Diarrea crónica por Lamblia intestinalis en un adulto / (Chronic diarrhea by Lamblia intestinalis in an adult)

ResumenSe presenta un caso de inmunodeficiencia común variable en un paciente masculino, joven con larga historia (9 años) de procesos infecciosos gastrointestinales y respiratorios recurrentes; a pesar de los diferentes esquemas terapéuticos, con evidencia diagnóstica de niveles bajos de inmunoglobulinas de las clases IgG, IgM e IgA; se pretende comparar su evolución a partir de su historia clínica y los resultados de sus exámenes complementarios, con la bibliografía revisada. La inmunodeficiencia común variable explica un déficit primario de IgG (al menos 2 desviaciones estándar por debajo de los valores de referencia para su edad), al menos otra de las Ig (IgA o IgM) y una reducción o ausencia de producción de anticuerpos. Esta entidad se considera poco frecuente en términos de incidencia, aunque cursa muchas veces inadvertida por el predominio de sus efectos. Clínicamente, se manifiesta por la presencia de infecciones recurrentes con preponderancia de las respiratorias y gastrointestinales. Desde el punto de vista etiológico, su génesis es controversial, pero se describen niveles bajos de inmunoglobulinas y una reducción o ausencia de producción de anticuerpos.

AbstractWe present a variable common immunodeficiency case in a young male patient with a long history (9 years) of recurrent gastrointestinal and respiratory infectious processes, despite the different therapeutic schemes, with diagnostic evidence of low levels of IgG, IgM And IgA; aiming to make a comparison of its evolution in function of its clinical history and the results of its complementary examinations, with the bibliography reviewed. Variablecommon immunodeficiency explains a primary IgG deficit (at least 2 standard deviations below the reference values for his age) and at least one other Ig (IgA or IgM) and a reduction or absence of antibody production. This entity is considered infrequent in terms of incidence, although it is often inadvertent due to the predominance of its effects. Clinically it is manifested by the presence of recurrent infections with preponderance of the respiratory and gastrointestinal. From an aetiological point of view, its genesis is controversial, but low levels of immunoglobulins and a reduction or absence of antibody production are all described.

Anticuerpos policlonales contra la proteína recombinante NS3 del virus del dengue / Polyclonal antibodies against recombinant dengue virus NS3 protein

Resumen Introducción: El dengue es una enfermedad causada por uno de los cuatro serotipos del virus del dengue (DENV) y es endémica en, aproximadamente, 130 países. Su incidencia ha aumentado notablemente en las últimas décadas, así como la frecuencia y la magnitud de los brotes. A pesar de los esfuerzos, no existen tratamientos profilácticos ni terapéuticos contra la enfermedad y, en ese contexto, el estudio de los procesos que gobiernan el ciclo de infección del DENV es esencial para desarrollar vacunas o terapias antivirales. Una de las moléculas del DENV más prometedoras es la proteína no estructural 3 (NS3), la cual es indispensable para la replicación viral y es uno de los principales blancos inmunológicos durante la infección. Objetivo: Producir anticuerpos policlonales para contribuir a los futuros estudios sobre las interacciones entre la proteína NS3 y otras proteínas celulares. Materiales y métodos: Se expresaron dos proteínas recombinantes del dominio helicasa de NS3 del DENV de serotipo 2, las cuales se emplearon para inmunizar ratas y producir anticuerpos policlonales. Resultados: Los anticuerpos producidos fueron útiles en ensayos de Western blot e inmunofluorescencia y se reportó por primera vez un anticuerpo policlonal anti-NS3 que permitió la inmunoprecipitación de la proteína viral y la detecta con Western blot sin necesidad de inducir sobreexpresión de NS3 o de usar extractos de células marcados metabólicamente con radioisótopos. Conclusión: Las proteínas recombinantes expresadas y los anticuerpos producidos constituyen herramientas valiosas para estudiar procesos infecciosos del DENV que involucren a la proteína NS3 y evaluar pruebas dirigidas a interferir las funciones de esta proteína.

