Limits...
Self-tolerance to the murine homologue of a tyrosinase-derived melanoma antigen: implications for tumor immunotherapy.

Colella TA, Bullock TN, Russell LB, Mullins DW, Overwijk WW, Luckey CJ, Pierce RA, Restifo NP, Engelhard VH - J. Exp. Med. (2000)

Bottom Line: The murine peptide was processed and presented by AAD similarly to its human counterpart.Collectively, these data suggest that FMDGTMSQV is naturally processed and presented in vivo, and that this presentation leads to substantial but incomplete self-tolerance.The relevance of this model to an understanding of the human immune response to tyrosinase is discussed.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and the Beirne Carter Center for Immunology Research, University of Virginia, Charlottesville, Virginia 22908, USA.

ABSTRACT
The human tyrosinase-derived peptide YMDGTMSQV is presented on the surface of human histocompatibility leukocyte antigen (HLA)-A*0201(+) melanomas and has been suggested to be a tumor antigen despite the fact that tyrosinase is also expressed in melanocytes. To gain information about immunoreactivity and self-tolerance to this antigen, we established a model using the murine tyrosinase-derived homologue of this peptide FMDGTMSQV, together with transgenic mice expressing the HLA-A*0201 recombinant molecule AAD. The murine peptide was processed and presented by AAD similarly to its human counterpart. After immunization with recombinant vaccinia virus encoding murine tyrosinase, we detected a robust AAD-restricted cytotoxic T lymphocyte (CTL) response to FMDGTMSQV in AAD transgenic mice in which the entire tyrosinase gene had been deleted by a radiation-induced mutation. A residual response was observed in the AAD(+)tyrosinase(+) mice after activation under certain conditions. At least some of these residual CTLs in AAD(+)tyrosinase(+) mice were of high avidity and induced vitiligo upon adoptive transfer into AAD(+)tyrosinase(+) hosts. Collectively, these data suggest that FMDGTMSQV is naturally processed and presented in vivo, and that this presentation leads to substantial but incomplete self-tolerance. The relevance of this model to an understanding of the human immune response to tyrosinase is discussed.

Show MeSH

Related in: MedlinePlus

Characteristics of residual tyrosinase-specific response in AAD+tyrosinase+ mice. 3 wk after immunization with either rvv-mu tyr or rvv-hu tyr, splenocytes from AAD+tyrosinase+ mice or AAD+ albino mice were cultured in vitro with an irradiated feeder layer that had been pulsed with 1 μg/ml of either synthetic FMDGTMSQV (murine) or YMDGTMSQV (human) peptides. 7 d later, cultures were harvested, stimulated for 5 h with C1R-AAD cells that had been pulsed with synthetic FMDGTMSQV or YMDGTMSQV, and assayed for the intracellular accumulation of IFN-γ.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2193167&req=5

Figure 6: Characteristics of residual tyrosinase-specific response in AAD+tyrosinase+ mice. 3 wk after immunization with either rvv-mu tyr or rvv-hu tyr, splenocytes from AAD+tyrosinase+ mice or AAD+ albino mice were cultured in vitro with an irradiated feeder layer that had been pulsed with 1 μg/ml of either synthetic FMDGTMSQV (murine) or YMDGTMSQV (human) peptides. 7 d later, cultures were harvested, stimulated for 5 h with C1R-AAD cells that had been pulsed with synthetic FMDGTMSQV or YMDGTMSQV, and assayed for the intracellular accumulation of IFN-γ.

Mentions: The preceding results demonstrated that there was no detectable FMDGTMSQV-specific CTL response from immunized AAD+tyrosinase+ mice either at the peak of the primary response or after secondary in vitro stimulation. These results appeared at odds with our previous demonstration that CTL lines elicited against the human tyrosinase epitope recognize the murine tyrosinase-derived peptide in a cross-reactive manner (Fig. 2 A). To further investigate the immunogenicity of murine tyrosinase in AAD+tyrosinase+ mice, spleen cells from AAD+tyrosinase+ and AAD+ albino mice immunized with either rvv-mu tyr or rvv-hu tyr were cultured with stimulators pulsed with either the murine or human peptides and analyzed for their ability to make IFN-γ in response to either peptide (Fig. 6). As expected, AAD+ albino mice responded well to both rvv-mu tyr and rvv-hu tyr. The T cells elicited were largely cross-reactive, based on their activation and differentiation in response to either peptide during 7 d of in vitro culture, as well as their subsequent ability to be induced to produce IFN-γ in a 5-h incubation. In contrast, AAD+tyrosinase+ mice failed to respond to rvv-mu tyr based on the lack of any detectable response in spleen cells cultured for 7 d with either the murine or human peptides. Consistent with our previous observations (Fig. 5, and reference 44), spleen cells from AAD+tyrosinase+ mice immunized with rvv-hu tyr and cultured in vitro for 7 d with the human peptide led to their activation and differentiation as measured by their ability to produce IFN-γ when induced with the human peptide for 5 h. Significantly, many of these T cells also recognized the murine tyrosinase-derived peptide as judged by their ability to make IFN-γ when induced with the murine peptide for 5 h. However, in parallel cultures of T cells from these same rvv-hu tyr–immunized AAD+tyrosinase+ mice, this murine tyrosinase-derived peptide did not cause activation and differentiation during a 7-d in vitro culture. These data establish that there are residual murine tyrosinase-specific T cells in AAD+ tyrosinase+ mice but that their ability to respond to the murine peptide is impaired.


