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1.
Vet Pathol ; 57(6): 915-925, 2020 11.
Artículo en Inglés | MEDLINE | ID: mdl-33016243

RESUMEN

Mouse kidney parvovirus (MKPV), also known as murine chapparvovirus (MuCPV), is an emerging, highly infectious agent that has been isolated from laboratory and wild mouse populations. In immunocompromised mice, MKPV produces severe chronic interstitial nephropathy and renal failure within 4 to 5 months of infection. However, the course of disease, severity of histologic lesions, and viral shedding are uncertain for immunocompetent mice. We evaluated MKPV infections in CD-1 and Swiss Webster mice, 2 immunocompetent stocks of mice. MKPV-positive CD-1 mice (n = 30) were identified at approximately 8 weeks of age by fecal PCR (polymerase chain reaction) and were subsequently housed individually for clinical observation and diagnostic sampling. Cage swabs, fecal pellets, urine, and blood were evaluated by PCR at 100 and 128 days following the initial positive test, which identified that 28 of 30 were persistently infected and 24 of these were viremic at 100 days. Histologic lesions associated with MKPV in CD-1 (n = 31) and Swiss mice (n = 11) included lymphoplasmacytic tubulointerstitial nephritis with tubular degeneration. Inclusion bodies were rare; however, intralesional MKPV mRNA was consistently detected via in situ hybridization within tubular epithelial cells of the renal cortex and within collecting duct lumina. In immunocompetent CD-1 mice, MKPV infection resulted in persistent shedding of virus for up to 10 months and a mild tubulointerstitial nephritis, raising concerns that this virus could produce study variations in immunocompetent models. Intranuclear inclusions were not a consistent feature of MKPV infection in immunocompetent mice.


Asunto(s)
Nefritis Intersticial , Infecciones por Parvoviridae , Parvovirinae , Enfermedades de los Roedores , Animales , Riñón , Ratones , Ratones Endogámicos , Nefritis Intersticial/veterinaria , Infecciones por Parvoviridae/veterinaria , Parvovirinae/patogenicidad
2.
J Am Assoc Lab Anim Sci ; 63(1): 49-56, 2024 01 01.
Artículo en Inglés | MEDLINE | ID: mdl-38191146

RESUMEN

Alfaxalone is a commonly used injectable anesthetic in dogs and cats due to its minimal cardiovascular side effects. Data for its use in mice are limited and demonstrate strain- and sex-associated differences in dose-response relationships. We performed a dose-comparison study of alfaxalone-xylazine-buprenorphine (AXB) in Crl: CFW (SW) mice. Subcutaneous injection of 50 mg/kg alfaxalone-10 mg/kg xylazine-0.1 mg/kg buprenorphine HCl consistently achieved a surgical plane of anesthesia (loss of toe pinch) for 48.6 ± 4.7 and 60.8 ± 9.6 min in females and males, respectively. The same dose and route of AXB induced a surgical plane of anesthesia in C57Bl/6NCrl (females: 42.3 ± 11.2 min; males: 51.6 ± 12.3 min), NCr-Foxn1nu (females: 76.8 ± 32.5 min; males: 80.0 ± 1.2 min), and NOD. Cg-Prkdc SCID Il2rg tm1Wjl /SzJCr (females: 56.0 ± 37.2 min and males: 61.2 ± 10.2 min) mice. We found no significant difference in the duration of the surgical plane of anesthesia between males and females within the mouse strains Crl: CFW (SW), C57Bl/6NCrl, NCr-Foxn1nu, and NOD. Cg-PrkdcSCID Il2rgtm1Wjl /SzJCr. We next performed an echocardiography study (n = 5 per group) of Crl: CFW (SW) mice ( n = 5 per group) to compare subcutaneous AXB anesthesia with that produced by intraperitoneal injection of 100 mg/kg ketamine and 10 mg/kg xylazine (KX). AXB induced significantly less bradycardia (295.4 ± 29 bpm) than KX (185.8 ± 38.9 bpm) did, with no significant differences in cardiac output, ejection fraction, end-diastolic volume, end-systolic volume, or fractional shortening. These results suggest that subcutaneous administration of AXB is a viable alternative to KX for inducing a surgical plane of anesthesia in Crl: CFW (SW), C57Bl/6NCrl, NCr-Foxn1nu, and NOD. Cg-PrkdcSCID Il2rgtm1Wjl /SzJCr mice, regardless of sex. AXB may also be a better injectable anesthetic option as compared with KX for avoiding adverse cardiac effects in mice.


