Leukocyte composition of human breast cancer

Department of Pathology and Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, CA 94143, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 08/2011; 109(8):2796-801. DOI: 10.1073/pnas.1104303108
Source: PubMed


Retrospective clinical studies have used immune-based biomarkers, alone or in combination, to predict survival outcomes for women with breast cancer (BC); however, the limitations inherent to immunohistochemical analyses prevent comprehensive descriptions of leukocytic infiltrates, as well as evaluation of the functional state of leukocytes in BC stroma. To more fully evaluate this complexity, and to gain insight into immune responses after chemotherapy (CTX), we prospectively evaluated tumor and nonadjacent normal breast tissue from women with BC, who either had or had not received neoadjuvant CTX before surgery. Tissues were evaluated by polychromatic flow cytometry in combination with confocal immunofluorescence and immunohistochemical analysis of tissue sections. These studies revealed that activated T lymphocytes predominate in tumor tissue, whereas myeloid lineage cells are more prominant in "normal" breast tissue. Notably, residual tumors from an unselected group of BC patients treated with neoadjuvant CTX contained increased percentages of infiltrating myeloid cells, accompanied by an increased CD8/CD4 T-cell ratio and higher numbers of granzyme B-expressing cells, compared with tumors removed from patients treated primarily by surgery alone. These data provide an initial evaluation of differences in the immune microenvironment of BC compared with nonadjacent normal tissue and reveal the degree to which CTX may alter the complexity and presence of selective subsets of immune cells in tumors previously treated in the neoadjuvant setting.

