RESUMO
Four monkeys were exposed to a retroviral vector and replication-competent murine amphotropic retrovirus in a bone marrow transplantation/gene transfer protocol (Kantoff et al., 1987). We have studied these animals 2 and 3 years post-transplantation and did not detect replicating virus in serum, peripheral blood mononuclear cells, or bone marrow cells. Amphotropic envelope sequences could not be detected in blood or bone marrow cells by Southern blotting or the polymerase chain reaction. Antibodies directed against the p30 and gp70 viral antigens were detected by Western blot and immunoprecipitation. The animals remain alive and well. Our findings suggest that primates can clear murine amphotropic retroviruses even when exposure occurs during a time of severe immunosuppression.
Assuntos
Engenharia Genética , Vetores Genéticos , Vírus da Leucemia Murina/isolamento & purificação , Macaca fascicularis/microbiologia , Retroviridae , Adenosina Desaminase/genética , Animais , Anticorpos Antivirais/sangue , Sequência de Bases , Sangue/microbiologia , Southern Blotting , Western Blotting , Medula Óssea/microbiologia , Transplante de Medula Óssea , DNA Viral/análise , Genes env , Hospedeiro Imunocomprometido , Vírus da Leucemia Murina/genética , Vírus da Leucemia Murina/imunologia , Vírus da Leucemia Murina/fisiologia , Leucócitos Mononucleares/microbiologia , Macaca fascicularis/sangue , Macaca fascicularis/imunologia , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Proteínas Recombinantes/genética , Estudos Retrospectivos , Retroviridae/genética , Viremia , Replicação ViralRESUMO
A patient with a localized HTLV-I-associated lymphoproliferative disease that was misdiagnosed as Hodgkin's disease is presented. The patient's serum was negative for HTLV-I antibodies by enzyme-linked immunosorbent assay (ELISA), Western blot, and radioimmunoprecipitation. Tumor tissue DNA was negative for HTLV-I by Southern blotting but was positive for distinct HTLV-I sequences when subjected to DNA amplification using the polymerase chain reaction. We conclude that the clinical and pathologic diagnosis of HTLV-I-related lymphoma can be difficult and can be confused with Hodgkin's disease. Extremely sensitive molecular biological techniques may be required to establish a diagnosis of HTLV-I-induced lymphoma.
Assuntos
DNA de Neoplasias/genética , DNA Polimerase Dirigida por DNA/genética , Infecções por Deltaretrovirus/genética , Deltaretrovirus/genética , Doença de Hodgkin/genética , Adulto , Sequência de Bases , Análise Mutacional de DNA , Infecções por Deltaretrovirus/enzimologia , Infecções por Deltaretrovirus/patologia , Diagnóstico Diferencial , Amplificação de Genes , Doença de Hodgkin/enzimologia , Doença de Hodgkin/patologia , Humanos , MasculinoRESUMO
The APAAP technique is a sensitive and relatively easy immunocytochemical method to perform. It requires only a modest amount of laboratory space and no expensive equipment with the exception of a light microscope. Staining can be performed on peripheral blood and bone marrow films as well as cryostat and paraffin-embedded tissue sections. Prior to staining peripheral blood and bone marrow films as well as cryostat sections, slides can be stored at -70 degrees C, thus allowing for batch processing of specimens. Stained slides can be kept at room temperature for prolonged periods of time without loss of label. Surface or cytoplasmic determinants can be stained with the APAAP method. Because the preparations are usually counterstained, the identification of the labeled cells can be observed and photographed. The APAAP method is a reliable procedure and a significant addition to the routine hematology and histopathology laboratory.
