RESUMEN
The ability of nematodes to manipulate the immune system of their host towards a Th2 and T regulatory responses has been proposed to suppress the inflammatory response. Clinical trials have proposed a useful effect of helminth infections on improvement of inflammatory disorders. In this study, we investigated the immunomodulatory effect of Syphacia obvelata infection to induce intestinal tolerance in C57BL/6 mice. Mice were infected through the cagemates with self-infected BALB/c mice. Four weeks post-infection, expression levels of IFN-γ, TNF-α, IL-17, and IL-10 were assessed in the supernatant of mesenteric lymph node (MLN) culture. Foxp3+Treg were measured in MLN cells by flow cytometry. In the S. obvelata-infected group, the percentage of Tregs (5.2±0.4) was significantly higher than the control (3.6±0.5) (P<0.05). The levels of IL-10 (55.3±2.2 vs 35.2±3.2), IL-17 (52.9±3.8 vs 41±1.8), IFN-γ (44.8±4.8 vs 22.3±2.3) and TNF-α (71.1±5.8 vs 60.1±3.3) were significantly increased in infected mice compared to the control group (P<0.05). The above results showed the potential effects of S. obvelata to induce intestinal tolerance. Therefore, it seems that S. obvelata may increase the immunological suppressive function in the intestinal tract.
Asunto(s)
Interacciones Huésped-Parásitos/inmunología , Tolerancia Inmunológica/inmunología , Intestinos/inmunología , Oxiuriasis/inmunología , Oxyuroidea/inmunología , Animales , Células Cultivadas , Citocinas/metabolismo , Inmunomodulación/inmunología , Inflamación/inmunología , Ganglios Linfáticos/inmunología , Ratones Endogámicos C57BL , Linfocitos T Reguladores/inmunología , Células Th2/inmunologíaRESUMEN
One of the problems frequently faced in laboratory facilities is the possibility of the natural parasitic infection of lab animals, which can interfere with biomedical research results. The present study aimed to evaluate cross-reactivity among serum samples from Wistar rats (Rattus norvegicus) naturally infected with Syphacia muris and experimentally infected with Strongyloides venezuelensis. Forty rats were divided into four groups of ten animals each. Parasite load was evaluated by quantifying the adult worms from both helminthes species recovered from the intestines and the S. venezuelensis eggs eliminated in feces. Serological cross-reactivity by parasite-specific IgG detection was tested via enzyme linked immunosorbent assay (ELISA), immunofluorescence antibody test (IFAT) and immunoblotting. The results demonstrated that the quantity of S. venezuelensis eliminated eggs and parthenogenetic females decreased significantly in cases of co-infection with S. muris. ELISA revealed 100% cross-reactivity of serum samples from both species against the opposing antigen. IgG cross-reactivity was confirmed by IFAT using tissue sections of S. venezuelensis larvae and adult S. muris. Immunoblotting showed that IgG antibodies from the sera of animals infected with S. muris recognized eight antigenic bands from S. venezuelensis saline extract and that IgG antibodies from the sera of animals infected with S. venezuelensis recognized seven bands from S. muris saline extract. These results demonstrate the serological cross-reactivity between S. muris and S. venezuelensis in infected rats.
