Impact of CD68/(CD3+CD20) ratio at the invasive front of primary tumors on distant metastasis development in breast cancer

PLoS One. 2012;7(12):e52796. doi: 10.1371/journal.pone.0052796. Epub 2012 Dec 26.


Tumors are infiltrated by macrophages, T and B-lymphocytes, which may favor tumor development by promoting angiogenesis, growth and invasion. The aim of this study was to investigate the clinical relevance of the relative amount of macrophages (CD68⁺), T-cells (CD3⁺ and B-cells (CD20⁺) at the invasive front of breast carcinomas, and the expression of matrix metalloproteases (MMPs) and their inhibitors (TIMPs) either at the invasive front or at the tumor center. We performed an immunohistochemical study counting CD3, CD20 and CD68 positive cells at the invasive front, in 102 breast carcinomas. Also, tissue sections were stained with MMP-2, -9, -11, -14 and TIMP-2 antibodies, and immunoreactivity location, percentage of reactive area and intensity were determined at the invasive front and at the tumor center. The results showed that an increased CD68 count and CD68/(CD3+CD20) ratio were directly associated with both MMP-11 and TIMP-2 expression by mononuclear inflammatory cells at the tumor center (p = 0.041 and p = 0.025 for CD68 count and p = 0.001 and p = 0.045 for ratio, respectively for MMP-11 and TIMP-2). In addition, a high CD68/(CD3+CD20) ratio (>0.05) was directly associated with a higher probability of shortened relapse-free survival. Multivariate analysis revealed that CD68/(CD3+CD20) ratio was an independent factor associated with distant relapse-free survival (RR: 2.54, CI: (1.23-5.24), p<0.01). Therefore, CD68/(CD3+CD20) ratio at the invasive front could be used as an important prognostic marker.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD / metabolism*
  • Antigens, CD20 / metabolism*
  • Antigens, Differentiation, Myelomonocytic / metabolism*
  • B-Lymphocytes / metabolism
  • B-Lymphocytes / pathology
  • Biomarkers, Tumor / metabolism
  • Breast Neoplasms / metabolism
  • Breast Neoplasms / pathology*
  • CD3 Complex / metabolism*
  • Carcinoma, Ductal, Breast / metabolism
  • Carcinoma, Ductal, Breast / secondary*
  • Disease-Free Survival
  • Female
  • Humans
  • Macrophages / metabolism
  • Macrophages / pathology
  • Matrix Metalloproteinases, Secreted / metabolism
  • Multivariate Analysis
  • Neoplasm Invasiveness
  • Prognosis
  • Proportional Hazards Models
  • Survival Analysis
  • T-Lymphocytes / metabolism
  • T-Lymphocytes / pathology
  • Tissue Array Analysis
  • Tissue Inhibitor of Metalloproteinase-2 / metabolism


  • Antigens, CD
  • Antigens, CD20
  • Antigens, Differentiation, Myelomonocytic
  • Biomarkers, Tumor
  • CD3 Complex
  • CD68 antigen, human
  • TIMP2 protein, human
  • Tissue Inhibitor of Metalloproteinase-2
  • Matrix Metalloproteinases, Secreted

Grant support

This work was supported by grants from Fondo de Inversión Sanitaria-Instituto Carlos III (FIS-PI07/0306, FIS-PI10/02106), from Fundación para el Fomento en Asturias de la Investigación Científica Aplicada y la Tecnología (FICYT IB08-170), and Fundación para la Investigación con Células Madre Uterinas (FICEMU). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.