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Identification of TRP-2 as a human tumor antigen recognized by cytotoxic T lymphocytes.

Wang RF, Appella E, Kawakami Y, Kang X, Rosenberg SA - J. Exp. Med. (1996)

Bottom Line: This epitope peptide was capable of sensitizing target cells for lysis by a CTL clone at 1 nM peptide concentration.Although some modified peptides could be recognized by the CTL clone, none were found to be better recognized by T cells than the parental peptide.Like other melamona differentiation antigens, TRP-2 was only expressed in melanoma, melanocytes, and retina, but not in other human tissues tested.

View Article: PubMed Central - PubMed

Affiliation: National Cancer Institute, Bethesda, Maryland 20892, USA.

ABSTRACT
The infusion of TIL586 along with interleukin-2 into the autologous patient with metastatic melanoma resulted in the objective regression of tumor. A gene encoding a tumor antigen recognized by TIL586 was previously isolated and shown to encode gp75 or TRP-1. Here we report that TRP-2 was identified as a second tumor antigen recognized by a HLA-A31-restricted CTL clone derived from the TIL586 cell line. The peptide LLPGGRPYR epitope was subsequently identified from the coding region of TRP-2 based on studies of the recognition of truncated TRP-2 cDNAs and the HLA-A31 binding motif. This epitope peptide was capable of sensitizing target cells for lysis by a CTL clone at 1 nM peptide concentration. Although some modified peptides could be recognized by the CTL clone, none were found to be better recognized by T cells than the parental peptide. Like other melamona differentiation antigens, TRP-2 was only expressed in melanoma, melanocytes, and retina, but not in other human tissues tested.

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Characterization of the antigenic peptide recognized by CTL clone 4. (A) GM-CSF release by T cells at different peptide concentrations.  586EBV (A31+) were pulsed with the TRP197–205 peptide ( ) and T2 (non-A31) cells were pulsed with the TRP197–205 ( ) at various peptide concentrations for 90 min. ORF3P as a control peptide was pulsed onto 586EBV B cells (--▵--). GM-CSF release by CTL clone 4 was determined after coincubation with 586EBV B cells pulsed with TRP197–205 and ORF3P, and T2 cells pulsed with TRP197–205. (B) Sensitization of the target cells for lysis by  CTL clone 4 at different peptide concentrations. 586EBV B cells were incubated with TRP197–205 ( ), an irrelevant peptide ORF3P (--▵--), and T2  cells pulsed with TRP197–205 ( ) at various peptide concentrations. After peptide incubation, target cells were labeled for 30 min. Following washes, cytolytic activity of CTL clone 4 at an E/T of 40:1 was measured after a 4 h incubation of T cells with target cells. (C) Lysis of the target cells by CTL clone  4 at different E/T. Target 586EBV cells were separately incubated with TRP197–205 ( ) or the irrelevant peptides ORF3P (--▵--), and target T2 cells  were incubated with the TRP197–205 peptide ( ) for 90 min. 586mel ( ) and 397mel ( ) were used as positive and negative controls, respectively.
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Figure 5: Characterization of the antigenic peptide recognized by CTL clone 4. (A) GM-CSF release by T cells at different peptide concentrations. 586EBV (A31+) were pulsed with the TRP197–205 peptide ( ) and T2 (non-A31) cells were pulsed with the TRP197–205 ( ) at various peptide concentrations for 90 min. ORF3P as a control peptide was pulsed onto 586EBV B cells (--▵--). GM-CSF release by CTL clone 4 was determined after coincubation with 586EBV B cells pulsed with TRP197–205 and ORF3P, and T2 cells pulsed with TRP197–205. (B) Sensitization of the target cells for lysis by CTL clone 4 at different peptide concentrations. 586EBV B cells were incubated with TRP197–205 ( ), an irrelevant peptide ORF3P (--▵--), and T2 cells pulsed with TRP197–205 ( ) at various peptide concentrations. After peptide incubation, target cells were labeled for 30 min. Following washes, cytolytic activity of CTL clone 4 at an E/T of 40:1 was measured after a 4 h incubation of T cells with target cells. (C) Lysis of the target cells by CTL clone 4 at different E/T. Target 586EBV cells were separately incubated with TRP197–205 ( ) or the irrelevant peptides ORF3P (--▵--), and target T2 cells were incubated with the TRP197–205 peptide ( ) for 90 min. 586mel ( ) and 397mel ( ) were used as positive and negative controls, respectively.


