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Identification of tyrosinase-related protein 2 as a tumor rejection antigen for the B16 melanoma.

Bloom MB, Perry-Lalley D, Robbins PF, Li Y, el-Gamil M, Rosenberg SA, Yang JC - J. Exp. Med. (1997)

Bottom Line: In spite of this, little is known about the optimal ways to use these antigens to treat patients with cancer.A peptide conforming to the predicted MHC class I H2-Kb binding motif, TRP-2181-188, was identified as the major reactive epitope within TRP-2 recognized by these anti-B16 CTLs.By site-directed mutagenesis, it was shown that alteration of this epitope eliminated recognition of TRP-2.

View Article: PubMed Central - PubMed

Affiliation: Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.

ABSTRACT
Recently, major advances have been made in the identification of antigens from human melanoma which are recognized by T cells. In spite of this, little is known about the optimal ways to use these antigens to treat patients with cancer. Progress in this area is likely to require accurate preclinical animal models, but the availability of such models has lagged behind developments in human tumor immunology. Whereas many of the identified human melanoma antigens are normal tissue differentiation proteins, analogous murine tumor antigens have not yet been identified. In this paper we identify a normal tissue differentiation antigen, tyrosinase-related protein 2 (TRP-2), expressed by the murine B16 melanoma which was found by screening a cDNA library from B16 with tumor-reactive cytotoxic T lymphocytes (CTL). A peptide conforming to the predicted MHC class I H2-Kb binding motif, TRP-2181-188, was identified as the major reactive epitope within TRP-2 recognized by these anti-B16 CTLs. By site-directed mutagenesis, it was shown that alteration of this epitope eliminated recognition of TRP-2. It was further demonstrated that a CTL line raised from splenocytes by repeated stimulation in vitro with this peptide could recognize B16 tumor and was therapeutic against 3-d-old established pulmonary metastases. The use of TRP-2 in a preclinical model of tumor immunotherapy may be helpful in suggesting optimal vaccination strategies for cancer therapy in patients.

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Related in: MedlinePlus

Treatment of 3-d-old pulmonary metastases from B16 by the  intravenous administration of T cells reactive with TRP-2181–188 generated by in vitro peptide stimulation of splenocytes from normal or B16immunized mice. Low-dose systemic IL-2 was given with adoptive cell  transfer. Therapeutic effects were seen from anti–TRP-2181–188 CTLs  from normal mice (P = 0.0004 vs. IL-2 alone, and P = 0.014 vs.  ovalbumin257–264 stimulated cultures), as well as from anti-B16 immunized  mice (P = 0.0003 vs. IL-2 alone, and P = 0.010 vs. ovalbumin257–264  stimulated cultures).
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Figure 4: Treatment of 3-d-old pulmonary metastases from B16 by the intravenous administration of T cells reactive with TRP-2181–188 generated by in vitro peptide stimulation of splenocytes from normal or B16immunized mice. Low-dose systemic IL-2 was given with adoptive cell transfer. Therapeutic effects were seen from anti–TRP-2181–188 CTLs from normal mice (P = 0.0004 vs. IL-2 alone, and P = 0.014 vs. ovalbumin257–264 stimulated cultures), as well as from anti-B16 immunized mice (P = 0.0003 vs. IL-2 alone, and P = 0.010 vs. ovalbumin257–264 stimulated cultures).

Mentions: The therapeutic efficacy of CTL lines generated by in vitro stimulation with TRP-2181–188 was also examined. T cell lines generated from normal or B16 immune mice by stimulation with the TRP-2181–188 peptide were used to treat 3-d-old B16 lung metastases by intravenous administration of the CTL in combination with low dose IL-2. A highly significant therapeutic effect was seen (P <0.001) with cultures from normal or B16-immunized mice when stimulated with TRP-2181–188, but not with ova257–264 peptide (when compared to treatment with IL-2 alone) (Fig. 4).


