RESUMO
OBJECTIVES: Our aim was to investigate the effect of PEGylation on the uptake of osteoprotegerin/osteoclastogenesis inhibitory factor (OPG/OCIF) into rat liver, kidney and spleen, and human liver. METHODS: Copolymer of polyethyleneglycol allylmethylether and maleamic acid sodium salt with OCIF (poly(PEG)-OCIF) (0.5 mg/kg) was administered to rats and the concentrations of poly(PEG)-OCIF in the liver, kidney and spleen at 15 min after administration were measured by ELISA. For human liver uptake, the liver perfusion of OCIF and (3)H-labelled poly(PEG)-OCIF was conducted using fresh human liver block. KEY FINDINGS: The tissue uptake of poly(PEG)-OCIF in rats was significantly lower compared with that of OCIF. In fresh human liver perfusion, (3)H-poly(PEG)-OCIF was rarely taken up into the liver. On the other hand, more than 50% of the perfused OCIF was taken up. CONCLUSIONS: PEGylation of OCIF using poly(PEG) dramatically suppressed the uptake of OCIF into human liver as well as into rat liver and could be a promising approach for improving the pharmacokinetic and pharmacological effects of OCIF in the clinical setting.
Assuntos
Conservadores da Densidade Óssea/farmacocinética , Fígado/metabolismo , Osteoprotegerina/farmacocinética , Polietilenoglicóis/química , Animais , Transporte Biológico , Conservadores da Densidade Óssea/administração & dosagem , Conservadores da Densidade Óssea/sangue , Conservadores da Densidade Óssea/química , Células Cultivadas , Química Farmacêutica , Regulação para Baixo , Ensaio de Imunoadsorção Enzimática , Feminino , Heparina/metabolismo , Humanos , Injeções Intravenosas , Rim/metabolismo , Maleatos/química , Camundongos , Osteoclastos/efeitos dos fármacos , Osteoprotegerina/administração & dosagem , Osteoprotegerina/sangue , Osteoprotegerina/química , Ovariectomia , Perfusão , Ratos , Ratos Sprague-Dawley , Baço/metabolismo , Distribuição TecidualRESUMO
To assess whether the antigenic properties of H5 hemagglutinin (HA) change over time due to antigenic drift, we produced a panel of monoclonal antibodies (mAbs) against the HA of the index H5N1 human influenza A virus, A/Hong Kong/156/97. By immunizing mice with a plasmid expressing this HA and boosting the initial immunization with cell lysates transfected with the plasmid, a total of six hybridomas producing HA-specific mAbs were established: four to the HA1 subunit with hemadsorption-inhibiting activity and two to the HA2 subunit. None of the mAbs to HA1 could bind to the HA of a recent human isolate, A/Hong Kong/213/2003, indicating that there are substantial antigenic differences between the H5N1 human influenza virus isolated in 1997 and that isolated in 2003.
