RESUMEN
Due to their inability to generate a complete immune response, mice knockout for type I interferon (IFN) receptors (Ifnar-/-) are more susceptible to viral infections, and are thus commonly used for pathogenesis studies. This mouse model has been used to study many diseases caused by highly pathogenic viruses from many families, including the Flaviviridae, Filoviridae, Arenaviridae, Bunyaviridae, Henipaviridae, and Togaviridae. In this review, we summarize the findings from these animal studies, and discuss the pros and cons of using this model versus other known methods for studying pathogenesis in animals.
Asunto(s)
Modelos Animales de Enfermedad , Brotes de Enfermedades , Ratones Noqueados/genética , Receptor de Interferón alfa y beta/genética , Virosis/epidemiología , Animales , Ratones , Ratones Noqueados/inmunología , Receptor de Interferón alfa y beta/fisiología , Virosis/genética , Virosis/inmunología , Virosis/virologíaRESUMEN
This study aimed to evaluate aspects of host immune response using an experimental infection model of Corynebacterium pseudotuberculosis (CP) in C57/Black6 wild-type and knockout for nitric oxide (KO-NO) mice. 28 mice were evaluated: 4 wild-type controls; 10 wild-type infected with CP; 4 KO-NO controls; 10 KO-NO infected with CP. Infection procedures were carried out by intraperitoneal inoculation using 107. Infected C57/Black6 KO-NO mice began to die after the 5° day post-inoculation, up until the 14º day. Neutrophils were found in increased numbers in the infiltrate of KO-NO murine peritoneal cavities. Examination of splenic tissue revealed an accumulation of lymphocytes, predominantly CD8 T-cells, in experimental animal groups. KO-NO animals were found to have a predominance of granulomas 7 days post-inoculation, primarily in the lymph nodes. In addition, greater amounts of bacteria were recovered from the mesenteric lymph nodes of KO-NO mice. There was no statistically significant difference in the levels of total IgG and its subclasses 14 days post-inoculation between KO-NO and wild groups. The results suggest the importance of nitric oxide in the process of controlling CP infection, as KO-NO animals were observed to be markedly more affected by infection with this bacterium.(AU)
O objetivo deste estudo foi avaliar os aspectos da resposta imune do hospedeiro, mediante o uso de um modelo experimental de infecção de Corynebacterium pseudotuberculosis (CP) em camundongos C57/Black6 do tipo selvagem e em C57/Black6 knockout para o óxido nítrico (KO-NO). Foram avaliados 28 camundongos: quatro controles de tipo selvagem; 10 do tipo selvagem infectados com CP; quatro controles KO-NO; e 10 KO-NO infectados com CP. A infecção foi realizada via intraperitoneal, usando-se 107. Os animais C57/Black6 KO-NO infectados começaram a vir a óbito no quinto dia pós-inoculação, o que aconteceu até o 14º dia. Um número maior de neutrófilos foi encontrado na sua cavidade peritoneal. O exame do baço revelou um acúmulo de linfócitos, predominantemente células T CD8, nos grupos de animais experimentais. Nos animais KO-NO, foi observada a presença de granulomas, sete dias pós-inoculação, principalmente nos gânglios linfáticos. Além disso, uma maior quantidade de bactérias foi detectada dos linfonodos mesentéricos desses animais. Não houve diferença estatisticamente significante nos níveis séricos IgG total e em suas subclasses aos 14 dias pós-inoculação nos grupos KO-NO e selvagem. Os resultados obtidos sugerem a importância do óxido nítrico no processo de controle da infecção por CP.(AU)
Asunto(s)
Animales , Ratones , Corynebacterium pseudotuberculosis/inmunología , Linfadenitis/prevención & control , Ratones Noqueados/inmunología , Óxido Nítrico/uso terapéutico , Linfadenitis/veterinariaRESUMEN
NOTCH1 plays an important role in epithelial differentiation and carcinogenesis. To investigate the impact of Notch1 inactivation in oroesophageal epithelium, we generated conditional knockout (cKO) mice, using a combined construct which induces the expression of single guide RNA targeting Notch1 and Cas9 by the KRT14 promoter. The cKO mice exhibited patchy hair loss and multiple NOTCH1-negative areas in the tongue epithelium, indicative of heterogeneous knockout. The cKO mice showed susceptibility to esophageal tumorigenesis, underscoring Notch1 as a tumor suppressor. Our one-step strategy for generation of cKO mice provides a versatile method to examine a gene function in vivo.
