RESUMO
The role of innate cells and their receptors within the male genital tract remains poorly understood. Much less is known about the relative contribution of different genital tract cells such as epithelial/stromal cells and resident leucocytes. In this study, we examined innate immune responses to Chlamydia trachomatis by prostate epithelial/stromal cells and prostate resident leucocytes. Murine prostate primary cultures were performed and leucocyte and epithelial/stromal cells were sorted based on surface protein expression of CD45 by magnetism-activated cell sorting or fluorescence-activated cell sorting. Prostate derived CD45- and CD45+ cells were infected with C. trachomatis and chemokine secretion assayed by ELISA. Similar experiments were performed using prostate CD45+ and CD45- cells from myeloid differentiation factor 88 (Myd88(-/-)) mice or toll-like receptor (Tlr2(-/-)) and Tlr4(mutant) double-deficient mice. Moreover, a TLR-signalling pathway array was used to screen changes in different genes involved in TLR-signalling pathways by real-time PCR. Prostate derived CD45- and CD45+ cells responded to chlamydial infection with the production of different chemokines. Both populations expressed genes involved in TLR signalling and required to respond to pathogen-associated molecular patterns and to C. trachomatis infection. Both populations required the adaptor molecule MYD88 to elicit chemokine response against C. trachomatis. TLR2-TLR4 was essential for chemokine production by CD45+ prostate derived cells, but in their absence, CD45- cells still produced significant levels of chemokines. We demonstrate that C. trachomatis is differentially recognised by prostate derived CD45+ and CD45- cells and suggest that diverse strategies are taking place in the local microenvironment of the host in response to the infection.
Assuntos
Quimiocinas/metabolismo , Infecções por Chlamydia/patologia , Chlamydia trachomatis/fisiologia , Antígenos Comuns de Leucócito/metabolismo , Próstata/metabolismo , Próstata/patologia , Animais , Células Cultivadas , Quimiocina CXCL1/metabolismo , Infecções por Chlamydia/genética , Infecções por Chlamydia/metabolismo , Regulação da Expressão Gênica , Mediadores da Inflamação/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C3H , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Fator 88 de Diferenciação Mieloide/genética , Fator 88 de Diferenciação Mieloide/fisiologia , Cultura Primária de Células , Próstata/microbiologia , Regulação para CimaRESUMO
Grape (Vitis vinifera L.) is an important commercial crop in the temperate regions of Bolivia where it has been grown for hundreds of years. In October of 2001, diseased canes of grape (cv. Muscat of Alexandria) were collected in a vineyard in Yotala, Department of Chuquisaca in southern Bolivia. In this planting of more than 1,000 plants, more than 75% were exhibiting cane dieback symptoms and many were dead or dying. No disease was observed on grape berries. Symptoms of the disease were similar to those reported for Diplodia cane dieback (1). Cankers ranging from 2 to 10 cm long and 0.5 to 3 cm wide were observed. When diseased canes were placed in a moist chamber, conidia oozed from pycnidia in black cirri. Immature conidia were hyaline and one-celled, but mature conidia were dark brown (20 to 30 × 10 to 15 µm) with one median septum and longitudinal striations. The pathogen was tentatively identified as Lasiodiplodia theobromae (Pat.) Griffon & Maubl. (synonyms Diplodia natalensis Pole-Evans and Botryodiplodia theobromae Pat.), teleomorph Botryosphaeria rhodina (Cooke) Arx) (2). Fungi were isolated from cankers on diseased canes by surface disinfestation in 0.25% NaOCl for 5 min and placing small pieces of tissue on 2% water agar or potato dextrose agar (PDA). L. theobromae was isolated from these tissues. Koch's postulates were fulfilled by inoculating grape berries and canes with the pathogen. Five grape berries were surface disinfested and inoculated by wounding with a sterile scalpel and inserting a piece of fungal mycelium on PDA in the wounded sites. The same number of control berries was similarly treated with sterile PDA. Inoculated and control berries were placed in plastic, moist chambers in the laboratory at ambient temperature (15 to 28°C) in the dark. Five canes on two potted plants were inoculated with the same isolate of the pathogen in a similar manner as the berries. The inoculated and control sites on canes were wrapped with masking tape. Plants were placed in a moist chamber for 5 days. After 8 days, inoculated berries were rotting and the inoculated sites were covered with grayish mycelium. Within 12 days, cankers as much as 3 cm long developed on the inoculated canes, and in some lesions, black pycnidia were observed. No lesions developed in the wounded control canes. The pathogen was reisolated from inoculated berries and canes, but not from control berries or canes. The teleomorph was not observed on any naturally infected canes or on those inoculated with the anamorph. The pathogen was identified as L. theobromae based on symptoms (1), cultural and morphological characteristics (2), and pathogenicity tests. The disease poses a potential threat to the cultivation of grapevine in southern Bolivia. To our knowledge, this is the first report of Diplodia cane dieback of grapevine in Bolivia. References: R. C. Pearson and A. C. Goheen. Compendium of Grape Diseases. The American Phytopathological Society, St. Paul, MN, 1988. (2). E. Punithalingam. No. 519 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, England, 1976.