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Available from: Alfred Au, Jan 14, 2015
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    • "Insufficient attention was given to the intratumoral heterogeneity of TAM subtypes, in particular in relation to the different intratumoral structures. In human breast cancer following intratumoral compartments can be defined: (1) areas with soft fibrous stroma characterized by pronounced inflammatory infiltrates that are beneficial for invasive cell growth (Ham and Moon, 2013); (2) areas with coarse fibrous stroma containing collagen fibers and characterized by impaired synthesis of extracellular matrix proteins (ECM) (Campbell et al., 2011; Eiro et al., 2012; Ruffell et al., 2012; Tang, 2013); (3) areas of maximum stromaland-parenchymal relationship revealing certain similarities with soft fibrous stroma (Mahmoud et al., 2012); (4) parenchymal elements ; (5) gaps of ductal tumor structures (Pinder, 2010). All five stromal subtypes demonstrate functionally distinct areas of tumor mircoenvironment with not yet identified mechanistic role in metastatic spread. "
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    ABSTRACT: Tumor associated macrophages (TAM) support tumor growth and metastasis in several animal models of breast cancer, and TAM amount is predictive for efficient tumor growth and metastatic spread via blood circulation. However, limited information is available about intratumoral TAM heterogeneity and functional role of TAM subpopulations in tumor progression. The aim of our study was to examine correlation of TAM presence in various morphological segments of human breast cancer with clinical parameters. Thirty six female patients with nonspecific invasive breast cancer T1-4N0-3M0 were included in the study. Morphological examination was performed using Carl Zeiss Axio Lab.A1 and MiraxMidiZeiss. Immunohistochemical and immunofluorescence/confocal microcopy analysis was used to detect CD68 and stabilin-1 in 5 different tumor segments: (1) areas with soft fibrous stroma; (2) areas with coarse fibrous stroma; (3) areas of maximum stromal-and-parenchymal relationship; (4) parenchymal elements; (5) gaps of ductal tumor structures. The highest expression of CD68 was in areas with soft fibrous stroma or areas of maximum stromal-and-parenchymal relationship (79%). The lowest expression of CD68 was in areas with coarse fiber stroma (23%). Inverse correlation of tumor size and expression of CD68 in gaps of tubular tumor structures was found (R=-0.67; p=0.02). In case of the lymph node metastases the average score of CD68 expression in ductal gaps tumor structures was lower (1.4±0.5) compared to negative lymph nodes case (3.1±1.0; F=10.9; p=0.007). Confocal microscopy identified 3 phenotypes of TAM: CD68(+)/stabilin-1(-); CD68(+)/stabilin-1(+) (over 50%); and CD68(-)/stabilin-1(+). However, expression of stabilin-1 did not correlate with lymph node metastasis. We concluded, that increased amount of CD68+TAM in gaps of ductal tumor structures is protective against metastatic spread in regional lymph nodes.
    Immunobiology 09/2015; DOI:10.1016/j.imbio.2015.09.011 · 3.04 Impact Factor
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    • "In this study, we identified a Th2 biased gene expression of the intratumoral CD4+ T cells, which has previously been shown to suppress the anti-tumor immunity of the host [10] [11] [12] [33]. By integrating the DE genes of the intratumoral CD4+ T cells (CD4 DEGs) with the DE genes of the bulk BC tissues (BC DEGs), we identified five dysregulated cytokines as the predictor to reflect the Th2 polarization of the intratumoral CD4+ T cells, which in turn could be used to predict the immune-polarization side effects of tamoxifen treatment. "
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    ABSTRACT: Factors within the tissue of breast cancer (BC) may shift the polarization of CD4+ T cells towards Th2 direction. This tendency can promote tumor development and be enhanced by the use of tamoxifen during the treatment. Thus, the patients with low levels of tumor-induced Th2 polarization prior to tamoxifen treatment may better endure the immune-polarizing side effects (IPSE) of tamoxifen and have better prognoses. Estimation of Th2 polarization status should help predict the IPSE among tamoxifen-treated patients and guide the use of tamoxifen among all BC patients before the tamoxifen therapy. Here, we report profiling of differentially expressed (DE) intratumoral cytokines as a signature to evaluate the IPSE of tamoxifen. The DE genes of intratumoral CD4+ T cells (CD4 DEGs) were identified by gene expression profiles of purified CD4+ T cells from BC patients and validated by profiling of cultured intratumoral CD4+ T cells. Functional enrichment analyses showed a directed Th2 polarization of intratu-moral CD4+ T cells. To find the factors inducing the Th2 polarization of CD4+ T cells, we identified 995 common DE genes of bulk BC tissues (BC DEGs) by integrating five independent datasets. Five DE cytokines observed in bulk BC tissues with dysregulated receptors in the intratumoral CD4+ T cells were selected as the predictor of the IPSE of tamoxifen. The patients predicted to suffer low IPSE (low Th2 polarization) had a significantly lower distant relapse risk than the patients predicted to suffer high IPSE in independent datasets (n = 608; HR = 4.326, P = 0.000897; HR = 2.014, P = 0.0173; HR = 2.72, P = 0.04077). Patients predicted to suffer low IPSE would benefit from tamoxi-fen treatment (HR = 2.908, P = 0.03905). The DE intratumoral cytokines identified in this study may help predict the IPSE of tamoxifen and justify the use of tamoxifen in BC treatment.
    American Journal of Cancer Research 02/2015; 5(2):726-737. · 4.17 Impact Factor
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    • "Firstly, cellular markers that distinguish the phenotypes of macrophages during inflammatory diseases need further studies to confirm. For example, some recent studies found that it was not appropriate to identify TAM through the expression of CD68 as it was also expressed by other stromal populations, so the conclusions drawn from CD68 as the marker of TAM should be further evaluated 77, 90. Secondly, it is usually not convenient to acquire fresh macrophages of human, so most of human studies now are conducted to perform from cell lines, which are not exactly perfect and hinder the transformation to the clinical trials. "
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    ABSTRACT: Diversity and plasticity are two hallmarks of macrophages. M1 macrophages (classically activated macrophages) are pro-inflammatory and have a central role in host defense against infection, while M2 macrophages (alternatively activated macrophages) are associated with responses to anti-inflammatory reactions and tissue remodeling, and they represent two terminals of the full spectrum of macrophage activation. Transformation of different phenotypes of macrophages regulates the initiation, development, and cessation of inflammatory diseases. Here we reviewed the characters and functions of macrophage polarization in infection, atherosclerosis, obesity, tumor, asthma, and sepsis, and proposed that targeting macrophage polarization and skewing their phenotype to adapt to the microenvironment might hold great promise for the treatment of inflammatory diseases.
    International journal of biological sciences 05/2014; 10(5):520-529. DOI:10.7150/ijbs.8879 · 4.51 Impact Factor
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