Identification of TRP-2 as a human tumor antigen recognized by cytotoxic T lymphocytes.

Wang RF, Appella E, Kawakami Y, Kang X, Rosenberg SA - J. Exp. Med. (1996)

Characterization of the antigenic peptide recognized by CTL clone 4. (A) GM-CSF release by T cells at different peptide concentrations.  586EBV (A31+) were pulsed with the TRP197–205 peptide ( ) and T2 (non-A31) cells were pulsed with the TRP197–205 ( ) at various peptide concentrations for 90 min. ORF3P as a control peptide was pulsed onto 586EBV B cells (--▵--). GM-CSF release by CTL clone 4 was determined after coincubation with 586EBV B cells pulsed with TRP197–205 and ORF3P, and T2 cells pulsed with TRP197–205. (B) Sensitization of the target cells for lysis by  CTL clone 4 at different peptide concentrations. 586EBV B cells were incubated with TRP197–205 ( ), an irrelevant peptide ORF3P (--▵--), and T2  cells pulsed with TRP197–205 ( ) at various peptide concentrations. After peptide incubation, target cells were labeled for 30 min. Following washes, cytolytic activity of CTL clone 4 at an E/T of 40:1 was measured after a 4 h incubation of T cells with target cells. (C) Lysis of the target cells by CTL clone  4 at different E/T. Target 586EBV cells were separately incubated with TRP197–205 ( ) or the irrelevant peptides ORF3P (--▵--), and target T2 cells  were incubated with the TRP197–205 peptide ( ) for 90 min. 586mel ( ) and 397mel ( ) were used as positive and negative controls, respectively.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2211562&req=5

Figure 5: Characterization of the antigenic peptide recognized by CTL clone 4. (A) GM-CSF release by T cells at different peptide concentrations. 586EBV (A31+) were pulsed with the TRP197–205 peptide ( ) and T2 (non-A31) cells were pulsed with the TRP197–205 ( ) at various peptide concentrations for 90 min. ORF3P as a control peptide was pulsed onto 586EBV B cells (--▵--). GM-CSF release by CTL clone 4 was determined after coincubation with 586EBV B cells pulsed with TRP197–205 and ORF3P, and T2 cells pulsed with TRP197–205. (B) Sensitization of the target cells for lysis by CTL clone 4 at different peptide concentrations. 586EBV B cells were incubated with TRP197–205 ( ), an irrelevant peptide ORF3P (--▵--), and T2 cells pulsed with TRP197–205 ( ) at various peptide concentrations. After peptide incubation, target cells were labeled for 30 min. Following washes, cytolytic activity of CTL clone 4 at an E/T of 40:1 was measured after a 4 h incubation of T cells with target cells. (C) Lysis of the target cells by CTL clone 4 at different E/T. Target 586EBV cells were separately incubated with TRP197–205 ( ) or the irrelevant peptides ORF3P (--▵--), and target T2 cells were incubated with the TRP197–205 peptide ( ) for 90 min. 586mel ( ) and 397mel ( ) were used as positive and negative controls, respectively.
Bottom Line: This epitope peptide was capable of sensitizing target cells for lysis by a CTL clone at 1 nM peptide concentration.Although some modified peptides could be recognized by the CTL clone, none were found to be better recognized by T cells than the parental peptide.Like other melamona differentiation antigens, TRP-2 was only expressed in melanoma, melanocytes, and retina, but not in other human tissues tested.

View Article: PubMed Central - PubMed

Affiliation: National Cancer Institute, Bethesda, Maryland 20892, USA.

ABSTRACT
The infusion of TIL586 along with interleukin-2 into the autologous patient with metastatic melanoma resulted in the objective regression of tumor. A gene encoding a tumor antigen recognized by TIL586 was previously isolated and shown to encode gp75 or TRP-1. Here we report that TRP-2 was identified as a second tumor antigen recognized by a HLA-A31-restricted CTL clone derived from the TIL586 cell line. The peptide LLPGGRPYR epitope was subsequently identified from the coding region of TRP-2 based on studies of the recognition of truncated TRP-2 cDNAs and the HLA-A31 binding motif. This epitope peptide was capable of sensitizing target cells for lysis by a CTL clone at 1 nM peptide concentration. Although some modified peptides could be recognized by the CTL clone, none were found to be better recognized by T cells than the parental peptide. Like other melamona differentiation antigens, TRP-2 was only expressed in melanoma, melanocytes, and retina, but not in other human tissues tested.

Show MeSH
Related in: MedlinePlus