RESUMO
The prostate is one of the main male sex accessory glands and the target of many pathological conditions affecting men of all ages. Pathological conditions of the prostate gland range from infections, chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) of a still unknown aetiology to benign hyperplasia and cancer. CP/CPPS is one of the most prevalent diseases in the urologic clinic and affects men younger than 50 years old. A significant advance in the understanding of CP/CPPS was made when an autoimmune response against prostate antigens was revealed in a considerable number of patients. During the last 30 years, extensive work has been done regarding the development and characterization of different rodent models of experimental autoimmune prostatitis (EAP). It has been demonstrated that tolerance to prostate antigens can be disrupted in some strains of rats and mice and cellular and humoral responses to prostate antigens are elicited. A Th1 pattern has been described and the cellular response seems to be the major pathogenic mechanism involved. Immune cells infiltrate the gland and induce prostate lesions. The genetic background and hormonal imbalance are factors that could contribute to the onset of the disease in susceptible young males. Moreover, spontaneous autoimmune prostatitis could also occur with advanced age in susceptible strains. In this review, we summarize the current knowledge regarding rodent models of EAP and the immunological alterations present in CP/CPPS patients. We also discuss the reliability of these experimental approaches as genuine tools for the study of human disease.
Assuntos
Doenças Autoimunes/imunologia , Prostatite/imunologia , Animais , Doenças Autoimunes/patologia , Doença Crônica , Modelos Animais de Doenças , Humanos , Masculino , Prostatite/patologiaRESUMO
OBJECTIVE: Miliary tuberculosis (MTB) is difficult to diagnose. When prompt diagnosis is necessary, the polymerase chain reaction (PCR) to detect mycobacterial DNA may be valuable. SETTING: Tuberculosis clinic in an academic tertiary-level hospital in Mexico. DESIGN: Bone marrow (BM) aspiration samples from 30 consecutive clinically suspected MTB patients and 58 non-tuberculosis hematologic patients were evaluated by in-house PCR using a fragment of the insertion sequence IS6110; results were compared with those obtained by acid-fast-stained smears, culture in Löwenstein-Jensen medium, histology, and serology. RESULTS: Tuberculosis diagnosis was confirmed in all MTB suspects, 28 by microscopy and culture in pulmonary or extra-pulmonary samples other than BM, and two by clinical and radiologic improvement after antituberculosis treatment. In fresh BM specimens, in-house PCR was positive in 21/30 (70%) suspects, contrasting with only one positive (3.3%) in staining and culture, and four with compatible histologic findings (13.3%). BM samples from the control group showed negative results in bacteriologic and histologic studies, except in nine who had positive PCR results. These nine control cases had malignant processes. CONCLUSION: PCR in aspirates of BM is a useful diagnostic assay in cases of MTB, mainly when bacteriological results are negative.
Assuntos
Medula Óssea/microbiologia , Medula Óssea/patologia , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/isolamento & purificação , Reação em Cadeia da Polimerase , Tuberculose Miliar/genética , Tuberculose Miliar/microbiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Elementos de DNA Transponíveis/genética , DNA Bacteriano/genética , DNA Bacteriano/isolamento & purificação , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Teste Tuberculínico , Tuberculose Miliar/diagnósticoRESUMO
To establish the frequency of infectious aetiology in Mexican adult patients with cervical lymphadenopathies (CLAs), 87 consecutive patients with enlarged cervical lymphatic nodes, HIV negative and without anti-tuberculous treatment, were selected from a tertiary-level speciality concentration hospital. Histopathological studies, investigation of acid-fast bacilli, cultures in Löwenstein Jensen and Mycobacterium growth indicator tube (MGIT) media, and in-house polymerase chain reaction (PCR) with IS6110-based primers for Mycobacterium tuberculosis complex were performed in resected lymphatic nodes. Non-infectious aetiology corresponded to 45 cases (52 %). Tuberculosis was suspected in 42 cases (48%) by histology and confirmed positive results were obtained by staining in 8 (19%), by culture in 23 (55%), and by PCR in 34 (81 %) patients. All were confirmed after therapeutic success. In addition to the epidemiological transition process occurring in Mexico, tuberculosis remains an important cause of CLA. Histopathology with confirmatory studies including PCR can detect tuberculous aetiology.