Asunto(s)
Carcinogénesis/metabolismo , Modelos Animales de Enfermedad , Neoplasias Esofágicas/metabolismo , Ratones Noqueados/metabolismo , Receptor Notch1/metabolismo , Animales , Carcinogénesis/patología , Línea Celular Tumoral , Neoplasias Esofágicas/genética , Neoplasias Esofágicas/patología , Ratones , Ratones Noqueados/genética , Ratones Noqueados/inmunología , Receptor Notch1/genéticaRESUMEN
Estrogens, predominantly 17ß-estradiol, exert diverse effects throughout the body in both normal and pathophysiology, during development and in reproductive, metabolic, endocrine, cardiovascular, nervous, musculoskeletal and immune systems. Estrogen and its receptors also play important roles in carcinogenesis and therapy, particularly for breast cancer. In addition to the classical nuclear estrogen receptors (ERα and ERß) that traditionally mediate predominantly genomic signaling, the G protein-coupled estrogen receptor GPER has become recognized as a critical mediator of rapid signaling in response to estrogen. Mouse models, and in particular knockout (KO) mice, represent an important approach to understand the functions of receptors in normal physiology and disease. Whereas ERα KO mice display multiple significant defects in reproduction and mammary gland development, ERß KO phenotypes are more limited, and GPER KO exhibit no reproductive deficits. However, the study of GPER KO mice over the last six years has revealed that GPER deficiency results in multiple physiological alterations including obesity, cardiovascular dysfunction, insulin resistance and glucose intolerance. In addition, the lack of estrogen-mediated effects in numerous tissues of GPER KO mice, studied in vivo or ex vivo, including those of the cardiovascular, endocrine, nervous and immune systems, reveals GPER as a genuine mediator of estrogen action. Importantly, GPER KO mice have also demonstrated roles for GPER in breast carcinogenesis and metastasis. In combination with the supporting effects of GPER-selective ligands and GPER knockdown approaches, GPER KO mice demonstrate the therapeutic potential of targeting GPER activity in diseases as diverse as obesity, diabetes, multiple sclerosis, hypertension, atherosclerosis, myocardial infarction, stroke and cancer.
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Ratones Noqueados/genética , Receptores Acoplados a Proteínas G/genética , Animales , Enfermedades Cardiovasculares/genética , Enfermedades Cardiovasculares/inmunología , Enfermedades Cardiovasculares/metabolismo , Diabetes Mellitus/genética , Diabetes Mellitus/inmunología , Diabetes Mellitus/metabolismo , Modelos Animales de Enfermedad , Humanos , Inmunidad , Ratones , Ratones Noqueados/inmunología , Ratones Noqueados/fisiología , Neoplasias/genética , Neoplasias/inmunología , Neoplasias/metabolismo , Obesidad/genética , Obesidad/inmunología , Obesidad/metabolismo , Receptores de Estrógenos/genética , Receptores Acoplados a Proteínas G/inmunología , Receptores Acoplados a Proteínas G/metabolismoRESUMEN
In malaria, the evidence concerning the nucleotide-binding, oligomerization domain (NOD) 2 (NOD2) receptor is fragmented and the stimuli that might activate NOD2 are not well characterized. We investigated the role of NOD2 in vitro in the response of macrophages to Plasmodium falciparum products. Immortalized or primary bone marrow derived macrophages from wild type C57Bl/6 mice, or knockout mice for NOD2 or its adaptor proteins, were either primed with interferon gamma or left untreated, and stimulated with parasite products. Both lysates of infected erythrocytes or hemozoin induced higher levels of nitric oxide in primed than in unprimed wild type macrophages. When stimulated with hemozoin, primed macrophages knockout for NOD2, or for its adaptor proteins, produced significantly lower nitric oxide levels compared to wild type cells. Differently from hemozoin, the use of ß-hematin (synthetic hemozoin) as stimulus showed that NOD2 is dispensable. Furthermore, the production of inflammatory cytokines by wild type cells treated with hemozoin was not dependent on NOD2. These data indicate that parasite components present in the hemozoin, differently from ß-hematin, induce the production of nitric oxide through the activation of NOD2, whereas the production of inflammatory cytokines, like TNF-α or MIP-2 (CXCL2), seems to be NOD2 independent.
