RESUMO
A series of plasmids containing the HSP70 gene of Mycobacterium tuberculosis fused to the hemagglutinin (H5) gene of H5N1 avian influenza virus (AIV) (H5-HSP70 (heat shock protein 70) vaccine) or individual H5 gene (H5 vaccine) or HSP70 gene (HSP70 vaccine) were constructed based on the plasmid pcDNA3.1. Expression of H5 gene in Vero cells in vitro and in chickens in vivo was confirmed following their transfection and immunization with H5 or H5-HSP70 vaccines. Controls consisted of HSP70 vaccine, empty plasmid pcDNA3.1 and co-administered H5 and HSP70 vaccines. H5-HSP70 vaccine produced in chicken higher hemagglutination inhibition (HI) antibody titer than H5 vaccine. However, the increase was not statistically significant. We have demonstrated for the first time that the H5 DNA vaccine with fused HSP70 gene may produce an enhanced induction of humoral immune response to AIV in chickens.
Assuntos
Anticorpos Antivirais/sangue , Proteínas de Bactérias , Proteínas de Choque Térmico HSP70 , Glicoproteínas de Hemaglutininação de Vírus da Influenza , Virus da Influenza A Subtipo H5N1/imunologia , Proteínas Recombinantes de Fusão , Vacinas de DNA , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/metabolismo , Galinhas , Chlorocebus aethiops , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/imunologia , Proteínas de Choque Térmico HSP70/metabolismo , Testes de Inibição da Hemaglutinação , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Glicoproteínas de Hemaglutininação de Vírus da Influenza/metabolismo , Humanos , Vacinas contra Influenza/administração & dosagem , Vacinas contra Influenza/imunologia , Influenza Aviária/imunologia , Influenza Aviária/prevenção & controle , Influenza Aviária/virologia , Plasmídeos , Doenças das Aves Domésticas/imunologia , Doenças das Aves Domésticas/prevenção & controle , Doenças das Aves Domésticas/virologia , Proteínas Recombinantes de Fusão/administração & dosagem , Proteínas Recombinantes de Fusão/imunologia , Vacinação/veterinária , Vacinas de DNA/administração & dosagem , Vacinas de DNA/imunologia , Células VeroRESUMO
OBJECTIVE: The aim of this prospective study was to determine whether human cytomegalovirus (HCMV)-DNA detected by polymerase chain reaction (PCR) analysis in the plasma of bone marrow transplant (BMT) patients is a predictor of HCMV disease progression. METHODS: Plasma samples were collected from 15 patients who received allogenic BMTs. Each individual was sampled 1 week before and then weekly for 17 weeks after transplantation. The 270 plasma specimens were processed with a PCR method for detecting HCMV-DNA. Patients were also physically examined for signs or symptoms of HCMV-related disease. RESULTS: Eight (53.5%) of the 15 patients tested positive for HCMV-DNA. Two (25%) of these 8 individuals also had positive PCR findings before transplantation. Six (75%) of the 8 HCMV-DNA-positive patients had positive plasma-PCR results a week before clinical symptoms developed. The other 2 (25%) remained asymptomatic throughout their hospital stay. All 6 symptomatic cases were treated with ganciclovir, and 4 converted to negative plasma-PCR status at a median of 21 days. There was a significant correlation between PCR-detection of HCMVDNA in plasma and presence of HCMV-related symptoms (P < 0.01). CONCLUSION: Qualitative plasma-PCR analysis before and after bone marrow transplantation is a valuable way to screen for HCMV infection in BMT patients. Plasma-PCR monitoring of HCMV activity in this patient group might make it possible to administer an antiviral drug and thus reduce mortality. However, quantitative PCR is still considered the best way to accurately identify active HCMV infection and monitor treatment.