RESUMO
With the long-term goal of developing a gene-based treatment for osteoarthritis (OA), we performed studies to evaluate the equine joint as a model for adeno-associated virus (AAV)-mediated gene transfer to large, weight-bearing human joints. A self-complementary AAV2 vector containing the coding regions for human interleukin-1-receptor antagonist (hIL-1Ra) or green fluorescent protein was packaged in AAV capsid serotypes 1, 2, 5, 8 and 9. Following infection of human and equine synovial fibroblasts in culture, we found that both were only receptive to transduction with AAV1, 2 and 5. For these serotypes, however, transgene expression from the equine cells was consistently at least 10-fold higher. Analyses of AAV surface receptor molecules and intracellular trafficking of vector genomes implicate enhanced viral uptake by the equine cells. Following delivery of 1 × 10(11) vector genomes of serotypes 2, 5 and 8 into the forelimb joints of the horse, all three enabled hIL-1Ra expression at biologically relevant levels and effectively transduced the same cell types, primarily synovial fibroblasts and, to a lesser degree, chondrocytes in articular cartilage. These results provide optimism that AAV vectors can be effectively adapted for gene delivery to large human joints affected by OA.
Assuntos
Dependovirus/genética , Técnicas de Transferência de Genes , Proteína Antagonista do Receptor de Interleucina 1/genética , Osteoartrite/genética , Animais , Cartilagem Articular/metabolismo , Cartilagem Articular/patologia , Cartilagem Articular/virologia , Vetores Genéticos , Proteínas de Fluorescência Verde/genética , Cavalos , Humanos , Interleucina-1/genética , Articulações/metabolismo , Articulações/patologia , Articulações/virologia , Osteoartrite/terapia , Membrana Sinovial/metabolismo , Membrana Sinovial/patologia , Membrana Sinovial/virologiaRESUMO
Serial hippuran renograms were performed in pediatric allograft recipients in an effort to detect the onset of rejection following the initial 3 month post transplant period. The plots of the normalized effective renal plasma flows (NERPF) were compared with reciprocal of the serum creatinines. A strong correlation was noted between both values. NERPF could be predicted from the serum creatinine utilizing a logarithmic equation. However, there was no detectable difference between the onset of changes in the slopes of either variable. Therefore, serial hippuran renograms were no better than monitoring serum creatinine in detecting allograft dysfunction when performed at the frequency of the study.
