Generation of mammaglobin-A-specific CD4 T cells and identification of candidate CD4 epitopes for breast cancer vaccine strategies

Carsten T. Viehl, Daniel M. Frey, Chanpheng Phommaly, Tingting Chen, Timothy P. Fleming, William E. Gillanders, Timothy J. Eberlein, Peter S. Goedegebuure

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Background: Mammaglobin-A (MGB) is a breast cancer-associated antigen that is an attractive target for immune intervention. MGB has been shown to induce a specific CD8 T cell response in breast cancer patients, but little is known about a possible MGB-specific CD4 T cell response. Methods: Peripheral blood-derived CD4+CD25- T cells were stimulated in vitro with MGB-pulsed antigen-presenting cells (APC). The MGB and human leukocyte antigen (HLA) class II specificity of the CD4 T cell lines was confirmed by cytokine release following restimulation with autologous and allogenic APC pulsed with MGB from different sources. Candidate HLA class II-restricted epitopes were identified by computer algorithm and validated in cytokine release assays. Results: MGB-specific CD4 T cells were successfully generated in cultures from six of seven donors. Restimulation of MGB-specific CD4 T cells with MGB-pulsed APC induced significantly higher levels of interferon (IFN)-γ release than APC pulsed with an irrelevant protein (P = 0.0004). Cultures from five of seven donors showed a pure Th1 type response as evidenced by the absence of interleukin (IL)-4. MGB-specific CD4 T cells recognized both recombinant and naturally processed MGB presented by APC. This recognition was HLA class II-restricted, as HLA-DR mismatched APC were not recognized. MGB-specific CD4 T cells from three of four donors recognized MGB-derived, HLA class II-restricted peptides pulsed onto APC. Conclusions: We have successfully generated MGB-specific CD4 T cell cultures and identified candidate MGB HLA class II epitopes. These studies should facilitate study of the CD4 T cell response to MGB, and the development and monitoring of vaccine strategies targeting this unique antigen.

Original languageEnglish
Pages (from-to)305-314
Number of pages10
JournalBreast Cancer Research and Treatment
Volume109
Issue number2
DOIs
StatePublished - May 2008

Keywords

  • Antigen presenting cells
  • Breast cancer vaccine
  • CD4 T cells
  • Epitopes
  • Mammaglobin-A
  • T helper type 1 response
  • Tat fusion protein

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