Characterization of CD8 + cytotoxic T lymphocyte/tumor cell interactions reflecting recognition of an endogenously expressed murine wild-type p53 determinant

Ryan, Mary H.; Abrams, Scott I.

doi:10.1007/s002620000156

Cited by 38 publications

(34 citation statements)

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“…To extend the observation that irradiation sensitizes tumor cells to Ag-specific CTL-mediated killing, we tested another CD8 ϩ CTL line with specificity for a wild-type p53 determinant previously found to be functionally expressed by MC38 cells, but not EL-4 cells (40). Since MC38 CEA ϩ cells were derived from MC38 cells, we assayed them here for sensitivity to p53-specific CTL.…”

Section: Irradiation Increases Tumor Cell Sensitivity To Lysis By a Smentioning

confidence: 99%

“…EL-4 cells were prepared for use as targets in a standard cytotoxicity assay using 51 Cr as previously described (40). These radiolabeled cells (5 ϫ 10 3 cells/well) were incubated with 1 g/ml CEA 526 peptide or vesicular stomatitis virus-N 52-59 (RGYVYQGL) (42) as a negative control and coincubated with CEA 526 CTL at various E:T cell ratios for 4 h at 37°C with 5% CO 2 .…”

Section: Cytotoxicity Assaysmentioning

confidence: 99%

Section: T Cell Linesmentioning

confidence: 99%

“…The H-2D b -restricted, p53 peptide-specific CD8 ϩ CTL line, designated p53 232 , was generated from C57BL/6 mice as previously described (40) and recognizes the peptide epitope p53 232-240 (KYMCNS SCM). The H-2D b -restricted, FluNP peptide-specific CD8 ϩ CTL line, designated FluNP 366 , was generated from C57BL/6 mice as previously described (40) and recognizes the peptide epitope FluNP 366 -374 . The CTL lines were maintained by weekly in vitro stimulation with irradiated syngeneic naive splenocytes in complete medium (RPMI 1640 with FCS (10%); L-glutamine (2 mM), sodium pyruvate (1 mM), HEPES (7 mM), gentamicin (50 g/ml), 2-ME (50 M), and nonessential amino acids (0.1 mM; Biofluids, Rockville, MD)), supplemented with 1 g/ml specific peptide and 10 IU/ml murine IL-2 (Roche, Indianapolis, IN).…”

Section: T Cell Linesmentioning

confidence: 99%

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Irradiation of Tumor Cells Up-Regulates Fas and Enhances CTL Lytic Activity and CTL Adoptive Immunotherapy

Chakraborty¹,

Abrams²,

Camphausen

et al. 2003

The Journal of Immunology

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417

304

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CD8+ CTL play important roles against malignancy in both active and passive immunotherapy. Nonetheless, the success of antitumor CTL responses may be improved by additional therapeutic modalities. Radiotherapy, which has a long-standing use in treating neoplastic disease, has been found to induce unique biologic alterations in cancer cells affecting Fas gene expression, which, consequently, may influence the overall lytic efficiency of CTL. Here, in a mouse adenocarcinoma cell model, we examined whether exposure of these tumor cells to sublethal doses of irradiation 1) enhances Fas expression, leading to more efficient CTL killing via Fas-dependent mechanisms in vitro; and 2) improves antitumor activity in vivo by adoptive transfer of these Ag-specific CTL. Treatment of carcinoembryonic Ag-expressing MC38 adenocarcinoma cells with irradiation (20 Gy) in vitro enhanced Fas expression at molecular, phenotypic, and functional levels. Furthermore, irradiation sensitized these targets to Ag-specific CTL killing via the Fas/Fas ligand pathway. We examined the effect of localized irradiation of s.c. growing tumors on the efficiency of CTL adoptive immunotherapy. Irradiation caused up-regulation of Fas by these tumor cells in situ, based on immunohistochemistry. Moreover, localized irradiation of the tumor significantly potentiated tumor rejection by these carcinoembryonic Ag-specific CTL. Overall, these results showed for the first time that 1) regulation of the Fas pathway in tumor cells by irradiation plays an important role in their sensitization to Ag-specific CTL; and 2) a combination regimen of tumor-targeted irradiation and CTL promotes more effective antitumor responses in vivo, which may have implications for the combination of immunotherapy and radiation therapy.

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Section: Irradiation Increases Tumor Cell Sensitivity To Lysis By a Smentioning

confidence: 99%

Section: Cytotoxicity Assaysmentioning

confidence: 99%

Section: T Cell Linesmentioning

confidence: 99%

Section: T Cell Linesmentioning

confidence: 99%

See 2 more Smart Citations

Irradiation of Tumor Cells Up-Regulates Fas and Enhances CTL Lytic Activity and CTL Adoptive Immunotherapy

Chakraborty¹,

Abrams²,

Camphausen

et al. 2003

The Journal of Immunology

Self Cite

417

304

View full text Add to dashboard Cite

show abstract

“…To evaluate CD8 + T-cell responses specific for tumor antigens, splenocytes from vaccinated animals were pooled and dispersed into single-cell suspensions, and stimulated with the H-2D b -restricted peptide CEA 526-533 (10 μg/mL, EAQNTTYL) (24,30), the H-2D b -restricted peptide p53 232-240 (2 μg/mL, KYMCNSSCM) (31) or the H-2K b -restricted peptide p15E 604-611 (1 μg/mL, KSPWFTTL, referred as gp70 peptide) (32). Six days later, bulk lymphocytes were separated by centrifugation through a Ficoll-Hypaque gradient.…”