Assuntos
Países em Desenvolvimento , Doenças Linfáticas/epidemiologia , Doenças Linfáticas/microbiologia , Tuberculose Pulmonar/complicações , Tuberculose Pulmonar/epidemiologia , Adolescente , Adulto , Idoso , DNA Bacteriano/análise , Estudos Epidemiológicos , Feminino , Humanos , Incidência , Doenças Linfáticas/etiologia , Masculino , México/epidemiologia , Pessoa de Meia-Idade , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/patogenicidade , Pescoço/patologia , Reação em Cadeia da PolimeraseRESUMO
Gala and Winter Banana apples are important commercial crops in Azurduy and Lima Bamba, which are located in the Department (state) of Chuquisaca, Bolivia. White or bot rot (causal agent Botryosphaeria dothidea (Moug.:Fr.) Ces. De. Not. [anamorph Fusicoccum aesculi Corda]) and black rot (causal agent B. obtusa (Schwein.) Shoemaker [anamorph Sphaeropsis malorum Berk.]) have not been reported previously from Bolivia. Both fungi were isolated from apple fruit and branch cankers in Azurduy, but only B. dothidea was isolated from rotted fruit and limb cankers in Lima Bamba. Both fungi also were isolated from rotted Gala and Winter Banana fruit purchased in the markets in Sucre, Bolivia. Symptoms on fruit consisted of light-to-dark brown lesions that ranged from 3- to 8-cm in diameter. Cankers on limbs were sunken and reddish brown and ranged from 2 to 25+ cm in length and 0.5 to 3 cm in diameter. Neither pathogen produced pycnidia in lesions on rotted fruit, but they often developed in branch cankers. Pseudothecia of B. dothidea and B. obtusa were not observed. Identification of both pathogens was based on descriptions of their anamorphic stages (1). To fulfill Koch's postulates, four healthy Gala apple fruit were inoculated with two isolates of each pathogen by wounding the opposite faces of surface-disinfected fruit with a 5-mm-diameter cork borer and inserting mycelial plugs of the pathogens. Plugs were obtained from the margins of cultures growing on potato dextrose agar (PDA). Wounds were made on the opposite sides of each fruit, a mycelial plug of one of the pathogens was inserted in one wound, and on the opposite side, a plug of sterile PDA was inserted as a control. Each plug containing fungal mycelium or sterile PDA was covered with a plug of trimmed apple tissue, and the apple fruit were incubated in a moist chamber at 17 to 20°C for 10 days. Six branches on two young apple trees growing outdoors in a nursery were inoculated in a similar manner with one isolate of each pathogen: bark was wounded with a 5-mm-diameter cork borer, and the wounded area was inoculated with a plug of PDA containing the pathogen or a plug of sterile PDA for the control. The inoculated sites were wrapped with masking tape to prevent dehydration. Within 10 days, all fruit wounds inoculated with isolates of each pathogen developed brown lesions up to 5 cm in diameter. Each pathogen was reisolated from tissues in which it had been inoculated, but not from any of the noninoculated control sites. Within 6 to 8 weeks, all but one wound on branches inoculated with each pathogen developed depressed canker lesions up to 2 cm in length. Each pathogen was reisolated from the canker produced by inoculation with that pathogen, but not from any of the control sites. Reference: (1) T. B. Sutton. White rot and black rot. Pages 16-20 in: Compendium of Apple and Pear Diseases, A. L. Jones and H. S. Aldwinckle, eds. The American Phytopathological Society, St. Paul, MN, 1991.
RESUMO
Prostatic steroid binding protein (PSBP) is the major protein produced ( approximately 20% of the total cytosolic protein) and secreted into the seminal fluid by the rat ventral prostate but its physiological function has not been elucidated yet. Since PSBP is secreted into the seminal fluid (which is itself a potent immunosuppressor) and has strong homology with uteroglobin (which possess an important anti-inflammatory function) our aim was to determine what effect, if any, PSBP would have on the immune system. With that purpose in mind we performed mononuclear cell cultures in the presence or absence of purified PSBP and analysed the effect of this protein on different functional parameters. PSBP inhibits the mitogen-induced proliferation of normal rat spleen mononuclear cells (MNC) specifically and in a dose-dependent manner. It reduces the production of IL-2 and the expression of its receptor (analysed by flow cytometry) which are important events for lymphocyte proliferation. Also, PSBP was able to inhibit OVA-specific proliferation of lymph node cells from previously primed animals. The immunosuppressive effect of PSBP is not due to an inherent toxic effect to the cells, since the cell viability was kept intact at the different times of culture studied. We also analysed the effect of rat PSBP on mitogen-induced proliferation of mouse spleen and human blood MNC. The proliferation was strongly abolished in a dose-dependent and non-species specific fashion. Moreover, PSBP strongly inhibits the human mixed lymphocyte reaction. Taken together, the present data support evidence for a new type of function for PSBP. We report that PSBP is a potent immunosuppressor factor and we describe its effect on the immune function in vitro. Here, we discuss the possible implications of these findings in the protection of sperm from immunologic damage in the feminine reproductive tract.