Identification of tyrosinase-related protein 2 as a tumor rejection antigen for the B16 melanoma.

Bloom MB, Perry-Lalley D, Robbins PF, Li Y, el-Gamil M, Rosenberg SA, Yang JC - J. Exp. Med. (1997)

Treatment of 3-d-old pulmonary metastases from B16 by the  intravenous administration of T cells reactive with TRP-2181–188 generated by in vitro peptide stimulation of splenocytes from normal or B16immunized mice. Low-dose systemic IL-2 was given with adoptive cell  transfer. Therapeutic effects were seen from anti–TRP-2181–188 CTLs  from normal mice (P = 0.0004 vs. IL-2 alone, and P = 0.014 vs.  ovalbumin257–264 stimulated cultures), as well as from anti-B16 immunized  mice (P = 0.0003 vs. IL-2 alone, and P = 0.010 vs. ovalbumin257–264  stimulated cultures).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196033&req=5

Figure 4: Treatment of 3-d-old pulmonary metastases from B16 by the intravenous administration of T cells reactive with TRP-2181–188 generated by in vitro peptide stimulation of splenocytes from normal or B16immunized mice. Low-dose systemic IL-2 was given with adoptive cell transfer. Therapeutic effects were seen from anti–TRP-2181–188 CTLs from normal mice (P = 0.0004 vs. IL-2 alone, and P = 0.014 vs. ovalbumin257–264 stimulated cultures), as well as from anti-B16 immunized mice (P = 0.0003 vs. IL-2 alone, and P = 0.010 vs. ovalbumin257–264 stimulated cultures).
Mentions: The therapeutic efficacy of CTL lines generated by in vitro stimulation with TRP-2181–188 was also examined. T cell lines generated from normal or B16 immune mice by stimulation with the TRP-2181–188 peptide were used to treat 3-d-old B16 lung metastases by intravenous administration of the CTL in combination with low dose IL-2. A highly significant therapeutic effect was seen (P <0.001) with cultures from normal or B16-immunized mice when stimulated with TRP-2181–188, but not with ova257–264 peptide (when compared to treatment with IL-2 alone) (Fig. 4).

Bottom Line: In spite of this, little is known about the optimal ways to use these antigens to treat patients with cancer.A peptide conforming to the predicted MHC class I H2-Kb binding motif, TRP-2181-188, was identified as the major reactive epitope within TRP-2 recognized by these anti-B16 CTLs.By site-directed mutagenesis, it was shown that alteration of this epitope eliminated recognition of TRP-2.

View Article: PubMed Central - PubMed

Affiliation: Surgery Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.

ABSTRACT
Recently, major advances have been made in the identification of antigens from human melanoma which are recognized by T cells. In spite of this, little is known about the optimal ways to use these antigens to treat patients with cancer. Progress in this area is likely to require accurate preclinical animal models, but the availability of such models has lagged behind developments in human tumor immunology. Whereas many of the identified human melanoma antigens are normal tissue differentiation proteins, analogous murine tumor antigens have not yet been identified. In this paper we identify a normal tissue differentiation antigen, tyrosinase-related protein 2 (TRP-2), expressed by the murine B16 melanoma which was found by screening a cDNA library from B16 with tumor-reactive cytotoxic T lymphocytes (CTL). A peptide conforming to the predicted MHC class I H2-Kb binding motif, TRP-2181-188, was identified as the major reactive epitope within TRP-2 recognized by these anti-B16 CTLs. By site-directed mutagenesis, it was shown that alteration of this epitope eliminated recognition of TRP-2. It was further demonstrated that a CTL line raised from splenocytes by repeated stimulation in vitro with this peptide could recognize B16 tumor and was therapeutic against 3-d-old established pulmonary metastases. The use of TRP-2 in a preclinical model of tumor immunotherapy may be helpful in suggesting optimal vaccination strategies for cancer therapy in patients.

Show MeSH
Related in: MedlinePlus