RESUMEN
The compound named Histidine-pyridine-histidine (HPH) is an oxygen-activating ligand derived from the structure of bleomycin. We synthesized HPH derivatives, namely HPH-1 to -8, and investigated their antiviral activities against herpes simplex virus type 1. HPH-8 showed potent antiviral activity with an EC50 of 15 microM, and relatively high cytotoxicity with a CC50 of 37 microM. In contrast, HPH-4 indicated a weaker antiviral activity with an EC50 of 79 microM, but exhibited a far more less cytotoxicity (CC50 500 microM). Other HPH derivatives showed no effects against antiviral activities and cytotoxicities.
Asunto(s)
Antivirales/química , Antivirales/farmacología , Herpesvirus Humano 1/efectos de los fármacos , Histidina/análogos & derivados , Piridinas/química , Piridinas/farmacología , Animales , Chlorocebus aethiops , Histidina/farmacología , Humanos , Ratones , Relación Estructura-Actividad , Células VeroRESUMEN
We previously reported the isolation of a novel subtype of SRV/D-Tsukuba (SRV/D-T) from two cynomolgus monkeys (Macaca facicularis) in the breeding colony of Tsukuba Primate Research Center (TPRC). We surveyed for SRV/D infection in the TPRC cynomolgus colony using SRV/D-specific PCR primer sets designed based on the entire gag region sequence. The only SRV/D subtype detected in the colony was SRV/D-T with a positive infection rate of 22.4% (n = 49). It has been reported that the mode of transmission of SRV/D is via contact with virus shed in the body fluids. In this report, to investigate the infection route of SRV/D-T in monkeys at TPRC, we performed virus isolation and PCR for detection of the SRV/D genome from peripheral blood mononuclear cells (PBMCs), plasma, saliva, urine, and feces. Virus isolation and PCR detection were positive in plasma, saliva, urine, and fecal samples from all monkeys on which virus was isolated from PBMCs. This suggests that the spread of SRV/D-T infection in TPRC is via contact with virus shed in saliva, urine, and/or feces. Also, comparison of sequences of gp70 on multiple SRV/D-T isolates revealed that there was little intra- and inter-monkey variation, suggesting that SRV/D-T is fairly stable.
Asunto(s)
Líquidos Corporales/virología , Glicoproteínas/sangre , Glicoproteínas/orina , Fragmentos de Péptidos/sangre , Fragmentos de Péptidos/orina , Retrovirus de los Simios/fisiología , Proteínas Virales/sangre , Proteínas Virales/orina , Esparcimiento de Virus/fisiología , Animales , Secuencia de Bases , Heces/virología , Femenino , Glicoproteínas/genética , Transmisión Vertical de Enfermedad Infecciosa , Macaca fascicularis , Masculino , Datos de Secuencia Molecular , Infecciones por Retroviridae/transmisión , Infecciones por Retroviridae/virología , Retrovirus de los Simios/clasificación , Retrovirus de los Simios/aislamiento & purificación , Saliva/virología , Infecciones Tumorales por Virus/transmisión , Infecciones Tumorales por Virus/virología , Proteínas Virales/genéticaRESUMEN
The exogenous simian type D retroviruses (SRV/Ds) are prevalent in macaque monkeys and sometimes cause immunodeficiency with anemia, weight loss, and persistent unresponsive diarrhea. SRV/D isolates are classified as subtypes 1 to 6, and the entire sequences of the gag region of SRV/D-1, -2, and -3 and SRV/D-Tsukuba (SRV/D-T) have been determined. We designed specific primers in the gag region of SRV/D-T that enabled us to directly detect by polymerase chain reaction (PCR) SRV/D-T proviral DNA sequences in DNA extracted from whole blood. Using this assay and another PCR assay that detects multiple SRV/D subtypes, we performed a survey for SRV/D infection in our specific pathogen-free (SPF) and conventional colonies at Tsukuba Primate Center (TPC). In the SPF colony, no SRV/D signal was detected in any animal. On the other hand, SRV/D-T was detected in 11 of 49 animals (22.5%) in the conventional colony. SRV/D-T was the only SRV/D subtype detected. Consequently, SRV/D-T is the major SRV/D subtype present in cynomolgus monkeys at TPC.
Asunto(s)
Macaca fascicularis/virología , Enfermedades de los Monos/virología , Reacción en Cadena de la Polimerasa/veterinaria , Infecciones por Retroviridae/veterinaria , Retrovirus de los Simios/aislamiento & purificación , Infecciones Tumorales por Virus/veterinaria , Animales , ADN Viral/análisis , Femenino , Genes gag , Encuestas Epidemiológicas , Japón/epidemiología , Masculino , Enfermedades de los Monos/epidemiología , Enfermedades de los Monos/patología , Infecciones por Retroviridae/epidemiología , Infecciones por Retroviridae/patología , Retrovirus de los Simios/genética , Organismos Libres de Patógenos Específicos , Infecciones Tumorales por Virus/epidemiología , Infecciones Tumorales por Virus/patologíaRESUMEN
Exogenous type D simian retroviruses (SRV/D) are prevalent in captive and feral populations of various macaque monkeys. Thus far, five subtypes of SRV/Ds have been reported, three of which (SRV-1, -2 and -3) have been molecularly characterized. Two SRV/D strains (N27 and T150) were isolated from seropositive cynomolgus macaques at the Tsukuba Primate Center (TPC) in Japan, showing clinical signs of SRV/D infection, including anemia and persistent unresponsive diarrhea. Electron microscopy demonstrated that both SRV/D isolates have a virion morphology typical of type D retrovirus. The SRV/D N27 and T150 isolates were essentially the same based on sequence analysis. From homology analysis of the entire gag sequence, the N27 isolate is closely related to the other known SRV/Ds but is distinct from the three molecularly characterized SRV/Ds. Thus, we have tentatively designated the N27 and T150 viruses isolated from TPC cynomolgus macaques as SRV/D-Tsukuba (SRV/D-T).
