Dendritic cell maturation with CpG for tumor immunotherapy (original) (raw)
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99 Dendritic Cell Maturation with CpG for Tumor
2015
Background: Bacterial DNA has immunostimulatory effects on different types of immune cells such as dendritic cells (DCs). Application of DCs as a cellular adjuvant represents a promising approach in the immunotherapy of infectious disease and can-cers. Objectives: To investigate the effect of tumor antigen pulsed DCs in the pres-ence of CpG-1826 in treatment of a murine model of cancer. Methods: WEHI-164 cells (Balb/c derived fibrosarcoma cell line) were injected subcutaneously in the right flank of mice. Bone marrow cells were cultured in the presence of GM-CSF and IL-4. After 5 days, tumor lysate, CpG-1826, and oligodeoxynucleosides, as control, were added to the culture media and incubated for 2 days. Cytokine production in DCs culture media was measured by ELISA. Then DCs were injected subcutane-ously around the tumor site in the right flank of mice. Tumor growth rate was moni-tored in case and control groups. Two weeks after DCs immunotherapy, cytotoxic assay was conducted usin...
Cellular Immunology, 2011
CpG motifs have been advanced as agents that stimulate the maturation of DCs for immunotherapy. The present study tested the hypothesis that multiple doses of CpG-matured DC vaccine would be efficacious for complete eradication of experimentally-induced tumor. Accordingly, WEHI164 cells were implanted subcutaneously in the flank of BALB/c mice. During DC culture, tumor lysate was added to immature DCs followed by addition of CpG or non-CpG control 4-6 h later. A total of three doses of CpG or non-CpG control-matured DCs were injected around tumors. The results showed that multiple doses of CpG-matured DCs led to considerable decrease in cytotoxicity of lymphocytes and significantly increased tumor growth rate compared to a single dose. Further, mice which received three doses of the vaccine also displayed significant FoxP3 in tumor tissue. In conclusion, multiple doses of CpG-matured DCs exhibited decreased anti-tumor immunity in association with increased expression of FoxP3.
Cellular Immunology, 1997
numbers in lymphoid and nonlymphoid organs, as well as in the circulation. They play a crucial role in the Dendritic cells (DC) are highly efficient antigen-preinitiation of immune responses, including the sensitisenting cells able to capture, process, and present antization of T-cells restricted by MHC molecules and the gens to naive and primed T-cells. In this study, we have development of T-cell-dependent antibodies (1-6). DC investigated the ability of DC, derived from murine bone marrow and pulsed with tumor cell extracts, to originate in bone marrow and migrate to many noninduce regression of preexisting tumors. In an experilymphoid tissues, such as skin (Langerhans cells), tumental model of B16 melanoma in B6 mice, a signifimors, and mucosa (7, 8). After capturing antigens, DC cant reduction in metastatic nodules in the lungs was migrate through afferent lymph or the bloodstream to observed in tumor-bearing animals treated with either lymphoid organs where, in the context of MHC gene DC alone or DC pulsed with tumor extracts. Kinetic products, they efficiently present antigens to T-cells studies demonstrate that the efficacy of these tumor (2). In addition to expressing high levels of MHC antivaccines is inversely related to tumor burden. In this gens, DC express high levels of accessory molecules model, tumor-specific cytotoxic T-cells (CTL) could including ICAM-1, LFA-3, and B7, which are required also be induced in vitro from spleen cells derived from for T-cell activation (9-11). Thus, the high efficiency tumor-bearing animals treated with DC pulsed with of DC in antigen presentation is attributed to the high tumor extracts. Untreated mice had no CTL. Furtherexpression of MHC and accessory molecules, as well as more, DC alone elicited tumor-specific CTL responses their capacity to deliver costimulatory signals (2, 4, 12). in tumor-bearing mice, but not in naive mice. Immune DC pulsed with intact proteins (13) or peptides (14) cell depletion experiments show that the therapeutic are able to elicit primary CTL responses in vitro. These effects of DC are primarily mediated by CD8 / T-cells, while CD4 / T-cells and NK cells are involved in DC-cells are capable of priming naive T-cells and inducing mediated antitumor immunity to a limited extent. potent CTL following injection into mice . These These results illustrate the potential use of DC and results have led investigators to propose using DC DC pulsed with tumor extracts as potent therapeutic pulsed with tumor antigens or peptides to induce antireagents for cancer and provide a rationale for using tumor immunity in vivo or to induce tumor antigen-DC in vivo to eliminate disseminated tumors or residspecific CTL in vitro .
