RESUMEN
OBJECTIVE: To evaluate the performance of an automated DNA-image-cytometry system as a tool to detect cervical carcinoma. METHODS: Of 384 liquid-based cervical cytology samples with available biopsy follow-up were analyzed by both the Imager System and a high-risk HPV test (Cobas). RESULTS: The sensitivity and specificity of Imager System for detecting biopsy proven high-grade squamous intraepithelial lesion (HSIL, cervical intraepithelial neoplasia [CIN]2-3) and carcinoma were 89.58% and 56.25%, respectively, compared to 97.22% and 23.33% of HPV test but additional HPV 16/18 genotyping increased the specificity to 69.58%. The sensitivity and specificity of the Imager System for predicting HSIL+ (CIN2-3+) lesions among atypical squamous cells of undetermined significance samples were 80.00% and 70.53%, respectively, compared to 100% and 11.58% of HPV test whilst the HPV 16/18 genotyping increased the specificity to 77.89%. Among atypical squamous cells-cannot exclude HSIL, the sensitivity and specificity of Imager System for predicting HSIL+ (CIN2-3+) lesions upon follow up were 82.86% and 33.33%%, respectively, compared to 97.14% and 4.76% of HPV test and the HPV 16/18 genotyping increased the specificity to 19.05%. Among low-grade squamous intraepithelial lesion cases, the sensitivity and specificity of the Imager System for predicting HSIL+ (CIN2-3+) lesions were 66.67% and 35.71%%, respectively, compared to 66.67% and 29.76% of HPV test while HPV 16/18 genotyping increased the specificity to 79.76%. The overall results of imager and high-risk HPV test agreed in 69.43% (268) of all samples. CONCLUSIONS: The automated imager system and HPV 16/18 genotyping can enhance the specificity of detecting HSIL+ (CIN2-3+) lesions.
Asunto(s)
Cuello del Útero/patología , ADN Viral/genética , Lesiones Intraepiteliales Escamosas de Cuello Uterino/patología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Células Escamosas Atípicas del Cuello del Útero/patología , Células Escamosas Atípicas del Cuello del Útero/virología , Biopsia/métodos , Cuello del Útero/virología , Colposcopía/métodos , Femenino , Pruebas de ADN del Papillomavirus Humano/métodos , Papillomavirus Humano 16/genética , Papillomavirus Humano 18/genética , Humanos , Citometría de Imagen/métodos , Persona de Mediana Edad , Infecciones por Papillomavirus/patología , Infecciones por Papillomavirus/virología , Sensibilidad y Especificidad , Lesiones Intraepiteliales Escamosas de Cuello Uterino/virología , Neoplasias del Cuello Uterino/patología , Neoplasias del Cuello Uterino/virología , Frotis Vaginal/métodos , Adulto Joven , Displasia del Cuello del Útero/patología , Displasia del Cuello del Útero/virologíaRESUMEN
Cellular immunity against cancer can be achieved with viral vector- and DNA-based immunizations. In preclinical studies, cancer vaccines are very potent, but in clinical trials these potencies are not achieved yet. Thus, a rational approach to improve cancer vaccines is warranted. We previously demonstrated that the relatively low intrinsic immunogenicity of DNA vaccines could be enhanced by inclusion of endoplasmic reticulum (ER) targeting and universal helper epitopes within the vaccine. We now evaluated whether an optimal antigen format, as defined in DNA vaccines, can further enhance the effectiveness of recombinant Semliki Forest virus (rSFV) vaccines. To this purpose, we generated, characterized and evaluated the efficacy of rSFV replicon particles expressing human papillomavirus E6 and/or E7 proteins fused to several helper T-cell epitopes and an ER targeting signal. Here, we show that inclusion of a helper cassette and an ER targeting signal enhanced protein stability and markedly augmented the frequencies of human papillomavirus-specific T cells. Even at an immunization dose of as low as 10(5) replicon particles, this novel vaccine achieved tumor regression and protection. Thus, even highly effective viral vector vaccines can benefit from an improved antigen format, based on the inclusion of defined helper epitopes and ER targeting.
