A comparative study of the JAM test and 51Cr-release assay to assess the cytotoxicity of dendritic cells on hematopoietic tumor cells

Immunol Invest. 2003 Nov;32(4):219-27. doi: 10.1081/imm-120025102.

Abstract

Dendritic cells (DCs) are potent antigen presenting cells and possess a direct anti-tumor cytotoxic ability. Nevertheless, the mechanism of anti-tumor cytotoxicity by DCs and the methods for its evaluation are not fully elucidated. In order to clarify this mechanism of cytotoxicity, we examined the ability of DCs 1) to suppress [3H] thymidine (3H-TdR) uptake by tumor cells; 2) to induce cytolysis on 51Cr-labeled tumor cells; 3) and to induce DNA fragmentation on 3H-TdR labeled tumor cells (JAM test). Cytolysis and DNA fragmentation are markers of necrotic and apoptotic mechanisms of cytotoxicity in vitro, respectively. DCs inhibited approximately 38.6% to 54.8% of the growth of B4D6, NB4, U937, and Daudi cells as evaluated by the uptake of 3H-TdR. However no cytolysis was verified by 51Cr-release assay. On the other hand, cytotoxicity rates found using the JAM test ranged from 3 to 81% depending on the cell line and the effector to target cell ratio. The discrepancy of cytotoxicity between 51Cr-release assay and the JAM test may be due to the phagocytosis of apoptotic tumor cells or the absorption of released 51Cr by DCs surrounding the target cells. In conclusion, the JAM test was more sensitive than the 4-h and the 10-h 51Cr-release assay to investigate cytotoxicity mediated by DCs toward hematopoietic tumor cell lines in vitro.

Publication types

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

MeSH terms

  • Antigens, CD / analysis
  • Cell Death / immunology
  • Cell Differentiation / drug effects
  • Cell Differentiation / immunology
  • Cell Division / immunology
  • Cell Line, Tumor
  • Chromates / metabolism
  • Chromium Radioisotopes
  • Coculture Techniques
  • Cytotoxicity Tests, Immunologic / methods*
  • DNA Fragmentation / immunology
  • Dendritic Cells / chemistry
  • Dendritic Cells / cytology
  • Dendritic Cells / immunology*
  • Granulocyte-Macrophage Colony-Stimulating Factor / pharmacology
  • Hematologic Neoplasms / immunology*
  • Hematologic Neoplasms / pathology
  • Hematopoietic Stem Cells / cytology
  • Hematopoietic Stem Cells / immunology
  • Humans
  • Interferon-gamma / pharmacology
  • Interleukin-4 / pharmacology
  • Sodium Compounds / metabolism
  • Thymidine / metabolism
  • Tritium
  • U937 Cells

Substances

  • Antigens, CD
  • Chromates
  • Chromium Radioisotopes
  • Sodium Compounds
  • Tritium
  • Interleukin-4
  • sodium chromate(VI)
  • Interferon-gamma
  • Granulocyte-Macrophage Colony-Stimulating Factor
  • Thymidine