RESUMO
Several studies have recently shown the use of recombinant rabies virus as potential vector-viral vaccine for HIV-1. The sequence homology between gp 120 and rabies virus glycoprotein has been reported. The McCoy cell line has therefore been used to show CD4+ or CD4+ like receptors. Samples of HIV-1 were isolated, when plasma of HIV-1 positive patients was inoculated in the McCoy cell line. The virus infection was then studied during successive virus passages. The proteins released in the extra cellular medium were checked for protein activity, by exposure to SDS Electrophoresis and blotting to nitro-cellulose filter, then reacting with sera of HIV positive and negative patients. Successive passages were performed, and showed viral replication, membrane permeabilization, the syncytium formation, and the cellular lysis (cytopathic effect). Flow cytometry analysis shows clear evidence that CD4+ receptors are present in this cell line, which enhances the likelihood of easy isolation and replication of HIV. The results observed allow the use of this cell line as a possible model for isolating HIV, as well as for carrying out studies of the dynamics of viral infection in several situations, including exposure to drugs in pharmacological studies, and possibly studies and analyses of the immune response in vaccine therapies.
Assuntos
Antígenos CD4/análise , Linfócitos T CD4-Positivos/química , Linhagem Celular , HIV-1/fisiologia , Replicação Viral/fisiologia , Animais , Western Blotting , Linhagem Celular/virologia , Citometria de Fluxo , Técnica Indireta de Fluorescência para Anticorpo , Vetores Genéticos , Humanos , CamundongosRESUMO
Several studies have recently shown the use of recombinant rabies virus as potential vector-viral vaccine for HIV-1. The sequence homology between gp 120 and rabies virus glycoprotein has been reported. The McCoy cell line has therefore been used to show CD4+ or CD4+ like receptors. Samples of HIV-1 were isolated, when plasma of HIV-1 positive patients was inoculated in the McCoy cell line. The virus infection was then studied during successive virus passages. The proteins released in the extra cellular medium were checked for protein activity, by exposure to SDS Electrophoresis and blotting to nitro-cellulose filter, then reacting with sera of HIV positive and negative patients. Successive passages were performed, and showed viral replication, membrane permeabilization, the syncytium formation, and the cellular lysis (cytopathic effect). Flow cytometry analysis shows clear evidence that CD4+ receptors are present in this cell line, which enhances the likelihood of easy isolation and replication of HIV. The results observed allow the use of this cell line as a possible model for isolating HIV, as well as for carrying out studies of the dynamics of viral infection in several situations, including exposure to drugs in pharmacological studies, and possibly studies and analyses of the immune response in vaccine therapies
Assuntos
Humanos , HIV-1 , Replicação Viral , Antígenos CD4 , Western Blotting , Linfócitos T CD4-Positivos , Linhagem Celular , Citometria de Fluxo , Técnica Indireta de Fluorescência para Anticorpo , Replicação ViralRESUMO
A cistinose nefropática é uma doença genética autossômica recessiva, sistêmica e progressiva, caracterizada pelo acúmulo intrtalisossomal de cistina.Para o tratamento específico da doença, o estudo objetivou utilizar uma droga depletora de estoques intracelulares de cistina, o bitartrato de cisteamine. O estudo avalia também os efeitos da droga em seis crianças, por meio de dosagem pré e pós-tratamento de CISTIeuco, assim como benefícios e efeitos adversos da droga. A adequaçäo da dose de bitartrato de cisteamine deve ser feita de acordo com a medida do contúdo intraleucocitário de cistina (CISTIeuco), para o qual o estudo desenvolveu um método para sua realizaçäo. Neste estudo, descreve-se o método de dosagem utilizado e as vantagens desse método em relaçäo a outros existentes. O estudo também avaliou os efeitos da droga em seis crianças, por meio de dosagem pré e pós-tratamento de CISTIeuco, assim como benefícios e feitos adversos da droga. Após período médio de observaçäo de quatro meses (um a nove meses), ocorreram uma melhora no ganho pôndero-estatural e reduçäo no número de internaçöes e reduçäo nos níveis de CISTIeuco de 60,7 porcento em média com a medicaçäo. No curto período de observaçäo, o estudo confirma pela casuística utilizada que existem benefícios reais com a medicaçäo, além do método utilizado no estudo ser confável e poder ser utilizado no diagnóstico da doença e do estado de portador. Espera-se que essa droga seja em breve utilizada por todos os portadores de cistinose no Brasil.(au)
