In order to use retroviral-mediated gene transfer technology in clinical application, retroviral vector must be of high titer and free of detectable replication-competent retroviruses (RCR). The aim of this study was to optimize methods of defective retroviral vector production. Study was conducted using a LXSN vector inserted with human tumor necrosis factor-? gene and an amphotropic retrovirus packaging cell line-PA317. The results indicated that viral titer was influenced by volume of medium and concentration of fetal calf serum. Inactivation of retroviral vector was greater at 37癈 than at 32癈. In experiment of transfection of PA317 and transduction of 3T3, integration of retroviral vector into genome of packaging cells and target cells, and free of RCR were detected by polymerase chain reaction analysis. Viral vector with high titer and free of RCR is able to use in clinical trial

Purpose To study the reversal effect on multidrug resistance (MDR) by TNF-α gene combined with verapamil (VRP) or tamoxifen (TAM). Methods By using recombinant retrovirus vector, TNF-α gene was transfected into multidrug-resistant human breast cancer cell line MCF7/ADR. The TNF-α secreting cell clone MCF7/ADR-TNF was obtained by G418 screening. The integrating and secreting of TNF-α were analyzed by PCR and ELISA. MTT assay and formula"I = d/D1 + d/D2" were used to evaluate the reversal effect of multidmg resistance with TNF-α gene combined with verapamil or tamoxifen. ResultsThe level of TNF-α secreted by MCF7/ADR-TNF was 1 737 pg/ml (106cells/48 h). Compared with control,the resistance to ADR of MCF7/ADR-TNF was reversed by 1.6 times. The reversal effect produced by combination of TNF-α gene and VRP was antagonistic. The combination of TNF-α gene and TAM produced synergic effect (interaction index I = 0.64). ConclusionsTNF-α gene combined with TAM has synergic effect on reversing MDR.

Over the past few years, the human virome and its complex interactions with microbial communities and the immune system have gained recognition as a crucial factor in human health. Individuals with compromised immune function encounter distinctive challenges due to their heightened vulnerability to a diverse range of infectious diseases. This review aims to comprehensively explore and analyze the growing evidence regarding the role of the virome in immunocompromised disease status. By surveying the latest literature, we present a detailed overview of virome alterations observed in various immunodeficiency conditions. We then delve into the influence and mechanisms of these virome changes on the pathogenesis of specific diseases in immunocompromised individuals. Furthermore, this review explores the clinical relevance of virome studies in the context of immunodeficiency, highlighting the potential diagnostic and therapeutic gains from a better understanding of virome contributions to disease manifestations.
Humans ; Viruses ; Virome ; Microbiota ; Immunologic Deficiency Syndromes