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Tumor eradication by wild-type p53-specific cytotoxic T lymphocytes.

Vierboom MP, Nijman HW, Offringa R, van der Voort EI, van Hall T, van den Broek L, Fleuren GJ, Kenemans P, Kast WM, Melief CJ - J. Exp. Med. (1997)

Bottom Line: The tumor suppressor protein p53 is overexpressed in close to 50% of all human malignancies.The p53 protein is therefore an attractive target for immunotherapy.Importantly, this occurred in the absence of any demonstrable damage to normal tissue.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunohematology and Blood Bank, University Hospital Leiden, 2300 RC Leiden, the Netherlands.

ABSTRACT
The tumor suppressor protein p53 is overexpressed in close to 50% of all human malignancies. The p53 protein is therefore an attractive target for immunotherapy. Cytotoxic T lymphocytes (CTLs) recognizing a murine wild-type p53 peptide, presented by the major histocompatibility complex class I molecule H-2Kb, were generated by immunizing p53 gene deficient (p53 -/-) C57BL/6 mice with syngeneic p53-overexpressing tumor cells. Adoptive transfer of these CTLs into tumor-bearing p53 +/+ nude mice caused complete and permanent tumor eradication. Importantly, this occurred in the absence of any demonstrable damage to normal tissue. When transferred into p53 +/+ immunocompetent C57BL/6 mice, the CTLs persisted for weeks in the absence of immunopathology and were capable of preventing tumor outgrowth. Wild-type p53-specific CTLs can apparently discriminate between p53-overexpressing tumor cells and normal tissue, indicating that widely expressed autologous molecules such as p53 can serve as a target for CTL-mediated immunotherapy of tumors.

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Related in: MedlinePlus

p53 specificity and Kb restriction of bulk culture of tumor-specific CTLs. (A) Recognition of 4J cells (filled squares) and not of Koko cells  (open circles) by a 2-wk-old CTL bulk culture, named line 8, in a Eu3+ release cytotoxicity assay (46; Genzyme, Cambridge, MA). (B) CTL bulk culture line 8, specifically recognizing 4J, recognizes COS-7 cells transfected  with plasmids containing the genes of H-2Kb and mutant p53 (37).
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Figure 1: p53 specificity and Kb restriction of bulk culture of tumor-specific CTLs. (A) Recognition of 4J cells (filled squares) and not of Koko cells (open circles) by a 2-wk-old CTL bulk culture, named line 8, in a Eu3+ release cytotoxicity assay (46; Genzyme, Cambridge, MA). (B) CTL bulk culture line 8, specifically recognizing 4J, recognizes COS-7 cells transfected with plasmids containing the genes of H-2Kb and mutant p53 (37).

Mentions: wt p53-specific CTLs were generated by immunizing C57BL/6 p53-deficient (p53 −/−) mice with p53-overexpressing 4J tumor cells. The tumor cell 4J expresses high levels of mutant p53 at the RNA level as tested on a Northern blot, and at the protein level p53 as shown by cytospin staining (data not shown). Spleen cells of mice immunized and boosted with irradiated IFN-γ–treated 4J cells were restimulated in vitro with 4J, and the antigen specificity of the resulting bulk CTL cultures was analyzed. The CTL specifically lysed 4J, but not the tumor cell line Koko derived from a p53 −/− mouse (Fig. 1 A). Specific recognition of p53 was tested by incubating these CTLs with COS-7 cells transiently transfected with cDNAs encoding one of the two MHC class I molecules, H-2Kb or H-2Db with or without mutant p53. Specific recognition was assayed by TNF-α production. Fig. 1 B shows that the CTL bulk cultures recognize COS-7 cells transfected with mutant p53 in an MHC class I H-2Kb–restricted manner. Several CTL clones, displaying the same specificity, were isolated from this bulk culture by limiting dilution. Subsequent experiments were performed with CTL clone 1H11, which is a representative of the clones recognizing p53.


