RESUMEN
In recent years, dendritic cells (DCs)-based vaccines have been developed to combat HIV-1 infection in preclinical and clinical trials. In this study, mice bone marrow cells-derived DCs were pulsed with the recombinant Nef, heat shock protein 27 (Hsp27) and Hsp27-Nef proteins, and also green fluorescent protein (GFP) as a positive control. Then, new platforms of DCs loaded with HIV-1 Nef and Hsp27-Nef proteins (i.e., DC prime/DC boost, DNA prime/DC boost, and DC prime/protein boost) were used to evaluate immune responses in BALB/c mice. Finally, the potency of splenocytes exposed to single-cycle replicable (SCR) HIV-1 virions was investigated to secret cytokines in vitro. Our data indicated that the recombinant Nef (â¼30 kDa), Hsp27 (â¼27 kDa), GFP (â¼27 kDa), and Hsp27-Nef (â¼53 kDa) proteins were greatly generated in E. coli. Moreover, the modified DCs with the recombinant proteins were prepared in large scale. The results of mice immunization showed the highest levels of antibodies, cytokines, and Granzyme B in heterologous DC prime/protein boost regimen using Hsp27-Nef antigen (DCHsp27-Nef prime/ protein Hsp27-Nef boost regimen). The levels of IFN-γ and IL-10 cytokines in splenocytes isolated from mice immunized with DCHsp27-Nef prime/ protein Hsp27-Nef boost regimen were higher than those in other regimens after exposure to SCR virions. These findings demonstrated the importance of Hsp27 as an adjuvant and heterologous DC prime/ protein boost regimen in improvement of immune responses. Indeed, DC Hsp27-Nef prime/ protein Hsp27-Nef boost regimen can be utilized as a promising candidate for HIV-1 vaccine development.
Asunto(s)
VIH-1 , Vacunas , Animales , Ratones , Citocinas , Proteínas de Choque Térmico HSP27 , Escherichia coli , Bazo , Virión , Antígenos Virales , Células DendríticasRESUMEN
The present study was conducted to analyze the genotypic diversity of circulating species A rotavirus (RVA) strains in Iran and also to investigate comparative analysis between the genotypes of VP4 and VP7 of cocirculating RVA and vaccine strains before the vaccine is introduced in the national immunization program. The G3-lineage I was found in this study as the most common G genotype which was followed by G9-lineage III, G1-lineages I, II, G12-lineage III, G2-lineage IV, and G4-lineage I. Also, P[8]-lineages III, IV was found as the predominant P genotype which was followed by P[4]-lineage V, and P[6]-lineage I. Overally, G3P[8] was determined as the most common combination. Moreover, the analysis of the VP7 antigenic epitopes showed that several amino acid differences existed between circulating Iranian and the vaccine strains. The comparison of genotype G1 of Iranian and vaccine strains (RotaTeq and Rotarix), and genotypes G2, G3, and G4 of Iranian and RotaTeq vaccine strains revealed three to five amino acids differences on the VP7 antigenic epitopes. Furthermore, analyzing of the VP8* epitopes of Iranian P[8] strains indicated that they contained up to 11 and 14 amino acid differences with Rotarix and RotaTeq, respectively. Based on different patterns of amino acid substitutions in circulating and vaccine strains, the emergence of antibody escaping mutants and potentially the decrease of immune protection might ensue in vaccinated children. However, considering the broad cross-protective activity of RVA vaccines, their efficacy should be monitored after the introduction in Iran.
Asunto(s)
Antígenos Virales/genética , Proteínas de la Cápside/genética , Vacunas contra Rotavirus/análisis , Rotavirus/genética , Sustitución de Aminoácidos , Preescolar , Heces/virología , Femenino , Variación Genética , Genotipo , Hospitalización/estadística & datos numéricos , Humanos , Lactante , Irán/epidemiología , Masculino , Vacunación Masiva , Filogenia , Rotavirus/clasificación , Infecciones por Rotavirus/epidemiología , Infecciones por Rotavirus/virología , Vacunas Atenuadas/análisisRESUMEN
Cluster of Differentiation 90 (CD90, Thy-1) has been proposed as one of the most important biomarkers in several cancer cells including cancer stem cells (CSCs). CD90 is considered as a potential normal stem cell and CSCs biomarker and also has been identified in lung cancer stem cells, hepatocellular carcinoma cells and high-grade gliomas. Using eukaryotic host systems involves complex procedures and frequently results in low protein yields. The expression of recombinant proteins in Escherichia coli is comparatively easier than eukaryotic host cells. The potential of large scale production of recombinant protein has made this system an economic production platform. In this study we expressed the extra-membrane domain of human CD90 (exCD90) antigen (Gln15-Cys130) in E. coli expression host cells. The epitope integrity of purified recombinant antigen was confirmed by antibody-antigen interaction using 5E10 anti-CD90 monoclonal antibody and binding study through ELISA and florescent staining of CD90(+) cells in a flow cytometry experiment.
Asunto(s)
Escherichia coli/genética , Antígenos Thy-1/genética , Antígenos Thy-1/inmunología , Animales , Anticuerpos/inmunología , Reacciones Antígeno-Anticuerpo , Secuencia de Bases , Clonación Molecular , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Datos de Secuencia Molecular , Estructura Terciaria de Proteína , Conejos , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Antígenos Thy-1/químicaRESUMEN
BACKGROUND: Human rotavirus has now been established as the most important cause of childhood gastroenteritis worldwide. The RNA genomic electrophoretic diversity of group A rotavirus strains in Tehran, Iran, during April 2002 through March 2005, was investigated in 1250 stool specimens. METHODS: Stool samples were obtained from young children with acute gastroenteritis. RNA electrophoretype of group A rotavirus strains was determined using RNA polyacrylamide gel electrophoresis in those samples found to be positive for rotavirus by enzyme immunoassay. RESULTS: The predominant RNA profile detected was the long electrophoretype (90%) followed by the short profile (8.8%). Four patients (1.1%) had patterns of mixed infection. Fourteen different electrophoretic RNA patterns, eight of long and six of short, were detected in the study area. A long RNA electrophoretype persisted during the years of the study with peak incidence in cool seasons. The long pattern occurred throughout the study period and in patients aged one to 60 months, but the short profile identified only in infants at varying intervals. CONCLUSION: Our data demonstrate epidemiologic differences between the incidence of long and short electrophoretypes. The long profiles appear to circulate continuously in the area, whereas the short patterns appear in an episodic fashion.