Cloning and characterization of the genes encoding the murine homologues of the human melanoma antigens MART1 and gp100

Yifan Zhai, James C. Yang, Paul Spiess, Michael I. Nishimura, Willem W. Overwijk, Bruce Roberts, Nicholas P. Restifo, Steven A. Rosenberg

Research output: Contribution to journalArticle

Abstract

The recent identification of genes encoding melanoma-associated antigens has opened new possibilities for the development of cancer vaccines designed to cause the rejection of established tumors. To develop a syngeneic animal model for evaluating antigen-specific vaccines in cancer therapy, the murine homologues of the human melanoma antigens MART1 and gp100, which were specifically recognized by tumor-infiltrating lymphocytes from patients with melanoma, were cloned and sequenced from a murine B16 melanoma cDNA library. The open reading frames of murine MART1 and gp100 encode proteins of 113- and 626-amino acids with 68.8 and 77% identity to the respective human proteins. Comparison of the DNA sequences of the murine MART1 genes, derived from normal melanocytes, the immortalized nontumorgenic melanocyte line Melan-a and the B16 melanoma, showed all to be identical. Northern and Western blot analyses confirmed that both genes encoded products that were melanocyte lineage proteins. Mice immunized with murine MART1 or gp100 using recombinant vaccinia virus failed to produce any detectable T-cell responses or protective immunity against B16 melanoma. In contrast, immunization of mice with human gp100 using recombinant adenoviruses elicited T cells specific for hgp100, but these T cells also cross reacted with B16 tumor in vitro and induced significant but weak protection against B16 challenge. Immunization with human and mouse gp100 together [adenovirus type 2 (Ad2)-hgp100 plus recombinant vaccinia virus (rVV)-mgp100], or immunization with human gp100 (Ad2-hgp100) and boosting with heterologous vector (rVV-hgp100 or rVV- mgp100) or homologous vector (Ad2-hgp100), did not significantly enhance the protective response against B16 melanoma. These results may suggest that immunization with heterologous tumor antigen, rather than self, may be more effective as an immunotherapeutic reagent in designing antigen-specific cancer vaccines.

Original languageEnglish (US)
Pages (from-to)15-25
Number of pages11
JournalJournal of Immunotherapy
Volume20
Issue number1
DOIs
StatePublished - 1997
Externally publishedYes

Fingerprint

gp100 Melanoma Antigen
Experimental Melanomas
Vaccinia virus
Cancer Vaccines
Organism Cloning
Melanocytes
Immunization
Adenoviridae
T-Lymphocytes
Genes
Heterophile Antigens
Melanoma-Specific Antigens
Tumor-Infiltrating Lymphocytes
Antigens
Human Adenoviruses
Proteins
Neoplasm Antigens
Gene Library
Northern Blotting
Open Reading Frames

Keywords

  • Adenovirus
  • B16 melanoma
  • Cancer vaccine
  • In vivo model
  • Vaccinia virus

ASJC Scopus subject areas

  • Cancer Research
  • Pharmacology
  • Immunology

Cite this

Zhai, Y., Yang, J. C., Spiess, P., Nishimura, M. I., Overwijk, W. W., Roberts, B., ... Rosenberg, S. A. (1997). Cloning and characterization of the genes encoding the murine homologues of the human melanoma antigens MART1 and gp100. Journal of Immunotherapy, 20(1), 15-25. https://doi.org/10.1097/00002371-199701000-00002

Cloning and characterization of the genes encoding the murine homologues of the human melanoma antigens MART1 and gp100. / Zhai, Yifan; Yang, James C.; Spiess, Paul; Nishimura, Michael I.; Overwijk, Willem W.; Roberts, Bruce; Restifo, Nicholas P.; Rosenberg, Steven A.

In: Journal of Immunotherapy, Vol. 20, No. 1, 1997, p. 15-25.

Research output: Contribution to journalArticle

Zhai, Y, Yang, JC, Spiess, P, Nishimura, MI, Overwijk, WW, Roberts, B, Restifo, NP & Rosenberg, SA 1997, 'Cloning and characterization of the genes encoding the murine homologues of the human melanoma antigens MART1 and gp100', Journal of Immunotherapy, vol. 20, no. 1, pp. 15-25. https://doi.org/10.1097/00002371-199701000-00002
Zhai, Yifan ; Yang, James C. ; Spiess, Paul ; Nishimura, Michael I. ; Overwijk, Willem W. ; Roberts, Bruce ; Restifo, Nicholas P. ; Rosenberg, Steven A. / Cloning and characterization of the genes encoding the murine homologues of the human melanoma antigens MART1 and gp100. In: Journal of Immunotherapy. 1997 ; Vol. 20, No. 1. pp. 15-25.
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abstract = "The recent identification of genes encoding melanoma-associated antigens has opened new possibilities for the development of cancer vaccines designed to cause the rejection of established tumors. To develop a syngeneic animal model for evaluating antigen-specific vaccines in cancer therapy, the murine homologues of the human melanoma antigens MART1 and gp100, which were specifically recognized by tumor-infiltrating lymphocytes from patients with melanoma, were cloned and sequenced from a murine B16 melanoma cDNA library. The open reading frames of murine MART1 and gp100 encode proteins of 113- and 626-amino acids with 68.8 and 77{\%} identity to the respective human proteins. Comparison of the DNA sequences of the murine MART1 genes, derived from normal melanocytes, the immortalized nontumorgenic melanocyte line Melan-a and the B16 melanoma, showed all to be identical. Northern and Western blot analyses confirmed that both genes encoded products that were melanocyte lineage proteins. Mice immunized with murine MART1 or gp100 using recombinant vaccinia virus failed to produce any detectable T-cell responses or protective immunity against B16 melanoma. In contrast, immunization of mice with human gp100 using recombinant adenoviruses elicited T cells specific for hgp100, but these T cells also cross reacted with B16 tumor in vitro and induced significant but weak protection against B16 challenge. Immunization with human and mouse gp100 together [adenovirus type 2 (Ad2)-hgp100 plus recombinant vaccinia virus (rVV)-mgp100], or immunization with human gp100 (Ad2-hgp100) and boosting with heterologous vector (rVV-hgp100 or rVV- mgp100) or homologous vector (Ad2-hgp100), did not significantly enhance the protective response against B16 melanoma. These results may suggest that immunization with heterologous tumor antigen, rather than self, may be more effective as an immunotherapeutic reagent in designing antigen-specific cancer vaccines.",
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