RESUMO
This study was designed to see if immunosuppression achieved using local application of cyclosporine A (Cs. A) or CD4 and CD8 antibodies would improve bone formation following intramuscular injections of human BMP-4 and BMP-9 adenoviral vectors (ADhBMP4 and ADhBMP9) in Sprague-Dawley rats. Cs. A was injected into the thigh muscle. After 2 days, ADhBMP4, ADhBMP9, and the antibodies were separately injected into the left and right rear legs. At this time, the number of CD4+/CD3+ cells was significantly lower and the number of CD8+/CD3+ cells higher in the Cs. A group than in the control group (P < 0.01). The total number of white blood cells 3 days following injection of CD4 and CD8 antibodies was significantly lower than that before the injection (P < 0.01). At 4 weeks after the viral and antibody injections, mean bone volumes at the ADhBMP9 treatment sites were 0.29 +/- 0.01 cm3 in the viral control group, 0.17 +/- 0.03 cm3 in the Cs. A-ADhBMPs group, and 0.59 +/- 0.07 cm3 in the antibodies-ADhBMPs group. ADhBMP4 did not induce new bone formation in any group. This study demonstrates that local immunomodulation may improve the osteogenic potential of bone morphogenetic protein gene therapy in the clinical setting.
Assuntos
Antígenos CD/administração & dosagem , Autoanticorpos/administração & dosagem , Proteínas Morfogenéticas Ósseas/genética , Terapia Genética/métodos , Terapia de Imunossupressão/métodos , Osteogênese/imunologia , Adenoviridae/genética , Animais , Autoanticorpos/imunologia , Proteína Morfogenética Óssea 4 , Proteínas Morfogenéticas Ósseas/uso terapêutico , Complexo CD3/imunologia , Antígenos CD4/imunologia , Antígenos CD8/imunologia , Terapia Combinada , Ciclosporina/administração & dosagem , Vetores Genéticos/administração & dosagem , Fator 2 de Diferenciação de Crescimento , Fatores de Diferenciação de Crescimento , Membro Posterior , Imunossupressores/administração & dosagem , Injeções Intramusculares , Contagem de Leucócitos , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Transdução Genética/métodosRESUMO
Circular dichroic (CD) spectra of two turnip isoperoxidases, P1 and P7, and of their derivatives were measured over the wavelength range of 200 to 650 nm. For the two isoenzymes, it was observed that although the visible and Soret bands are located at similar wavelengths, their ellipticities are different. These results suggest that the active sites are similar but that differences do exist. The results are compared with those reported for Japanese radish peroxidase a and horseradish peroxidase. It appears that a common property of plant peroxidases is the presence of negative CD Soret bands for the reduced forms and their inversion upon cyanide binding. The CD spectra in the far UV region indicate an appreciable helical content for both native enzymes and their various derivatives. The calculated contents of unordered structure are very high (greater than 50% for either P1 or P7), in agreement with other studies on glycoproteins.