Assuntos
Proteína de Ligação a Androgênios/farmacologia , Imunossupressores/farmacologia , Próstata/imunologia , Proteína de Ligação a Androgênios/imunologia , Proteína de Ligação a Androgênios/isolamento & purificação , Animais , Antígenos/administração & dosagem , Regulação para Baixo/efeitos dos fármacos , Feminino , Humanos , Imunossupressores/isolamento & purificação , Técnicas In Vitro , Interleucina-2/metabolismo , Ativação Linfocitária/efeitos dos fármacos , Teste de Cultura Mista de Linfócitos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Mitógenos/farmacologia , Ovalbumina/imunologia , Proteína de Ligação a Fosfatidiletanolamina , Prostateína , Ratos , Ratos Wistar , Receptores de Interleucina-2/metabolismo , Secretoglobinas , Espermatozoides/imunologia , UteroglobinaRESUMO
Lewis (Lw) rats are susceptible and Wistar (Wr) rats are usually resistant to the induction of experimental allergic encephalomyelitis (EAE). In this study we analyze the humoral response to myelin antigens, providing evidence for different B cell response to myelin basic protein (MBP) and other myelin proteins between these two strains of rats with different susceptibility to EAE. In fact, IgG anti-MBP titers in Wr rats were markedly higher than in Lw ones. Moreover, an inverse relationship between the amount of antigen injected to induced EAE and the level of anti-MBP antibodies was observed in Wr rats, while IgG anti-MBP varied in a positive dose-depending manner in sera from Lw rats. Also, sera from Wr rats analyzed by immunoblotting showed a strong reactivity with MBP and other myelin proteins, but sera from Lw rats reacted only with MBP. Evaluation of IgA and IgM against MBP in Wr rats showed again higher titers of these isotypes when compared with the titers observed in Lw rats. The distribution of IgG subclasses in sera from both strains indicated that Wr produced low titers of specific IgG1, while Lw rats did not produce specific IgG1. However, Wr rats showed high levels of IgG2a, IgG2b and IgG2c subclasses while lesser titers of these isotypes were observed in Lw animals. These findings indicate that both strains have the capacity to develop antibodies against portions of the MBP molecule, but antibody production is greater in the resistant Wistar rats suggesting a B cell activation in these animals, that could be related to their lower susceptibility.
Assuntos
Autoanticorpos/biossíntese , Encefalomielite Autoimune Experimental/imunologia , Proteína Básica da Mielina/imunologia , Animais , Formação de Anticorpos , Feminino , Imunoglobulina G/classificação , Masculino , Ratos , Ratos Endogâmicos Lew , Ratos Wistar , Especificidade da EspécieRESUMO
Suppression of Experimental Autoimmune Encephalomyelitis (EAE) can be achieved by i.p. administration of soluble myelin basic protein (MBP) in adult Wistar rats before the immunization. In the present work, we analyze the role of peritoneal antigen-presenting cells (APC) in the induction of tolerance to EAE. Peritoneal cells (PC) pulsed in vivo with MBP were obtained from rats that had been intraperitoneally injected 2 h previously with soluble MBP (MBP-PC) and then inoculated in recipient rats before the induction of EAE. Our findings show that the i.p. treatment of the animals with MBP-PC before the immunization was able to diminish the incidence and severity of the disease, reduce the histological alterations, abrogate the proliferative response against MBP and change the pattern of the humoral response to MBP. Moreover, when spleen mononuclear cells (MNC) from tolerant animals were cultured together with spleen MNC from sick animals, a dose-dependent inhibition of the proliferative response was observed, arguing for the presence of a regulatory cell population in the tolerant animals. It is also demonstrated that the MBP-PC are activated and their capability of inducing suppression of EAE is highly associated with the enhanced expression of MHC class II IA molecule. Our results show that peritoneal cells pulsed in vivo with MBP are able to induce tolerance and suggest that the up-regulation of MHC class II on MBP-PC is a necessary event for tolerance induction in our model.
