A targeting method for lentiviral vectors relying on the use of singlechain antibodies recognizing cellsurface antigens is applied to generate lentiviral vectors specific for endothelial cells. This work was supported by the intramural research program of the national. Spleenectomy definition of spleenectomy by the free. Thirdgeneration lentiviral sin vectors were produced using a fourplasmid split packaging system as described before and stored at. Male balbc mice received intravenous injections of appropriately diluted friend polycythemia virus fvp. L wolff and s ruscetti laboratory of genetics, national cancer institute, bethesda, maryland 20892. Rearrangements due to sffv integration were found in 95% of the erythroid tumors studied. The friend spleen focusforming virus sffv encodes a unique envelope. In this study, we tested various bdd constructs in the context of either wildtype or codonoptimized cdna sequences expressed under control of the strong, ubiquitous spleen focus forming virus promoter within a selfinactivating hivbased lentiviral vector. To enhance the level of gene expression in hematopoietic cells, we also generated a series of lentiviral vectors incorporating the spleen focus forming virus. Pyrogens can be nonbacterial as well as bacterial in origin 14.
Spleen cells from the diseased mice expressed the sffv env gene product but not. Spleen focusforming virus how is spleen focusforming virus abbreviated. Benjamin eberts lab contains the inserts spcas9, sp sgrna scaffold, sffv, and p2atrfp and is published in nat biotechnol. Promoter optimisation of lentiviral vectors for efficient. However, the main pyrogen encountered in the pharmaceutical industry is. Sffv is composed of genetic sequences homologous to friend type c helper virus fmulv and sffvspecific sequences not present in fmulv. Mink cell focus inducing viruses is a descriptor in the national library of medicines controlled vocabulary thesaurus, mesh medical subject headings. Linemeyer dl, ruscetti sk, scolnick em, evans lh, duesberg ph 1981 biological activity of the spleen focus forming virus is encoded by a molecularly cloned subgenomic fragment of spleen focus forming virus dna. Silvia monticellis lab contains the insert shrna no. Sffv is defined as spleen focus forming virus frequently. Spleenectomy synonyms, spleenectomy pronunciation, spleenectomy translation, english dictionary definition of spleenectomy. Quantitation of friend spleen focusforming virus by a. Similar results were observed when the cmv promoter was replaced with human. Cellspecific and efficient expression in mouse and human.
A strain of murine leukemia virus associated with mouse tumors similar to those caused by the friend murine leukemia virus. Glycoprotein encoded by the friend spleen focusforming virus. The human immunodeficiency virus hivspleen focusforming virus. Selection of an optimal promoter for gene transfer in normal b cells. Nonidet p40 np40, and centrifuged for 5 min at 3,000.
The following deletion constructs of sfstk were generated by pcr using. This gene encodes an etsdomain transcription factor that activates gene expression during myeloid and blymphoid cell development citation needed. Spi1 spleen focus forming virus sffv proviral integration oncogene spi1, authors. Lentiviral transduction of mammalian cells for fast. Spleen necrosis virus how is spleen necrosis virus. Codon optimization of human factor viii cdnas leads to. As the primary cellular targets of ebov infection are macrophages and conventional dendritic cells cdcs, followed by hepatocytes, we titrated virus in both the spleen and liver by focus forming assay ffa and quantitative realtime reverse transcriptase polymerase chain reaction qrtpcr. A previously described 3day 59fe assay for quantitation of friend spleen focus forming virus has been modified to produce a 200fold more sensitive 9day 59fe assay. This protocol describes a suite of lentiviral transfer plasmids that can be used for highyield, time and costefficient, and constitutive or inducible production of soluble and membrane proteins. Highlevel transduction and gene expression in hematopoietic. Production of spleen focus forming virus and murine leukemia virus by early erythroblasts after friend virus infection t. Words that match the pattern sple onelook dictionary. Spleen focus forming viruses is a descriptor in the national library of medicines controlled vocabulary thesaurus, mesh medical subject headings.
The friend spleen focusforming virus fsffv released from cultured erythroleukemia cells cell line f46k was cloned free of its helper lymphatic leukemia virus fmulv. Pyrogens and endotoxins are a heterogeneous group of chemical entities that share the characteristic of when injected being able to cause fever. The spleen focus forming virus promoter conferred the strongest transcriptional. The spleen focus forming virus sffv envelope gene, when introduced into mice in the absence of other sffv genes, induces acute erythroleukemia. It can act as a helper virus when complexing with a defective transforming component, rauscher spleen focus forming virus. Expression of the selectable marker in these viruses can be regulated by the homologous transcriptional promoter of the gene, by the promoter contained within the friend spleen focus forming virus. Two recombinant retroviruses expressing eporr129c, spleen focus forming virus sffvcepor and myeloproliferative sarcoma virus mpsvcepor, were used to infect fetal liver cells that served as a source of hematopoietic progenitors.
