A comparative study of the JAM test and 51Cr-release assay to assess the cytotoxicity of dendritic cells on hematopoietic tumor cells.
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.
Ayres
Flávio M
FM
First Department of Internal Medicine, School of Medicine, Niigata University, Niigata, Japan.
Narita
Miwako
M
Takahashi
Masuhiro
M
Alldawi
Louie
L
Liu
Aichun
A
Osman
Yasser
Y
Abe
Takashi
T
Yano
Toshio
T
Sakaue
Minori
M
Toba
Ken
K
Furukawa
Tatsuo
T
Aizawa
Yoshifusa
Y
D015703:Antigens, CD / Q000032:analysis; D016923:Cell Death / Q000276:immunology; D002454:Cell Differentiation / Q000187:drug effects / Q000276:immunology; D002455:Cell Division / Q000276:immunology; D045744:Cell Line, Tumor; D002840:Chromates / Q000378:metabolism; D002860:Chromium Radioisotopes; D018920:Coculture Techniques; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods*; D053938:DNA Fragmentation / Q000276:immunology; D003713:Dendritic Cells / Q000737:chemistry / Q000166:cytology / Q000276:immunology*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D019337:Hematologic Neoplasms / Q000276:immunology* / Q000473:pathology; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D015847:Interleukin-4 / Q000494:pharmacology; D017670:Sodium Compounds / Q000378:metabolism; D013936:Thymidine / Q000378:metabolism; D014316:Tritium; D020298:U937 Cells
D015703:Antigens, CD; D002840:Chromates; D002860:Chromium Radioisotopes; D017670:Sodium Compounds; D014316:Tritium; D015847:Interleukin-4; C028982:sodium chromate(VI); D007371:Interferon-gamma; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D013936:Thymidine