Genetic immunization with LYVE-1 cDNA yields function-blocking antibodies against native protein.

Published

Journal Article

LYVE-1 is a surface bound hyaluronic acid (HA) receptor that is preferentially expressed by lymphatic endothelial cells (LEC). cDNA encoding full-length human LYVE-1 was coated onto gold particles that were then delivered via helium-assisted jet propulsion (gene gun) into the skin of Balb/C mice. LYVE-1 antisera, but not control pre-immune sera, recognized LYVE-1-transfected 293T cells by flow cytometry. While 40-70% of cultured human dermal microvascular endothelial cells (HMEC) were positive for LYVE-1 staining, human lung microvascular endothelial cells (LMEC) were negative. LYVE-1 antisera was used to effectively separate HMEC into LYVE-1 (hi) and LYVE-1(lo) populations that were enriched or depleted, respectively, for podoplanin, another LEC marker. By immunohistochemistry, LYVE-1 antisera detected CD31(lo) podoplanin(hi) lymphatic channels in normal and psoriatic human skin as well as in human tonsil. LYVE-1 antisera also blocked binding of FITC-labeled HA to HMEC (but not LMEC), demonstrating that these antibodies recognized regions of LYVE-1 required for HA binding. In summary, gene gun-assisted delivery of cDNA encoding LYVE-1 into skin resulted in reliable production of antisera that specifically and functionally recognized native LYVE-1 protein.

Full Text

Duke Authors

Cited Authors

  • Cardones, AR; Leitner, WW; Fang, L; Murakami, T; Kapoor, V; Udey, MC; Hwang, ST

Published Date

  • January 2006

Published In

Volume / Issue

  • 71 / 1

Start / End Page

  • 32 - 39

PubMed ID

  • 16257423

Pubmed Central ID

  • 16257423

International Standard Serial Number (ISSN)

  • 0026-2862

Digital Object Identifier (DOI)

  • 10.1016/j.mvr.2005.09.002

Language

  • eng

Conference Location

  • United States