Asunto(s)
Hemoproteínas/inmunología , Malaria/inmunología , Proteína Adaptadora de Señalización NOD2/inmunología , Animales , Citocinas/metabolismo , Humanos , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados/inmunologíaRESUMEN
We have recently shown that wild type mice are highly tolerant, whereas thyrotropin receptor (TSHR) knockout (KO) mice are susceptible to immunization with the mouse TSHR, the autoantigen in Graves' disease. However, because TSHR KO mice lack the endogenous TSHR, Graves-like hyperthyroidism cannot be expected to occur in these mice. We therefore performed adoptive transfer of splenocytes from TSHR KO mice into nude mice expressing the endogenous TSHR. Anti-TSHR autoantibodies were detected in approximately 50 % recipient mice 4 wk after adoptive transfer of splenocytes (5 × 107/mouse) from TSHR KO mice immunized with adenovirus expressing mTSHR A subunit and persisted for 24 wk. Depletion of regulatory T cells by anti-CD25 antibody in the donor mice increased successful transfer rates without increasing antibody levels. Some recipient mice showed transient increases in thyroid-stimulating antibodies and T4 levels 4-8 wk after transfer, but many became thyroid-blocking antibody positive and hypothyroid 24 wk later. Adoptive transfer of splenocytes from naïve TSHR KO mice transiently induced very low antibody titers when the recipient mice were treated with anticytotoxic lymphocyte antigen 4 and antiprogrammed cell death 1 ligand 1 antibodies for 8 wk after transfer. Histologically, macrophages infiltrated the retrobulbar adipose tissues and extraocular muscles in a small fraction of the recipients. Our findings demonstrate successful adoptive transfer of anti-TSHR immune response from TSHR KO mice to nude mice. Although the recipient mice developed only transient and infrequent hyperthyroidism, followed by eventual hypothyroidism, induction of orbital inflammation suggests the possible role of anti-TSHR immune response for Graves' orbitopathy.
Asunto(s)
Traslado Adoptivo/métodos , Autoinmunidad/inmunología , Ratones Noqueados/inmunología , Ratones Desnudos/inmunología , Receptores de Tirotropina/deficiencia , Receptores de Tirotropina/inmunología , Bazo/inmunología , Animales , Anticuerpos Antiidiotipos/metabolismo , Modelos Animales de Enfermedad , Femenino , Enfermedad de Graves/complicaciones , Enfermedad de Graves/inmunología , Enfermedad de Graves/fisiopatología , Ratones , Ratones Endogámicos BALB C , Enfermedades Orbitales/etiología , Enfermedades Orbitales/inmunología , Enfermedades Orbitales/fisiopatología , Receptores de Tirotropina/genética , Bazo/citología , Glándula Tiroides/fisiopatología , TrasplantesRESUMEN
BACKGROUND: We wished to develop alternate production strategies to generate antibodies against traditionally problematic antigens. As a model we chose butyrylcholinesterase (BChE), involved in termination of cholinergic signaling, and widely considered as a poor immunogen. METHODOLOGY/PRINCIPAL FINDINGS: Jettisoning traditional laborious in silico searching methods to define putative epitopes, we simply immunized available BChE knock-out mice with full-length recombinant BChE protein (having been produced for crystallographic analysis). Immunization with BChE, in practically any form (recombinant human or mouse BChE, BChE purified from human serum, native or denatured), resulted in strong immune responses. Native BChE produced antibodies that favored ELISA and immunostaining detection. Denatured and reduced BChE were more selective for antibodies specific in Western blots. Two especially sensitive monoclonal antibodies were found capable of detecting 0.25 ng of BChE within one min by ELISA. One is specific for human BChE; the other cross-reacts with mouse and rat BChE. Immunization of wild-type mice served as negative controls. CONCLUSIONS/SIGNIFICANCE: We examined a simple, fast, and highly efficient strategy to produce antibodies by mining two expanding databases: namely those of knock-out mice and 3D crystallographic protein-structure analysis. We conclude that the immunization of knock-out mice should be a strategy of choice for antibody production.