Anticancer research
Dendritic cells (DC) express class I/II MHC-determinants and co-stimulatory molecules required for T cell activation. In a mouse model of squamous cell carcinoma (SCC), we compared the immunogenic properties of allogeneic DNA-based fibroblast vaccines, which are taken up and processed by DC of the host, and syngeneic DNA-based DC vaccines, which present antigens directly. The incentive was the important practical advantages of using fibroblasts rather than DC in generating vaccines for the immunotherapy of SCC. The fibroblast vaccine was prepared by transfer of genomic DNA-fragments (25 kb) from relatively small numbers (10(7) = 64 mm3 tumor) of SCCVII/SF cells into LM cells, a mouse fibroblast cell line (H-2(k)). SCCVII/SF cells are a highly aggressive squamous carcinoma cell line of C3H/He mouse origin (H-2(k)). As the transferred DNA spontaneously integrates into the genome of the recipient cells, and is replicated as the cells divide, the number of transfected fibroblasts could ...
Surgery, 1997
Background. Dendritic cells (DCs) are potent antigenpresenting cells regarded as crucial in the priming of an immune response. The goal of our study was to test whether bone marrow-generated DCs are capable of inducingjnotective immunity against a murine breast carcinoma (4Tl). Methods. DCs were grown from Balb/c mice by culturing lymphocyte-immunodepleted bone marrow in murine granulocyte-macrophage colony-stimulating factor containin medium fm 10 days. Balb/c mice (five to eight per group) were immunized intradermally with 1 x 1 of DCs mixed with 2 x 106 Lethally irradiated 4Tl cells on day 0. Mice in control groups were given intradermal inoculations of phosphatebuffered saline solution, 1 x 106 DCs, or lethally irradiated 4Tl cells alone. Booster intraperitoneal immunizations of 2 x 106 lethally irradiated 4Tl cells were given on days 7 and 14. All mice were challenged with 5 x ld 4Tl cells subcutaneously 7 days after the final immunization. Animals were examined daily, and tumor volume was recorded twice weekly with calipers. Results. At 21 days there was a signaficant reduction in tumor growth in mice immunized with DCs mixed with irradiated 4Tl cells as compared with the control groups (p = 0.0005, Kruskal-Wallis test). Conclusions. These results suggest that DCs mixed with tumor cells as a source of undefined tumor antigen can induce an effective antitumor immune response. This finding provides a rationale for the use of cultured DCs in immunotherah of breast and other cancers.
Clinical & Experimental Immunology, 2009
Carcinoembryonic antigen (CEA) is over-expressed on various human cancer cells and has been the target of immunotherapies using dendritic cells (DCs) pulsed with CEA-specific RNA or peptides, or transduced by CEA-expressing adenovirus or vaccinia virus. Because activated DCs do not phagocytose soluble protein antigens efficiently and pure immature DCs are not obtained easily ex vivo, an efficacious whole CEA protein-loaded DC vaccine has not been reported. To improve the antigen delivery into DCs, we utilized CEA conjugated to a protein-transduction domain, human immunodeficiency virus transactivating Tat. Furthermore, we purified the truncated nonglycosylated CEA from Escherichia coli to overcome the safety concerns and immunosuppressive functions associated with the native CEA protein. Using confocal microscopy and fluorescence activating cell sorter analysis, we demonstrated that the Tat-CEA protein entered the cytoplasm of DCs efficiently within 10 min of co-culture, compared with the negligible amount of CEA into DCs 30 min later. CEA-specific T cell proliferation and cytotoxic T cell responses were enhanced significantly in mice immunized with Tat-CEApulsed DCs [DC (Tat-CEA)] compared with those immunized with CEApulsed DCs [DC (CEA)]. T helper type 1 responses were more prominent in the DC (Tat-CEA) immunized mice whose splenocytes secreted more interferon-g and less interleukin-4 than those from DC (CEA) immunized mice. In vivo, the DC (Tat-CEA) vaccine delayed tumour growth significantly and prolonged survival of tumour-bearing mice. These results suggest that protective epitopes are well preserved on bacteria-derived recombinant Tat-CEA. This strategy may provide a basic platform for DC-based anti-CEA vaccines that could be utilized in combination with advanced immuneenhancing therapeutics.