Self-tolerance to the murine homologue of a tyrosinase-derived melanoma antigen: implications for tumor immunotherapy.

Colella TA, Bullock TN, Russell LB, Mullins DW, Overwijk WW, Luckey CJ, Pierce RA, Restifo NP, Engelhard VH - J. Exp. Med. (2000)

Characteristics of residual tyrosinase-specific response in AAD+tyrosinase+ mice. 3 wk after immunization with either rvv-mu tyr or rvv-hu tyr, splenocytes from AAD+tyrosinase+ mice or AAD+ albino mice were cultured in vitro with an irradiated feeder layer that had been pulsed with 1 μg/ml of either synthetic FMDGTMSQV (murine) or YMDGTMSQV (human) peptides. 7 d later, cultures were harvested, stimulated for 5 h with C1R-AAD cells that had been pulsed with synthetic FMDGTMSQV or YMDGTMSQV, and assayed for the intracellular accumulation of IFN-γ.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2193167&req=5

Figure 6: Characteristics of residual tyrosinase-specific response in AAD+tyrosinase+ mice. 3 wk after immunization with either rvv-mu tyr or rvv-hu tyr, splenocytes from AAD+tyrosinase+ mice or AAD+ albino mice were cultured in vitro with an irradiated feeder layer that had been pulsed with 1 μg/ml of either synthetic FMDGTMSQV (murine) or YMDGTMSQV (human) peptides. 7 d later, cultures were harvested, stimulated for 5 h with C1R-AAD cells that had been pulsed with synthetic FMDGTMSQV or YMDGTMSQV, and assayed for the intracellular accumulation of IFN-γ.
Mentions: The preceding results demonstrated that there was no detectable FMDGTMSQV-specific CTL response from immunized AAD+tyrosinase+ mice either at the peak of the primary response or after secondary in vitro stimulation. These results appeared at odds with our previous demonstration that CTL lines elicited against the human tyrosinase epitope recognize the murine tyrosinase-derived peptide in a cross-reactive manner (Fig. 2 A). To further investigate the immunogenicity of murine tyrosinase in AAD+tyrosinase+ mice, spleen cells from AAD+tyrosinase+ and AAD+ albino mice immunized with either rvv-mu tyr or rvv-hu tyr were cultured with stimulators pulsed with either the murine or human peptides and analyzed for their ability to make IFN-γ in response to either peptide (Fig. 6). As expected, AAD+ albino mice responded well to both rvv-mu tyr and rvv-hu tyr. The T cells elicited were largely cross-reactive, based on their activation and differentiation in response to either peptide during 7 d of in vitro culture, as well as their subsequent ability to be induced to produce IFN-γ in a 5-h incubation. In contrast, AAD+tyrosinase+ mice failed to respond to rvv-mu tyr based on the lack of any detectable response in spleen cells cultured for 7 d with either the murine or human peptides. Consistent with our previous observations (Fig. 5, and reference 44), spleen cells from AAD+tyrosinase+ mice immunized with rvv-hu tyr and cultured in vitro for 7 d with the human peptide led to their activation and differentiation as measured by their ability to produce IFN-γ when induced with the human peptide for 5 h. Significantly, many of these T cells also recognized the murine tyrosinase-derived peptide as judged by their ability to make IFN-γ when induced with the murine peptide for 5 h. However, in parallel cultures of T cells from these same rvv-hu tyr–immunized AAD+tyrosinase+ mice, this murine tyrosinase-derived peptide did not cause activation and differentiation during a 7-d in vitro culture. These data establish that there are residual murine tyrosinase-specific T cells in AAD+ tyrosinase+ mice but that their ability to respond to the murine peptide is impaired.

Bottom Line: The murine peptide was processed and presented by AAD similarly to its human counterpart.Collectively, these data suggest that FMDGTMSQV is naturally processed and presented in vivo, and that this presentation leads to substantial but incomplete self-tolerance.The relevance of this model to an understanding of the human immune response to tyrosinase is discussed.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and the Beirne Carter Center for Immunology Research, University of Virginia, Charlottesville, Virginia 22908, USA.

ABSTRACT
The human tyrosinase-derived peptide YMDGTMSQV is presented on the surface of human histocompatibility leukocyte antigen (HLA)-A*0201(+) melanomas and has been suggested to be a tumor antigen despite the fact that tyrosinase is also expressed in melanocytes. To gain information about immunoreactivity and self-tolerance to this antigen, we established a model using the murine tyrosinase-derived homologue of this peptide FMDGTMSQV, together with transgenic mice expressing the HLA-A*0201 recombinant molecule AAD. The murine peptide was processed and presented by AAD similarly to its human counterpart. After immunization with recombinant vaccinia virus encoding murine tyrosinase, we detected a robust AAD-restricted cytotoxic T lymphocyte (CTL) response to FMDGTMSQV in AAD transgenic mice in which the entire tyrosinase gene had been deleted by a radiation-induced mutation. A residual response was observed in the AAD(+)tyrosinase(+) mice after activation under certain conditions. At least some of these residual CTLs in AAD(+)tyrosinase(+) mice were of high avidity and induced vitiligo upon adoptive transfer into AAD(+)tyrosinase(+) hosts. Collectively, these data suggest that FMDGTMSQV is naturally processed and presented in vivo, and that this presentation leads to substantial but incomplete self-tolerance. The relevance of this model to an understanding of the human immune response to tyrosinase is discussed.

Show MeSH
Related in: MedlinePlus