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

Adela R. Cardones, Wolfgang W. Leitner, Lei Fang, Takashi Murakami, Veena Kapoor, Mark C. Udey, Samuel T Hwang

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

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 CD31lo podoplaninhi 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.

Original languageEnglish (US)
Pages (from-to)32-39
Number of pages8
JournalMicrovascular Research
Volume71
Issue number1
DOIs
StatePublished - Jan 2006
Externally publishedYes

Keywords

  • Angiogenesis
  • Immunization
  • Lymphatics

ASJC Scopus subject areas

  • Biochemistry
  • Cardiology and Cardiovascular Medicine
  • Cell Biology

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