title: Quantification and phenotypic characterization of circulating tumor cells for monitoring response to a preventive HER2/neu vaccine-based immunotherapy for breast cancer: a pilot study.
abstract: BACKGROUND
  Ongoing cancer vaccine trials are limited by the inability of immunologic assays to monitor clinically relevant surrogates of response. Recent advances in the ability to quantify and phenotype circulating tumor cells (CTCs) in breast cancer patients may lead to a role for CTCs in monitoring response to vaccine-based immunotherapy.
  
  
  METHODS
  The CellSearch System (Veridex-LLC, Warren, NJ) was used to enumerate total and HER2/neu+ CTCs in 20 mL of blood from all 16 node-positive (NP) breast cancer patients active in our NP HER2/neu E75 peptide vaccine trial at the initiation of this pilot study. These patients were vaccinated with E75 (1000 microg)/GM-CSF (250 microg) monthly x 6 after completion of multimodality therapy. Mean (+/-SEM) number of CTCs and HER2/neu+ CTCs were compared in unmatched (n = 16) and matched (n = 9) prevaccination and postvaccination cases.
  
  
  RESULTS
  CTCs were detected in 14 of 16 (88%) patients (mean: 3.4 +/- 0.2 CTC/20 mL). After vaccination, a reduction in CTC/20 mL (prevaccination 3.9 +/- 1.5 vs postvaccination 0.7 +/- 0.4, P = .077) and HER2/neu+ CTC/20 mL (prevaccination 2.8 +/- 1.0 vs postvaccination 0.5 +/- 0.2, P = .048) was demonstrated. A significant delayed-type hypersensitivity (DTH) response suggesting that vaccination was effective in eliciting a peptide-specific immune response was confirmed (22.3 +/- 4.1 vs 3.0 +/- 2.2 [controls] mm, P < .01). All nine patients followed throughout the vaccination series also showed significant reduction in CTCs (4.8 +/- 1.5 vs 0.3 +/- 0.2, P < .01) and HER2/neu+ CTCs (3.0 +/- 0.9 vs 0.4 +/- 0.2, P = .013).
  
  
  CONCLUSIONS
  CTCs are readily demonstrated in posttreatment, clinically disease-free NP breast cancer patients. E75+GM-CSF vaccination appears to reduce the number of CTCs. These data suggest a potential role for this clinically validated CTC assay in assessing response to preventive vaccine-based immunotherapy, and further validation studies are underway.
authors: Department of Surgery, Walter Reed Army Medical Center, Washington, DC, USA. alexander.stojadinovic@amedd.army.mil
authors: Alexander
authors: A
authors: Stojadinovic
authors: Elizabeth A
authors: EA
authors: Mittendorf
authors: Jarrod P
authors: JP
authors: Holmes
authors: Asna
authors: A
authors: Amin
authors: Matthew T
authors: MT
authors: Hueman
authors: Sathibalan
authors: S
authors: Ponniah
authors: George E
authors: GE
authors: Peoples
mesh_terms: D000328:Adult; D000368:Aged; D001943:Breast Neoplasms / Q000097:blood* / Q000628:therapy*; D019496:Cancer Vaccines / Q000008:administration & dosage*; D002986:Clinical Trials as Topic; D005260:Female; D006801:Humans; D006968:Hypersensitivity, Delayed; D007167:Immunotherapy; D008875:Middle Aged; D009367:Neoplasm Staging; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D010446:Peptide Fragments / Q000276:immunology*; D010641:Phenotype; D010865:Pilot Projects; D011379:Prognosis; D011446:Prospective Studies
chemical_list: D019496:Cancer Vaccines; C474513:HER-2 peptide E75 (369-377), human; D010446:Peptide Fragments