Asunto(s)
Antígenos Helmínticos/inmunología , Inmunoglobulina G/sangre , Oxiuriasis/inmunología , Oxyuroidea/inmunología , Strongyloides/inmunología , Estrongiloidiasis/inmunología , Animales , Coinfección , Reacciones Cruzadas , Ensayo de Inmunoadsorción Enzimática , Heces/parasitología , Femenino , Immunoblotting , Intestinos/parasitología , Larva , Oxiuriasis/complicaciones , Oxiuriasis/parasitología , Oxiuriasis/veterinaria , Carga de Parásitos , Ratas Wistar , Pruebas Serológicas , Estrongiloidiasis/complicaciones , Estrongiloidiasis/parasitologíaRESUMEN
Syphacia muris (Nematoda: Oxyuridae) is a ubiquitous nematode that commonly infects rats in the laboratory and can interfere with the development of unrelated biological assays. In this context, we analysed the effect of a patent S. muris infection in Wistar rats on a superimposed infection with the intestinal trematode, Echinostoma caproni (Trematoda: Echinostomatidae). The results indicate that in the rats, infection with S. muris induces an immunity against a subsequent infection with E. caproni, although each parasite occupies different niches in the host. Echinostoma caproni worm recovery was significantly decreased in the rats primarily infected with S. muris and, at 3 and 4 weeks post-primary infection, the rats primarily infected with S. muris were refractory to the challenge infection with E. caproni. We observed that the main alterations induced by S. muris in the niche of E. caproni (ileum) that may be the cause of the resistance are: (i) a local antibody response with elevated levels of mucosal IgA, IgE, IgG, IgG1 and IgG2a that cross-react with E. caproni antigens; (ii) development of a biased Th17/Th2 phenotype; and (iii) changes in the glycosylation of ileal mucins. This indicates that S. muris induces distant alterations to the ileum of rats affecting the development of other helminth species. Apart from the interest of these results in the study of the interactions between helminths in a single host, it has been demonstrated that pinworm infections may interfere in non-related experiments.
Asunto(s)
Resistencia a la Enfermedad , Echinostomatidae/inmunología , Oxiuriasis/inmunología , Oxyuroidea/inmunología , Infecciones por Trematodos/inmunología , Animales , Anticuerpos Antihelmínticos/inmunología , Modelos Animales de Enfermedad , Femenino , Íleon/inmunología , Inmunidad Mucosa , Mucosa Intestinal/inmunología , Oxiuriasis/parasitología , Ratas , Ratas Wistar , Infecciones por Trematodos/parasitologíaRESUMEN
Mites of 3 genera-Myobia, Myocoptes, and Radfordia -continue to plague laboratory mouse facilities, even with use of stringent biosecurity measures. Mites often spread before diagnosis, predominantly because of detection difficulty. Current detection methods have suboptimal sensitivity, are time-consuming, and are costly. A sensitive serodiagnostic technique would facilitate detection and ease workload. We evaluated whether total IgE increases could serve as a serodiagnostic marker to identify mite infestations. Variables affecting total IgE levels including infestation duration, sex, age, mite species, soiled-bedding exposure, and ivermectin treatment were investigated in Swiss Webster mice. Strain- and pinworm-associated effects were examined by using C57BL/6 mice and Swiss Webster mice dually infested with Syphacia obvelata and Aspiculuris tetraptera, respectively. Mite infestations led to significant increases in IgE levels within 2 to 4 wk. Total IgE threshold levels and corresponding sensitivity and specificity values were determined along the continuum of a receiver-operating characteristic curve. A threshold of 81 ng/mL was chosen for Swiss Webster mice; values above this point should trigger screening by a secondary, more specific method. Sex-associated differences were not significant. Age, strain, and infecting parasite caused variability in IgE responses. Mice exposed to soiled bedding showed a delayed yet significant increase in total IgE. Treatment with ivermectin reduced total IgE levels within 2 wk. Our data suggest that increases in total IgE in Swiss Webster and C57BL/6 mice warrant investigation, especially because mite infestations can rapidly elevate total IgE levels. We propose that using total IgE levels routinely in serologic panels will enhance biosecurity.