Asunto(s)
Anestesia , Anestésicos , Buprenorfina , Enfermedades de los Gatos , Enfermedades de los Perros , Pregnanodionas , Enfermedades de los Roedores , Masculino , Femenino , Ratones , Animales , Gatos , Perros , Xilazina/farmacología , Enfermedades de los Gatos/tratamiento farmacológico , Ratones Endogámicos NOD , Ratones SCID , Enfermedades de los Perros/tratamiento farmacológico , Anestésicos/farmacología , Anestesia/veterinaria , Ecocardiografía/veterinaria , Enfermedades de los Roedores/tratamiento farmacológico
3.
Vet Parasitol ; 148(3-4): 219-30, 2007 Sep 30.
Artículo en Inglés | MEDLINE | ID: mdl-17651896

RESUMEN

Neospora caninum, an obligate intracellular protozoan parasite, is the causative agent of bovine neosporosis, an important disease affecting the reproductive performance of cattle worldwide. Currently there is no effective vaccine available to prevent N. caninum infection in cattle. In this study, we examined the feasibility of developing a live, recombinant N. caninum vaccine using Brucella abortus vaccine strain RB51 as the expression and delivery vector. We generated two recombinant RB51 strains each expressing SRS2 (RB51/SRS2) or GRA7 (RB51/GRA7) antigens of N. caninum. BALB/c mice immunized by single intraperitoneal inoculation of the recombinant RB51 strains developed IgG antibodies specific to the respective N. caninum antigen. In vitro stimulation of splenocytes from the vaccinated mice with specific antigen resulted in the production of interferon-gamma, but not IL-5 or IL-10, suggesting the development of a Th1 type immune response. Upon challenge with N. caninum tachyzoites, mice vaccinated with strain RB51/SRS2, but not RB51/GRA7, showed significant resistance to cerebral infection when compared to the RB51 vaccinated mice, as determined by the tissue parasite load using a real-time quantitative TaqMan assay. Interestingly, mice vaccinated with either strain RB51 or RB51/GRA7 also contained significantly lower parasite burden in their brains compared to those inoculated with saline. Mice vaccinated with strain RB51/SRS2 or RB51/GRA7 were protected to the same extent as the strain RB51 vaccinated mice against challenge with B. abortus virulent strain 2308. These results suggest that a recombinant RB51 strain expressing an appropriate protective antigen(s), such as SRS2 of N. caninum, can confer protection against both neosporosis and brucellosis.


Asunto(s)
Antígenos de Protozoos/inmunología , Antígenos de Superficie/inmunología , Vacuna contra la Brucelosis/inmunología , Brucella abortus/genética , Coccidiosis/inmunología , Neospora/inmunología , Proteínas Protozoarias/inmunología , Vacunas Antiprotozoos/inmunología , Animales , Anticuerpos Antiprotozoarios/sangre , Antígenos de Protozoos/genética , Antígenos de Superficie/genética , Coccidiosis/parasitología , Coccidiosis/prevención & control , Escherichia coli/genética , Femenino , Interferón gamma/metabolismo , Ratones , Ratones Endogámicos BALB C , Neospora/genética , Proteínas Protozoarias/genética , Distribución Aleatoria , Proteínas Recombinantes/inmunología , Bazo/citología , Bazo/inmunología , Vacunas de ADN/inmunología
4.
Health Phys ; 111(2): 160-8, 2016 08.
Artículo en Inglés | MEDLINE | ID: mdl-27356060