Asunto(s)
Betaretrovirus/aislamiento & purificación , Macaca fascicularis/virología , Enfermedades de los Monos/virología , Infecciones por Retroviridae/veterinaria , Retrovirus de los Simios/aislamiento & purificación , Infecciones Tumorales por Virus/veterinaria , Secuencia de Aminoácidos , Animales , Betaretrovirus/clasificación , Betaretrovirus/genética , Femenino , Genes gag , Japón , Datos de Secuencia Molecular , Enfermedades de los Monos/patología , Filogenia , Infecciones por Retroviridae/patología , Retrovirus de los Simios/clasificación , Retrovirus de los Simios/genética , Alineación de Secuencia , Homología de Secuencia de Ácido Nucleico , Infecciones Tumorales por Virus/patologíaRESUMEN
A basic understanding of the major histocompatibility complex (MHC) class I, which, together with T-cell receptors, is a key player in antigen recognition by cytotoxic T lymphocytes, is necessary to study the cellular immune response to intracellular pathogens. The MHC has hardly been reported in cynomolgus monkeys ( Macaca facicularis), although cynomolgus monkeys have been frequently used as the surrogate animal model. We attempted to determine the nucleotide sequences of the MHC class I A locus of cynomolgus monkeys ( Mafa-A) and eventually 34 independent sequences of Mafa-A were obtained from 29 cynomolgus monkeys. These 34 sequences were classified into 14 Mafa-A alleles according to the results of phylogenetic analyses using the neighbor-joining method. One to three Mafa-A alleles were obtained from a single animal. We also tried to establish a multiplex PCR-SSP method for convenient typing of Mafa-A alleles. cDNA from a family of cynomolgus monkeys, which is composed of four sirs and four dams, were examined by multiplex PCR-SSP. The result of multiplex PCR-SSP showed that an individual cynomolgus monkey had two or three Mafa-A alleles, suggesting that the A locus of cynomolgus monkeys might be duplicated.
Asunto(s)
Alelos , Genes MHC Clase I , Antígenos de Histocompatibilidad Clase I/genética , Macaca fascicularis/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Femenino , Antígenos de Histocompatibilidad Clase I/química , Antígenos de Histocompatibilidad Clase I/metabolismo , Masculino , Datos de Secuencia Molecular , Linaje , Filogenia , Reacción en Cadena de la Polimerasa , Alineación de SecuenciaRESUMEN
We previously reported that peripheral lymphocytes from about 12% of cynomolgus monkeys lacked reactivity with anti-rhesus monkey CD3 monoclonal antibody (FN18). The nucleotide sequence analysis of the genes encoding CD3 component proteins revealed that a single amino acid substitutions found in the CD3epsilon chain determined the phenotype. In this study, we attempted to develop a method based on the restriction fragment length polymorphism (RFLP) and apply it for determination of the genotypes of individual monkeys. Comparison of the phenotype determined by fluorescence-activated cell sorter analysis with the genotype determined by RFLP analysis revealed that the FN18 -positive trait was dominant over the FN18-negative trait. It was also revealed that allele frequency was significantly different among macaques depending on the geographical region where their ancestors were derived from.
Asunto(s)
Anticuerpos Monoclonales/inmunología , Complejo CD3/genética , Macaca fascicularis/genética , Polimorfismo de Longitud del Fragmento de Restricción , Animales , Cartilla de ADN , Fluorescencia , Frecuencia de los Genes , Geografía , Leucocitos Mononucleares/inmunología , Macaca fascicularis/inmunología , Linaje , Reacción en Cadena de la Polimerasa , ARN Mensajero/genéticaRESUMEN
A Japanese encephalitis (JE) vaccine candidate encoding JE virus premembrane (prM) and envelope (E) genes, designated pNJEME, was evaluated for safety and immunogenicity in non-human primate, cynomolgus monkeys. pNJEME was constructed using a vector (pNGVL4a) designed to address some of the safety concerns of DNA vaccine. In two different experiments, two immunizations with 300 microg of pNJEME by intramuscular (i.m.) injection, and 3 microg of pNJEME using a gene gun, and three immunizations by i.m. injection with 500 microg of pNJEME were performed. All the three protocols induced low to high levels of neutralizing antibody, indicating an ability of pNJEME to induce neutralizing antibody in monkeys with a wide individual variation in response to pNJEME. In one experiment designed to compare the DNA vaccine with a commercial inactivated JE vaccine, three immunizations by i.m. inoculation with 300 microg of pNJEME or by gene gun administration with 3 microg of pNJEME induced similar levels of neutralizing antibody to those induced by three immunizations with a human dose of the inactivated vaccine in most monkeys. After intranasal challenge with the Beijing P3 or JaTH160 strain of JE virus, pNJEME-immunized monkeys showed anamnestic neutralizing antibody responses, indicating that pNJEME induced memory B cells which were responsive to infection with JE virus. No systemic and local reactions were observed in any monkeys after i.m. or gene gun inoculations with plasmid DNAs.