Asunto(s)
Anticuerpos/inmunología , Técnicas Inmunológicas , Ratones Noqueados/inmunología , Animales , Formación de Anticuerpos , Butirilcolinesterasa/genética , Butirilcolinesterasa/inmunología , Células COS , Chlorocebus aethiops , Femenino , Humanos , Inmunización , Masculino , Ratones , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunologíaRESUMEN
Sustained intratumoral delivery of interleukin-12 (IL-12) and granulocyte macrophage colony-stimulating factor induces tumor regression via restoration of tumor-resident CD8+ T-effector/memory cell cytotoxicity and subsequent repriming of a secondary CD8+ T-effector cell response in tumor-draining lymph nodes (TDLN). However, treatment-induced T-effector activity is transient and is accompanied with a CD4+ CD25+ Foxp3+ T-suppressor cell rebound. Molecular and cellular changes in posttherapy tumor microenvironment and TDLN were monitored to elucidate the mechanism of counterregulation. Real-time PCR analysis revealed a 5-fold enhancement of indoleamine 2,3-dioxygenase (IDO) expression in the tumor and the TDLN after treatment. IDO induction required IFNgamma and persisted for up to 7 days. Administration of the IDO inhibitor D-1-methyl tryptophan concurrent with treatment resulted in a dramatic enhancement of tumor regression. Enhanced efficacy was associated with a diminished T-suppressor cell rebound, revealing a link between IDO activity and posttherapy regulation. Further analysis established that abrogation of the regulatory counterresponse resulted in a 10-fold increase in the intratumoral CD8+ T-cell to CD4+ Foxp3+ T-cell ratio. The ratio of proliferating CD8+ T-effector to CD4+ Foxp3+ T-suppressor cells was prognostic for efficacy of tumor suppression in individual mice. IFNgamma-dependent IDO induction and T-suppressor cell expansion were primarily driven by IL-12. These findings show a critical role for IDO in the regulation of IL-12-mediated antitumor immune responses.
Asunto(s)
Indolamina-Pirrol 2,3,-Dioxigenasa/inmunología , Interferón gamma/inmunología , Interleucina-12/inmunología , Ratones Noqueados/inmunología , Subgrupos de Linfocitos T/inmunología , Linfocitos T/inmunología , Animales , Citometría de Flujo , Tolerancia Inmunológica/inmunología , Linfocitos Infiltrantes de Tumor/inmunología , Ratones , Ratones Endogámicos BALB C , Neoplasias Experimentales , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Physiologic anti-inflammatory mechanisms are selected by evolution to control the immune system and to prevent infectious and inflammatory disorders. Central-acting alpha2-agonists attenuate systemic inflammation and improve survival in experimental sepsis. This anti-inflammatory and therapeutic mechanism of central sympatholytics appears to be mediated by an unexpected vagomimetic potential of the alpha2-agonists to activate the vagus nerve. Recent studies, however, rule out a cholinergic anti-inflammatory mechanism based on a direct cholinergic interaction between the vagus nerve and the immune system. Since the nervous system is the principal regulator of the immune system, physiologic studies understanding the neuroimmune connections can provide major advantages to design novel therapeutic strategies for sepsis.
Asunto(s)
Neuroinmunomodulación/fisiología , Sepsis/inmunología , Agonistas de Receptores Adrenérgicos alfa 2 , Animales , Ratones , Ratones Noqueados/inmunología , Receptores Adrenérgicos alfa 2/uso terapéutico , Sepsis/fisiopatología , Nervio Vago/efectos de los fármacos , Nervio Vago/inmunologíaRESUMEN
The risk of blindness caused by ocular toxoplasmosis supports efforts to improve our understanding for control of this disease. In this study, the involvement of CD8(+), CD4(+), B cell, and IL-10 gene in the immune response of primary ocular infection with the temperature-sensitive mutant (ts-4) of the RH Toxoplasma gondii strain, and in the protective immunity of ocular ts-4 vaccination and challenge with RH strain was investigated in murine models utilizing inbred C57BL/6 mice-deficient in CD4(+), CD8(+), B cells (microMT), or IL-10 gene. Compared to naive mice, all WT and mutant mice had different degree of ocular pathological changes after ts-4 ocular infection, in which both CD8 KO and IL-10 KO mice showed the most severe ocular lesions. Immunized by ts-4 intracameral (i.c.) inoculation, all mutant mice had partially decreased vaccine-induced resistance associated with increased ocular parasite burdens after RH strain challenge. A significant increase of the percentages of B cells and CD8(+) T cells in the draining lymph nodes were observed in WT and IL-10 KO mice after either infection or challenge. The levels of specific anti-toxoplasma IgG in both eye fluid and serum from all the mice were significantly increased after ts-4 i.c. immunization, except microMT mice. These results suggest that the avirulent ts-4 of T. gondii inoculated intracamerally can induce both ocular pathology and ocular protective immunity; CD4(+), CD8(+), B cell, and IL-10 gene are all necessary to the vaccine-induced resistance to ocular challenge by virulent RH strain, in which CD8(+) T cells are the most important component.