Tumor eradication by wild-type p53-specific cytotoxic T lymphocytes.

Vierboom MP, Nijman HW, Offringa R, van der Voort EI, van Hall T, van den Broek L, Fleuren GJ, Kenemans P, Kast WM, Melief CJ - J. Exp. Med. (1997)

p53 specificity and Kb restriction of bulk culture of tumor-specific CTLs. (A) Recognition of 4J cells (filled squares) and not of Koko cells  (open circles) by a 2-wk-old CTL bulk culture, named line 8, in a Eu3+ release cytotoxicity assay (46; Genzyme, Cambridge, MA). (B) CTL bulk culture line 8, specifically recognizing 4J, recognizes COS-7 cells transfected  with plasmids containing the genes of H-2Kb and mutant p53 (37).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199025&req=5

Figure 1: p53 specificity and Kb restriction of bulk culture of tumor-specific CTLs. (A) Recognition of 4J cells (filled squares) and not of Koko cells (open circles) by a 2-wk-old CTL bulk culture, named line 8, in a Eu3+ release cytotoxicity assay (46; Genzyme, Cambridge, MA). (B) CTL bulk culture line 8, specifically recognizing 4J, recognizes COS-7 cells transfected with plasmids containing the genes of H-2Kb and mutant p53 (37).
Mentions: wt p53-specific CTLs were generated by immunizing C57BL/6 p53-deficient (p53 −/−) mice with p53-overexpressing 4J tumor cells. The tumor cell 4J expresses high levels of mutant p53 at the RNA level as tested on a Northern blot, and at the protein level p53 as shown by cytospin staining (data not shown). Spleen cells of mice immunized and boosted with irradiated IFN-γ–treated 4J cells were restimulated in vitro with 4J, and the antigen specificity of the resulting bulk CTL cultures was analyzed. The CTL specifically lysed 4J, but not the tumor cell line Koko derived from a p53 −/− mouse (Fig. 1 A). Specific recognition of p53 was tested by incubating these CTLs with COS-7 cells transiently transfected with cDNAs encoding one of the two MHC class I molecules, H-2Kb or H-2Db with or without mutant p53. Specific recognition was assayed by TNF-α production. Fig. 1 B shows that the CTL bulk cultures recognize COS-7 cells transfected with mutant p53 in an MHC class I H-2Kb–restricted manner. Several CTL clones, displaying the same specificity, were isolated from this bulk culture by limiting dilution. Subsequent experiments were performed with CTL clone 1H11, which is a representative of the clones recognizing p53.

Bottom Line: The tumor suppressor protein p53 is overexpressed in close to 50% of all human malignancies.The p53 protein is therefore an attractive target for immunotherapy.Importantly, this occurred in the absence of any demonstrable damage to normal tissue.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunohematology and Blood Bank, University Hospital Leiden, 2300 RC Leiden, the Netherlands.

ABSTRACT
The tumor suppressor protein p53 is overexpressed in close to 50% of all human malignancies. The p53 protein is therefore an attractive target for immunotherapy. Cytotoxic T lymphocytes (CTLs) recognizing a murine wild-type p53 peptide, presented by the major histocompatibility complex class I molecule H-2Kb, were generated by immunizing p53 gene deficient (p53 -/-) C57BL/6 mice with syngeneic p53-overexpressing tumor cells. Adoptive transfer of these CTLs into tumor-bearing p53 +/+ nude mice caused complete and permanent tumor eradication. Importantly, this occurred in the absence of any demonstrable damage to normal tissue. When transferred into p53 +/+ immunocompetent C57BL/6 mice, the CTLs persisted for weeks in the absence of immunopathology and were capable of preventing tumor outgrowth. Wild-type p53-specific CTLs can apparently discriminate between p53-overexpressing tumor cells and normal tissue, indicating that widely expressed autologous molecules such as p53 can serve as a target for CTL-mediated immunotherapy of tumors.

Show MeSH
Related in: MedlinePlus