Dendritic cells, pulsed with lysate of allogeneic tumor cells, are capable of stimulating MHC-restricted antigen-specific antitumor T cells.
A variety of approaches have been used to deliver tumor-associated antigens (TAA) in conjunction with dendritic cells (DC) as cellular adjuvants. DC derived from monocytic precursors have been pulsed with whole tumor antigen using a variety of strategies and have been demonstrated to induce CD4+ and CD8+ antitumor responses. In the present study, monocyte-derived DC have been pulsed with lysate from an allogeneic melanoma cell line, A-375, and used to repeatedly stimulate T cells. The resultant T cells were examined for cytotoxic activity against A-375 targets as well as the HLA A2-positive melanoma cell line DFW. Uptake of FITC-labeled melanoma lysate by DC established that lysate of melanoma cells was efficiently endocytosed. Stimulation with lysate-pulsed DC resulted in strong proliferative responses by T cells, which could be inhibited by antibodies against both MHC class I and class II. T cells stimulated in vitro with lysate-pulsed DC demonstrated potent cytotoxicity against the melanoma targets which were blocked by antibodies against MHC class I. Lysate-pulsed DC also elicited IFN-gamma secretion by T cells as measured in an ELISPOT assay. We have also examined the ability of lysate-pulsed DC to present melanoma-associated antigens to T cells. ELISPOT assays with synthetic peptides of melanoma-associated antigens, such as gp100, mage1, NY-ESO, and MART-1, revealed that lysate-pulsed DC could stimulate T cells in an antigen-specific manner. The results demonstrate that lysate from allogeneic tumor cells may be used as a source of antigens to stimulate tumor-specific T cells in melanoma.
Mahdian
Reza
R
Immune and Gene Therapy Lab, Cancer Center Karolinska, Karolinska University Hospital, Stockholm, Sweden.
Kokhaei
Parviz
P
Najar
Hossein Motieian
HM
Derkow
Katja
K
Choudhury
Aniruddha
A
Mellstedt
Håkan
H
D017951:Antigen Presentation / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000276:immunology* / Q000494:pharmacology; D019496:Cancer Vaccines / Q000276:immunology*; D045744:Cell Line, Tumor; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000276:immunology*; D006801:Humans; D007519:Isoantigens / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology; D020014:K562 Cells; D008213:Lymphocyte Activation / Q000276:immunology*; D008545:Melanoma / Q000737:chemistry / Q000276:immunology*; D009000:Monocytes; D013601:T-Lymphocytes / Q000276:immunology*
D000951:Antigens, Neoplasm; D019496:Cancer Vaccines; D007519:Isoantigens