Definition of spleen focus forming viruses in the dictionary. A previously described 3day 59fe assay for quantitation of friend spleen focusforming virus has been modified to produce a 200fold more sensitive 9day 59fe assay. Specific gene transfer to neurons, endothelial cells and. Information and translations of spleen focus forming viruses in the most comprehensive dictionary definitions resource on the web.
Spleen removal process, free spleen removal process software downloads, page 2. Loxp sites were inserted in intron 3 and a loxpflanked neomycin selection cassette was inserted into intron 5. Spleen focusforming virus how is spleen focusforming. The genomic locus was called spi1, for sffv proviral integration. Transduced 293t cells in vitro demonstrated detectable fviii activity. A recent report described two xcgd patients treated by autologous transplantation of hsc transduced with a murine leukemia virus derived retroviral vectorexpressing gp91 phox from the spleen. Among hematopoietic cell lines, iw2 and 745a are erythroleukemic cell lines derived from friend murine leukemia virus fmulv or spleen focus forming virus sffvinduced erythroid tumor cells, 921 is an hs2 erythroleukemic cell line from spi1transgenic mice, n and n11 are macrophagic cell lines, fdcp1 is a multipotent progenitor. They have also attracted considerable attention as a potential tool for. Prt perlovga removal tool removes the so called temp2. Friend spleen focusforming virus glycoprotein gp55. It is a replicationcompetent murine leukemia virus. Immunization of rats with syngeneic cells infected with spleen focus forming virus sffv but not with its helper, friend murine leukemia virus fmulv, produces antisera which specifically neutralize sffv, and not fmulv, in the friend virus complex. Bondurant division of hematology, vanderbilt university school of medicine and veterans administration medical center, nashville, usa.
A viral antigen specified by spleen focus forming virus and differentiation antigen controlled by the fv2 locus. After allowing adsorption to sc1 fibroblasts at a low multiplicity of infection. Rescue of spleen focus forming activity was obtained on i fection of these cells with mulvf or mulvmol, but not with mulvgross. Looking for online definition of splc or what splc stands for.
Spi1 spleen focus forming virus sffv proviral integration oncogene house mouse gene id. The nuclear protein binds to a purinerich sequence known as the pubox found on enhancers of target genes, and regulates their expression in. Transcriptional correlates of tolerance and lethality in. Viruses free fulltext friend spleen focusforming virus. Promoter optimisation of lentiviral vectors for efficient insulin gene. Friend spleen focusforming virus activates the tyrosine kinase sfstk and the transcription factor pu. Umbilical cord as a longterm source of activatable. Benjamin eberts lab contains the inserts spcas9, sp sgrna scaffold, sffv, and p2aegfp and is published in nat biotechnol. Infection of mice with friend spleen focusforming virus sffv results in a. Removal of the downstream ires gfp from the lentiviral vector did not. Spleenectomy article about spleenectomy by the free.
Differential susceptibility of spleen focusforming virus. Envelope gene of the friend spleen focusforming virus. It contains a 32s, presumably defective, genome, which most likely is responsible for spleen focus formation, and a 35s helper virus genome. Descriptors are arranged in a hierarchical structure, which enables searching at various levels of specificity. Spleen removal process software free download spleen. Interleukin 3dependent lymphoid cell lines that have been stably transfected with the epor. Various studies have shown that blood loss was minimal with laparoscopic splenectomy compared with open spleenectomy.
The spleen focusforming virus sffv envelope gene, when introduced into. Splc is listed in the worlds largest and most authoritative dictionary database of abbreviations and acronyms the free dictionary. Erythroblast transformation by the friend spleen focusforming. The spleen focus forming virus sffv promoter was replaced by the chromatinopening element cbx3 in combination with the elongation factor 1. The envelope glycoprotein of friend spleen focusforming virus. The friend spleen focusforming virus sffv is a highly pathogenic retrovirus. Sple is listed in the worlds largest and most authoritative dictionary database of abbreviations and acronyms the free dictionary.
Gene transduction efficiency in cells of different species. The rna of the rv complex resembles closely that of friend virus fv. Production of spleen focusforming virus and murine. Thus, the drugs might be useful in distinguishing other types of oncornaviruses. This work was supported by the intramural research program of the. Our findings indicate a remarkable biological selectivity of ansamycins, as well as nonansamycin components of svcx, against the transforming and defective spleen focus forming virus as compared to mulv. Lineage and stagerestricted lentiviral vectors for the. To determine the biological significance of the 585basepair deletion in the env gene of friend spleen focusforming virus sffv encoding a leukemogenic. Abstractgp55p is a dimeric membrane protein with a single transmembrane helix that is coded by the env gene of the polycythemic strain of the spleen focus forming virus.