RESUMEN

Decisions on whether to start a therapeutic intervention for management of the Acute Radiation Syndrome (ARS) should be made early after exposure, and it should be based on readily available clinical signs and laboratory parameters. Here, the authors use the minipig to assess if early prediction of the later developing clinical outcome and necessity of therapeutic interventions can be determined within the first 3 d after exposure and whether it is comparable to human data. Retrospective analysis of data accumulated in the period 2009-2012 was used. Male Göttingen minipigs (age 4-5 mo, weight 9-10 kg) were irradiated (or sham-irradiated) bilaterally with gamma-photons (Co, 0.5-0.6 Gy min) in the dose range of 1.6-12 Gy. Complete blood counts, serum chemistry, and clinical symptoms were collected up to 60 d after irradiation in untreated minipigs. Changes in these early parameters (up to 3 d after exposure) were correlated with later occurrence (10-60 d after irradiation) of (1) hematological severity scores, (2) severe thrombocytopenia, (3) severe neutropenia, as well as need for (4) therapeutic intervention, (5) administration of cytokines/antibiotics, or (6) thrombocyte transfusions. Binary endpoints were analyzed using logistic regression analysis and calculating receiver operating characteristic (ROC) curves. Most predictive were decreased lymphocyte counts and increases in body temperature at 3 h after irradiation. These data corroborate earlier findings performed on human radiation victims suffering from severe hematological syndrome and provide further evidence for the suitability of the minipig model as a potential alternative non-rodent animal model.


Asunto(s)
Síndrome de Radiación Aguda/diagnóstico por imagen , Síndrome de Radiación Aguda/terapia , Diagnóstico Precoz , Evaluación de Resultado en la Atención de Salud/métodos , Evaluación de Síntomas/métodos , Índices de Gravedad del Trauma , Síndrome de Radiación Aguda/sangre , Animales , Biomarcadores/sangre , Recuento de Células Sanguíneas , Sistemas de Apoyo a Decisiones Clínicas , Masculino , Pronóstico , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Porcinos , Porcinos Enanos , Irradiación Corporal Total
5.
PLoS One ; 11(8): e0161079, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-27579862

RESUMEN

Acute radiation sickness (ARS) following exposure to ionizing irradiation is characterized by radiation-induced multiorgan dysfunction/failure that refers to progressive dysfunction of two or more organ systems, the etiological agent being radiation damage to cells and tissues over time. Radiation sensitivity data on humans and animals has made it possible to describe the signs associated with ARS. A mouse model of total-body irradiation (TBI) has previously been developed that represents the likely scenario of exposure in the human population. Herein, we present the Mouse Intervention Scoring System (MISS) developed at the Veterinary Sciences Department (VSD) of the Armed Forces Radiobiology Research Institute (AFRRI) to identify moribund mice and decrease the numbers of mice found dead, which is therefore a more humane refinement to death as the endpoint. Survival rates were compared to changes in body weights and temperatures in the mouse (CD2F1 male) TBI model (6-14 Gy, 60Co γ-rays at 0.6 Gy min-1), which informed improvements to the Scoring System. Individual tracking of animals via implanted microchips allowed for assessment of criteria based on individuals rather than by group averages. From a total of 132 mice (92 irradiated), 51 mice were euthanized versus only four mice that were found dead (7% of non-survivors). In this case, all four mice were found dead after overnight periods between observations. Weight loss alone was indicative of imminent succumbing to radiation injury, however mice did not always become moribund within 24 hours while having weight loss >30%. Only one survivor had a weight loss of greater than 30%. Temperature significantly dropped only 2-4 days before death/euthanasia in 10 and 14 Gy animals. The score system demonstrates a significant refinement as compared to using subjective assessment of morbidity or death as the endpoint for these survival studies.