Asunto(s)
Ratones Noqueados/inmunología , Mutación , Vacunas Antiprotozoos/inmunología , Temperatura , Toxoplasma/genética , Toxoplasma/inmunología , Toxoplasmosis Ocular/parasitología , Animales , Linfocitos B , Linfocitos T CD4-Positivos , Linfocitos T CD8-positivos , Modelos Animales de Enfermedad , Ojo/inmunología , Ojo/parasitología , Fibroblastos , Humanos , Interleucina-10 , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados/parasitología , Toxoplasmosis Ocular/inmunología , Toxoplasmosis Ocular/fisiopatologíaAsunto(s)
Inmunidad Innata/inmunología , Ratones Noqueados , Receptores Toll-Like/inmunología , Animales , Historia del Siglo XX , Historia del Siglo XXI , Humanos , Inmunidad Innata/genética , Japón , Ratones , Ratones Noqueados/genética , Ratones Noqueados/inmunología , Receptores Toll-Like/genética , Receptores Toll-Like/historiaRESUMEN
The culmination of decades of research on humanized mice is leading to advances in our understanding of human haematopoiesis, innate and adaptive immunity, autoimmunity, infectious diseases, cancer biology and regenerative medicine. In this Review, we discuss the development of these new generations of humanized mice, how they will facilitate translational research in several biomedical disciplines and approaches to overcome the remaining limitations of these models.
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Quimera/inmunología , Ratones Noqueados/inmunología , Ratones SCID/inmunología , Ratones Transgénicos/inmunología , Animales , Quimera/genética , Enfermedades Transmisibles/inmunología , Modelos Animales de Enfermedad , Hematopoyesis/inmunología , Humanos , Ratones , Ratones Noqueados/genética , Ratones SCID/genética , Ratones Transgénicos/genética , Neoplasias/inmunologíaRESUMEN
In this study, p75NTREXONIII knockout mice were used as immune-naive hosts to produce functional antibodies to human p75NTR. Three monoclonal antibodies were produced and named MLR1, MLR2 and MLR3, and isotyped as IgG1, IgG2a and IgG2a, respectively. MLR1 and MLR2 bound to human p75NTR with higher affinity than the well-characterized ME20.4 in ELISA and also recognized p75NTR present on neurons in both rat and mouse. MLR1 and MLR2 bound to nerves known to express p75NTR following injection into Balb/C mice but not p75NTREXONIII knockout mice, indicating the antibodies are directed against the ligand binding extracellular region absent in knockout mice. Both MLR1 and MLR2 partially blocked NGF induced cell death in a mouse cell-line that expresses p75NTR but not TrKA. Importantly, intracerebroventricular injections indicated MLR2 was internalized within the cell bodies of mouse basal forebrain neurons, further demonstrating that this antibody is biologically active.
