Breast Cancer Research and Treatment

, Volume 138, Issue 1, pp 109–118 | Cite as

Mammaglobin-A cDNA vaccination of breast cancer patients induces antigen-specific cytotoxic CD4+ICOShi T cells

  • Venkataswarup Tiriveedhi
  • Timothy P. Fleming
  • Peter S. Goedegebuure
  • Michael Naughton
  • Cynthia Ma
  • Craig Lockhart
  • Feng Gao
  • William E. Gillanders
  • T. MohanakumarEmail author
Clinical Trial


Mammaglobin-A (Mam-A) is a 10 kDa secretory protein that is overexpressed in 80 % of primary and metastatic human breast cancers. Previous studies from our laboratory demonstrated that Mam-A cDNA vaccine can induce Mam-A-specific CD8 T cell responses and mediate regression of human breast cancer xenografts in NOD/SCID mice. In this article, we present our results on a phase I clinical trial of a Mam-A cDNA vaccination in breast cancer patients with stage-IV metastatic disease, including the impact of vaccination on the expression of the inducible co-stimulator molecule (ICOS) on CD4 T cells. Specimens from seven patients with stage-IV metastatic cancer were available for these analyses. Patients were vaccinated with a Mam-A cDNA vaccine on days 0, 28, and 56, and immune responses were assessed at serial time points following vaccination. At 6 months following the first vaccination, flow cytometric analysis demonstrated a significant increase in the frequency of CD4+ICOShi T cells from 5 ± 2 % pre-vaccination to 23 ± 4 % (p < 0.001), with a concomitant decrease in the frequency of CD4+FoxP3+ T cells (regulatory T cells [Treg]) from 19 ± 6 to 10 ± 5 % (p < 0.05). ELISpot analysis of CD4+ICOShi sorted T cells demonstrated that following vaccination the cytokines produced by Mam-A-specific T cells switched from IL-10 (78 ± 21 spm pre-vaccination to 32 ± 14 spm 5 months post-vaccine p < 0.001) to IFN-γ (12 ± 6 spm pre-vaccination to 124 ± 31 spm 5 months post-vaccine p < 0.001). The ratio of CD4+ICOShi T cells to CD4+FoxP3+ T cells increased from 0.37 ± 0.12 before vaccination to 2.3 ± 0.72 (p = 0.021) following vaccination. Further, these activated CD4+ICOShi T cells induced preferential lysis of human breast cancer cells expressing Mam-A protein. We conclude that Mam-A cDNA vaccination is associated with specific expansion and activation of CD4+ICOShi T cells, with a concomitant decrease in Treg frequency. These encouraging results strongly suggest that Mam-A cDNA vaccination can induce antitumor immunity in breast cancer patients.


DNA vaccine Mammaglobin-A Breast cancer T cells ICOS 



Antigen presenting cells


Inducible co-stimulatory molecule




Monoclonal antibody


Peripheral blood mononuclear cells


Regulatory T cells



The authors would like to thank Ms. Billie Glasscock for her assistance in submitting this manuscript. This project was funded by DOD/CDMRP-BCRP W81XWH-06-1-0677 (WG). TM is funded by the BJC Foundation.

Conflict of Interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Science+Business Media, LLC. 2012

Authors and Affiliations

  • Venkataswarup Tiriveedhi
    • 1
  • Timothy P. Fleming
    • 1
  • Peter S. Goedegebuure
    • 1
  • Michael Naughton
    • 2
  • Cynthia Ma
    • 2
  • Craig Lockhart
    • 2
  • Feng Gao
    • 2
  • William E. Gillanders
    • 1
  • T. Mohanakumar
    • 1
    • 3
    Email author
  1. 1.Department of SurgeryWashington University School of MedicineSaint LouisUSA
  2. 2.Department of MedicineWashington University School of MedicineSaint LouisUSA
  3. 3.Department of Pathology and ImmunologyWashington University School of MedicineSaint LouisUSA

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