Assuntos
Encefalomielite Autoimune Experimental/imunologia , Encefalomielite Autoimune Experimental/prevenção & controle , Macrófagos Peritoneais/imunologia , Proteína Básica da Mielina/imunologia , Medula Espinal/imunologia , Animais , Anticorpos/análise , Apresentação de Antígeno/imunologia , Bovinos , Divisão Celular/imunologia , Transplante de Células , Células Cultivadas , Encefalomielite Autoimune Experimental/tratamento farmacológico , Feminino , Citometria de Fluxo , Antígenos de Histocompatibilidade Classe II/análise , Tolerância Imunológica , Imunoglobulina G/análise , Terapia de Imunossupressão , Injeções Intraperitoneais , Macrófagos Peritoneais/química , Macrófagos Peritoneais/citologia , Masculino , Proteína Básica da Mielina/análise , Proteína Básica da Mielina/farmacologia , Fluxo Pulsátil , Ratos , Ratos Wistar , Medula Espinal/química , Baço/citologia , Baço/imunologiaRESUMO
Experimental autoimmune prostatitis (EAP) is a disease that could be considered an experimental model of human non-bacterial prostatitis. In this experimental model, male rats are intradermally immunized with a saline extract of male sex accessory glands (RAG) in an adequate adjuvant. The prostatitis observed in the immunized animals develops as a consequence of the immune response against RAG antigens, and the histological lesion is strikingly similar to the pattern of prostatic inflammation observed in the human disease. In this study, we purified one of the prostatic autoantigens recognized by the autoantibodies in our model. Amino acid sequence analysis identified the purified protein as prostatein or rat prostatic steroid binding protein, a member of the uteroglobin superfamily. Prostatein was recognized not only by the humoral autoimmune response, but also by the cellular autoimmune response. Certainly, the DTH response and lymph node cell proliferative assays against prostatein in immunized animals yielded positive results. Prostatein is not only the target of the autoimmune response in animals immunized with the whole extract, but also an inducing antigen of the disease. Purified prostatein, when incorporated to an adequate adjuvant, elicited cellular and humoral autoimmune response and lesion in the prostate gland. The identification of one of the target antigens in autoimmune prostatitis has provided a further refinement and characterization of our model, which could serve for a better understanding of the aetiology, pathogenesis and pathophysiology of non-bacterial prostatitis.
Assuntos
Proteína de Ligação a Androgênios/imunologia , Autoantígenos/imunologia , Prostatite/imunologia , Animais , Citosol/imunologia , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Humanos , Imunidade Celular , Masculino , Peso Molecular , Proteína de Ligação a Fosfatidiletanolamina , Próstata/imunologia , Prostateína , Ratos , Ratos Wistar , Secretoglobinas , UteroglobinaRESUMO
Stress disturbs homeostasis by altering the equilibrium of various hormones which have a significant impact on immune responses. Few studies have examined the influence of stressors on autoimmune disease in animal models. In our work, we studied the effects of long-term exposure (14 days) to chronic varied stress (CVS) in a model of experimental autoimmune encephalomyelitis (EAE) in Wistar rats. We studied whether the exposure to CVS before or after the immune challenge would correlate with differences in the clinical course of the disease. We also examined whether the CVS would modulate the magnitude of the cellular or the humoral immune response. We observed opposite effects on the clinical signs in animals stressed before or after the immune challenge. The clinical signs of the disease were attenuated in animals stressed before but not after the immune challenge. Relationships were found in the modulation of the clinical severity related to the time of exposure to the CVS, the histological alterations and the proliferative results. Stressed animals with milder clinical signs presented an exacerbated humoral response against myelin antigens while stressed animals with more severe clinical symptoms exhibited a significantly diminished one. Besides, we detected the presence of specific IgG1 associated with the exposure to CVS before the induction of EAE. Our results show that, depending on the timing of the exposure of Wistar rats to the CVS, the neuroendocrine disbalance favors a more pronounced humoral or cellular profile of the response.
Assuntos
Encefalomielite Autoimune Experimental/fisiopatologia , Estresse Fisiológico/fisiopatologia , Animais , Anticorpos/imunologia , Formação de Anticorpos/fisiologia , Antígenos/imunologia , Bovinos , Doença Crônica , Encefalomielite Autoimune Experimental/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Imunoglobulina G/análise , Imunoglobulina G/classificação , Proteína Básica da Mielina/imunologia , Bainha de Mielina/imunologia , Ratos , Ratos WistarRESUMO
Intraperitoneal (i.p.) treatment of Wistar rats with bovine myelin (BM) or myelin basic protein (MBP) previously to immunization with BM-CFA showed a diminished incidence and severity of experimental autoimmune encephalomyelitis (EAE) (2/13 and 0/7, respectively) when compared with rats immunized with BM-CFA (11/17) or i.p. treated with ovalbumin (2/4). Concomitantly, animals treated with BM or MBP exhibited a marked reduction of proliferative response to MBP which was highly positive when spleen mononuclear cells from nontreated and ovalbumin treated animals were assayed. Rats that were treated with MBP before immunization produce IgA, IgM, total IgG and subclasses of IgG, IgG2a, IgG2b, IgG2c specific for MBP in similar levels than those observed in nontreated immunized animals. However, a higher incidence and level of IgG1 was observed in MBP treated rats, meanwhile rats i.p. treated with total BM showed a highly reduced humoral response. The herein presented results show that i.p. treatment with low amounts of soluble forms of myelin antigens markedly reduced the clinical symptoms of the disease, the histological alterations, the cellular proliferative response to MBP, and produced changes in the autoimmune humoral response.
