Human tumor-derived genomic DNA transduced into a recipient cell induces tumor-specific immune responses<i>ex</i><i>vivo</i>

Whiteside, Theresa L.; Gambotto, Andrea; Albers, Andreas E.; Stanson, Joanna; Cohen, Edward P.

doi:10.1073/pnas.142302399

Cited by 19 publications

(16 citation statements)

References 23 publications

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“…In prior reports, [17][18][19][20] we described the results of studies in tumor-bearing mice including mice with SCC treated by immunization with a unique vaccine prepared by transfer of sheared genomic tumor-DNA-fragments into LM cells, a mouse fibroblast cell line. As the transferred DNA integrates spontaneously into the genome of the recipient cells, replicates as the cells divide and is expressed, the number of vaccine cells could be conveniently expanded as desired for multiple rounds of immunization.…”

Section: Introductionmentioning

confidence: 99%

T-regulatory cells are relatively deficient in squamous carcinomas undergoing regression in mice immunized with a squamous carcinoma vaccine enriched for immunotherapeutic cells

et al. 2007

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In a prior report (Int J Cancer 2006; 119: 339-348), we described a new vaccination strategy for squamous cell carcinoma (SCC). The vaccine was prepared by transfer of unfractionated DNA-fragments (25 kb) from KLN205 cells, a squamous carcinoma cell line (DBA/2 origin; H-2 d ) into LM cells, a highly immunogenic mouse fibroblast cell line (C3H/He origin; (H-2 k )). As only a small proportion of the transfected cell population was expected to have incorporated DNA segments that included genes specifying antigens associated with the squamous carcinoma cells, we devised a novel strategy to enrich the vaccine for immunotherapeutic cells. Enhanced immunity to squamous carcinoma was induced in tumor-bearing mice treated solely by immunization with the enriched vaccine, which translated into prolonged survival without toxicity. Here, we describe the characteristics of the cell populations infiltrating established squamous carcinomas undergoing regression in mice immunized with vaccines enriched for immunotherapeutic cells. The results indicated that CD8 þ T cells were predominant and that T-regulatory cellshigh , CD4/CTLA-4e) were relatively deficient in the regressing tumors. Inflammatory infiltrates were not detected in various organs and tissues of mice immunized with the DNA-based vaccine.

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Section: Introductionmentioning

confidence: 99%

T-regulatory cells are relatively deficient in squamous carcinomas undergoing regression in mice immunized with a squamous carcinoma vaccine enriched for immunotherapeutic cells

et al. 2007

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“…5,6 Thus, autologous DC are either pulsed with known MHC class I-or class II-specific peptides or "fed" whole tumor lysates or apoptotic tumor cells when the identity of TAA is unknown. [6][7][8] Alternatively, tumor-derived RNA can be transferred to DC and expressed as immunogenic proteins. 3 Still another strategy involves transfer of cDNA specifying a known tumor antigen or epitope in the form of viral or nonviral plasmids.…”

Section: Introductionmentioning

confidence: 99%

“…8 It has been successfully used by others for in vivo delivery of tumor vaccines produced by genomic tumor DNA transfer to a recipient master cell to achieve tumor regression and prolonged survival of mice in several different murine models of tumor growth. [12][13][14][15][16] Based on these encouraging results, we reasoned that an effective vaccine could be prepared by transfer of genomic DNA from a tumor to DC, using human cells.…”

Section: Introductionmentioning

confidence: 99%

“…In order to evaluate nucleofection efficiency, genomic DNA was obtained from the melanoma cell line, previously transfected with the EGFP gene and selected (Mel/EGFP), so that all cells were green by flow cytometry or fluorescent microscopy, as previously described. 8 hMoDC were expected to appear green upon successful transfection of genomic DNA containing the EGFP gene. However, examination by flow cytometry consistently demonstrated expression of EGFP in fewer than 1% of hMoDC after nucleofection (data not shown).…”

Section: Introductionmentioning

confidence: 99%

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Transfection of human monocyte-derived dendritic cells with native tumor DNA induces antigen-specific T-Cell responses in vitro.