Asunto(s)
Síndrome de Radiación Aguda , Rayos gamma/efectos adversos , Traumatismos Experimentales por Radiación , Irradiación Corporal Total , Síndrome de Radiación Aguda/patología , Síndrome de Radiación Aguda/fisiopatología , Animales , Relación Dosis-Respuesta en la Radiación , Masculino , Ratones , Traumatismos Experimentales por Radiación/patología , Traumatismos Experimentales por Radiación/fisiopatología
6.
Comp Med ; 66(2): 150-3, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-27053570

RESUMEN

This case report describes a rhesus macaque (Macaca mulatta; male; age, 5 y; weight, 6.7 kg) with anorexia, dehydration, lethargy, ataxia, and generalized skin rashes that occurred 30 d after total-body irradiation at 6.5 Gy ((60)Co γ-rays). Physical examination revealed pale mucus membranes, a capillary refill time of 4 s, heart rate of 180 bpm. and respirations at 50 breaths per minute. Diffuse multifocal maculopapulovesicular rashes were present on the body, including mucocutaneous junctions. The CBC analysis revealed a Hct of 48%, RBC count of 6.2 × 10(6)/µL, platelet count of 44 × 10(3)/µL, and WBC count of 25 × 10(3)/µL of WBC. The macaque was euthanized in light of a grave prognosis. Gross examination revealed white foci on the liver, multifocal generalized petechiation on serosal and mucosal surfaces of the gastrointestinal tract, hemorrhagic lymph nodes, and hemorrhagic fluid in the thoracic cavity. Microscopic examination revealed cutaneous vesicular lesions with intranuclear eosinophilic viral inclusions within the epithelial cells, consistent with herpesvirus. Immunohistochemistry was positive for herpesvirus. The serum sample was negative for antibodies against Macacine herpesvirus 1 and Cercopithecine herpesvirus 9 (simian varicella virus, SVV). Samples submitted for PCR-based identification of the etiologic agent confirmed the presence of SVV DNA. PCR analysis, immunohistochemistry, and histology confirmed that lesions were attributed to an active SVV infection in this macaque. This case illustrates the importance of screening for SVV in rhesus macaques, especially those used in studies that involve immunosuppressive procedures.


Asunto(s)
Infecciones por Herpesviridae/veterinaria , Herpesvirus Cercopitecino 1/efectos de la radiación , Macaca mulatta , Enfermedades de los Monos/patología , Irradiación Corporal Total/efectos adversos , Animales , Infecciones por Herpesviridae/patología , Infecciones por Herpesviridae/virología , Inmunohistoquímica , Macaca mulatta/virología , Masculino , Enfermedades de los Monos/virología
7.
Radiat Res ; 185(3): 285-98, 2016 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-26930378

RESUMEN

The search for treatments to counter potentially lethal radiation-induced injury over the past several decades has led to the development of multiple classes of radiation countermeasures. However, to date only granulocyte colony-stimulating factor (G-CSF; filgrastim, Neupogen)and pegylated G-CSF (pegfilgrastim, Neulasta) have been approved by the United States Food and Drug Administration (FDA) for the treatment of hematopoietic acute radiation syndrome (ARS). Gamma-tocotrienol (GT3) has demonstrated strong radioprotective efficacy in the mouse model, indicating the need for further evaluation in a large animal model. In this study, we evaluated GT3 pharmacokinetics (PK) and efficacy at different doses of cobalt-60 gamma radiation (0.6 Gy/min) using the nonhuman primate (NHP) model. The PK results demonstrated increased area under the curve with increasing drug dose and half-life of GT3. GT3 treatment resulted in reduced group mean neutropenia by 3-5 days and thrombocytopenia by 1-5 days. At 5.8 and 6.5 Gy total-body irradiation, GT3 treatment completely prevented thrombocytopenia. The capability of GT3 to reduce severity and duration of neutropenia and thrombocytopenia was dose dependent; 75 mg/kg treatment was more effective than 37.5 mg/kg treatment after a 5.8 Gy dose. However, the higher GT3 dose (75 mg/kg) was associated with higher frequency of adverse skin effects (small abscess) at the injection site. GT3 treatment of irradiated NHPs caused no significant difference in animal survival at 60 days postirradiation, however, low mortality was observed in irradiated, vehicle-treated groups as well. The data from this pilot study further elucidate the role and pharmacokinetics of GT3 in hematopoietic recovery after irradiation in a NHP model, and demonstrate the potential of GT3 as a promising radioprotector.