Abstract Introduction: Dengue is a disease caused by one of four serotypes of the dengue virus (DENV) and is endemic in approximately 130 countries. The incidence of dengue has increased dramatically in recent decades, as well as the frequency and magnitude of outbreaks. Despite all efforts, there are no prophylactic or therapeutic treatments for the disease. Accordingly, research on the processes governing the DENV infection cycle is essential to develop vaccines or antiviral therapies. One of the most attractive DENV molecules to investigate is nonstructural protein 3 (NS3), which is essential for viral replication and a major immune target for infection. Objective: To produce antibodies to support future studies on NS3 and its cellular interactions with other proteins. Materials and methods: Two recombinant proteins of the helicase domain of DENV NS3 serotype 2 were expressed, and used to immunize mice and produce polyclonal antibodies. Results: The antibodies produced were useful in Western blot and immunofluorescence tests. We report an NS3 antibody that immunoprecipitates the viral protein and detects it in Western blot with no need to over-express it or use cell extracts with metabolic radiolabeling.

Preferential production of IgM-secreting hybridomas by immunization with DNA vaccines coding for Ebola virus glycoprotein: use of protein boosting for IgG-secreting hybridoma production

PURPOSE: The goal of this study was to investigate the utility of DNA vaccines encoding Ebola virus glycoprotein (GP) as a vaccine type for the production of GP-specific hybridomas and antibodies. MATERIALS AND METHODS: DNA vaccines were constructed to express Ebola virus GP. Mice were injected with GP DNA vaccines and their splenocytes were used for hybridoma production. Enzyme-linked immunosorbent assays (ELISAs), limiting dilution subcloning, antibody purification methods, and Western blot assays were used to select GP-specific hybridomas and purify monoclonal antibodies (MAbs) from the hybridoma cells. RESULTS: Twelve hybridomas, the cell supernatants of which displayed GP-binding activity, were selected by ELISA. When purified MAbs from 12 hybridomas were tested for their reactivity to GP, 11 MAbs, except for 1 MAb (from the A6-9 hybridoma) displaying an IgG2a type, were identified as IgM isotypes. Those 11 MAbs failed to recognize GP. However, the MAb from A6-9 recognized the mucin-like region of GP and remained reactive to the antigen at the lowest tested concentration (1.95 ng/mL). This result suggests that IgM-secreting hybridomas are predominantly generated by DNA vaccination. However, boosting with GP resulted in greater production of IgG-secreting hybridomas than GP DNA vaccination alone. CONCLUSION: DNA vaccination may preferentially generate IgM-secreting hybridomas, but boosting with the protein antigen can reverse this propensity. Thus, this protein boosting approach may have implications for the production of IgG-specific hybridomas in the context of the DNA vaccination platform. In addition, the purified monoclonal IgG antibodies may be useful as therapeutic antibodies for controlling Ebola virus infection.

Characterization of an IgM-like immunoglobulin from silver catfish (Rhamdia quelen) serum and its use for the production of polyclonal antibodies and development of immunoassays / Caracterização de imunoglobulina similar a IgM em soros de jundiás (Rhamdia quelen) e sua utilização para a produção de soro policlonal e desenvolvimento de imunoensaios

Knowledge on fish immunoglobulin (Ig) characteristics and the availability of monoclonal or polyclonal antibodies to fish Igs are essential to evaluate the humoral immune response and the Ig distribution on leukocyte cells. We demonstrated that silver catfish serum Ig is composed of one immunodominant H chain with approximately 75k Da and one L chain with approximately 28 kDa, similar to human IgM. Rabbit polyclonal antibodies to the catfish IgM-like Ig recognized both the H and L chain and were useful in developing an indirect ELISA to measure the production of antibodies in fish immunized with bovine serum albumin. Dot blot and western blot cross-reactivity studies indicated a wide degree of epitope sharing amongst Ig from several Siluriformes and Characiformes fish indigenous to Brazilian rivers. In these fish species, polyclonal antibodies reacted mostly with the H chain. The results presented here are central to the development of tools and strategies to investigate the antibody production to inoculated antigens and tissue distribution of Ig molecules in native fish species. Furthermore, because of the wide range of cross-reactivity, polyclonal antibodies to silver catfish IgM-like Ig might be used to develop immunoassays to measure the humoral immune response in other fish species.(AU)