Assuntos
Encefalomielite Autoimune Experimental/tratamento farmacológico , Encefalomielite Autoimune Experimental/imunologia , Proteína Básica da Mielina/farmacologia , Animais , Formação de Anticorpos/efeitos dos fármacos , Autoimunidade/efeitos dos fármacos , Encéfalo/citologia , Encéfalo/imunologia , Bovinos , Relação Dose-Resposta a Droga , Ensaio de Imunoadsorção Enzimática , Feminino , Imunização , Imunoglobulina A/sangue , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Injeções Intraperitoneais , Masculino , Ratos , Ratos Wistar , Solubilidade , Medula Espinal/química , Baço/citologia , Baço/imunologiaRESUMO
A comprehensive biochemical, immunological, and histological study was undertaken during suppression of experimental autoimmune encephalomyelitis (EAE) induced by antigen-specific inhibition of the immune response. Pretreatment of Wistar rats by intraperitoneal administration of low doses of saline-soluble bovine myelin or myelin basic protein (MBP) but not with ovalbumin suppresses the appearance of the clinical symptoms of EAE induced by sensitization with bovine myelin in complete Freund's adjuvant. Analysis of the central nervous system (CNS) of animals pretreated with MBP or whole myelin shows inhibition of the diminution of MBP and 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNPase) activity observed in the EAE animals or in rats pretreated with ovalbumin. With respect to the CNS lipid content, these suppressive treatments abolish the increase in esterified cholesterol and partially revert the diminution in the content of cerebrosides and total cholesterol characteristic of the acute stage of the disease. Concomitantly, meningeal and parenchymal infiltration with mononuclear cells and deposits of immunoglobulins in the infiltrated regions as well as in spinal cord motor neurons were reduced. Analysis of the humoral response to myelin antigens shows that all EAE as well as treated animals developed antibodies to MBP and other myelin proteins. However, a higher incidence and level of these antibodies was observed in nontreated EAE animals and MBP- and ovalbumin-treated rats, while rats treated with total bovine myelin showed a highly reduced humoral response. The present results indicate that intraperitoneal treatment with soluble forms of myelin antigens, concomitant with the suppression of the clinical symptoms of the disease, markedly reduces the biochemical and histological alterations occurring in EAE animals and produces changes in the autoimmune humoral response.
Assuntos
Alérgenos/imunologia , Encefalomielite Autoimune Experimental/imunologia , Animais , Formação de Anticorpos/imunologia , Encefalomielite Autoimune Experimental/metabolismo , Epitopos , Feminino , Imuno-Histoquímica , Injeções Intraperitoneais , Masculino , Proteína Básica da Mielina/farmacologia , Proteínas da Mielina/farmacologia , Fármacos Neuroprotetores/farmacologia , Ratos , Ratos WistarRESUMO
Although autoimmune response against most tissues in the body has been extensively described, very little is known about autoimmune response against prostate antigens either in humans or animals. In this work we studied the autoimmune response elicited against rat prostate (RP) in Wistar rats immunized with a chemically modified extract of syngeneic male sex accessory glands (MRAG). We observed that one immunization was enough for the induction of positive delayed-type hypersensitivity test (DTH), which was higher on day 15 than on day 30. It was also enough to induce IgG autoantibodies to RP although a second injection was necessary to obtain a more frequent occurrence and a greater reactivity. The autoantibodies against RP were directed mainly to the cytosolic fraction and reacted at least with two molecules of 43 and 20 KD. Sera obtained on days 30 and 45 showed presence of specific IgG, IgA and IgM. Specific IgG2b and IgG2c were found on both days. On day 30 none of the sera presented IgG2a anti-RP, while on day 45 only 38% of the sera were considered positive for this isotype. No IgG1 anti-RP was detected in any serum of either bleeding. Direct immunofluorescence staining showed intense immunofluorescence in prostate epithelium, mainly in the apical zone of the gland, in animals that had received two immunizations with MRAG-CFA. No positive staining was seen in prostates obtained on day 30 after just one immunization, in sections of normal prostates or in sections of other rat organs. Our data indicate that the main isotypes involved in this autoimmune phenomenon are IgG2b and IgG2c. A strong association between the cellular autoimmune response measured by the DTH response and the IgG2b and IgG2c isotype was found at early stages of the disease. Since the DTH response and the IgG2b isotype have been previously associated with Th1-like activity in rats, our results suggest that Th1-like cells could be playing an active role in early stages of this disease.