BioImpacts
Introduction: Ovarian cancer is one of the most lethal gynecologic cancers. Relapses after remission are common, hence novel strategies are urgently needed. Our group has previously developed a vaccination approach based on dendritic cells pulsed with HOCl-oxidized tumor lysates. Here we investigate the improvement of this vaccine strategy using squaric acid treatment of cancer cells during tumor lysate preparation and by differentiating dendritic cells in the presence of GM-CSF and IFNα. Methods: Induction of cell death by squaric acid treatment was assessed with propidium iodide (PI) and Annexin V in ID8 tumor cells. High mobility group box 1 (HMGB1) immunogenic status was analyzed using a western blot-based method, as previously described. For immunological tests, ID8 cells expressing ovalbumin (ova-ID8) were treated with squaric acid before cell lysis. DCs prepared with the canonical GM-CSF and IL-4 differentiation cocktail or IFNα and GM-CSF were pulsed with tumor cell lysates and further matured in the presence of IFNγ and LPS (4-DCs and α-DCs respectively). DCs were then used in co-culture assays with ova-specific T cells and IFNγ and IL-4 secretion measured by ELISA. DC phenotypes were characterized by FACS. Finally, DCs were tested in an ovarian cancer mouse model measuring body weight and animal survival. Results: Squaric acid treatment of mouse ovarian cancer cells induced tumor cell death as well as preserve HMGB1, a crucial Damage-associated molecular pattern (DAMP) signal, in its active reduced form. Squaric acid treatment of ID8-ova cells increased IFNγ and decreased IL-4 production from ova-specific T cells in co-culture experiments, promoting a more immunogenic cytokine secretion pattern. DCs differentiated in the presence of IFNα induced a considerable decrease in IL-4 production compared to canonical 4-DCs, without affecting IFNγ release. DC phenotyping demonstrated a more mature and immunogenic phenotype for IFNα-differentiated DCs. Vaccination in tumor-bearing mice showed that IFNα-differentiated DCs pulsed with squaric acid-treated lysates were the most potent at delaying tumor growth, improving animal survival. Conclusion: We identified squaric acid as a novel immunogenic treatment of tumor cells for cancer vaccines particularly efficient in prolonging animal survival when used in combination with IFNαdifferentiated DCs. These promising results support future efforts for the clinical translation of this approach.
Cancer Gene Therapy, 2005
Immunotherapy of squamous cell carcinoma (SCC) at an early stage of the disease increases the likelihood of success. We report a new vaccination strategy designed to prepare SCC vaccines from microgram amounts of tumor tissue, enabling the treatment of patients with minimal residual disease. The vaccine was prepared by transfer of sheared genomic DNA-fragments (25 kb) from KLN205 cells, an SCC cell line of DBA/2 mouse origin, into syngeneic bone marrow-derived mature dendritic cells (DCs). More than 90% of the transfected DCs took up DNA from the neoplasm and transferred genes were expressed as protein. The DCs expressed CD11c, CD11b, and the costimulatory molecules CD40, CD80 and CD86, characteristic of mature DCs. Syngeneic DBA/2J mice, highly susceptible to the growth of KLN205 cells, were injected intravenously (i.v.) with the transfected DCs, followed by a subcutaneous (s.c.) injection of the tumor cells. The strong immunogenic properties of the transfected cells were indicated by the finding that the survival of the tumor-bearing mice was prolonged (Po.001), relative to that of mice in various control groups. Enzyme-linked immuno spot (ELISPOT IFN-g) assays revealed the activation of cell-mediated immunity directed toward the SCC in mice immunized with the transfected DCs. Two independent in vitro cytotoxicity assays indicated the presence of robust cell-mediated immunity directed toward the SCC in mice immunized with the transfected cells.
Folia histochemica et cytobiologica / Polish Academy of Sciences, Polish Histochemical and Cytochemical Society, 2007
The recognition, internalization and intracellular processing of antigen are the main functions of dendritic cells (DCs). In the course of these processes, DCs differentiate and acquire the ability to produce cytokines responsible for polarization of the immunological response. Therefore, vaccination with tumor antigen-loaded DCs is one of the most promising approaches to induce tumor-specific immune response. The purpose of this study was to analyze the migratory abilities, from an injection site to tumor-draining lymph nodes (tLN), of DCs applied as an anti-tumor vaccine and their capacity for immune response activation. Mouse DCs of the established JAWS II cell line transduced with EGFP gene or ex vivo bone marrow-isolated DCs (BM-DCs) stained with intravital CFDA dye were loaded with MC38 colon carcinoma tumor lysate (TAg) and then administered peritumorally to MC38 tumor-bearing C57BL/6 mice. On the first, third, fifth and seventh days after injection the tumors, tLNs and splee...