Mature CD83(+) dendritic cells infected with recombinant gp100 vaccinia virus stimulate potent antimelanoma T cells.
BACKGROUND
Mature dendritic cells (DCs) are potent antigen-presenting cells that activate naive T lymphocytes and initiate cellular immune responses. The ability of CD83(+) mature DCs infected with vaccinia virus encoding the gp100 melanoma transgene (rV-gp100) to stimulate an antimelanoma CD8(+) T-cell response was investigated.


METHODS
Monocyte-derived immature or CD83(+) mature DCs were infected with rV-gp100. The activation state of the DCs and the expression of gp100 protein were evaluated by flow cytometry. The reactivity of antimelanoma CD8(+) T cells was confirmed by measuring specific interferon gamma secretion by using enzyme-linked immunosorbent assay in a mixed-tumor lymphocyte culture.


RESULTS
Both immature and CD83(+) mature DCs expressed gp100 protein when the DCs were infected with rV-gp100. Calcium-signaling agents were required to induce maturation of both infected and noninfected immature DCs. Only rV-gp100-infected CD83(+) DCs induced CD8(+) T cells, after a single stimulation that recognized both peptide-pulsed target cells to multiple gp100 epitopes and a melanoma cell line that endogenously expressed gp100 antigen.


CONCLUSIONS
CD83(+) DCs transduced with rV-gp100 are capable of generating a strong CD8(+) T-cell response against melanoma tumor cells. Expression of melanoma antigens by mature DCs offers the potential advantage of presenting multiple endogenously processed T-cell epitopes and using multiple HLA restriction elements for antimelanoma vaccine therapy.
Prabakaran
Indira
I
Department of Surgery, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Menon
Chandrakala
C
Xu
Shuwen
S
Gómez-Yafal
Alicia
A
Czerniecki
Brian J
BJ
Fraker
Douglas L
DL
D017951:Antigen Presentation / Q000235:genetics*; D015703:Antigens, CD; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D003602:Cytotoxicity, Immunologic / Q000235:genetics; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism; D005434:Flow Cytometry; D015789:HLA-A2 Antigen / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics* / Q000276:immunology; D008545:Melanoma / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000276:immunology; D009363:Neoplasm Proteins / Q000235:genetics* / Q000276:immunology; D014407:Tumor Cells, Cultured; D014616:Vaccinia virus / Q000235:genetics* / Q000276:immunology; D058951:gp100 Melanoma Antigen; D000097802:CD83 Antigen
D015703:Antigens, CD; D015789:HLA-A2 Antigen; D007136:Immunoglobulins; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; D058951:gp100 Melanoma Antigen; D000097802:CD83 Antigen; C548637:PMEL protein, human