Informações sobre as características das imunoglobulinas (Ig) de peixes e a disponibilidade de anticorpos mono ou policlonais são essenciais para avaliar a resposta imune humoral e a distribuição leucocitária de Igs. Nesse trabalho nós demonstramos que a Ig do soro de jundiás é composta por uma cadeia pesada (H) imunodominante, de aproximadamente 75kDA e de uma cadeia leve (L) de aproximadamente 28 kDa, similar à IgM humana. Anticorpos policlonais produzidos contra a Ig do jundiá reconheceram a cadeia H e L e permitiram o desenvolvimento de um ELISA indireto para mensurar a produção de anticorpos em peixes imunizados com albumina sérica bovina. Estudos de reatividade cruzada, por meio de Dot blot e western blot, indicaram um alto grau de compartilhamento de epitopos entre as Igs de diversos peixes Siluriformes e Caraciformes nativos do Brazil. Nestas espécies de peixes, os anticorpos policlonais reconheceram principalmente a cadeia H. Os resultados deste estudo são fundamentais para o desenvolvimento de ferramentas e estratégias para investigar a produção de anticorpos subsequente à imunização e a distribuição tecidual de Igs em peixes nativos. Além disso, devido ao compartilhamento de epitopos entre as espécies de peixes avaliadas, os anticorpos policlonais anti Ig do jundiá poderão ser usados para desenvolver ensaios imunoenzimáticos para avaliar a resposta imune humoral nestas espécies.(AU)

Avaliação do perfil dos linfócitos B de pacientes com Imunodeficiência Comun Variável antes a após administração de antígenos protéicos e polissacarídicos / Evaluation of B lymphocyte profile of Common Variable Immunodeficiency patients before and after immunization with protein and polysaccharide antigens

Introdução: A Imunodeficiência Comum Variável (ICV) faz parte de um grupo de imunodeficiências primárias na qual os pacientes apresentam defeitos na maturação e diferenciação dos linfócitos B (LB), resultando em distúrbios funcionais além de alterações na distribuição de seus subtipos. Consequentemente, estes pacientes apresentam hipogamaglobulinemia, susceptibilidade a infecções e ausência de produção de anticorpos a antígenos específicos. Na tentativa de reduzir os episódios de infecções recorrentes, alguns trabalhos têm recomendado a vacinação com patógenos mortos ou subunidades e em trabalho anterior demonstramos a eficácia clínica da vacinação de pacientes com ICV, porém, a experiência com a administração de vacinas em imunocomprometidos é limitada. Objetivos: Avaliar a cinética da distribuição das subpopulações de linfócitos B antes e após a vacinação com antígenos proteicos e polissacarídicos em pacientes com ICV acompanhados no Ambulatório de Imunodeficiências Primárias do Hospital das Clínicas, FMUSP, além da produção de anticorpos específicos aos antígenos vacinais. Pacientes e Métodos: Um grupo de 35 pacientes com ICV e 16 controles foram vacinados contra Influenza, H1N1 e S. pneumoniae. Após as coletas nos tempos pré e pós 1, 3 e 6 meses foram realizados a separação de PBMC e cultura de linfócitos com lisado viral e hemaglutinina de Influenza, além da citometria de fluxo para identificação das subpopulações de LB naive, zona marginal (MZB), memória com troca de isotipo (SMB) e plasmoblastos (PBL). Foram dosados os anticorpos específicos e no grupo dos pacientes foi aplicado um score de sintomas antes e após a imunização. Resultados: Apesar da redução significativa na pontuação do score de sintomas, a maioria dos pacientes não produziu anticorpos específicos para Influenza, H1N1 e S. pneumoniae...

