Overexpression of CCL-21/secondary lymphoid tissue chemokine in human dendritic cells augments chemotactic activities for lymphocytes and antigen presenting cells

Mol Cancer. 2003 Nov 2:2:35. doi: 10.1186/1476-4598-2-35.

Abstract

Background: Ex vivo generated dendritic cells (DC) genetically modified to express secondary lymphoid tissue chemokine (CCL-21/SLC) have been shown to stimulate potent antitumor responses in murine models. When injected intratumorally, CCL-21 colocalizes DC and lymphocyte effector cells at the tumor site. This may improve tumor antigen presentation and T cell activation by utilizing the tumor as an in vivo source of antigen for DC. In order to develop DC-based cancer therapies for intratumoral injection that could promote tumor antigen uptake and presentation in situ, we constructed and characterized an adenoviral vector that expresses human CCL-21 (AdCCL-21).

Results: Human monocyte derived DC were cultured in GM-CSF and IL-4 for 6 days. Following AdCCL-21 transduction, CCL-21 protein production was assessed by ELISA on day 8. DC transduced with AdCCL-21 at multiplicities of infection (MOIs) of 50:1 or 100:1 produced up to 210 +/- 9 ng/ml and 278 +/- 6.5 ng/ml /106 cells/48 hours, respectively. Following transduction, an immature DC phenotype was maintained and an upregulation of the costimulatory molecule, CD86 was noted. In addition, supernatant from AdCCL-21-DC caused significant chemotaxis of peripheral blood lymphocytes and mature DC.

Conclusions: These studies demonstrate that AdCCL-21-DC generate functional levels of CCL-21 without adversely altering DC phenotype. These findings strengthen the rationale for further investigation of AdCCL-21-DC as a DC-based therapy in cancer treatment.

Publication types

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

MeSH terms

  • Adenoviridae / genetics
  • Antibodies, Monoclonal / pharmacology
  • Antigen-Presenting Cells / cytology
  • Antigen-Presenting Cells / physiology*
  • Cell Movement / drug effects
  • Chemokine CCL21
  • Chemokines, CC / genetics
  • Chemokines, CC / immunology
  • Chemokines, CC / metabolism*
  • Chemotaxis / drug effects
  • Culture Media, Conditioned / pharmacology
  • Dendritic Cells / drug effects
  • Dendritic Cells / immunology
  • Dendritic Cells / metabolism*
  • Flow Cytometry
  • Genetic Vectors / genetics
  • Humans
  • Immunophenotyping
  • Interferon-gamma / pharmacology
  • Interleukin-10 / biosynthesis
  • Interleukin-12 / biosynthesis
  • Lipopolysaccharides / pharmacology
  • Lymphocytes / cytology
  • Lymphocytes / physiology*
  • Transfection

Substances

  • Antibodies, Monoclonal
  • CCL21 protein, human
  • Chemokine CCL21
  • Chemokines, CC
  • Culture Media, Conditioned
  • Lipopolysaccharides
  • Interleukin-10
  • Interleukin-12
  • Interferon-gamma