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Tumor-specific immunity in MUC1.Tg mice induced by immunization with peptide vaccines from the cytoplasmic tail of CD227 (MUC1)

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Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Purpose: CD227 (MUC1), a membrane-associated glycoprotein expressed by many types of ductal epithelia, including pancreas, breast, lung, and gastrointestinal tract, is overexpressed and aberrantly glycosylated by malignant cells. We sought to define epitopes on MUC1 recognized by the different cell-mediated immune responses by an in vivo assay. Epitopes identified by this assay were evaluated for efficacy to protect mice transgenic for human MUC1 (MUC1.Tg) against MUC1-expressing tumor growth. Methods: We investigated contributions of the tandem repeat (TR) and the cytoplasmic tail (CT) of MUC1 to the MUC1-specific immunological rejection of tumor cells. MUC1 cDNA constructs, in which the TR region was deleted or the CT was truncated, were transfected into two different murine tumor cell lines (B16 and Panc02), which were used to challenge mice and evaluate immunological rejection of the tumors. We used tumor rejection in vivo to define epitopes on the TR and CT of MUC1 recognized by T cell–mediated immune responses in a preclinical murine model. Results: Our findings demonstrated that the TR and a portion of the MUC1 CT contributed to CD4+ T cell rejection of MUC1-expressing B16 tumor cells, but not rejection of MUC1-expressing Panc02 tumor cells. A separate epitope in the CT of MUC1 was necessary for CD8+ T cell rejection of Panc02 tumor cells. Based on these studies, we sought to evaluate the efficacy of immunizing mice transgenic for (and immunologically tolerant to) human MUC1 with peptides derived from the amino acid sequence of the CT of MUC1. Results showed that survival can be significantly prolonged in vaccinated MUC1.Tg mice challenged with MUC1-expressing tumor cells, without induction of autoimmune responses. Conclusions: These studies demonstrated that MUC1 peptides may be utilized as an effective anticancer immunotherapeutic, and confirmed the importance of immunogenic epitopes outside of the TR.

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Abbreviations

aa:

Amino acid

CT:

Cytoplasmic tail

IFN-γ:

Interferon gamma

MUC1.Tg:

MUC1 transgenic mice

TR:

Tandem repeat

wt:

Wild-type C57BL/6 mice

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Acknowledgements

We would like to thank Dr Richard M. Tempero, Dr Michael D. Burdick, Dr Naoaki Akisawa, and Hēth Turnquist for their technical assistance and helpful discussions with Dr Joyce Solheim. We thank Dr Charles A. Kuszynski and Linda M. Wilkie, of the UNMC Cell Analysis Facility and Dr R. Lee Mosley for providing assistance with flow cytometry analysis. We thank Dr Rakesh Singh and Michelle Varney for their assistance with immunohistochemistry. We also thank the Eppley Animal Facility for maintaining our animal colonies. We also acknowledge the generous gifts of anti-MUC1 antibodies HMFG-2 and CT-2, and the cDNA constructs that encode CT3, CT33, and CT45 from Dr Sandra J. Gendler, Mayo Clinic, Scottsdale, AZ, USA.

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Authors and Affiliations

  1. Eppley Institute for Research in Cancer and Allied Diseases, and the Department of Pathology and Microbiology, University of Nebraska Medical Center, 986805 Nebraska Medical Center, Omaha, NE 68198-6805, USA

    Karl G. Kohlgraf, Andrew J. Gawron, Michiyo Higashi, Michelle L. VanLith, XiaoLing Shen, Thomas C. Caffrey, Judy M. Anderson & Michael A. Hollingsworth

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  1. Karl G. Kohlgraf
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  2. Andrew J. Gawron
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  3. Michiyo Higashi
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  4. Michelle L. VanLith
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  5. XiaoLing Shen
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  6. Thomas C. Caffrey
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  7. Judy M. Anderson
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  8. Michael A. Hollingsworth
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Corresponding author

Correspondence to Michael A. Hollingsworth.

Additional information

This work was supported by National Institutes of Health grants CA72712 and CA57362 (M.A.H.), National Institutes of Health training grant CA09476 (K.G.K., A.J.G, and M.L.V.), and fellowship awards from the University of Nebraska Medical Center (to K.G.K. and M.L.V).

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Kohlgraf, K.G., Gawron, A.J., Higashi, M. et al. Tumor-specific immunity in MUC1.Tg mice induced by immunization with peptide vaccines from the cytoplasmic tail of CD227 (MUC1). Cancer Immunol Immunother 53, 1068–1084 (2004). https://doi.org/10.1007/s00262-004-0557-1

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