Asunto(s)
Síndrome de Radiación Aguda/tratamiento farmacológico , Cromanos/administración & dosificación , Primates , Protectores contra Radiación/administración & dosificación , Vitamina E/análogos & derivados , Síndrome de Radiación Aguda/sangre , Síndrome de Radiación Aguda/patología , Animales , Cromanos/sangre , Cromanos/farmacocinética , Radioisótopos de Cobalto , Modelos Animales de Enfermedad , Relación Dosis-Respuesta en la Radiación , Rayos gamma , Humanos , Macaca mulatta , Protectores contra Radiación/farmacocinética , Trombocitopenia/etiología , Trombocitopenia/patología , Estados Unidos , Vitamina E/administración & dosificación , Vitamina E/sangre , Vitamina E/farmacocinética , Irradiación Corporal Total
8.
Radiat Prot Dosimetry ; 172(1-3): 174-191, 2016 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-27473690

RESUMEN

An assessment of multiple biomarkers from radiation casualties undergoing limited- or full-supportive care including treatment with filgrastim is critical to develop rapid and effective diagnostic triage strategies. The efficacy of filgrastim with full-supportive care was compared with results with limited-supportive care by analyzing survival, necropsy, histopathology and serial blood samples for hematological, serum chemistry and protein profiles in a non-human primate (Macaca mulatta, male and female) model during 60-d post-monitoring period following sham- and total-body irradiation with 6.5 Gy 60Co gamma-rays at 0.6 Gy min-1 Filgrastim (10 µg kg-1) was administered beginning on Day 1 post-exposure and continued daily until neutrophil counts were ≥2,000 µL-1 for two consecutive days. Filgrastim and full-supportive care significantly decreased the pancytopenia duration and resulted in improved animal survival and recovery compared to animals with a limited-supportive care. These findings also identified and validated a multiparametric biomarker panel to support radiation diagnostic device development.


Asunto(s)
Bioensayo/métodos , Modelos Animales de Enfermedad , Filgrastim/uso terapéutico , Traumatismos por Radiación/diagnóstico , Traumatismos por Radiación/terapia , Monitoreo de Radiación/métodos , Irradiación Corporal Total/métodos , Animales , Biomarcadores/sangre , Femenino , Macaca mulatta , Masculino , Dosis de Radiación , Exposición a la Radiación/análisis , Traumatismos por Radiación/sangre , Protectores contra Radiación/uso terapéutico , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Resultado del Tratamiento
9.
Health Phys ; 106(6): 727-33, 2014 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-24776906

RESUMEN

The minipig is emerging as a potential alternative non-rodent animal model. Several biological markers (e.g., blood counts, laboratory parameters, and clinical signs) have been proposed for rapid triage of radiation victims. Here, the authors focus on the significance of bio-indicators for prediction of survivors after irradiation and compare it with human data; the relationship between these biomarkers and radiation dose is not part of this study. Male Göttingen minipigs (age 4-5 mo, weight 9-10 kg) were irradiated (or sham-irradiated) bilaterally with gamma-photons (6°Co, 0.5-0.6 Gy min⁻¹) in the dose range of 1.6-12 Gy. Peripheral blood cell counts, laboratory parameters, and clinical symptoms were collected up to 10 d after irradiation and analyzed using logistic regression analysis and calculating ROC curves. In moribund pigs, parameters such as decreased lymphocyte/granulocyte counts, increased C-reactive protein, alkaline phosphatase values, as well as increased citrulline values and body temperature, significantly (p < 0.002 up to p < 0.0001) discriminated non-survivors from survivors with high precision (ROC > 0.8). However, most predictive within the first 3 d after exposure was a combination of decreased lymphocyte counts and increased body temperature observed as early as 3 h after radiation exposure (ROC: 0.93-0.96, p < 0.0001). Sham-irradiated animals (corresponding to "worried wells") could be easily discriminated from dying pigs, thus pointing to the diagnostic significance of this analysis. These data corroborate with earlier findings performed on human radiation victims suffering from severe hematological syndrome and provide further evidence for the suitability of the minipig model as a potential alternative non-rodent animal model.