Assuntos
Hipersensibilidade Tardia/imunologia , Imunoglobulina G/imunologia , Próstata/imunologia , Animais , Especificidade de Anticorpos , Complexo Antígeno-Anticorpo/análise , Antígenos/imunologia , Autoanticorpos/imunologia , Autoantígenos/análise , Autoimunidade , Immunoblotting , Cinética , Masculino , Ratos , Ratos Wistar , Frações Subcelulares/imunologiaRESUMO
We studied the histological modifications in the accessory glands of autoimmune rats. Adult male Wistar rats were id immunized three times with saline extract of rat male accessory glands (RAG) chemically modified (MRAG) in complete Freund's adjuvant (CFA) (groups 1, 2, and 3). Prior to the first immunization with MRAG-CFA groups 2 and 3 received peritoneal cells obtained from rats that had been injected 2 or 24 hr previously with low doses of RAG. Furthermore, an additional group (group 4) ip immunized with liposome-associated-RAG was incorporated. The delayed-type hypersensitivity response studied 10 or 15 days after first immunization was positive for rats of groups 1, 3, and 4, while it was negative for rats of group 2. Serum samples obtained on Day 45 and studied by ELISA showed high levels of autoantibodies in groups 1 and 2 and lower levels of autoantibodies in group 3, but did not show autoantibodies in group 4. The histological studies performed 10 days after the last immunization showed organ-specific lesions in the accessory glands in animals of groups 1, 3, and 4. Infiltration of mononuclear cells was the main alteration in group 1, while infiltration of mast cells and polymorphonuclear leukocytes were present in specimens of groups 3 and 4. The main finding of this study was a significant increase (P < 0.0005) in the extent of mast cell degranulation in the specimens of accessory glands stained with toluidine blue. Our results suggest that mast cells are activated in our experimental model of autoimmune disease.
Assuntos
Doenças Autoimunes/imunologia , Genitália Masculina/imunologia , Genitália Masculina/patologia , Mastócitos/imunologia , Prostatite/imunologia , Animais , Formação de Anticorpos/imunologia , Autoantígenos/imunologia , Doenças Autoimunes/patologia , Adjuvante de Freund/farmacologia , Imunidade Celular/imunologia , Imunização , Lipossomos , Masculino , Prostatite/patologia , Ratos , Ratos Wistar , Extratos de Tecidos/farmacologiaRESUMO
El propósito del estudio fué analizar las relaciones entre alimentación natural, estado nutricional y morbilidad en el primer semestre de vida. Se estudió durante seis meses una cohorte de 128 recién nacidos de familias de bajos ingresos económicos, atendidos en dos consultorios del sector norte de Santiago. Se evaluó tipo de alimentación, peso/edad y talla/edad según tablas NCHS/OMS e incidencia de morbilidad en el primer semestre. Las frecuencias de lactancia natural absoluta fueron 73, 48 y 20 por ciento al segundo, cuarto y sexto mes respectivamente. El riesgo relativo de déficit de peso fue 4,9 a 9,8 (p<0,02) y de déficit de talla 1,6 a 2,7 veces mayor (p<0,05) dependiendo de la edad de introducción de alimentos diferentes a la leche materna. El riesgo relativo de diarrea fue 5,5 a 21,7 veces mayor con la introducción precoz de otros alimentos (p<0,02). El efecto protector en relación a infecciones respiratorias y hospitalizaciones fue también significativo. Los resultados apoyan la importancia de mantener la lactancia absoluta durante los primeros seis meses de vida
Assuntos
Humanos , Masculino , Feminino , Recém-Nascido , Lactente , Aleitamento Materno/estatística & dados numéricos , Nutrição do Lactente , Apoio Nutricional/estatística & dados numéricos , Doenças Transmissíveis/epidemiologia , Diarreia Infantil/epidemiologia , Crescimento , Comportamento Alimentar , Aleitamento Materno Parcial , Morbidade , Estudos Prospectivos , Doenças Respiratórias/epidemiologia , Fatores Socioeconômicos , Peso-Idade , Peso-EstaturaRESUMO
1. The immunization of Wistar rats with 5 mg of chemically modified rat male accessory glands saline extract (MRAG) incorporated into complete Freund's adjuvant (CFA) induced a delayed type hypersensitivity (DTH) response to rat male accessory glands (RAG). Pretreatment with peritoneal cells (PC) obtained from rats 2 h after an intraperitoneal (ip) injection of a partially purified fraction (FI) of RAG (FI-PC2h) suppressed the DTH response to MRAG after immunization with MRAG-CFA, while pretreatment with PC obtained 24 h after an ip injection of FI-RAG (FI-PC24h) induced potentiation of the DTH response to MRAG. 2. The injection of spleen mononuclear cells (SpM), obtained from rats rendered unresponsive to MRAG by pretreatment with FI-PC2h, into normal syngeneic recipients markedly prevented the DTH reaction to MRAG. The transfer of SpM cells from animals injected with FI-PC24h (potentiated animals) to suppressed recipients (recipients of FI-PC2h on days -10 and -3, prior to immunization with MRAG-CFA) showed that SpM cells did not eliminate the suppression state in these recipients, but when they were transferred to normal recipients, they were able to induce a positive response to RAG (P < 0.005). 3. The study of the phenotypic characteristics of the SpM cells prior to transfer showed similar numbers of CD4 and IL-2R SpM cells in both potentiated and normal animals. However, the number of CD8 cells was significantly reduced in SpM cells from potentiated animals compared to that observed in SpM cells from normal animals (P < 0.05).