Asunto(s)
Anticuerpos Monoclonales/fisiología , Especificidad de Anticuerpos , Ratones Noqueados/inmunología , Receptor de Factor de Crecimiento Nervioso/genética , Receptor de Factor de Crecimiento Nervioso/inmunología , Animales , Reacciones Antígeno-Anticuerpo , Vasos Sanguíneos/metabolismo , Western Blotting/métodos , Células Cultivadas , Ensayo de Inmunoadsorción Enzimática , Femenino , Citometría de Flujo/métodos , Humanos , Inmunohistoquímica/métodos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratas , Ratas Sprague-DawleyRESUMEN
IL-1 receptor antagonist (IL-1Ra)-deficient mice spontaneously develop several inflammatory diseases, resembling rheumatoid arthritis, aortitis, and psoriasis in humans. As adoptive T cell transplantation could induce arthritis and aortitis in recipient mice, it was suggested that an autoimmune process is involved in the development of diseases. In contrast, as dermatitis developed in scid/scid-IL-IRa-deficient mice and could not be induced by T cell transfer, a T cell-independent mechanism was suggested. The expression of proinflammatory cytokines was augmented at the inflammatory sites. The development of arthritis and aortitis was significantly suppressed by the deficiency of TNFalpha or IL-17. The development of dermatitis was also inhibited by the deficiency of TNFalpha. These observations suggest that TNFalpha and IL-17 play a crucial role in the development of autoimmunity downstream of IL-1 signaling, and excess IL-1 signaling-induced TNFalpha also induces skin inflammation in a T cell-independent manner.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Interleucina-17/inmunología , Interleucina-1/inmunología , Ratones Noqueados/inmunología , Receptores de Interleucina-1/metabolismo , Transducción de Señal/fisiología , Factor de Necrosis Tumoral alfa/inmunología , Animales , Aortitis/inmunología , Aortitis/patología , Artritis Experimental/inmunología , Artritis Experimental/patología , Dermatitis/inmunología , Dermatitis/patología , Humanos , Ratones , Receptores de Interleucina-1/genéticaRESUMEN
Introducing lpr mutation prevents early mortality associated with IL-2Ralpha knockout (KO) mice, prompting us to determine the role of Fas in the immune system biology of IL-2Ralpha KO mice. Consistent with a defect in CD4+CD25+ regulatory T (Treg) cell expression, spontaneous lymphocyte activation in lymphoid organs was observed in 6-wk-old mice. In 16- to 22-wk-old mice, infiltration of leukocytes was observed in bone marrow, colon, lung, pancreas, lacrimal gland, and salivary gland, but not in heart, thyroid, liver, stomach, small intestine, ovary, and kidney. In the lymphocytes-infiltrated bone marrow, B cell lymphopoiesis was blocked at pro-B to pre-B/immature B stage, culminating in an age-dependent B cell loss in the periphery. These phenotypes were also observed in IL-2Ralpha KO mice bearing the lpr mutation (DM mice), indicating Treg cell function and the phenotypes attributed directly to Treg cell abnormality are largely Fas-independent. However, anemia and body weight loss were partially prevented, tissue cell apoptosis was inhibited, and lifespan was improved in the DM mice, demonstrating Fas-dependent elements in these processes. Our age-dependent, lifelong analysis of IL-2Ralpha KO and DM mice supports a CD4+CD25+ Treg cell-based mechanism for the abnormal immune system biology observed in IL-2Ralpha KO mice and provides a global view of the interplays among Treg cells, multiorgan inflammation, hemopoiesis, and apoptosis.
Asunto(s)
Apoptosis/inmunología , Hematopoyesis/inmunología , Ratones Noqueados/inmunología , Receptores de Interleucina/deficiencia , Receptores de Interleucina/genética , Linfocitos T Reguladores/inmunología , Receptor fas/fisiología , Animales , Apoptosis/genética , Subgrupos de Linfocitos B/inmunología , Subgrupos de Linfocitos B/patología , Células de la Médula Ósea/inmunología , Células de la Médula Ósea/patología , Diferenciación Celular/genética , Diferenciación Celular/inmunología , Colon/inmunología , Colon/metabolismo , Colon/patología , Femenino , Hematopoyesis/genética , Deficiencia de IgG/genética , Deficiencia de IgG/inmunología , Deficiencia de IgG/patología , Inflamación/genética , Inflamación/inmunología , Inflamación/prevención & control , Subunidad alfa del Receptor de Interleucina-2 , Longevidad/genética , Longevidad/inmunología , Pulmón/inmunología , Pulmón/metabolismo , Pulmón/patología , Ganglios Linfáticos/inmunología , Ganglios Linfáticos/metabolismo , Ganglios Linfáticos/patología , Activación de Linfocitos/genética , Recuento de Linfocitos , Linfopenia/genética , Linfopenia/inmunología , Linfopenia/patología , Trastornos Linfoproliferativos/genética , Trastornos Linfoproliferativos/inmunología , Trastornos Linfoproliferativos/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos MRL lpr , Ratones Noqueados/genética , Receptores de Interleucina/fisiología , Análisis de Supervivencia , Linfocitos T Reguladores/patología , Receptor fas/genéticaRESUMEN
We generated a novel tumor necrosis factor (TNF) null mutation using Cre-loxP technology. Mice homozygous for this mutation differ from their "conventional" counterparts; in particular, they completely lack Peyer's patches (PP) but retain all lymph nodes. Our analysis of these novel TNF-knockout mice supports the previously disputed notion of the involvement of TNF-TNFR1 signaling in PP organogenesis. Availability of TNF-knockout strains both with and without PP enables more definitive studies concerning the roles of TNF and PP in various immune functions and disease conditions. Here, we report that systemic ablation of TNF, but not the presence of PP per se, is critical for protection against intestinal Listeria infection in mice.
