criança , Humanos , Anticorpos Heterófilos/imunologia , Anticorpos Antiprotozoários/imunologia , Eritrócitos/imunologia , Schistosoma mansoni/imunologia , Esquistossomose mansoni/imunologia , Anticorpos Heterófilos/sangue , Fezes/parasitologia , Hemólise , Temperatura Alta , Helmintos/imunologia , Imunoensaio/métodos , Modelos Estatísticos , Contagem de Ovos de Parasitas , Ovinos , Schistosoma mansoni/metabolismo , TemperaturaRESUMO
Mild splenomegaly is common in patients with Epstein-Barr virus-associated infectious mononucleosis. Massive splenomegaly, however, is rare and requires further evaluation to exclude other causes. We report an adolescent girl with previously undiagnosed type 1 Gaucher disease who presented with massive splenomegaly. The diagnosis of her underlying condition was hampered by the presence of a positive heterophile antibody test for infectious mononucleosis.
Assuntos
Anticorpos Heterófilos/sangue , Anticorpos Antivirais/sangue , Erros de Diagnóstico , Doença de Gaucher/diagnóstico , Herpesvirus Humano 4/imunologia , Mononucleose Infecciosa/diagnóstico , Esplenomegalia/etiologia , Adolescente , Especificidade de Anticorpos , Exame de Medula Óssea , Reações Falso-Positivas , Fadiga/etiologia , Feminino , Febre/etiologia , Doença de Gaucher/complicações , Doença de Gaucher/imunologia , Glucosilceramidase/sangue , Glucosilceramidase/deficiência , Hepatomegalia/etiologia , Humanos , Mononucleose Infecciosa/imunologia , Faringite/etiologia , Esplenectomia , Esplenomegalia/cirurgiaRESUMO
The occurrence of cases of yellow fever (YF) and also the extensive distribution of A. aegypti in Brazil, inspired a study about the estimate of immunity against vaccinal virus (17D) among the residents at two cities of the Bahia State, Ipupiara (n = 461) and Prado (n = 228). At this non-endemic area of YF, the search for serology antibody against 17D (Ab17D) and 18 another arbovirus was made thereby hemagglutination inhibition (HI). Only 1.2% (8/689) showed Ab17D, six of those with monotypic sort. The heterotypic sort for flavivirus (FLV) was interpreted as associated to immunity against 17D too, being much frequent in Prado (30.3%) than in Ipupiara (23.2%). The age > or = 50 years and residence in another states were related to seropositive for FLV, the same way that vaccination's history (17D). However, the history of vaccination presented low percentages of sensibility (< or = 45.4%) and predictive-positive value (< or = 38.4%), but high specificity (> or = 70.8%) and predictive-negative value (> or = 78.8%). Therefore, the frequency of residents with Ab17D was low (1.2%), although the higher frequency (25.5%) of antibody FLV carrier's, what signifies that 26.7% of the studied population should present protection against the YF virus.
Assuntos
População Urbana , Vacinas Virais/imunologia , Vírus da Febre Amarela/imunologia , Anticorpos Heterófilos/sangue , Anticorpos Antivirais/sangue , Formação de Anticorpos , Brasil , Feminino , Humanos , Masculino , População Urbana/estatística & dados numéricos , Febre Amarela/prevenção & controleRESUMO
Several oligosaccharides containing the terminal structure Gal(alpha)1-3Gal (alphaGal) and different side chains were tested in vitro for their ability to block natural anti(alpha)Gal antibodies. A di-and a trisaccharide (di(alpha)Gal and tri(alpha)Gal) were selected. A blood group B baboon, having IgG and IgM natural antipig titers of 1:256 and 1:1024 and a hemolytic titer (to pig red blood cells, RBCs) of 1:8, was chosen to measure pharmacokinetic parameters of the saccharides and to assess the extent of in vivo neutralization of the antibodies. Three grams each of the di(alpha)Gal and the tri(alpha)Gal dissolved in saline were administered by bolus intravenous (i.v.) injection. Blood samples were collected at various times and urine was collected at 8 and 24 h. Plasma and urine concentrations of the alphaGal saccharides were estimated by an ELISA specially developed for this study. A fast distribution phase followed by equilibrium and excretion phases were observed, indicating a T1/2 in the order of 1 h. Fifty-eight per cent of the saccharides were recovered in the urine within 24 h. Determination of antipig antibody binding by FACS analysis and of serum cytotoxicity titers for pig endothelial cells demonstrated that a 70% reduction in binding and cytotoxicity could be achieved with plasma saccharide levels of 300-400 microg/ml. Six months later, a pig heart was transplanted heterotopically into the baboon. A 3-g bolus of the saccharide mixture (1.5 g of each saccharide) was given i.v. before allowing blood reperfusion of the transplanted heart, followed by an i.v. infusion of 1 g/hr for 1 hr and 0.5 g/hr for the 3 succeeding hours. Blood concentrations of the saccharides, CH50, hematology and cytotoxicity for PK15 cells were estimated in blood samples taken at various times. Heart function was observed to be satisfactory for 8 h, but was found to have ceased at 18 h. Myocardial biopsies taken at 3 and 5 h showed congestion only, suggestive of minimal vascular rejection, but by 18 h demonstrated severe vascular rejection. In conclusion, alphaGal saccharide therapy given for a period of 4 h delayed, but did not totally prevent, the development of vascular rejection in the pig-to-baboon heart transplant model. alphaGal saccharide therapy may be one of several useful approaches for the prevention of hyperacute rejection in pig-to-primate organ transplantation.
Assuntos
Rejeição de Enxerto/prevenção & controle , Transplante de Coração/imunologia , Oligossacarídeos/administração & dosagem , Doença Aguda , Animais , Anticorpos Heterófilos/sangue , Dissacarídeos/administração & dosagem , Dissacarídeos/imunologia , Dissacarídeos/farmacocinética , Eritrócitos/imunologia , Rejeição de Enxerto/imunologia , Transplante de Coração/patologia , Hemaglutinação , Hemólise , Oligossacarídeos/imunologia , Oligossacarídeos/farmacocinética , Papio , Segurança , Suínos , Transplante Heterólogo , Trissacarídeos/administração & dosagem , Trissacarídeos/imunologia , Trissacarídeos/farmacocinéticaRESUMO
An ensymatic immunoassay was developed in order to evaluate the statistical distribution of IgG serum antibodies against pooled pigeon ser antigen in 102 healthy blood donors (HBD). A non-normal distribution was obtained as demonstrated by abnormal values of skewness (2.02) and kurtosis (6.50). A cut-off point (0.120) was determined from the mean plus 2 standard derivations of the optical density values obtained in the HBD group. This value was able to segregate 94 percent of subjects. However, when calculation of the mean less 2 SD was performed to delimit 95 percent of the samples, an aberrant negative value was obtained. In contrast, when the nonparametric method of percentile calculation was applied, an optical density value of 0.130 discriminated 97.5 percent of samples. In addition, the interval between p97.5 and p2.5 delimited 95 percent of samples. We conclude that when reference values and cut-off point are determined from an enzymatic immunoassay, careful analysis of the statistical distribution of reference values is necesary in order to avoid the inappropiated in this study for antibodies against pigeon serum antigens