Phase I Trial Of Intravenous Peptide-Pulsed Dendritic Cells in Patients With Metastatic Melanoma

Abstract

Sixteen patients with metastatic stage IV melanoma were treated with use of intravenous infusions of dendritic cells (DC) derived by incubation of plastic-adherent peripheral blood mononuclear cells (PBMC) with IL-4 and GM-CSF for 8 days in serumless AIM-V medium, followed by overnight pulsing with peptides. The tyrosinase368-376 (370D) and gp100(209-217 (210M)) peptides restricted to HLA class I A*0201 each differed from wild type by one amino acid modified to increase HLA binding. Median age was 49, with nine men and seven women. All patients, except one, had visceral disease. Patients received escalating doses of peptide-pulsed DCs at 10e7, 3 x 10e7, and 10e8 cells/dose twice at 2 weeks apart, with toxicity and clinical and immune responses as the principal endpoints. The first infusion of DCs was fresh, and frozen DCs were given for the second infusion of each cycle. Mean DC purity by flow cytometry was 49%, with a mean HLA-DR level of 57%, CD86 of 41%, CD58 of 46%, and mean CD14 cells of 0.9%. Toxicity was minimal, with two patients having transient grade III DC-related toxicity. Ten patients received one cycle of treatment and six patients received two cycles of treatment. One patient had a complete remission (CR) of lung and pleural disease after two cycles of DC therapy. Two additional patients had stable disease and two patients had mixed responses. Overall immunity was assessed by recall skin testing with peptides, gamma interferon ELISA assays of peptide specific cytolytic T cell (CTL) stimulated twice with peptide, IL-2, and IL-7 over 24 days, and peptide-specific tetramer assays performed before and after vaccination. Five of 16 patients had an immune response to gp100 or tyrosinase by gamma interferon ELISA assay; four of five were clinically stable or had tumor regression. These data suggest that melanoma antigen peptide-pulsed DC given intravenously are not toxic, and regression or stability of tumor appeared to correlate with the detection of a peptide-specific immune response in the peripheral blood.

The Duration of Signaling through CD40 Directs Biological Ability of Dendritic Cells to Induce Antitumor Immunity

“…However, the appropriate conditioning of DCs to induce the most potent anticancer immune reaction in vivo is still unclear (18,19,(33)(34)(35)(36)(37)(38)(39)(40)(41)(42). One of the problems to be elucidated is how to obtain DCs that are capable of migrating into secondary lymphoid organs to prime T cells when injected.…”

Both Langerhans cells and interstitial DC cross‐present melanoma antigens and efficiently activate antigen‐specific CTL

“…Most clinical trials have been performed with DC loaded with synthetic HLA-class I peptide(s), aiming at the induction of tumor-antigen specific CTL [7][8][9][10][11][12][13][14][15]. We performed a clinical trial for metastatic melanoma with melanoma peptide-loaded DC generated from CD34 + -hematopoietic progenitor cells (CD34-HPC).…”

Induction of CAMEL/NY-ESO-ORF2-specific CD8+ T cells upon stimulation with dendritic cells infected with a modified Ad5 vector expressing a chimeric Ad5/35 fiber

“…T-cell cultures were weekly restimulated with autologous DC infected with Ad5F35-CAMEL or pulsed with rCAMEL protein and tested for the presence of CD8 þ T cells specific for the previously described HLA-A*0201-binding epitope CAMEL/NY-ESO-ORF2 [1][2][3][4][5][6][7][8][9][10][11] 33 by tetramer staining. In both donor 1 (Fig 1) and donor 2 (Fig 2), CAMEL/NY-ESO-ORF2 1-11 -specific CD8 þ T cells Generation of a CAMEL 46-54 -specific CD8 þ T-cell line by DC infected with Ad5F35-CAMEL The T-cell cultures at days 14 and 21 that were shown to contain CAMEL/NY-ESO-ORF2 1-11 -specific T cells after stimulation with DC infected with Ad5F35-CAMEL were also screened for T cells specific for unknown CAMEL epitopes in IFN-g ELISPOT assays.…”

“…The Ad5F35-CAMEL-stimulated T-cell cultures were tested against autologous DC infected with Ad5F35-CAMEL and, as a control, empty Ad5F35-vector and autologous DC pulsed with mixes of overlapping peptides covering the entire CAMEL protein sequence. Peptide mix 1 contained three 20-mer peptides (CAMEL [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20] , CA-MEL [14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30][31][32][33] and CAMEL [46][47][48][49][50][51][52][53][54][55][56][57]…”

“…To date, melanoma patients are vaccinated with HLA class I-binding peptides derived from melanocytic differentiation antigens like MART-1/Melan-A, 2-4 gp100 [3][4][5][6][7] and tyrosinase, 3,4,7,8 or cancer-testis antigens like MAGE-1, 4,9,10 MAGE-3 4,11,12 and NY-ESO-1. 13 These peptides are either injected alone 2,3,5,6,8,11,13 or pulsed on dendritic cells (DC), 4,7,9,10,12 which are believed to be the most potent antigen-presenting cells for the induction of T-cell responses in vivo. Recently, melanoma patients have been described to be treated by adoptive transfer of CD8 þ cytotoxic T lymphocytes (CTL), raised in vitro by autologous DC pulsed with HLA-A*0201-binding peptides derived from MART-1/Melan-A 14,15 or gp100.…”