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Although chemotherapy is still one of the best treatments for most cancers, immunotherapies such as dendritic cell (DC) vaccines have emerged as an alternative protocol for destroying residual tumors. In this study, we investigated antitumor effects of the combined therapy using DC vaccine and irinotecan plus infusional 5-fluorouracil and leucovorin (FOLFIRI) which have been clinically used for the treatment of colorectal cancer. A maximum tolerated dose of FOLFIRI was preliminarily determined for MC38/CEA2 colorectal cancer model. Vaccination with DC expressing carcinoembryonic antigen (CEA) enhanced antitumor effect after FOLFIRI treatment. The combined therapy also increased CEA-specific Th1 and cytotoxic T-cell responses. Interestingly, although FOLFIRI treatment rather showed a rebound in the number of myeloid-derived suppressor cells (MDSC) and regulatory T-cells (Treg) after 14 days, additional DC vaccine could inhibit the rebound of these immunosuppressive cells. Furthermore, mice cured by the combined therapy showed antigen-specific T-cell responses and resistance against challenge of MC38/CEA2 compared with mice cured with FOLFIRI. These results demonstrated that DC vaccine in addition to FOLFIRI regimen could improve antitumor effects through the inhibition of immunosuppressive tumor environments in murine colorectal cancer model, and may provide knowledge useful for the design of chemo-immunotherapeutic strategies for the treatment of colorectal carcinoma in clinical trials., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Because multiple tumor antigens, including carcinoembryonic antigen (CEA) and survivin (SVV), have been frequently observed in human colorectal cancer, we investigated whether the expression of both CEA and SVV by co-transduction of adenovirus vectors into dendritic cells (DCs) could improve anti-tumor immunity in a murine colorectal cancer model. The adaptor fusion protein of Coxsackie and adenovirus receptor and TAT-protein transduction domain (CAR-TAT) enhanced co-transduction of adenovirus vectors encoding CEA (AdCEA) and SVV (AdSVV) into DCs, and increased anti-tumor immunity. DCs expressing both CEA and SVV in the presence of CAR-TAT (DC-AdCEA/AdSVV+CAR-TAT) induced T-cell responses specific for CEA and SVV, and enhanced cytotoxic T-cell activity on MC38/CEA2 cells expressing CEA and SVV compared with DCs expressing either CEA or SVV alone. Particularly, DC-AdCEA/AdSVV+CAR-TAT induced higher number of CEA-specific IFN-gamma secreting T cells compared with DC-AdCEA+CAR-TAT. Vaccination with DC-AdCEA/AdSVV+CAR-TAT also more efficiently inhibited tumor growth compared with DCs expressing either CEA or SVV alone in therapeutic tumor models. These results suggest that efficient co-transduction of multiple adenovirus vectors by CAR-TAT could be used to develop various strategies for therapeutic DC vaccines., (Copyright (c) 2010 Elsevier B.V. All rights reserved.)

Rationale: Fas/Fas ligand (FasL)-mediated apoptosis has been implicated in various lung diseases, but whether Fas/FasL-mediated apoptosis in the lungs plays a critical role in the development of hypersensitivity pneumonitis (HP) is unclear., Objectives: To explore the functional roles of Fas/FasL-mediated apoptosis in HP., Methods: Fas-deficient (lpr/lpr), FasL-deficient (gld/gld), and B6 mice were challenged with Saccharopolyspora rectivirgula (SR) antigen intranasally., Measurements and Main Results: lpr/lpr and gld/gld mice exhibited attenuation of HP in terms of histological alterations, influx of immune cells in bronchoalveolar lavage fluid (BALF), and SR-specific immune responses compared with B6 mice, similar to the effects of SR in B6 mice given a caspase inhibitor. The lungs of lpr/lpr and gld/gld mice showed high IL-4 production and low IFN-gamma, IL-8, macrophage inflammatory protein-2, IL-1beta, and tumor necrosis factor-alpha production compared with those of B6 mice. Moreover, mice with chimeric B6 and lpr/lpr bone marrow revealed that apoptosis of nonhematopoietic and BALF immune cells of the lungs enhanced immune responses against SR antigen. Gr-1(+) granulocytes in BALF expressed annexin V and their depletion in B6 mice attenuated HP. Apoptosis of nonhematopoietic cells and Gr-1(+) granulocytes in the lungs enhanced the maturation of pulmonary CD11c(+) dendritic cells and their production of macrophage inflammatory protein-1alpha and monocyte chemoattractant protein-1, resulting in recruitment of immune cells into the lungs during HP., Conclusions: These results suggest that apoptosis in nonhematopoietic cells and Gr-1(+) granulocytes of the lungs promotes HP by enhancing maturation and chemokine production of CD11c(+) dendritic cells.

Carcinoembryonic antigen (CEA) is expressed on human colon carcinomas, is well characterized, and continues to be a promising target for cancer immunotherapy in humans. To enhance the immunogenecity of CEA, we developed a fusion gene (CRT-TAT-DeltaCEA) of the TAT protein transduction domain (PTD) and calreticulin (CRT) with human CEA devoid of its signal sequences (DeltaCEA) and evaluated anti-tumor immunity using RNA-pulsed dendritic cell (DC) vaccination. Mice vaccinated with DC by electroporation with mRNA encoding TAT-DeltaCEA (DC/TAT-DeltaCEA) and CRT-DeltaCEA (DC/CRT-DeltaCEA) had enhanced induction of tumor-specific cytotoxic T lymphocyte (CTL) and increased numbers of IFN-gamma-secreting T cells by ELISPOT, as compared to mice vaccinated with DC/DeltaCEA. DC/CRT-DeltaCEA and DC/TAT-DeltaCEA vaccines preferentially stimulated CD4+ and CD8+ T cells, respectively. The DC vaccine by electroporation with mRNA encoding CRT-TAT-DeltaCEA (DC/CRT-TAT-DeltaCEA) enhanced both CD4+ and CD8+ T cells. DC/CRT-TAT-DeltaCEA had the additional effects of CRT and TAT PTD and enhanced the anti-tumor effect against CEA-expressing tumors compared to DC/CRT-DeltaCEA or DC/TAT-DeltaCEA. These findings suggest that modification of CEA with both CRT and TAT PTD induces potent anti-tumor immune responses in RNA-pulsed DC vaccination and may be a useful approach for DC-based immunotherapy.

Although chemotherapy remains among the best treatment options for most cancers, adjuvant therapies such as dendritic cell (DC)-based immunotherapy have been added to treatment protocols to destroy residual tumour cells. Combination treatment with low-dose temozolomide (TMZ) chemotherapy followed by vaccination with TAT-survivin-pulsed DCs enhanced T-cell responses specific for survivin and improved survival rate, as compared with DC alone or TMZ alone. Moreover, antigen-specific immunity appears to be mediated by CD8(+) T cells, as determined by in vitro T-cell subset depletion. These studies demonstrated that a combination of low-dose TMZ chemotherapy and TAT-based DC immunotherapy may be a novel strategy for safe and effective treatment of malignant gliomas.