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J Immunol Methods. 2014 Nov;413:69-73. doi: 10.1016/j.jim.2014.07.008. Epub 2014 Jul 24.

Robust expansion of dendritic cells in vivo by hydrodynamic FLT3L-FC gene transfer.

Author information

1
LakePharma Inc., 530 Harbor Blvd., Belmont, CA 94002, USA.
2
Palo Alto Veterans Institute for Research & Veterans Affairs Palo Alto Health Care System, 3801 Miranda Ave., Palo Alto, CA 94304, USA.
3
Stanford University School of Medicine, Department of Pathology, 300 Pasteur Dr., Lane 235, Stanford, CA 94305, USA.
4
Palo Alto Veterans Institute for Research & Veterans Affairs Palo Alto Health Care System, 3801 Miranda Ave., Palo Alto, CA 94304, USA. Electronic address: bazabel@alum.mit.edu.

Abstract

Due to low numbers of endogenous dendritic cells (DCs) in vivo, exogenous DC-poietin Fms-like tyrosine kinase 3-ligand (FLT3L) is routinely used to generate DC for subsequent studies. We engineered a novel FLT3L-FC DNA construct that, when combined with hydrodynamic gene transfer (HDT), induced robust DC expansion in mice. DC generated in vivo by FLT3L-FC HDT produced cytokines in response to stimulation by an array of TLR agonists and promoted T cell proliferation. The FLT3L-FC protein produced in vivo spontaneously homodimerized to enable effective FLT signaling and the FC-domain enhanced its plasma half-life, providing an improved reagent and method to boost DC numbers.

KEYWORDS:

Cytokine; Pharmacodynamics; Pharmacokinetics; Progenitor

PMID:
25066631
PMCID:
PMC4253009
DOI:
10.1016/j.jim.2014.07.008
[Indexed for MEDLINE]
Free PMC Article
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