Recombinant human CD19L-sTRAIL effectively targets B cell precursor acute lymphoblastic leukemia

Low Dose Total Body Irradiation Combined With Recombinant CD19-Ligand×Soluble TRAIL Fusion Protein is Highly Effective Against Radiation-resistant B-precursor Acute Lymphoblastic Leukemia in Mice

“…The CD19-specific immunoreactivity of CD19L-sTRAIL was confirmed by demonstrating that fluorescent-labeled CD19L-sTRAIL binds to the surface membrane of CD19 + human leukemia cells, but not to CD19 receptor-negative human 293T cells (Uckun et al, 2015a). The binding of CD19L-sTRAIL to CD19 + human leukemia cells was mediated via its CD19L domain which was directed to the CD19 extracellular domain, because it could be blocked by an IgG1-Fc fusion protein of recombinant human CD19L used as a CD19 receptor-specific competitor as well as by a recombinant CD19 extracellular domain protein (but not by a recombinant CD19 intracellular domain protein) (Uckun et al, 2015a). CD19L-sTRAIL caused apoptosis in the CD19 receptor positive ALL-1 cell line in a concentration-dependent fashion with ~90% apoptosis at 210 fM and >99% apoptosis at 2.1 pM (Uckun et al, 2015a).…”

“…A stepwise molecular cloning strategy was employed using the commercially available pFUSE-hIgG1-Fc2 plasmid (Invivogen, CA) as the backbone vector to construct an expression cassette for CD19L-sTRAIL (Uckun et al, 2015a). CD19L-sTRAIL is a fusion protein of sTRAIL connected to the C-terminus of the CD19L protein through a flexible linker (Figure S1A).…”

“…CD19L-sTRAIL is a fusion protein of sTRAIL connected to the C-terminus of the CD19L protein through a flexible linker (Figure S1A). It was produced in a mammalian expression system, as recently described in detail (Uckun et al, 2015a). CD19L-sTRAIL was purified using anion exchange (AIE) chromatography and size exclusion (SE) chromatography (Figure S1B1).…”

“…Cells were analyzed for apoptosis at 48 h after initiation of treatment using the standard quantitative flow cytometric apoptosis assay with the Annexin V-FITC Apoptosis Detection Kit from Sigma (St. Louis, MO, as previously reported (Uckun et al, 2015a, Uckun et al, 2013, 2011b). The labeled cells were analyzed on a LSR II flow cytometer (Becton Dickinson, Lakes, NJ).…”

Value of Liberal Arts education in the Era of Precision Medicine

“…Data mining, bioinformatics, computational modeling and simulation have increasingly become integral to development of novel hypotheses in disease progression and potentially novel therapeutic innovations that target prominent receptors in leukemias (CD19 and CD22 receptors) to eradicate cancer cells. [14][15][16][17][18][19][20] Our project attempted to link data mining efforts that identify genetic signatures in high risk patient sub-groups to data explorations methods developed using R made available in Bioconductor (http://www.bioconductor. org/) by writing interactive, exploration based statistical analysis platform provided by Shiny applications (http://shiny.rstudio.com/).…”