Introduction: Common Variable Immunodeficiency (CVID) is a primary antibody deficiency characterized by defects in B lymphocyte maturation, resulting in disturbed differentiation, distribution and functional variations on its subtypes. As a result , CVID patients have hypogammaglobulinemia and poor antibody response to specific antigens with increased susceptibility to infections. In an effort to minimize the recurrent episodes of infections, some studies have recommended immunization with inactivated pathogens or subunits and in a former study we have shown the clinical improvement determined by immunization in CVID patients, but the experience with vaccines' administration to immunodeficient patients is limited. Objectives: To evaluate the changes in distribution of B cell subtypes before and after vaccination of CVID patients followed at the Division of Clinical Immunology and Allergy of University of São Paulo Medical School with protein and polysaccharide antigens, as well as specific antibody production . Methods: A group of 35 CVID patients and 16 controls were vaccinated against Influenza, H1N1 and S. pneumoniae vaccines. Blood samples were collected before and 1, 3 and 6 months post vaccination. PBMCs were stimulated with Influenza viral lysate and hemagglutinin peptide. Flow cytometry was performed to identify naïve B cells, marginal zone (MZB), switched memory B cells (SMB) and plasmablasts (PBL). Specific antibody production was measured and a symptoms score was applied for clinical evaluation before and after immunization. Results: In spite of the significant reduction in symptoms score after vaccination, most patients didn't produce specific antibodies to Influenza, H1N1 and S. pneumoniae...

Immune enhancement activities of silk lutein extract from Bombyx mori cocoons

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.

Saccharomyces uvarum mannoproteins stimulate a humoral immune response in mice

Yeasts discarded in industrial processes can be used as a nutritional supplement and to extract cellular components with biotechnological aims. In this study, the humoral immune response of Swiss mice treated with mannoproteins (MP) from the yeast Saccharomyces uvarum was evaluated. The mice were treated with MPs at different doses and times and inoculated with 2% sheep red blood cells. An increase in total Ig in mice treated with 100 μg of MP at the time of immunization or 24 h before was observed in the primary immune response; in the secondary immune response, an increase was observed in total Ig values for all groups, and an increase of IgG was observed in the mice treated with MPs (100 μg) at the time of immunization or 24 h before. These results show that S. uvarum MPs present an immunostimulatory action on the humoral immune response in mice.

Usefulness of a Microcolumn Agglutination Card for Neonates / 대한수혈학회지

BACKGROUND: Blood loss due to laboratory phlebotomy among neonates is correlated with anemia as well as transfusion. In this study, microcolumn agglutination cards for performing ABO & RhD blood typing and direct antiglobulin tests in neonates were evaluated and compared with other established systems. Also, the blood group antibody production rates according to the age were calculated to determine the upper age limit for the new method. METHODS: Eighty subjects were tested by using the DianaGel Neonatal cards (Diagnostic Grifols, Barcelona, Spain), and the results were compared with those of the slide methods for ABO and RhD blood typing, and the DiaMed-ID DC-Screening I test (DiaMed, Morat, Switzerland) for direct antiglobulin tests. A total of 546 subjects who were under 12 months old were tested for the ABO back-typing, and 58 subjects with the AB blood type were excluded. RESULTS: The results of the DianaGel Neonatal card were in agreement with those of the conventional methods for all the subjects. Only one subject showed a discrepant result for the DAT between the DianaGel and DiaMed methods. Blood group antibodies were detected in 29 out of 169 (17.2%) one-day-old neonates, in eight out of 34 (23.5%) infants between one and three months of age and in 81 out of 96 (84.4%) infants between six and twelve months of age. CONCLUSION: The DianaGel Neonatal card showed at least equivalent performance as compared to that of the conventional methods, and it showed advantages for a low blood volume requirement and stronger agglutination grades. The DianaGel card is a suitable alternative for blood typing and DAT in infants under the age of 3 months and who do not necessarily need back-typing of the blood groups due to the low production rate of antibodies.