Asunto(s)
Ratones Noqueados/inmunología , Ganglios Linfáticos Agregados/inmunología , Factor de Necrosis Tumoral alfa/deficiencia , Factor de Necrosis Tumoral alfa/genética , Animales , Citometría de Flujo , Hematopoyesis/inmunología , Inmunohistoquímica , Listeriosis/inmunología , Ratones , Ratones Noqueados/genética , Mutación , Ganglios Linfáticos Agregados/patología , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
The dopamine transporter (DAT) is responsible for the re-uptake of dopamine into presynaptic nerve terminals and thereby controls dopaminergic neurotransmission. Deletion of DAT results in a hyperdopaminergic phenotype and DAT(-/-) mice are characterized by pituitary hypoplasia, impaired maternal behavior, and increased locomotion. From earlier studies, we have evidence that the activity of the central dopaminergic system may play a role in determining immune reactivity and disease susceptibility. To further explore the functional relation between the dopaminergic system and the immune system, we investigated the activity of the immune system in DAT(-/-) mice. We show that in vitro, splenocytes from DAT(-/-) mice displayed reduced natural killer cell activity and reduced mitogen-induced cytokine responses. In contrast, LPS-induced cytokine production by macrophages was enhanced. In vivo, the cellular response to immunization with ovalbumine (OVA-induced delayed type hypersensitivity response) was significantly reduced. Interestingly, the OVA-induced humoral response (anti-OVA IgG) was increased in DAT(-/-) mice compared to wild-type animals. Plasma levels of catecholamines and corticosterone did not differ significantly between DAT(-/-) and wild-type animals. In conclusion, we show in the present study that interfering with the dopaminergic system has major consequences for both the acquired and the innate immune response.
Asunto(s)
Sistema Inmunológico/fisiología , Glicoproteínas de Membrana/deficiencia , Proteínas de Transporte de Membrana/deficiencia , Ratones Noqueados/inmunología , Proteínas del Tejido Nervioso/deficiencia , Análisis de Varianza , Animales , Antígenos CD/metabolismo , Catecolaminas/sangre , Proliferación Celular , Células Cultivadas , Corticosterona/sangre , Citocinas/metabolismo , Proteínas de Transporte de Dopamina a través de la Membrana Plasmática , Citometría de Flujo/métodos , Inflamación/inducido químicamente , Inflamación/genética , Interleucina-10/metabolismo , Células Asesinas Naturales/efectos de los fármacos , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/metabolismo , Lipopolisacáridos/farmacología , Macrófagos/inmunología , Macrófagos/metabolismo , Glicoproteínas de Membrana/genética , Proteínas de Transporte de Membrana/genética , Ratones , Proteínas del Tejido Nervioso/genética , Radioinmunoensayo/métodos , Bazo/citología , Linfocitos T/efectos de los fármacos , Linfocitos T/metabolismoRESUMEN
The thymus is the primary site of T cell lymphopoiesis. To undergo proper differentiation, developing T cells follow a well-ordered genetic program that strictly depends on the heterogeneous and highly specialized thymic microenvironment. In this study, we used microarray technology to extensively describe transcriptional events regulating alphabeta T cell fate. To get an integrated view of these processes, both whole thymi from genetically engineered mice together with purified thymocytes were analyzed. Using mice exhibiting various transcriptional perturbations and developmental blockades, we performed a transcriptional microdissection of the organ. Multiple signatures covering both cortical and medullary stroma as well as various thymocyte maturation intermediates were clearly defined. Beyond the definition of histological and functional signatures (proliferation, rearrangement), we provide the first evidence that such an approach may also highlight the complex cross-talk events that occur between maturing T cells and stroma. Our data constitute a useful integrated resource describing the main gene networks set up during thymocyte development and a first step toward a more systematic transcriptional analysis of genetically modified mice.
