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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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Recognition of various target cells and the antigenic  peptide by CTL clones derived  from TIL586. T cell clones were  generated by limiting dilution (1  cell/well) from the TIL586 cell  line and were further expanded  in AIM-V medium containing  6,000 IU/ml IL-2. GM-CSF secretion by CTL clone 586TILC1 (left) and clone 4 (right)  was measured after coculturing  with a normal melanocyte cell  line NHEM680 (HLA-A31+),  586EBV B cells pulsed with the  ORF3P peptide or irrelevant  peptide, 397mel or 586mel cells.
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Figure 1: Recognition of various target cells and the antigenic peptide by CTL clones derived from TIL586. T cell clones were generated by limiting dilution (1 cell/well) from the TIL586 cell line and were further expanded in AIM-V medium containing 6,000 IU/ml IL-2. GM-CSF secretion by CTL clone 586TILC1 (left) and clone 4 (right) was measured after coculturing with a normal melanocyte cell line NHEM680 (HLA-A31+), 586EBV B cells pulsed with the ORF3P peptide or irrelevant peptide, 397mel or 586mel cells.

Mentions: In previous studies, we isolated a number of T cell clones from the TIL586 cell line by the limiting dilution method (22). Among them, six clones recognized 586EBV B cells pulsed with the ORF3P peptide derived from a gene product translated from an alternative open reading of the TRP1/gp75 gene, and the autologous 586mel tumor cells, but did not recognize 586EBV B cells pulsed with an irrelevant peptide. TIL586-C1 was a representative of these T cell clones as shown in Fig. 1. However, several T cell clones isolated from the same TIL586 cell line recognized neither 586EBV B cells pulsed with the TRP-1 peptide ORF3P nor COS cells transfected with TRP-1 and HLA-A31 cDNAs, but were capable of recognizing 586mel as well as HLA-A31+ NHEM680 melanocyte line (Fig. 1). These results suggested that these T cell clones recognized additional tumor antigens on the 586mel tumor cells. These T cell clones were then expanded to obtain enough cells for screening cDNA libraries or testing other cDNAs for recognition by methods described in the Materials and Methods section. One of the clones, CTL clone 4, was successfully expanded and used for further studies as described below.


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)

Recognition of various target cells and the antigenic  peptide by CTL clones derived  from TIL586. T cell clones were  generated by limiting dilution (1  cell/well) from the TIL586 cell  line and were further expanded  in AIM-V medium containing  6,000 IU/ml IL-2. GM-CSF secretion by CTL clone 586TILC1 (left) and clone 4 (right)  was measured after coculturing  with a normal melanocyte cell  line NHEM680 (HLA-A31+),  586EBV B cells pulsed with the  ORF3P peptide or irrelevant  peptide, 397mel or 586mel cells.
© Copyright Policy
Related In: Results  -  Collection

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

Figure 1: Recognition of various target cells and the antigenic peptide by CTL clones derived from TIL586. T cell clones were generated by limiting dilution (1 cell/well) from the TIL586 cell line and were further expanded in AIM-V medium containing 6,000 IU/ml IL-2. GM-CSF secretion by CTL clone 586TILC1 (left) and clone 4 (right) was measured after coculturing with a normal melanocyte cell line NHEM680 (HLA-A31+), 586EBV B cells pulsed with the ORF3P peptide or irrelevant peptide, 397mel or 586mel cells.
Mentions: In previous studies, we isolated a number of T cell clones from the TIL586 cell line by the limiting dilution method (22). Among them, six clones recognized 586EBV B cells pulsed with the ORF3P peptide derived from a gene product translated from an alternative open reading of the TRP1/gp75 gene, and the autologous 586mel tumor cells, but did not recognize 586EBV B cells pulsed with an irrelevant peptide. TIL586-C1 was a representative of these T cell clones as shown in Fig. 1. However, several T cell clones isolated from the same TIL586 cell line recognized neither 586EBV B cells pulsed with the TRP-1 peptide ORF3P nor COS cells transfected with TRP-1 and HLA-A31 cDNAs, but were capable of recognizing 586mel as well as HLA-A31+ NHEM680 melanocyte line (Fig. 1). These results suggested that these T cell clones recognized additional tumor antigens on the 586mel tumor cells. These T cell clones were then expanded to obtain enough cells for screening cDNA libraries or testing other cDNAs for recognition by methods described in the Materials and Methods section. One of the clones, CTL clone 4, was successfully expanded and used for further studies as described below.

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