Artusio¹,

Hathaway²,

Stanson³

et al. 2006

Cancer Biology & Therapy

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Objective: Nucleofection of genomic tumor (Tu) DNA into human monocyte-derived dendritic cells (hMoDC) was evaluated for use in producing anti-tumor vaccines able to induce effective T-cell specific immune responses.Methods: Cultured hMoDC obtained from HLA-A2+ normal donors were nucleofected with genomic DNA extracted from an HLA-A2+gp100+ Mel 526 cell line and 3' end-labeled with biotinylated TdT nucleotides or from a genetically-modified Mel 526 expressing enhanced green fluorescent protein (EGFP). An Amaxa Nucleofector™ system was used for electroporation. Nucleofected hMoDC were matured in the presence of cytokines and examined in ELISPOT assays for the ability to present the gp100 [209][210][211][212][213][214][215][216][217] epitope to epitope-specific T cells or to prime autologous naïve T cells in culture.Results: The nucleofected hMoDC presented gp100 protein to HLA-A2+gp100-specific T cells as observed in IFN-g ELISPOT assays. Spot formation was inhibited by anti-HLA class I and HLA-A2 but not anti-HLA class II antibodies (Abs). Tu DNA-nucleofected hMoDC also primed naïve autologous peripheral blood T cells in culture to develop into Tu-reactive effector cells (CTL). These CTL recognized Tu cells which had donated genomic DNA, and these responses were MHC class I-and class II-restricted. The CTL recognized shared Tu antigens encoded in Tu-derived DNA.Conclusion: Nucleofection of hMoDC with genomic Tu-derived DNA is a useful strategy for Tu vaccine production: it is feasible, does not require Tu epitope isolation, can be used when few Tu cells are available, and avoids Tu-induced DC suppression.

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“…Poorly immunogenic TAA, characteristic of malignant cells, can become strongly antigenic if they are expressed by highly immunogenic cells. In animal tumor models, immunization with DNA-based vaccine was sufficient to deter tumor growth and to prolong the lives of tumorbearing mice [14,15].…”

Section: Introductionmentioning

confidence: 99%

Antigenic Differences Between Normal and Malignant Cells as a Basis for Treatment of Intracerebral Neoplasms Using a DNA-Based Vaccine

Lichtor¹,

Glick

O-Sullivan³

et al. 2006

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Antigenic differences between normal and malignant cells of the cancer patient form the rationale for clinical immunotherapeutic strategies. Because the antigenic phenotype of neoplastic cells varies widely among different cells within the same malignant cell-population, immunization with a vaccine that stimulates immunity to the broad array of tumor antigens expressed by the cancer cells is likely to be more efficacious than immunization with a vaccine for a single antigen. A vaccine prepared by transfer of DNA from the tumor into a highly immunogenic cell line can encompass the array of tumor antigens that characterize the patient's neoplasm. Poorly immunogenic tumor antigens, characteristic of malignant cells, can become strongly antigenic if they are expressed by highly immunogenic cells. A DNA-based vaccine was prepared by transfer of genomic DNA from a breast cancer that arose spontaneously in a C3H/He mouse into a highly immunogenic mouse fibroblast cell line, where genes specifying tumor-antigens were expressed. The fibroblasts were modified in advance of DNA-transfer to secrete an immune augmenting cytokine and to express allogeneic MHC class I-determinants. In an animal model of breast cancer metastatic to the brain, introduction of the vaccine directly into the tumor bed stimulated a systemic cellular anti-tumor immune response measured by two independent in vitro assays and prolonged the lives of the tumor-bearing mice. Furthermore, using antibodies against the various T-cell subsets, it was determined that the systemic cellular anti-tumor immunity was mediated by CD8 + , CD4 + and NK/LAK cells. The application of DNA-based genomic vaccines for the treatment of a variety of brain tumors is being explored.

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Human tumor-derived genomic DNA transduced into a recipient cell induces tumor-specific immune responsesexvivo

Cited by 19 publications

References 23 publications

T-regulatory cells are relatively deficient in squamous carcinomas undergoing regression in mice immunized with a squamous carcinoma vaccine enriched for immunotherapeutic cells

T-regulatory cells are relatively deficient in squamous carcinomas undergoing regression in mice immunized with a squamous carcinoma vaccine enriched for immunotherapeutic cells

Transfection of human monocyte-derived dendritic cells with native tumor DNA induces antigen-specific T-Cell responses in vitro.

Antigenic Differences Between Normal and Malignant Cells as a Basis for Treatment of Intracerebral Neoplasms Using a DNA-Based Vaccine

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