ABSTRACT
The correlation between human papillomavirus and induction of cervical cancer has been established since 1974, but the induction of transient idiopathic infertility, attributed to HPV, has also recently been addressed. Therefore, there is a clear need for a more effective combat against the virus, which has periodically been correlated with new diseases that indiscriminately affect men, women, and children of all social classes. Recent technological advances have contributed notably to the improvement of experimental work tools, a fact that provides scientists with new horizons to be explored in the development of new products, free from serious side effects, due to genetic manipulation in the construction of expression vectors directed to therapeutic targets of interest. Thus, this book aims to tell readers a little bit of the natural history about HPV and the advances in prophylaxis and minimally invasive therapy, aiming to eradicate the virus and treat infected patients, who have developed HPV-induced cancer. Periodic exams and vaccination are still the best options to prevent HPV infection and the development of cancers associated with the virus, as well as the possibility of transient infertility occurrence attributed to the HPV.
Desde 1974 foi estabelecida a correlação do papilomavírus humano com a indução de câncer cervical, mas recentemente tem sido abordada também a indução de infertilidade idiopática transiente, atribuída ao HPV. Portanto, fica evidente a necessidade de combate mais efetivo ao vírus, que periodicamente vem sendo correlacionado com novas doenças que afetam indiscriminadamente homens, mulheres e crianças de todas as classes sociais. Os avanços tecnológicos recentes têm contribuído de forma notável para o aprimoramento das ferramentas de trabalho experimental, fato que proporciona aos cientistas novos horizontes a serem explorados no desenvolvimento de novos produtos, isentos de efeitos colaterais graves, devido à manipulação genética na construção de vetores de expressão gênica direcionados a alvos terapêuticos de interesse. Assim, este livro pretende relatar aos leitores um pouco da história natural sobre HPV e os avanços em termos de profilaxia e de terapia minimamente invasiva, visando erradicar o vírus e tratar os pacientes infectados, que desenvolveram câncer induzido por HPV. Os exames periódicos e a vacinação ainda são as melhores opções para se evitar a infecção por HPV e o desenvolvimento de cânceres associados ao vírus, assim como a possibilidade de ocorrência da infertilidade transiente atribuída ao HPV.
ABSTRACT
El propósito de este estudio fue evaluar la inmunogenicidad de concentrados de factor VIII producidos en UNC-Hemoderivados antes y después del tratamiento térmico empleando un enzimoinmunoensayo (EIE) desarrollado en nuestro laboratorio. Materiales y método: Para ese propósito se obtuvieron anticuerpos contra factor VIII calentado y no-calentado por inmunización de conejos y la inmunoglobulina G específica fue aislada por afinidad a Proteína A-Sepharosa. El EIE fue realizado empleando como antígeno de captura el factor VIII con o sin tratamiento térmico (0.5 ug/pocillo), los sitios inespecíficos bloqueados con albúmina al 2 por ciento y el anticuerpo revelado con un anti-IgG de conejo conjugada a peroxidasa. La presencia de neoantígenos fue estudiada por incubar durante 2 hs a 37°C y luego a 4°C toda la noche, con cantidades crecientes de factor VIII antes y después del tratamiento térmico (0.03 a 600 ug de proteínas), con cantidades adecuadas de IgG anti-factor VIII calentado y no-calentado. Luego de centrifugar la muestra para la separación de los inmunocomplejos, se valoró la presencia de anticuerpos en el sobrenadante por EIE tanto para factor VIII calentado y no calentado. Resultados: Los resultados obtenidos permitieron observar que para ambos anticuerpos (calentado y no calentado) se neutralizaba la misma cantidad de factor VIII calentado y no calentado (0.30 ug) y además, se pudo comprobar que esta conducta se repetía cuando se empleaban en el EIE como antígeno de captura, factor VIII con y sin tratamiento térmico. Conclusiones: Los datos obtenidos de este estudio nos permiten concluir que el tratamiento térmico aplicado a los concentrados de factor VIII elaborados en la UNC-Hemoderivados no producen ninguna formación de neoantígenos. A juzgar por el sensible y específico EIE desarrollado en nuestro laboratorio.
Introduction: with the purpose of improving viral security, plasma-derived products are generally subjects of solvent/detergent and heating (100°C for 30 minutes) treatment which are introduced during the manufacturing process. The formation of neoantigens in factor VIII concentrates produced after the heat treatment could trigger an immune response against the modified protein and the native protein. Objetives: the purpose of this study was to evaluate the immunogenicity of factor VIII concentrates produced in UNC-Hemoderivados before and after heat treatment, using an EIA developed in our laboratory. Materials and methods: antibodies against factor VIII with and without heating treatment were obtained by rabbit immunization. The specific IgG was isolated by protein-A-Sepharose affinity chromatograpy. Two EIA plates were coated (0.5 ug/well) one with factor VIII heated (H) and the other one with factor VIII no-heat (no-H) and the antibodies detection were performed using rabbit anti-lgG peroxidase conjugated. The neoantigens were studied by incubation of increasing concentrations of factor VIII (0.03-600 ug of proteins) before and after heating treatment during 2 hours at 37 °C and overnight at 4 °C with adequate concentrations of anti-factor VIII IgG (with and without treatment). The immunocomplexes were removed by centrifugation and the free antibodies in the supernatant were measured by EIA in plates H and no-H. Results: the EIA showed a linear behavior between antibodies dilution ranged from 1/8000 to 1/512000. With these results we can conclude that the same concentration of factor VIII heated and unheated (0.30 ug) were neutralized with either antibodies (heated and unheated). These results were similar in both EIA plates coated with factor VIII H and no-H...
