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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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Recovery of wt p53-specific CTLs from nude mice after  successful tumor eradication. wt p53-specific CTL clone 1H11 was retrieved from the spleens of C57BL/6 nude mice 1 mo after tumor eradication as tested in a Eu3+ release assay. CTLs were administered intravenously (A) or intratumorally (B). A nontreated naive mouse (C) was taken  along as a negative control. Spleen cells were restimulated in vitro with 4J  cells and tested for their peptide specificity on p53 −/− Koko cells (open  circles), Koko cells pulsed with the p53 peptide AIYKKSQHM (filled circles), and recognition of 4J cells (filled squares) after 7 d of culture.
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Figure 6: Recovery of wt p53-specific CTLs from nude mice after successful tumor eradication. wt p53-specific CTL clone 1H11 was retrieved from the spleens of C57BL/6 nude mice 1 mo after tumor eradication as tested in a Eu3+ release assay. CTLs were administered intravenously (A) or intratumorally (B). A nontreated naive mouse (C) was taken along as a negative control. Spleen cells were restimulated in vitro with 4J cells and tested for their peptide specificity on p53 −/− Koko cells (open circles), Koko cells pulsed with the p53 peptide AIYKKSQHM (filled circles), and recognition of 4J cells (filled squares) after 7 d of culture.

Mentions: Since these wt p53-specific CTLs cross-reacted on nontransformed cells, we then assessed whether mice carrying a functional p53 gene would survive adoptive transfer of these potentially autoreactive CTLs and whether these CTLs could eradicate established 4J tumors in these mice without overt immunopathology. Adoptively transferred wt p53-specific CTLs were retrieved from the spleen of B6 p53 +/+ nude mice up to 3 mo after intravenous administration (Fig. 4, A and B), but not from splenocytes of untreated B6 nude mice (Fig. 4 C). No signs of autoimmune-induced damage were observed in mice that had recieved wt p53-specific CTLs. We subsequently tested the possibility of eradicating established 4J tumors by adoptive transfer of wt p53-specific CTLs. Tumors grew progressively in untreated mice (Fig. 5 A) and in mice injected with an irrelevant CTL clone recognizing an Ad5 E1B–derived epitope (5; Fig. 5 B). Adoptive transfer by intravenous infusion of the wt p53-specific clone 1H11 resulted in complete and permanent (>5 mo) tumor eradication in mice with small- (average size = 41 mm3; Fig. 5 C) and medium- (average size = 140 mm3; Fig. 5 D) sized tumors. Intratumoral injection of similar numbers of wt p53-specific CTLs in combination with rIL-2 also led to the complete eradication of medium-sized tumors (data not shown). Even large established 4J tumors (average size = 427 mm3) were eradicated in three out of six mice (Fig. 5 E) after intravenous infusion of the wt p53-specific clone 1H11. Mice that had rejected p53-induced tumors after treatment with wt p53-specific CTLs retained long-term tumor-specific CTL immunity, since wt p53-specific CTLs could be retrieved from spleens of these animals one month after CTL treatment (Fig. 6, A and B).


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)

Recovery of wt p53-specific CTLs from nude mice after  successful tumor eradication. wt p53-specific CTL clone 1H11 was retrieved from the spleens of C57BL/6 nude mice 1 mo after tumor eradication as tested in a Eu3+ release assay. CTLs were administered intravenously (A) or intratumorally (B). A nontreated naive mouse (C) was taken  along as a negative control. Spleen cells were restimulated in vitro with 4J  cells and tested for their peptide specificity on p53 −/− Koko cells (open  circles), Koko cells pulsed with the p53 peptide AIYKKSQHM (filled circles), and recognition of 4J cells (filled squares) after 7 d of culture.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 6: Recovery of wt p53-specific CTLs from nude mice after successful tumor eradication. wt p53-specific CTL clone 1H11 was retrieved from the spleens of C57BL/6 nude mice 1 mo after tumor eradication as tested in a Eu3+ release assay. CTLs were administered intravenously (A) or intratumorally (B). A nontreated naive mouse (C) was taken along as a negative control. Spleen cells were restimulated in vitro with 4J cells and tested for their peptide specificity on p53 −/− Koko cells (open circles), Koko cells pulsed with the p53 peptide AIYKKSQHM (filled circles), and recognition of 4J cells (filled squares) after 7 d of culture.
Mentions: Since these wt p53-specific CTLs cross-reacted on nontransformed cells, we then assessed whether mice carrying a functional p53 gene would survive adoptive transfer of these potentially autoreactive CTLs and whether these CTLs could eradicate established 4J tumors in these mice without overt immunopathology. Adoptively transferred wt p53-specific CTLs were retrieved from the spleen of B6 p53 +/+ nude mice up to 3 mo after intravenous administration (Fig. 4, A and B), but not from splenocytes of untreated B6 nude mice (Fig. 4 C). No signs of autoimmune-induced damage were observed in mice that had recieved wt p53-specific CTLs. We subsequently tested the possibility of eradicating established 4J tumors by adoptive transfer of wt p53-specific CTLs. Tumors grew progressively in untreated mice (Fig. 5 A) and in mice injected with an irrelevant CTL clone recognizing an Ad5 E1B–derived epitope (5; Fig. 5 B). Adoptive transfer by intravenous infusion of the wt p53-specific clone 1H11 resulted in complete and permanent (>5 mo) tumor eradication in mice with small- (average size = 41 mm3; Fig. 5 C) and medium- (average size = 140 mm3; Fig. 5 D) sized tumors. Intratumoral injection of similar numbers of wt p53-specific CTLs in combination with rIL-2 also led to the complete eradication of medium-sized tumors (data not shown). Even large established 4J tumors (average size = 427 mm3) were eradicated in three out of six mice (Fig. 5 E) after intravenous infusion of the wt p53-specific clone 1H11. Mice that had rejected p53-induced tumors after treatment with wt p53-specific CTLs retained long-term tumor-specific CTL immunity, since wt p53-specific CTLs could be retrieved from spleens of these animals one month after CTL treatment (Fig. 6, A and B).

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