Assuntos
Células Apresentadoras de Antígenos/imunologia , Autoimunidade/imunologia , Genitália Masculina/imunologia , Imunização , Baço/citologia , Animais , Hipersensibilidade Tardia , Injeções Intraperitoneais , Masculino , Fenótipo , Ratos , Ratos WistarRESUMO
1. The immunization of Wistar rats with 5 mg of chemically modified rat male accessory glands saline extract (MRAG) incorporated into complete Freund's adjuvant (CFA) induced a delayed type hypersensitivity (DTH) response to rate male accessory glands (RAG). Pretreatment with peritoneal cells (PC) obtained from rats 2 h after an intraperitoneal (ip) injection of a partially purified fraction (FI) of RAG (FI-PC2h) suppressed the DTH response to MRAG after immunization with MRAG-CFA, while pretreatment with PC obtained 24 h after an ip injection of FI-RAG (FI-PC24h) induced potentiation of the DTH response to MRAG. 2. The injection of spleen mononuclear cells (SpM), obtained from rats rendered unresponsive to MRAG by pretreatment with FI-PC2h, into normal syngeneic recipients markedly prevented the DTH reaction to MRAG. The transfer of SpM cells from animals injected with FI-PC24h (potentiated animals) to suppressed recipients (recipients of FI-PC2h on days -10 and -3, prior to immunization with MRAG-CFA) showed that SpM cells did not eliminate the suppression state in these recipients, but when they were transferred to normal recipients, they were able to induce a positive response to RAG (P<0.005). 3. The study of the phenotypic characteristics of the SpM cells prior to transfer showed similar numbers of CD4 and IL-2R SpM cells in both potentiated and normal animals. However, the number of CD8 cells was significantly reduced in SpM cells from potentiated animals compared to that observed in SpM cells from normal animals (P<0.05)
Assuntos
Animais , Masculino , Ratos , Células Apresentadoras de Antígenos/imunologia , Baço/citologia , Genitália Masculina/imunologia , Imunização , Hipersensibilidade Tardia , Injeções Intraperitoneais , Fenótipo , Ratos WistarRESUMO
Peritoneal cells (PC) obtained 2 h subsequent to intraperitoneal (i.p.) injection of low doses (200 micrograms) of a purified fraction of rat male accessory glands (FI-RAG) are phenotypically and functionally different from those obtained 24 h after i.p. injection. In fast, PC obtained 2 h after FI-RAG injection are mainly IE+ and are involved in inducing specific suppression to RAG. In contrast, PC obtained 24 h after FI-RAG injection are mainly IA+ and capable of inducing specific response to RAG. For their induction, IA+ PC require cells within or recently derived from bone marrow. In order to analyze the mechanisms involved in IA+ bone marrow-dependent cell generation in the peritoneum, we studied the distribution of FI-RAG following i.p. injection. It was established that FI-RAG is found mainly in the thymus 2 h after injection and remains there for at least 24 h. Subsequently, we analyzed, in 4 groups of rats, the influence of thymic culture supernatants on the phenotype of cells appearing in the peritoneal cavity 2 h after FI-RAG injection. An increase in IA+ PC was observed 2 h after i.p. injection of FI-RAG in animals that had received either supernatants from normal thymic cells cultured with FI-RAG or those from thymic cells taken from animals injected with FI-RAG 2 h prior to being killed. Supernatants of thymic cells from animals injected with FI-RAG 24 h prior to being killed or from normal thymic cells do not increase the percentage of IA+ PC.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Células Apresentadoras de Antígenos/imunologia , Genitália Masculina/imunologia , Timo/imunologia , Animais , Antígenos de Superfície/análise , Autoantígenos/imunologia , Autoimunidade , Antígenos de Histocompatibilidade Classe II/imunologia , Imunofenotipagem , Injeções Intraperitoneais , Linfonodos/imunologia , Masculino , Cavidade Peritoneal/citologia , Ratos , Ratos WistarRESUMO
Se describen los efectos de un programa de fomento de lactancia materna donde participaron monitoras de la misma comunidad y se dio atención conjunta a madres e hijos, en familias de nivel socioeconómico bajo, comparando sus resultados (grupo intervenido) con los del centro de salud municipal del mismo sector (grupo no intervenido), en dos cohortes de 62 y 66 lactantes respectivamente. La prevalencia de lactancia materna (absoluta y exclusiva) fue significativamente mayor durante todo el periodo en el grupo experimental. Al sexto mes la lactancia exclusiva era de 80,6 por ciento en el grupo intervenido y 4,5 por ciento en el grupo control (p<0,01) y la proporción de niños destetados 1,6 por ciento y 37,8 por ciento respectivamente. El crecimiento, en peso y talla, fue significativamente mejor en el grupo experimental entre el cuarto y el sexto mes y su tasa de desnutrición fue mas baja (p<0,05). Se concluye que un programa con participación activa de la comunidad puede ser muy efectivo para fomentar la lactancia materna exclusiva y mejorar el crecimiento infantil