RESUMO
Parvoviruses such as parvovirus H-1 (H-1PV) may selectively infect and lysis cancer cells. The parvoviruses also induce an immune system to eliminate the tumor cells through the formation of anti-cancer immunity. One of the possible mechanisms of antitumor activity is associated with the direct induction of apoptosis by parvoviral proteins NS1 and 11 kDa. Parvovirus-based vectors are promising for gene therapy of oncological diseases and genetic disorders in humans. Parvoviruses were successfully used for the experimental treatment on animal models of human glioma, neuroblastomas, lymphomas, pancreatic carcinoma, carcinomas and breast tumors. ParvOryx is the first oncolytic preparation constructed on the base of H-1PV; its phase I/IIa clinical trials in patients with glioblastoma multiforme are in process.
Assuntos
Parvovirus H-1 , Imunoterapia/métodos , Terapia Viral Oncolítica , Vírus Oncolíticos , Antineoplásicos/uso terapêutico , Citotoxicidade Imunológica , Feminino , Vetores Genéticos , Glioblastoma/imunologia , Glioblastoma/terapia , Parvovirus H-1/química , Humanos , Masculino , Neoplasias/imunologia , Neoplasias/terapia , Proteínas não Estruturais Virais/uso terapêuticoRESUMO
Analysis of published reports helped us single out the most potent antigens among HCMV proteins: phosphoproteins pp150(UL32) and p52(UL44). Theoretical computer analysis of p52 epitopes showed the main antigenic determinants not cross-reacting with antigens of other viruses. Virus-containing (strain AD169) material was obtained and genome DNA was isolated. Amplification of a site of gene UL44 coding for unique determinants detected a PCR fragment of required electrophoretic mobility. The fragment was cloned in vector pLBE. The specificity of cloning was confirmed by restriction analysis of theoretical sites. Nucleotide sequence of cloned fragment of UL44 gene was studied by Maxam-Gilbert's method. Cloning in expressing bacterial vectors helped obtain HCMV recombinant protein p52 in the pure form and fused with beta-galactosidase. Enzyme immunoassay with HCMV-positive and negative donor sera and ABBOTT HCMV sera showed that recombinant p52 increased the sensitivity and specificity of a previously obtained recombinant pp150 as an antigen to HCMV-IgG and HCMV-IgM. The sensitivity and specificity is 100% with 98-99% reliability.
Assuntos
Antígenos Virais/isolamento & purificação , Citomegalovirus/imunologia , Proteínas de Ligação a DNA/isolamento & purificação , Proteínas Virais/isolamento & purificação , Sequência de Aminoácidos , Antígenos Virais/química , Antígenos Virais/genética , Sequência de Bases , Clonagem Molecular , Reações Cruzadas , DNA Viral , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/genética , Técnicas Imunoenzimáticas , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas Virais/química , Proteínas Virais/genéticaRESUMO
The genes encoding the precursors of secretory glycoprotein gp57-65 (A antigen) and glycoprotein complex gp100, gp60, gp48 (B antigen) of virulent strain JM of Marek's disease virus (MDV) have been cloned using PCR of viral DNA. Nucleotide sequences of both genes have been determined and amino acid sequence of the precursor peptides predicted, and compared with the corresponding sequences of other MDV strains. The results will be used for developing methods for the diagnosis of Marek's disease and vaccination against it.
