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Breast Cancer Research and Treatment

, Volume 91, Issue 3, pp 271–278 | Cite as

Tat mammaglobin fusion protein transduced dendritic cells stimulate mammaglobin-specific CD4 and CD8 T cells

  • Carsten T. Viehl
  • Yoshiyuki Tanaka
  • Tingting Chen
  • Daniel M. Frey
  • Andrew Tran
  • Timothy P. Fleming
  • Timothy J. Eberlein
  • Peter S. Goedegebuure
Article

Summary

Proteins can be efficiently introduced into cells when fused to a protein transduction domain, such as Tat from the human immunodeficiency virus. We recently reported that dendritic cells transduced with a Tat fusion protein containing the extracellular domain of Her2/neu (Tat-Her2/neu) induced CD8 cytotoxic T lymphocytes (CTL) that specifically lysed Her2/neu-expressing breast and ovarian cancer cells. In the current study we further investigated the mechanism of protein transduction, utilizing the breast cancer-associated protein, mammaglobin-A, which is expressed in about 80% of breast cancers. Using a Tat-mammaglobin fusion protein, we tested the ability of Tat-mammaglobin transduced dendritic cells to stimulate antigen-specific CD4 and CD8 T cells. Low levels of serum considerably improved protein transduction as determined by Western blot, and also improved presentation of antigenic peptide as evidenced by functional studies using antigen-specific T cells. Confocal microscope analyses of antigen-presenting cells (APC) incubated with Tat-mammaglobin showed localized distribution in addition to diffuse distribution in the cytosol. In contrast, mammaglobin lacking Tat showed only a localized distribution. Simultaneous incubation with both proteins resulted in overlapping localized distributions, suggesting Tat fusion proteins are processed through both the MHC class I and class II pathways. Indeed, stimulation of T cells with Tat-mammaglobin transduced dendritic cells led to an expansion of mammaglobin-specific CD4 T helper-1 lymphocytes along with CD8 CTL. We conclude that Tat-mammaglobin transduced dendritic cells can induce both CD4 and CD8 mammaglobin-specific T cells. These findings could be further exploited for the development of a mammaglobin-based vaccine for breast cancer.

Keywords

antigen-presenting cell breast cancer vaccine mammaglobin protein transduction T cells Tat fusion protein 

Abbreviations

Tat

trans-activating

APC

antigen-presenting cells

HIV

human immunodeficiency virus

MHC

major histocompatibility complex

MGB

mammaglobin

CTL

cytotoxic T lymphocyte

IL

interleukin

FITC

fluorescence isothiocyanate

PBMC

peripheral blood mononuclear cells

E/T

effector/target

IFNγ

interferon gamma

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Copyright information

© Springer 2005

Authors and Affiliations

  • Carsten T. Viehl
    • 1
    • 2
  • Yoshiyuki Tanaka
    • 1
    • 3
  • Tingting Chen
    • 1
  • Daniel M. Frey
    • 1
    • 2
  • Andrew Tran
    • 1
  • Timothy P. Fleming
    • 1
  • Timothy J. Eberlein
    • 1
  • Peter S. Goedegebuure
    • 1
  1. 1.Department of SurgeryWashington University School of Medicine, and Alvin J. Siteman Cancer CenterMOUSA
  2. 2.Department of Surgery, Divisions of General Surgery and Surgical ResearchUniversity of BaselSwitzerland
  3. 3.Graduate School of MedicineTokyo Women’s Medical UniversityTokyoJapan

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