Asunto(s)
Traumatismos Experimentales por Radiación/diagnóstico , Estadística como Asunto/métodos , Porcinos Enanos , Animales , Biomarcadores , Análisis Discriminante , Modelos Animales de Enfermedad , Recuento de Linfocitos , Masculino , Pronóstico , Traumatismos Experimentales por Radiación/sangre , Reproducibilidad de los Resultados , Porcinos , Factores de Tiempo , Triaje
10.
Comp Med ; 64(6): 456-63, 2014 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-25527026

RESUMEN

In the absence of supportive care, exposing Göttingen minipigs to γ-radiation doses of less than 2 Gy achieves lethality due to hematopoietic acute radiation syndrome. Doses of 2 to 5 Gy are associated with an accelerated hematopoietic syndrome, characterized by villus blunting and fusion, the beginning of sepsis, and a mild transient reduction in plasma citrulline concentration. We exposed male Göttingen minipigs (age, 5 mo; weight, 9 to 11 kg) to γ-radiation doses of 5 to 12 Gy (total body; (60)Co, 0.6 Gy/min) to test whether these animals exhibit classic gastrointestinal acute radiation syndrome (GI-ARS). After exposure, the minipigs were monitored for 10 d by using clinical signs, CBC counts, and parameters associated with the development of the gastrointestinal syndrome. Göttingen minipigs exposed to γ radiation of 5 to 12 Gy demonstrate a dose-dependent occurrence of all parameters classically associated with acute GI-ARS. These results suggest that Göttingen minipigs may be a suitable model for studying GI-ARS after total body irradiation, but the use of supportive care to extend survival beyond 10 d is recommended. This study is the first step toward determining the feasibility of using Göttingen minipigs in testing the efficacy of candidate drugs for the treatment of GI-ARS after total body irradiation.


Asunto(s)
Síndrome de Radiación Aguda/patología , Modelos Animales de Enfermedad , Rayos gamma/efectos adversos , Enfermedades Gastrointestinales/patología , Porcinos Enanos , Animales , Citrulina/sangre , Relación Dosis-Respuesta en la Radiación , Determinación de Punto Final , Técnicas Histológicas , Modelos Lineales , Masculino , Porcinos
11.
J Virol Methods ; 162(1-2): 231-5, 2009 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-19729039

RESUMEN

A TaqMan probe-based real-time RT-PCR assay was developed for simultaneous detection of RNA of transmissible gastroenteritis virus (TGEV) in pig fecal samples and RNA of enhanced green fluorescent protein (EGFP) added exogenously as an internal amplification control. The TGEV primers and probe were designed to be specific to a portion of the S gene sequence conserved in all TGEV isolates, but absent in the closely related porcine respiratory coronaviruses. The optimized TaqMan assay detected a minimum of 2.8 copies of in vitro transcribed RNA of the target S gene and RNA extracted from 1 TCID50/ml of TGEV. Using 113 clinical samples received at our diagnostic laboratory over a 4-year period, the performance of the assay was tested and compared with that of a previously described nested RT-PCR assay. All the fecal samples which tested positive for TGEV by the nested RT-PCR assay also tested positive by the TaqMan assay. However, approximately 9% of the samples that tested negative by the nested RT-PCR assay tested positive by the TaqMan assay. These results indicate that the developed TaqMan assay is a highly sensitive diagnostic test for rapid detection of TGEV in pig fecal samples.