Asunto(s)
Ensayo de Inmunoadsorción Enzimática/métodos , Inmunoglobulina E/sangre , Infestaciones por Ácaros/veterinaria , Ácaros/inmunología , Enfermedades de los Roedores/diagnóstico , Pruebas Serológicas/métodos , Acaricidas/uso terapéutico , Animales , Anticuerpos Antihelmínticos/sangre , Antiparasitarios/uso terapéutico , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Cabello/parasitología , Ivermectina/uso terapéutico , Masculino , Ratones , Ratones Endogámicos C57BL , Infestaciones por Ácaros/diagnóstico , Infestaciones por Ácaros/tratamiento farmacológico , Infestaciones por Ácaros/inmunología , Oxiuriasis/diagnóstico , Oxiuriasis/tratamiento farmacológico , Oxiuriasis/inmunología , Oxiuriasis/veterinaria , Oxyuroidea/inmunología , Oxyuroidea/fisiología , Enfermedades de los Roedores/tratamiento farmacológico , Enfermedades de los Roedores/inmunología , Pruebas Serológicas/veterinaria , Especificidad de la Especie , Factores de TiempoRESUMEN
Some helminthic infections, especially nematode infections, may behave as allergens and induce allergic sensitization. In this study, the investigators explored whether infections with Syphacia muris and Aspiculuris tetraptera have any effect on the development of allergen-induced cytokine responses and serum total immunoglobulin E (IgE) levels in ovalbumin (OVA)-sensitized rats. Four groups of male Wistar rats were studied. OVA sensitization was generated in 2 groups of rats; the rats in 1 group were infected and those in the other group were not. On day 21 after sensitization, tumor necrosis factor-alpha, interleukin-10, and total IgE levels in serum samples of rats were measured by enzyme-linked immunosorbent assay. The results showed that average concentrations of tumor necrosis factor-a and interleukin-10 were significantly greater in the group of rats infected with parasites and sensitized to OVA compared with the group uninfected with parasites and sensitized to OVA (P=.043 and P=.046, respectively). Upon comparison of total IgE concentrations, the group of rats infected with parasites and given saline solution showed higher levels compared with the group uninfected with parasites and given saline (P=.004). In conclusion, the investigators were unable to show a protective effect of an existing parasitic infection against the development of allergic sensitization upon exposure to OVA.
Asunto(s)
Hipersensibilidad/inmunología , Ovalbúmina/inmunología , Oxiuriasis/inmunología , Oxyuroidea/inmunología , Alérgenos/inmunología , Animales , Hipersensibilidad/metabolismo , Inmunoglobulina E/biosíntesis , Interleucina-10/biosíntesis , Masculino , Ratas , Ratas Wistar , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
Pinworm parasites commonly infect laboratory mice with high prevalence even in well-managed animal colonies. Although often considered as irrelevant, these parasites if undetected may significantly interfere with the experimental settings and alter the interpretation of final results. There are a few reports documenting the effects of pinworms on research and the effects of pinworms on the host hematopoiesis have not yet been investigated. In this study we examined the changes within various hematopoietic cell lineages in the bone marrow, spleen, peripheral blood and peritoneal space during naturally acquired Syphacia obvelata infection in inbred CBA mice. The data obtained showed significant hematopoietic alterations, characterized by increased myelopoiesis and erythropoiesis in S. obvelata-infected animals. In order to additionally evaluate if this pinworm infection modifies hematopoietic cells' reactivity, we examined the effect of murine interleukin-17, T cell-derived cytokine implicated in the regulation of hematopoiesis and inflammation, on the growth of bone marrow progenitor cells and demonstrated that bone marrow myeloid and erythroid progenitors from S. obvelata-infected mice displayed altered sensitivity to IL-17 when compared to non-infected controls. Taken together the alterations presented pointed out that this rodent pinworm is an important environmental agent that might significantly modify the hosts' hematopoietic response, and therefore interfere with the experimental settings and alter the interpretation of the final results. However, the results obtained also contributed new data concerning the activity of IL-17 on bone marrow hematopoietic cells, supporting our previous reports that depending on physiological/pathological status of the organism IL-17 exerts differential effects on the growth of progenitor cells.