Asunto(s)
Antígenos Virales/inmunología , Vacunas contra el Cáncer/inmunología , Proteínas Oncogénicas Virales/inmunología , Proteínas E7 de Papillomavirus/inmunología , Vacunas contra Papillomavirus/inmunología , Animales , Antígenos Virales/administración & dosificación , Antígenos Virales/genética , Vacunas contra el Cáncer/administración & dosificación , Cricetinae , Epítopos de Linfocito T/química , Femenino , Humanos , Riñón/citología , Ratones Endogámicos C57BL , Proteínas Oncogénicas Virales/administración & dosificación , Proteínas Oncogénicas Virales/genética , Proteínas Oncogénicas Virales/metabolismo , Proteínas E7 de Papillomavirus/administración & dosificación , Proteínas E7 de Papillomavirus/genética , Proteínas E7 de Papillomavirus/metabolismo , Vacunas contra Papillomavirus/genética , Vacunas contra Papillomavirus/metabolismo , Virus de los Bosques Semliki/genética , Linfocitos T Citotóxicos/inmunología , Linfocitos T Citotóxicos/metabolismo , Neoplasias del Cuello Uterino/prevención & control , Vacunas de ADN/administración & dosificación , Vacunas de ADN/inmunologíaAsunto(s)
Cistadenocarcinoma Mucinoso/patología , Osteosarcoma/patología , Neoplasias Retroperitoneales/patología , Anciano , Antígenos CD/análisis , Antígenos de Diferenciación Mielomonocítica/análisis , Cistadenocarcinoma Mucinoso/metabolismo , Femenino , Humanos , Inmunohistoquímica , Queratina-7 , Queratinas/análisis , Neoplasias Retroperitoneales/metabolismoRESUMEN
AIMS: To assess, in tissue microarray (TMA), the proliferative activity of endometrial carcinoma using one of the minichromosome maintenance (MCM) proteins (MCM7), and to explore its potential value for prognosis. MCM proteins are essential for eukaryotic DNA replication and have recently been used to define the proliferative compartments in human tissues. METHODS AND RESULTS: Immunohistochemistry for MCM7 and Ki67 was performed on TMAs constructed from 212 cases of endometrial carcinoma. MCM7 and Ki67 expression was quantified according to the extent of nuclear staining. An analysis was carried out of the association between MCM7 expression and that of Ki67 and the clinicopathological characteristics of endometrial carcinoma. MCM7 and Ki67 immunoreactivity was clearly evident in the nuclei of tumour cells. MCM7 and Ki67 labelling indices in endometrial carcinomas correlated with each other (P < 0.001). A significant correlation existed between the MCM7 labelling index and histological grade (P = 0.008) and patients' age at diagnosis (P < 0.001). Well-differentiated carcinomas and younger patients had a lower MCM7 index. Poor survival was observed in patients with endometrial carcinoma with a high MCM7 index (P = 0.03) and MCM7 was found to be an independent prognostic factor by multivariate analysis (P = 0.04). The Ki67 labelling index correlated with histological grade (P = 0.01) but had no significant prognostic impact (P = 0.50). CONCLUSIONS: In this TMA study on endometrial carcinoma, MCM7 was found to be a more reliable and useful marker than Ki67 in assessing tumour proliferation and in the prognosis of patients.
Asunto(s)
Biomarcadores de Tumor/análisis , Proteínas de Ciclo Celular/análisis , Proteínas de Unión al ADN/análisis , Neoplasias Endometriales/patología , Proteínas Nucleares/análisis , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Proliferación Celular , Replicación del ADN , Neoplasias Endometriales/metabolismo , Femenino , Humanos , Inmunohistoquímica , Antígeno Ki-67/análisis , Persona de Mediana Edad , Componente 7 del Complejo de Mantenimiento de Minicromosoma , Análisis Multivariante , Estadificación de Neoplasias , Pronóstico , Análisis de Supervivencia , Análisis de Matrices TisularesRESUMEN
The short term effect of optic nerve ligature was studied by analysing the lateral genuiclate bodies. There was significant difference in total RNA amount per unit tissue between the contralateral and ipsilateral sides 1 week after ligation. There were, however, no observable difference in the acid phosphatase activity and the quantity of contacts between the two sides. The dopamine receptors on the membranes also were much higher in number in the contralateral side.
Asunto(s)
Cuerpos Geniculados/análisis , Nervio Óptico/fisiología , Fosfatasa Ácida/análisis , Animales , Uniones Intercelulares , Ligadura , Ratones , ARN/análisis , Ratas , Receptores Dopaminérgicos/análisisRESUMEN
The aging optic nerves and visual cortices were studied in mice. Changes in the morphology of neuroglia were evident in the optic nerve but no difference in the numbers of neurotubules or myelin sheaths optic nerve fibers in the aged were observed. Large myelinated fibers, however, begun to degenerate. In the aging visual cortices, a higher proportion of dendrodendritic contacts were present and microglia cells were found to be active.