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Reagent Information
Name Ad-hCMV-Flt3L. 
Reagent Type Vector 
Cell Line Origin  
Short Description  
Long Description This replication-defective, first generation adenoviral vector is a based on human adenovirus serotype 5, with deletions in its E1a and E3 viral encoding regions. This vector encodes the transgene for human soluble fms-like tyrosine kinase 3 ligand (Flt3L) under the control of the human cytomegalovirus (hCMV) promoter. The Ftl3L transgene was subcloned into the first generation adenoviral shuttle plasmid pAL119 containing the hCMV promoter to make the plasmid pAL119-hsFlt3L. The adenoviral vector genome plasmid pJM17 (Microbix) and pAL119-hsFlt3L were co-transfected in 293 cells (Microbix) and the adenoviral vector Ad-hCMV-Flt3L was rescued by homologous recombination. Ad-hCMV-Flt3L underwent three successive rounds of plaque purification and then amplified on 293 cells. Ad-hCMV-Flt3L was purified from the 293 cell lysate by cesium chloride purification followed by three rounds of dialysis in 10mM Tris, ph 8.0.  
Vector Map Ad vector backbone.pdf
Grade Research 
Depositor Cedars-Sinai Medical Center, Los Angeles, CA 
References Publications: 1. Ali S, Curtin JF, Zirger JM, Xiong W, King GD, Barcia C, Liu C, Puntel M, Goverdhana S, Lowenstein PR, Castro MG. Inflammatory and anti-glioma effects of an adenovirus expressing human soluble Fms-like tyrosine kinase 3 ligand (hsFlt3L): treatment with hsFlt3L inhibits intracranial glioma progression. Molecular Therapy . 2004 Dec;10(6):1071-84. PMID: 15564139. 2. Ali S, King GD, Curtin JF, Candolfi M, Xiong W, Liu C, Puntel M, Cheng Q, Prieto J, Ribas A, Kupiec-Weglinski J, van Rooijen N, Lassmann H, Lowenstein PR, Castro MG. Combined immunostimulation and conditional cytotoxic gene therapy provide long-term survival in a large glioma model . Cancer Research . 2005 Aug 15;65(16):7194-204. PMID: 16103070. 3. Curtin JF, King GD, Barcia C, Liu C, Hubert FX, Guillonneau C, Josien R, Anegon I, Lowenstein PR, Castro MG. Fms-like tyrosine kinase 3 ligand recruits plasmacytoid dendritic cells to the brain . Journal of Immunology . 2006 Mar 15;176(6):3566-77. PMID: 16517725. 4. King GD, Muhammad AK, Curtin JF, Barcia C, Puntel M, Liu C, Honig SB, Candolfi M, Mondkar S, Lowenstein PR, Castro MG. Flt3L and TK gene therapy eradicate multifocal glioma in a syngeneic glioblastoma model . Neuro-Oncology . 2008 Feb;10(1):19-31. PMID: 18079358 5. King GD, Kroeger KM, Bresee CJ, Candolfi M, Liu C, Manalo CM, Muhammad AK, Pechnick RN, Lowenstein PR, Castro MG. Flt3L in combination with HSV1-TK-mediated gene therapy reverses brain tumor-induced behavioral deficits. Molecular Therapy . 2008 Apr;16(4):682-90. PMID: 18283279 . 6. Curtin JF, Candolfi M, Fakhouri TM, Liu C, Alden A, Edwards M, Lowenstein PR, Castro MG. Treg depletion inhibits efficacy of cancer immunotherapy: implications for clinical trials . PLoS One . 2008 Apr 23;3(4):e1983. PMID: 18431473 7. Curtin JF, Liu N, Candolfi M, Xiong W, Assi H, Yagiz K, Edwards MR, Michelsen KS, Kroeger KM, Liu C, Muhammad AK, Clark MC, Arditi M, Comin-Anduix B, Ribas A, Lowenstein PR, Castro MG. HMGB1 mediates endogenous TLR2 activation and brain tumor regression . PLoS Medicine . 2009 Jan 13;6(1):e10. PMID: 19143470 8. Candolfi M, Yagiz K, Foulad D, Alzadeh GE, Tesarfreund M, Muhammad AK, Puntel M, Kroeger KM, Liu C, Lee S, Curtin JF, King GD, Lerner J, Sato K, Mineharu Y, Xiong W, Lowenstein PR, Castro MG. Release of HMGB1 in response to proapoptotic glioma killing strategies: efficacy and neurotoxicity. Clinical Cancer Research . 2009 Jul 1;15(13):4401-14. PMID: 19570774 9. Ghulam Muhammad AK, Candolfi M, King GD, Yagiz K, Foulad D, Mineharu Y, Kroeger KM, Treuer KA, Nichols WS, Sanderson NS, Yang J, Khayznikov M, Van Rooijen N, Lowenstein PR, Castro MG. Antiglioma immunological memory in response to conditional cytotoxic/immune-stimulatory gene therapy: humoral and cellular immunity lead to tumor regression . Clinical Cancer Research . 2009 Oct 1;15(19):6113-27. PMID: 19789315 10. Larocque D, Sanderson NSR, Bergeron J, Curtin JF, Girton J, Wibowo M, Bondale N, Kroeger KM, Yang J, Lacayo LM, Reyes KC, Farrokhi C, Pechnick, Castro MG, Lowenstein PR. Exogenous Flt3L overrides the brain’s immune privilege and facilitates the recognition of a neo-antigen: implications for brain immunotherapies . The Proceedings of the National Academy of Sciences, USA . 2010 (In Press)  
Biosafety The Indiana University Vector Production Facility works with this line under BL-2 containment but the level of containment required of any cell line or vector is at the descretion of the investigator's Institutional Biosafety Committee that may require a different level of containment. 
Availability 1 vial (10 uL). Titer ~1x10x10e13 vp/mL