Effects of Job's Tear(Yul-Moo) Extracts on Mouse Immune Cell Activation

Natural products are increasingly appreciated as a lead for drug discovery and development. A number of investigators have studied various activities of natural products and have found that they have not only nutritional effects but also beneficial properties to cure various diseases and to maintain good health. Job's Tear(Yul-Moo) is a grass crop that have long been used in traditional medicine and a nourishing food. Job's Tear has been reported to exhibit anti-inflammatory, stomachic, antiallergic activity, and antispastic effects and has been used in China for the treatment of warts, rheumatism, and neuralgia although its mechanism remains unclear. Previous results in our laboratory demonstrated that the ethanol extract and water extract of Job's Tear exerted an immune regulatory function on mice cells in vitro. The present study was performed to investigate the ex vivo effect of Job's Tear on immune function. Seven to eight weeks old mices(Balb/c) were fed ad libitum on chow diet and water extract of Job's Tear were orally administrated every other day for two or four weeks at two different concentrations (50 and 500mg/kg B.W.). Proliferation of mice spenocytes and antibody production to sheep red blood cells(SRBC) using hemolytic plague forming cell assay were used to indicate the immune activity. Splenocytes proliferation of Job's Tear with mitogen stimulation such as Con A and LPS was enhanced at 50 mg/kg B.W. concentrations compared to those of control group. In case of antibody production to sheep red blood cells, the number of antibody- secreting cells was increased by administration of 50mg/kg B.W. concentration in mice immunized as a T-dependent antigen. From the present study, Job's Tear water extracts may be suggested to stimulate the mice immune response by enhancing the splenocytes proliferation and the number of plague forming cells.

Effect of vernoniaanthelmintica wild on the immune function of mice / 中国免疫学杂志

Objective:To explore the effect of the vernoniaanthelmintica willd injection on the immune function of mice. Methods: The action of vernoniaanthelmintica willd injection on spleen T, B cell proliferation and antibody production response by [3H]-TdR incorporation and antibody forming cell assays, respectively. Results: Vernoniaanthelmintica willd injection inhibited splenocyte proliferation of normal mouse in vitro and in vivo, the antibody forming cell in vivo, and delayed type of hypersensitivity (DTH) Conclusion: Vernoniaanthelmintica willd injection could inhibit the cellular immune function and humoral immune function.

The effects of substance K and substance P on lymphocytes of spleen and thymus in mice / 中国免疫学杂志

effects of tachykinin SK, SP on mouse splenocyte and thymocyte have been studied.It was found that SK and SP can stimulate ConA or LPS-induced splenocyte prolifertion and antibody production (in cluding IgG, IgM, IgA).5 ?10~(-7)M/L SK can stimulate ConA-induced thymocyte proliferation but SP can't. The subsets of T lymphocytes in spleen and thymus have been changed after SK treatment by FACS analysis. These results show that SK and SP may play a significant regulatory role in immune response.

Kinetics of autoantibody induced by oral immunization with formalized campylobacter jejuni in mice / 中国免疫学杂志

KM mice were orally immunized with formalized C. jejuni (CJ-S131 vaccine) for 16 weeks. The production kinetics of autoanti bodies against ss-DNA, ds-DNA, histories and extractable nuclear antigens (ENA) was investigated and two peaks of autoantibody production were observed within 16 weeks after immunizalion. The first peak appeared at 6th week after immunization when the positive percentage of autoantibody against histones (83%) and ENA (67%) was higher than that against ds-DNA (33%) and ss-DNA ( 0 %), and the second peak at 16th week when the positive percentage of anti-ds-DNA (50%) and anti-ss-DNA (83%) antibody higher than that of anti-histones (33%) and anti-ENA (33%) anti body. Natural autoanti bodies against nuclear acid (ds-DNA and ss-DNA) in the controlled mice could be detected, too, by ELISA, which raised with aging but obviously demonstrating individual difference; however, autoanti bodies to nucleoproteins (histones and ENA) could not be detected at the matched times. The results suggested that autoantibody against histones or ENA was more specific for autoimmune diseases than that against DNA, and might be of significance for earlier diagnosis of the systemic autoimmune diseases, e. g. SLE, in clinic.

ROLE OF RETINOIC ACID RECEPTOR IN ANTIBODY SYNTHESIS OF CORD BLOOD LYMPHOCYTES / 营养学报

Objective: To further explore the mechanism by which retinoic acid (RA) promotes the antibody production of cord blood lymphocytes (CBL). [WT5FZ]Methods: The levels of serum vitamin A in cord blood were measured. The expression of retinoic acid receptor ? (RAR ?) gene was determined by RT PCR (reverse transcription polymerase chain reaction). The IgM concentrations in the supernatants of cell culture were measured by ELISA. [WT5FZ]Results: A positive correlation was found between serum vitamin A level and RAR ? expression level in uncultured CBL:r=0.50 (P