A model for antimicrobial gene therapy: demonstration of human beta-defensin 2 antimicrobial activities in vivo

Hum Gene Ther. 2002 Nov 20;13(17):2017-25. doi: 10.1089/10430340260395875.

Abstract

We transfected host cells with an antimicrobial peptide/protein-encoding gene as a way to enhance host defense mechanisms against infection. The human beta-defensin 2 (HBD-2) gene was chosen as a model because its protein does not require cell type-specific processing. Using a retroviral vector carrying HBD-2 cDNA, we treated several mouse or human cell lines and primary cell cultures including fibroblasts, salivary gland cells, endothelial cells, and T cells. All transduced cells produced detectable HBD-2. In Escherichia coli gel overlay experiments, secreted HBD-2 from selected cell lines showed potent antimicrobial activity electrophoretically identical to that of purified HBD-2. We then used a mouse model (nonobese diabetic/severely compromised immunodeficient [NOD/SCID]) to test HBD-2 antimicrobial activities in vivo. HT-1080 cells carrying HBD-2 or control vector were implanted subcutaneously into NOD/SCID mice to allow tumor formation. Escherichia coli was then injected into each tumor mass. Tumors were resected after 16 hr and homogenized for bacterial colony-forming unit analysis. Compared with control tumors, HBD-2-bearing tumors contained only 7.8 +/- 3.3% viable bacteria. On the basis of this demonstration of HBD-2 in vivo antimicrobial activity, enhancement of antibacterial host defense by HBD-2 gene therapy may be feasible.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Anti-Bacterial Agents
  • Anti-Infective Agents / therapeutic use*
  • Cell Line
  • Colony Count, Microbial
  • Escherichia coli / genetics
  • Genetic Therapy*
  • Humans
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Retroviridae / genetics
  • Transduction, Genetic
  • Transplantation, Heterologous
  • Tumor Cells, Cultured
  • beta-Defensins / genetics*

Substances

  • Anti-Bacterial Agents
  • Anti-Infective Agents
  • DEFB4A protein, human
  • beta-Defensins