Induction of antigen-specific antitumor immunity with adenovirus-transduced dendritic cells

Abstract

Transduction of dendritic cells (DC) can result in presenmixed lymphocyte reactions. Mice immunized with Adtation of tumor-associated antigens and induction of transduced DC develop cytotoxic T lymphocytes that are immunity against undefined epitopes. The present studies specific for the ␤-galactosidase or DF3/MUC1 antigens. demonstrate adenovirus (Ad)-mediated transduction of the The results also demonstrate that Ad.MUC1-transduced ␤-galactosidase gene in mouse DC. Similar transductions DC induce a specific response which inhibits the growth have been obtained with the gene encoding the of DF3/MUC1-positive tumors. These findings support the DF3/MUC1 tumor-associated antigen. We show that the usefulness of Ad-transduced DC for in vivo immunization Ad-transduced DC are functional in primary allogeneic against tumor-associated antigens. Results and discussionT cells. 1 Murine DC pulsed with peptides prime antigenspecific CD8 + cytotoxic T lymphocytes (CTLs) in vivo. 2 Flow cytometry was used to define the phenotype of DC Peptides derived from tumor-associated antigens have following transduction with recombinant adenovirus. DC similarly been used to pulse DC and induce antitumor derived from bone marrow expressed MHC class I and II immunity. [3][4][5] Other studies have employed soluble products, costimulatory molecules and ICAM-1 18 (Figure tumor-associated antigens for loading DC and generating 1a). Transduction with Ad.␤gal resulted in a similar patantitumor activity. 6 Whereas peptides pulsed on to DC tern of antigen expression ( Figure 1a). Moreover, transmay dissociate from MHC molecules, CD34 + cells have duction with Ad.MUC1 was associated with DF3/MUC1 been retrovirally transduced to stably express antigens expression and little if any effect on cell surface levels of after differentiation to DC. 7,8 In contrast to pulsing, trans-MHC, costimulatory or adhesion molecules ( Figure 1a). duction of DC can result in longer term antigen presenThe Ad.MUC1-transduced DC exhibited a typical mortation and induction of immunity against undefined phology with veiled dendrites (Figure 1b). Staining with MHC epitopes. Thus, transduced DC may be effective in MAb M5/114 (anti-MHC class II) and MAb DF3 demonimmunizing against known tumor-associated antigens. strated expression of DF3/MUC1 by the transduced DC The human DF3/MUC1 glycoprotein is aberrantly (Figure 1b). Immunoblot analysis of the Ad.MUC1 transoverexpressed in breast and other carcinomas. 9 The DF3 duced DC confirmed DF3/MUC1 expression ( Figure 1c). protein is one member of the MUC1 family of carcinomaWhereas MAb DF3 detects glycosylated MUC1, the findassociated antigens that contain variable numbers of ing that MAb DF3-P reacts with an approximately 55 kDa highly conserved (G+C)-rich 60 base pair tandem protein in the transduced DC also provides support for repeats. 10,11 A C-terminal region includes a transmemdetection of the unglycosylated protein core 19 ( Figure 1c). brane domain that anchors the antigen at the cell sur-DC are potent stimulators ...

Human PBMC-derived dendritic cells transduced with an adenovirus vector induce cytotoxic T-lymphocyte responses against a vector-encoded antigen in vitro

“…8,14,15 Antigen-loaded murine DC have been shown to induce antigen-specific cytotoxic T-lymphocytes (CTL) and concomitant therapeutic antitumour immunity in murine models. [16][17][18][19][20][21][22][23][24] Results from limited human clinical experimentation, are also encouraging. 25,26 Antigen loading has been achieved using a variety of strategies including direct delivery of exogenous peptide fragments or tumour extracts, [16][17][18]21 or by endogenous antigen expression and processing within DC after delivery of antigen-encoding RNA or DNA molecules.…”

“…25,26 Antigen loading has been achieved using a variety of strategies including direct delivery of exogenous peptide fragments or tumour extracts, [16][17][18]21 or by endogenous antigen expression and processing within DC after delivery of antigen-encoding RNA or DNA molecules. [19][20][21][22][23][24]27 The extent to which these different approaches influence the efficiency of antigen-specific T-lymphocyte activation remains to be defined, although the advantages of expressing antigen-encoding genes in DC are compelling. Endogenous antigen expression and processing circumvents the MHC restriction imposed by the use of pep-tides, allows multiple undefined epitopes to be selected and presented to the immune system and should facilitate more efficient class-I loading and more persistent MHC-peptide complex presentation.…”

“…28 In contrast, recombinant adenovirus vectors have been used to achieve efficient gene transfer and expression (transduction) in both murine and human DC. 19,[22][23][24][28][29][30] Moreover, murine DC genetically modified by adenovirus transduction have been shown to induce antigen-specific therapeutic antitumour immunity. 19,22,24 While murine studies using human-tropic adenovirus vectors are promising, these findings are not necessarily predictive of the outcome of experiments using human DC, because the adenovirus vectors used retained the capacity for viral gene expression [31][32][33][34] and the potential to have different effects on function in mouse and human cells.…”

Efficient transduction and long-term retroviral expression of the melanoma-associated tumor antigen tyrosinase in CD34+ cord blood-derived dendritic cells

“…[5][6][7][8][9][10] Primary activation of tumor-specific cytotoxic T cells has been achieved by loading DCs with recombinant protein, tumor-derived peptides, transfer of mRNA or viral DNA coding for TAAs. [11][12][13][14][15][16][17][18][19] Also vaccination with dendritic cell-derived exosomes and fusion of DCs with carcinoma cells has been shown to elicit an anti-tumor response. 20,21 The advantage of viral expression is seen in the activation of a broad spectrum of T cells with differ-…”

Natural killer cells play a critical role in the immune response following immunization with melanoma-antigen-engineered dendritic cells

“…3 However, in this model, genetically modified DC generated antigen-specific CTL independent of NK cells. Multiple additional murine models confirmed that tumor antigen gene-modified DC directly stimulate CD8 þ CTL, usually requiring CD4 T help, [10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26] which was shown to be mediated by CD40 crosslinking of the DC. 27 Two reports have shown that NK1.1 þ cells may be involved in the antitumor response generated by tumor antigen gene-modified DC.…”