Asunto(s)
Ratones Noqueados/genética , Ratones Noqueados/inmunología , Modelos Animales , Subgrupos de Linfocitos T/citología , Subgrupos de Linfocitos T/inmunología , Timo/citología , Timo/inmunología , Factores de Transcripción/genética , Animales , Diferenciación Celular/genética , Diferenciación Celular/inmunología , Línea Celular , Línea Celular Transformada , Proliferación Celular , ADN Helicasas , Perfilación de la Expresión Génica/métodos , Reordenamiento Génico de Linfocito T , Genes Codificadores de la Cadena alfa de los Receptores de Linfocito T/genética , Leucemia P388 , Ratones , Ratones Endogámicos C57BL , Familia de Multigenes/inmunología , Proteínas Nucleares/biosíntesis , Proteínas Nucleares/genética , Análisis de Secuencia por Matrices de Oligonucleótidos/métodos , Proteínas Proto-Oncogénicas/deficiencia , Proteínas Proto-Oncogénicas/genética , Receptor Notch1 , Receptores de Superficie Celular/biosíntesis , Receptores de Superficie Celular/genética , Receptores de Interleucina-2/biosíntesis , Células del Estroma/inmunología , Células del Estroma/metabolismo , Subgrupos de Linfocitos T/metabolismo , Timo/metabolismo , Factor de Transcripción ReIB , Factores de Transcripción/biosíntesis , Factores de Transcripción/deficiencia , Factores de Transcripción/fisiología , Regulación hacia Arriba/genética , Regulación hacia Arriba/inmunologíaRESUMEN
The innate immune response is a first-line defense system in which individual Toll-like receptors (TLRs) recognize distinct pathogen-associated molecular patterns (PAMPs) and exert subsequent immune responses against a variety of pathogens. TLRs are composed of an extracellular leucine-rich repeat (LRR) domain and a cytoplasmic domain that is homologous to that of the IL-IR family. Upon stimulation, TLR recruits a cytoplasmic adaptor molecule MyD88, then IL-IR-associated kinase (IRAK), and finally induces activation of NF-kappaB and MAP kinases. However, the responses to TLR ligands differ, indicating the diversity of TLR signaling pathways. Besides MyD88, several novel adaptor molecules have recently been identified. Differential utilization of these adaptor molecules may provide the specificity in the TLR signaling.
Asunto(s)
Glicoproteínas de Membrana/inmunología , Ratones Noqueados/inmunología , Receptores de Superficie Celular/inmunología , Proteínas Adaptadoras Transductoras de Señales , Animales , Antígenos de Diferenciación/genética , Antígenos de Diferenciación/inmunología , Glicoproteínas de Membrana/deficiencia , Glicoproteínas de Membrana/genética , Ratones , Factor 88 de Diferenciación Mieloide , Receptores de Superficie Celular/deficiencia , Receptores de Superficie Celular/genética , Receptores Inmunológicos/genética , Receptores Inmunológicos/inmunología , Receptores Toll-LikeRESUMEN
Metallothionein (MT) is a low molecular weight protein, rich in cystein residues, and induced by stimulation with several stresses such as heavy metals, chemical agents, oxygen radicals and irradiation. It is involved in metal metabolism, detoxification and radical scavenging and also participates in immune responses. In this communication, to study the role of MT in immune responses, we analyzed immune functions, especially macrophage functions in MT-knock out (MT-KO) mice in vitro. When compared with wild type counterpart (MT-WT) mice, macrophages from MT-KO mice showed a defect of phagocytic and antigen-presenting activity. Cytokine productions such as IL-1alpha, IL-6, IL-10 and IL-12 were reduced in macrophages from MT-KO mice. Furthermore, the expressions of CD80, CD86 and MHC class II molecules were also reduced in macrophages from MT-KO mice. No obvious dysfunction was observed in T cells and B cells. These results suggest that MT plays an important role in the regulation of immune responses, especially macrophage functions.