Section: T Cell Responsesmentioning

confidence: 99%

4-1BB ligand enhances tumor-specific immunity of poxvirus vaccines

Kudo-Saito

Hodge

Kwak

et al. 2006

Vaccine

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Purpose-Recombinant poxvirus vaccines have been explored as tumor vaccines. The immunogenicity of these vaccines can be enhanced by co-expressing costimulatory molecules and tumor-associated antigens. While the B7-CD28 interaction has been most comprehensively investigated, other costimulatory molecules utilize different signaling pathways and might provide further cooperation in T cell priming and survival. 4-1BB (CD137) is a TNF family member and is critical for activation and long-term maintenance of primed T-cells. This study was conducted to determine if a poxvirus expressing the ligand for 4-1BB (4-1BBL) could further improve the immune and therapeutic responses of a previously reported poxvirus vaccine expressing a triad of costimulatory molecules (B7.1, ICAM-1, and LFA-3).Experimental Design-A recombinant vaccinia virus expressing 4-1BBL was generated and characterized in an in vitro infection system. This vaccine was then used alone or in combination with a vaccinia virus expressing CEA, B7.1, ICAM-1, and LFA-3 in CEA-transgenic mice bearing established MC38 tumors. Tumor growth and immune responses against CEA and other tumorassociated antigens were determined. The level of anti-apoptotic proteins in responding T cells was determined by flow cytometry on tetramer selected T cells.Results-The combination of 4-1BBL with B7.1-based poxvirus vaccination resulted in significantly enhanced therapeutic effects against CEA-expressing tumors in a CEA transgenic mouse model. This was associated with an increased level of CEA-specific CD4 + and CD8 + T cell responses, induction of antigen spreading to p53 and gp70, increased accumulation of CEA-specific T cells in the tumor microenvironment, and increased expression of bcl-XL and bcl-2 in CD4 + and CD8 + T cells in vaccinated mice. Conclusion-4-1BBLcooperates with B7 in enhancing anti-tumor and immunologic responses using a recombinant poxvirus vaccine model. The inclusion of costimulatory molecules targeting distinct T cell signaling pathways provides a mechanism for enhancing the therapeutic effectiveness of tumor vaccines.

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Impact of p53-based immunization on primary chemically-induced tumors

et al. 2004

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In mice as well as humans, cytotoxic T lymphocytes (CTL) specific for wild-type-sequence (wt) p53 peptides have been shown to react against a wide range of tumors, but not normal cells. As such, they are attractive candidates for developing broadly applicable cancer vaccines. Of particular interest is the potential of using p53-based vaccines in high-risk individuals to prevent cancer. Methylcholanthrene, an immunosuppressive polycyclic hydrocarbon carcinogen implicated as a causative agent in human cancers, has long been used to induce murine tumors with a high incidence of genetic alterations and sensitivity to wt p53-specific CTL. To analyze the potential of p53-based vaccines on primary tumors, we evaluated the efficacy of DNA and dendritic cell vaccines targeting wt p53 peptides given to methylcholanthrene-treated mice in the protection or therapy settings. The results indicate that the efficacy of these vaccines relative to reducing tumor incidence were severely compromised by vaccine-induced tumor escape. As compared to tumors induced in non-immunized mice, a higher incidence of epitope-loss tumors was detected in tumors from the immunized mice. The increase in tumor escape arose as a consequence of either increased frequencies of mutations within/flanking p53 epitope-coding regions or downregulation of expression of the major histocompatibility complex Class I molecules that present these epitopes for T cell recognition These findings are consistent with current views of immunoselection occurring in patients receiving tumor peptide-based immunotherapy, and impact on the design and implementation of p53-based vaccines, in particular, those aimed at treating individuals at high risk for developing cancer. © 2004 Wiley-Liss, Inc.Key words: methylcholanthrene; mouse sarcomas; p53; vaccine; immune evasionThe immunotherapeutic targeting of p53 is a particularly active area of investigation. p53 was initially identified as a transformation-related antigen expressed at elevated levels in chemically induced sarcomas and other transformed cells of the mouse. 1 Its subsequent classification as a tumor suppressor gene product that is genetically altered frequently in human tumors 2,3 made it an attractive candidate for development of cancer vaccines. The most common type of genetic alteration in p53 involves a missense mutation. Although these mutations represent ideal targets for tumor specific vaccines, the constraints imposed by antigen presentation limit their use. 4 Frequent accumulation of mutant p53 molecules in tumors, however, suggested that enhanced presentation by these tumors of wild-type sequence (wt) epitopes derived from these molecules for T cell recognition might occur 5 Like most of the presently defined melanoma antigens, wt p53 epitopes can be considered as widely expressed tumor associated antigens, 6 and vaccines targeting them would have broad applicability. [7][8][9] The potential of using p53-based vaccines in high-risk individuals to prevent cancer is of particular interest.Accompanying the...

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Characterization of CD8 + cytotoxic T lymphocyte/tumor cell interactions reflecting recognition of an endogenously expressed murine wild-type p53 determinant

Cited by 38 publications

References 0 publications

Irradiation of Tumor Cells Up-Regulates Fas and Enhances CTL Lytic Activity and CTL Adoptive Immunotherapy

Irradiation of Tumor Cells Up-Regulates Fas and Enhances CTL Lytic Activity and CTL Adoptive Immunotherapy

4-1BB ligand enhances tumor-specific immunity of poxvirus vaccines

Impact of p53-based immunization on primary chemically-induced tumors

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