Influence of AAV-2 terminal repeats on expression of heterologous promoters

The adeno-associated virus is a human parvovirus that can be used as a vector system for gene transduction into mammalian cells. For integration into the host cell genome, replication and packaging of the viral DNA only terminal repeats are necessary in cis. We investigated the influence of the terminal repeats on the expression of genes under the control of heterologous promoters in transient transfection assays. The activity of HPV-18, HCMV and HIV-1 promoters in vectors with and without terminal repeats was compared using luciferase as a reporter gene. We observed a reduction of gene expression from all promoters tested in plasmids with terminal repeats. The inhibitory effect was abolished with the expression of one viral regulatory protein and infection with adenovirus 5. RESUMEN El virus adeno-asociado es un parvovirus humano que puede ser utilizado como vector para la transduccion genica en celulas de mamiferos. Para la integracion en la celula hospedera, la replicacion y el empaquetamiento del ADN viral, solo se requieren de las secuencias terminales repetidas en cis. Nosotros estudiamos la influencia de estas secuencias en la expresion de genes bajo el control de promotores heterologos en experimentos de transfecion transiente. La actividad de promotores de HPV-18, HCMV y HIV-1 en vectores con y sin terminales repetidos, se comparo usando la luciferasa como gen reportero. Se observo una disminucion de los niveles de expresion con todos los promotores usados en plasmidios que contienen la secuencia de los terminales repetidos. El efecto inhibitorio se elimino con la expresion de una de las proteinas reguladoras del virus y la infeccion con adenovirus 5

Haematopoietic stem cell transplantation for severe autoimmune diseases in children: A review of current literature, registry activity and future directions on behalf of the autoimmune diseases and paediatric diseases working parties of the European Society for Blood and Marrow Transplantation

Although modern clinical management strategies have improved the outcome of paediatric patients with severe autoimmune and inflammatory diseases over recent decades, a proportion will experience ongoing or recurrent/relapsing disease activity despite multiple therapies often leading to irreversible organ damage, and compromised quality of life, growth/development and long-term survival. Autologous and allogeneic haematopoietic stem cell transplantation (HSCT) have been used successfully to induce disease control and often apparent cure of severe treatment-refractory autoimmune diseases (ADs) in children. However, transplant-related outcomes are disease-dependent and long-term outcome data are limited in respect to efficacy and safety. Moreover, balancing risks of HSCT against AD prognosis with continually evolving non-transplant options is challenging. This review appraises published literature on HSCT strategies and outcomes in individual paediatric ADs. We also provide a summary of the European Society for Blood and Marrow Transplantation (EBMT) Registry, where 343 HSCT procedures (176 autologous and 167 allogeneic) have been reported in 326 children (<18 years) for a range of AD indications. HSCT is a promising treatment modality, with potential long-term disease control or cure, but therapy-related morbidity and mortality need to be reduced. Further research is warranted to establish the position of HSCT in paediatric ADs via registries and prospective clinical studies to support evidence-based interspeciality guidelines and recommendations