Asunto(s)
Hematopoyesis , Interleucina-17/sangre , Oxiuriasis/sangre , Oxyuroidea/inmunología , Animales , Animales de Laboratorio/parasitología , Células de la Médula Ósea , Masculino , Ratones , Ratones Endogámicos CBA , Oxiuriasis/inmunología , Distribución Aleatoria , Investigación/normas , Bazo/citologíaAsunto(s)
Anticuerpos Antihelmínticos/biosíntesis , Antígenos Helmínticos/inmunología , Ratones Endogámicos BALB C/inmunología , Ratones Endogámicos BALB C/parasitología , Oxiuriasis/inmunología , Oxyuroidea/inmunología , Animales , Animales de Laboratorio/inmunología , Animales de Laboratorio/parasitología , Ratones , Oxiuriasis/parasitologíaRESUMEN
Antibody response to Syphacia obvelata infection was observed in AKR/J mice by ELISA. Experimental infection with the pinworm eggs showed the presence of specific IgG against S. obvelata somatic antigens at 12 days postinfection, and that it increased steadily thereafter. Sera of S. obvelata-infected mice showed cross-reactivity with somatic antigens of other Syphacia species such as S. mesocriceti and S. muris, but not with Aspiculuris asiatica. Western blotting of S. obvelata antigen with sera of S. obvelata-infected mice showed a corresponding increase in the number of bands during the course of infection. Infected mice showed significantly higher antibody production to sheep red blood cells than the uninfected control mice. Thus, S. obvelata infection is shown to alter the humoral response to nonparasitic antigenic stimuli. These observations indicate that infection by helminths, which apparently do not produce clinical symptoms, might modulate the immune system of the host and, therefore, affect experimental results.
Asunto(s)
Anticuerpos Antihelmínticos/biosíntesis , Oxiuriasis/inmunología , Oxyuroidea/inmunología , Animales , Antígenos Helmínticos/inmunología , Western Blotting , Reacciones Cruzadas , Ensayo de Inmunoadsorción Enzimática , Sueros Inmunes/inmunología , Inmunoglobulina G/biosíntesis , Masculino , Ratones , Ratones Endogámicos AKR , Organismos Libres de Patógenos EspecíficosRESUMEN
Electrophoresis on SDS gel and analytical isoelectric focusing showed that a crude extract of Angiostrongylus cantonensis consisted of at least 40 protein components with molecular weights ranging from 13 000-70 000 and isoelectric points of pI values ranging from 3.7-10.0. Crossed-immunoelectrophoresis with a hyperimmune antiserum to A. cantonensis showed at least 40 different antigenic components in the crude worm extract which were cross-reactive with those of Ascaris suum, Metastrongylus apri and Toxocara canis. Using preparative isoelectric focusing, the somatic worm preparation was divided into 13 equal fractions, of which 3, 4 and 5, with pI values of 3.7, 4.0 and 4.45 respectively, were later shown by immunoelectrophoretic techniques and enzyme-linked immunosorbent assay to contain antigens specific to A. cantonensis.
Asunto(s)
Angiostrongylus/inmunología , Antígenos Helmínticos/aislamiento & purificación , Epítopos/aislamiento & purificación , Metastrongyloidea/inmunología , Animales , Anticuerpos/análisis , Antígenos Helmínticos/análisis , Antígenos Helmínticos/inmunología , Ascaris/inmunología , Contrainmunoelectroforesis , Reacciones Cruzadas , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Epítopos/análisis , Epítopos/inmunología , Femenino , Técnica del Anticuerpo Fluorescente , Inmunización , Inmunoelectroforesis , Focalización Isoeléctrica/métodos , Masculino , Oxyuroidea/inmunología , Conejos , Ratas , Ratas Endogámicas , Toxocara/inmunologíaRESUMEN
The effect of infestation with the nematode Syphacia oblevata on adjuvant arthritis was studied in the rat. Animals with an established infestation with Syphacia were found to have a reduced incidence of arthritis after injection of Freund's complete adjuvant. Infested animals developing adjuvant arthritis were found to suffer from a less severe form of the disease than animals in which infestation had been eliminated with piperazine before immunization.