Breast Tumor Angiogenesis and Tumor-Associated Macrophages: Histopathologist's Perspective
Table 2
Summary of clinical studies exploring the link between tumor-associated macrophages and other clinicopathological parameters in invasive breast carcinomas.
Macrophage marker: CD68 Macrophage index was determined by 25-point Chalkey graticule as the mean of three “hot spot” counts under 250x field
(1) High macrophage index correlated with high vascular grade (2) High macrophage index in poorly vascularized areas (3) High macrophage index predicted reduced relapse-free and overall survival
75 invasive breast carcinomas with lymphoplasmacytic infiltrates [67]
Macrophage marker: CD11c Macrophage was counted as percentage of total leukocyte infiltrate identified by CD45
(1) Macrophage predominance in leukocyte infiltrate correlated with high grade and c-erbB-2 expression
Inflammation was classified as diffuse, perivascular, and perilobular on H&E and also using markers. Intensity was qualitatively graded as from 0 (absent) to 3 (marked) Macrophage marker: CD68
(1) In diffuse inflammation pattern, macrophage intensity predominated other cell types and was associated with high-grade, large tumor size, tumor necrosis, and c-erbB-2 expression (2) Intensity of diffuse inflammation but not macrophage correlated with vascularity
Macrophage marker: CD68 (KP-1 antibody) Macrophages were counted in 40 hpf (20 hpf tumor cell zones and 20 hpf stromal zones) and graded from weak (<300) to intense (>500)
(1) Intensity of macrophage was higher in node-negative tumors (2) Intratumoral macrophage infiltration correlated with high tumor grade, absence of ER, and high mitotic grade
Macrophage marker: CD68 Macrophages were counted in 5 hot spots, and the mean of the highest three was determined (per mm2)
(1) High macrophage count correlated with high levels of macrophage chemoattractant protein-1 and thymidine phosphorylase in breast cancer by ELISA (2) High level of macrophage chemoattractant protein-1 had worsened relapse-free survival
Macrophage marker: CD68 Macrophage index was determined by 25-point Chalkey graticule as the mean of three “hot spot” counts under 250x field
(1) Macrophage index correlated with high VEGF and EGFR expression (2) In EGFR-negative cases, high VEGF correlated with increased macrophage index, high grade, presence of necrosis, and increased tumor p53 expression (3) No significant prognostic value of VEGF
24 invasive breast carcinomas (12 ductal, 12 lobular) [74]
Macrophage marker: CD68 Macrophage index was determined by 25-point Chalkey graticule or by absolute count as the mean of five VEGF positive areas under 200x field. In VEGF-negative areas, 5 most or least vascularized areas were chosen
(1) Macrophage count was higher in less vascularized areas
Macrophage marker: CD68 macrophages were counted in 5 hot spots, and the mean of the highest three was determined (per mm2). Graded from 0 (<50/mm2) to 2 (>100mm2)
(1) High macrophage count showed a tendency of correlation with high level of tumoral macrophage chemoattractant protein-1 by immunohistochemistry . (2) High level of tumoral macrophage chemoattractant protein-1 showed a tendency of correlation with high microvessel density grade
Macrophage marker: CD68 Macrophages were semiqualitatively graded as 1 = no macrophages, 2 = small foci of macrophages, 3 = large foci of macrophages 4 = diffuse macrophages infiltration in tumor stroma
(1) Higher macrophage grade associated with higher VEGF expression, higher microvessel density, and higher mitotic activity index
Macrophage density was assessed as average density of three hot spots at a magnification of 400x
(1) Macrophage density significantly correlated with both the VEGF expression and MVD (2) Macrophage density was associated with the nuclear grade, estrogen receptor status, and MIB-1 count (3) Patients with a high macrophage density had a significantly worse disease-free survival prognosis than those with a low density
Macrophage marker: CD68 Macrophages were semiqualitatively graded as 1 = no macrophages, 2 = small and large foci of macrophages, 3 = diffuse macrophages infiltration in tumor stroma
(1) Macrophage grade was not correlated with tumor chemoattractant protein-1
78 invasive breast carcinomas (48 ductal, 30 lobular) [79]
Macrophage marker: HAM56 antibody Macrophages were semiqualitatively graded as 1 = no macrophages, 2 = small foci of macrophages, 3 = large foci of macrophages, 4 = diffuse macrophages infiltration in tumor stroma
(1) Higher macrophages in invasive ductal carcinomas compared to invasive lobular carcinomas (2) In invasive ductal carcinomas, macrophage grade correlated with tumor size, lymph node metastasis, stage, microvessel density, VEGF, and tumor grade (3) In invasive ductal carcinomas, macrophage grade and clinical stage were predictive in disease-free survival rate
Macrophage marker: CD68 Macrophages were counted in 5 consecutive 400x fields in areas identified as “hot spots” under 100x
(1) Higher macrophage count associated with high tumor grade, p53 expression, absence of ER, high VEGF expression in macrophage, and postsurgical serum VEGF level
168 invasive primary breast cancer (142 ductal, 20 lobular, 6 others) [81]
Macrophage marker: CD68 Macrophages were counted using point counting method (expressed as percentage of volume occupied by a component out of total volume) using a 100-point ocular grid counting at 400x field over 30 fields and were grouped tertiles
(1) High tertile percentage of macrophage correlated with high tumor grade, high Ki-67 labelling, absence of hormonal receptors, high microvessel density, high CD4 and CD8 count
Macrophage marker: CD68 Macrophages were counted as mean of the 3 densest areas at 200x field (per mm2) following a brief scan at low power and separated into <320 or >320/mm2 groups
(1) Macrophage count correlated with stromal chemoattractant protein-1 (2) Stromal chemoattractant protein-1 correlated with lymphatic invasion and predicted worsened relapse-free survival
