Targeting tissue factor on tumor vascular endothelial cells and tumor cells for immunotherapy in mouse models of prostatic cancer

Hu, Zhiwei; Garen, Alan

doi:10.1073/pnas.201420298

Cited by 103 publications

(173 citation statements)

References 23 publications

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“…8 Plasmids: m fVII/IgG 1 Fc and mfVII-GFP The adenoviral vector encoding the mfVII immunoconjugate-PcDNA3-mIcon was obtained from the Garen laboratory. [17][18][19][20] The amplified mfVII cDNA, which contains the leader and coding sequences without a stop codon, was cloned into the HindIII and BamHI sites of the pcDNA3.1( þ ) vector (Invitrogen, Carlsbad, CA) in-frame with a cDNA encoding the human IgG1 Fc domain. The vector DNA was amplified in HB101 competent cells (Life Technologies, Grand Island, NY) and sequenced.…”

Section: Cell Linesmentioning

confidence: 99%

“…The vector DNA was amplified in HB101 competent cells (Life Technologies, Grand Island, NY) and sequenced. The active site of mfVII cDNA was mutated as described originally by Hu and Garen [17][18][19][20] substituting an alanine codon for Lys-341. The mutagenesis procedure was carried out as described in the QuickChange site-directed mutagenesis manual (Stratagene, La Jolla, CA).…”

Section: Cell Linesmentioning

confidence: 99%

“…[17][18][19][20] In vivo intravital microscopy (IVM) Dorsal skin fold chambers in the mouse were prepared as described previously. 8,21 Tumor spheroids were derived from N202-H2B-GFP and was plated at 1 Â 10 5 cells/well in 100 ml/well in a 96-well round-bottomed plates coated with 50 ml of 1% agarose.…”

Section: Cell Linesmentioning

confidence: 99%

“…[17][18][19][20] Fluorescence-activated cell sorting analysis of binding of the m fVII/IgG 1 Fc protein to tissue factor The tumor cells were harvested in nonenzymatic dissociation solution (Sigma-Aldrich, St. Louis, MO), washed and resuspended in tris-buffered saline/BSA/Ca 2 þ (10 mM, Tris Á HCl at pH 7.4/150 mM, NaCl/20 mM, CaCl 2 /1% BSA and 0.1% NaN 3 ). The mfVII/hIgG 1 Fc immunoconjugate protein was then added (5 mg/ml final concentration), and the cells were incubated for 30 min at 371C.…”

Section: Purification Of Mfvii-gfp Proteinmentioning

confidence: 99%

“…Testing of the effect of a single intravenous injection of the fVII/IgG 1 Fc immunoconjugate protein on the vessels of the N202 breast cancer spheroid in the dorsal skin fold chamber of live mice As mentioned above and published previously, [17][18][19] Abbreviation:GFP, green fluoroscent protein. We studied the effect of intravenous injections of the mfVII/hIgG 1 Fc protein which extended over 11 days.…”

Section: Distribution Of Receptors On Vascular Endothelial Cells Of Tmentioning

confidence: 99%

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Mapping of angiogenic markers for targeting of vectors to tumor vascular endothelial cells

et al. 2007

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The vasculature of mouse breast tumor spheroids grown on mammary fat pad tissue in an intravital microscopy (IVM) viewing chamber was shown to derive from infiltrating angiogenic mammary vessels. The receptors tissue factor (TF), a V b 3 integrin and Tie-2 were expressed on the vascular endothelium in the periphery but not in the center of the tumor spheroids nor in the mammary tissue nor in smooth muscle tissue, whereas Tie-1 and PCAM-1 were expressed extensively in the entire tumor and in the vascular endothelium of the entire tumor nodule and in normal mammary tissue. TF is a specific target for adenoviral vector-mediated cancer immunotherapy. Subcutaneous injection of the AdfVII/IgG 1 Fc vector leads to the release into the system circulation of a fVII/ IgG 1 Fc immunoconjugate molecule that binds specifically and tightly to TF on vascular endothelial cells and tumor cells, activating a cytolytic immune response against the targeted cells. We show that a single administration of the AdfVII/IgG 1 Fc vector destroys the peripheral but not the central vasculature of a tumor spheroid, causing partial tumor regression; additional administrations prevent regeneration of the peripheral vasculature and regrowth of the tumor. These findings indicate that a critical parameter for optimizing tumor damage is the schedule for successive administrations of the AdfVII/IgG 1 Fc, which should coincide with the regeneration of the peripheral vasculature and continue until the tumor is destroyed.

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Section: Cell Linesmentioning

confidence: 99%