Adenovirus vector-mediated gene transfer to regional lymph nodes. Academic Article uri icon

Overview

MeSH

  • Animals
  • Drainage
  • Gene Expression Regulation
  • Interleukin-12
  • Luciferases
  • Male
  • Mice
  • Mice, Inbred C3H
  • Transgenes

MeSH Major

  • Adenoviridae
  • Gene Transfer Techniques
  • Genetic Vectors
  • Lymph Nodes

abstract

  • Regional lymph nodes (RLNs) possess important immune functions and represent a major pathway of metastasis for solid tumors. Given these facts, the ability to transfer exogenous genes to the RLNs with the goal of manipulating the local immunological milieu would be desirable. On the basis of the hypothesis that a significant proportion of adenovirus (Ad) gene transfer vectors traffic through the lymphatics, E1-E3- Ad vectors were injected into the hind footpad of C3H/He mice and the RLNs assessed for vector trafficking and transgene expression. A low dose (10(9) particles) of an Ad vector encoding the firefly luciferase gene (Ad-CMV.Luc) resulted in luciferase expression only in the injection site and RLNs, with no detectable systemic (liver, spleen, lung) expression. At a higher dose (10(11) particles), some expression could be detected systemically in addition to the RLNs, but at levels in liver 14-fold less than in the RLNs. Transgene expression in the RLNs was transient, peaking at 1 day, decreasing markedly by 7 days. At high doses (10(11) particles), interruption of draining lymphatics decreased the amount of systemic dissemination 22-fold, suggesting that a large proportion of the vector trafficks through the lymphatics before reaching the systemic circulation. Administration of a vector encoding the jellyfish green fluorescent protein gene (AdCMV.GFP, 10(11) particles) showed that transgene expression in the RLNs was primarily in the cortical area. After footpad injection of a fluorescent-labeled Ad vector (Cy3-AdCMV.Null), fluorescent virions were visualized in the draining lymph. Regional lymph collected from animals injected in the footpad with AdCMV.Luc (10(11) particles) contained functional vector. Augmentation of local immune function in the RLNs was achieved by footpad administration of an Ad vector encoding murine IL-12, resulting in high mIL-12 and IFN-gamma levels in the regional, but not distant, nodes. These data demonstrate that expression of exogenous genes in RLNs is easily accomplished with Ad vectors, Ad vector dissemination occurs primarily via the lymphatics after footpad administration in mice, and basic immune functions in the RLNs can be manipulated by Ad-mediated gene transfer in vivo.

publication date

  • March 20, 2000

has subject area

  • Adenoviridae
  • Animals
  • Drainage
  • Gene Expression Regulation
  • Gene Transfer Techniques
  • Genetic Vectors
  • Interleukin-12
  • Luciferases
  • Lymph Nodes
  • Male
  • Mice
  • Mice, Inbred C3H
  • Transgenes

Research

keywords

  • Journal Article

Identity

Language

  • eng

Digital Object Identifier (DOI)

  • 10.1089/10430340050015653

PubMed ID

  • 10757355

Additional Document Info

start page

  • 759

end page

  • 769

volume

  • 11

number

  • 5