Asunto(s)
Heces/virología , Gastroenteritis Porcina Transmisible , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Enfermedades de los Porcinos , Polimerasa Taq , Virus de la Gastroenteritis Transmisible/aislamiento & purificación , Animales , Gastroenteritis Porcina Transmisible/diagnóstico , Gastroenteritis Porcina Transmisible/virología , Control de Calidad , ARN Viral/análisis , ARN Viral/aislamiento & purificación , Porcinos , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/virología , Factores de Tiempo , Virus de la Gastroenteritis Transmisible/genética
12.
Comp Med ; 59(6): 534-44, 2009 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-20034428

RESUMEN

Infection of mice with Helicobacter hepaticus is common in research colonies, yet little is known about how this persistent infection affects immunologic research. The goal of this study was to determine whether H. hepaticus infection status can modulate immune responses specific to herpes simplex virus type 1 (HSV1) and the phenotypic and functional characteristics of dendritic cells (DC) of mice. We compared virus-specific antibody and T cell-mediated responses in H. hepaticus-infected and noninfected mice that were inoculated intranasally with HSV1. The effect of H. hepaticus on the HSV1-specific antibody and T cell-mediated immune responses in superficial cervical and tracheobronchal lymph nodes (LN) did not reach statistical significance. Surface expression of the maturation-associated markers CD40, CD80, CD86, and MHC II and percentages of IL12p40- and TNFalpha-producing DC from spleen and colic LN in H. hepaticus-infected mice and noninfected mice were measured in separate experiments. Expression of CD40, CD86, and MHC II and percentages of IL12p40- and TNFalpha-producing DC from colic LN were decreased in H. hepaticus-infected mice. In contrast, H. hepaticus infection did not reduce the expression of these molecules by splenic DC. Expression of CD40, CD80, CD86, and MHC II on splenic DC from H. hepaticus-infected mice was increased after in vitro lipopolysaccharide stimulation. These results indicate that H. hepaticus infection can influence the results of immunologic assays in mice and support the use of H. hepaticus-free mice in immunologic research.


Asunto(s)
Células Dendríticas/inmunología , Infecciones por Helicobacter/inmunología , Helicobacter hepaticus/patogenicidad , Herpesvirus Humano 1/inmunología , Animales , Secuencia de Bases , Cartilla de ADN , Masculino , Ratones , Ratones Endogámicos C57BL
13.
Clin Diagn Lab Immunol ; 12(12): 1429-36, 2005 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-16339067

RESUMEN

Brucella abortus strain RB51 is an attenuated rough mutant used as the live vaccine against bovine brucellosis in the United States and other countries. We previously reported the development of strain RB51 as a bacterial vaccine vector for inducing Th1-type immune responses against heterologous proteins. Because safety concerns may preclude the use of strain RB51-based recombinant live vaccines, we explored the ability of a gamma-irradiated recombinant RB51 strain to induce heterologous antigen-specific immune responses in BALB/c mice. Exposure of strain RB51G/LacZ expressing Escherichia coli beta-galactosidase to a minimum of 300 kilorads of gamma radiation resulted in complete loss of replicative ability. These bacteria, however, remained metabolically active and continued to synthesize beta-galactosidase. A single intraperitoneal inoculation of mice with 10(9) CFU equivalents of gamma-irradiated, but not heat-killed, RB51G/LacZ induced a beta-galactosidase-specific Th1-type immune response. Though no obvious differences were detected in immune responses to B. abortus-specific antigens, mice vaccinated with gamma-irradiated, but not heat-killed, RB51G/LacZ developed significant protection against challenge with virulent B. abortus. In vitro experiments indicated that gamma-irradiated and heat-killed RB51G/LacZ induced maturation of dendritic cells; however, stimulation with gamma-irradiated bacteria resulted in more interleukin-12 secretion. These results suggest that recombinant RB51 strains exposed to an appropriate minimum dose of gamma radiation are unable to replicate but retain their ability to stimulate Th1 immune responses against the heterologous antigens and confer protection against B. abortus challenge in mice.


Asunto(s)
Vacuna contra la Brucelosis/inmunología , Brucella abortus/inmunología , Células TH1/inmunología , Animales , Brucella abortus/genética , Brucella abortus/fisiología , Brucelosis/prevención & control , Citocinas/biosíntesis , Células Dendríticas/metabolismo , Escherichia coli , Rayos gamma , Vectores Genéticos , Inmunoglobulina G/sangre , Ratones , Ratones Endogámicos BALB C , Dosis de Radiación , Replicación Viral/efectos de la radiación
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