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Updated: Mar 3, 2026
Indirect Immunofluorescence on Frozen Sections of Mouse Mammary Gland
Published on: December 1, 2015
Image analysis of immune cell patterns in the human mammary gland during the menstrual cycle refines lymphocytic
Nadine S Schaadt1, Juan Carlos López Alfonso2, Ralf Schönmeyer3
1Institute of Pathology, Neuropathology, Hannover Medical School, Carl-Neuberg-Straße 1, 30625, Hannover, Germany.
Insights
Advanced image analysis distinguishes normal immune cell patterns from inflammation in breast tissue. Hormonal variations, including menstrual cycle and oral contraceptives, influence CD8+ T-cell density, aiding oncoimmunology research.
Area of Science:
- Breast cancer oncoimmunology
- Mammary gland immune cell analysis
- Digital pathology
Background:
- Immune cell infiltration patterns are crucial in breast cancer oncoimmunology.
- Distinguishing normal immune cell distribution from inflammatory conditions like lymphocytic lobulitis is challenging.
- Hormonal fluctuations may influence immune cell presence in non-neoplastic breast tissue.
Purpose of the Study:
- To enhance microscopic evaluation of immune cells in breast cancer oncoimmunology.
- To differentiate normal immune cell infiltration from lymphocytic lobulitis using advanced image analysis.
- To investigate immune cell variations related to the menstrual cycle and oral contraceptive use.
Main Methods:
- Analysis of lymphocyte and macrophage distribution in 53 reduction mammoplasty specimens.
- Immunohistochemical staining and digital whole slide imaging of resting mammary gland tissue.
- Image analysis workflow: automated region detection, immune cell recognition, and co-registration.
Main Results:
- Normal lobular epithelium contains approximately 7 CD8+ T-cells per 100 epithelial cells, with 70% near the basal layer.
- CD8+ T-cell density increased 1.6-fold in the luteal phase and 1.4-fold with oral contraceptive use.
- CD4+ T-cells were infrequent; CD163+ macrophages were abundant and widespread with minor cycle-related variation.
Conclusions:
- Spatial immune cell patterns define normal versus inflammatory breast microenvironments.
- Advanced image analysis quantifies hormonal effects on immune cells.
- This approach refines lymphocytic lobulitis diagnosis and aids oncoimmunology biopsy evaluation.
Purpose:
To improve microscopic evaluation of immune cells relevant in breast cancer oncoimmunology, we aim at distinguishing normal infiltration patterns from lymphocytic lobulitis by advanced image analysis. We consider potential immune cell variations due to the menstrual cycle and oral contraceptives in non-neoplastic mammary gland tissue.
Methods:
Lymphocyte and macrophage distributions were analyzed in the anatomical context of the resting mammary gland in immunohistochemically stained digital whole slide images obtained from 53 reduction mammoplasty specimens. Our image analysis workflow included automated regions of interest detection, immune cell recognition, and co-registration of regions of interest.
Results:
In normal lobular epithelium, seven CD8[Formula: see text] lymphocytes per 100 epithelial cells were present on average and about 70% of this T-lymphocyte population was lined up along the basal cell layer in close proximity to the epithelium. The density of CD8[Formula: see text] T-cell was 1.6 fold higher in the luteal than in the follicular phase in spontaneous menstrual cycles and 1.4 fold increased under the influence of oral contraceptives, and not co-localized with epithelial proliferation. CD4[Formula: see text] T-cells were infrequent. Abundant CD163[Formula: see text] macrophages were widely spread, including the interstitial compartment, with minor variation during the menstrual cycle.
Conclusions:
Spatial patterns of different immune cell subtypes determine the range of normal, as opposed to inflammatory conditions of the breast tissue microenvironment. Advanced image analysis enables quantification of hormonal effects, refines lymphocytic lobulitis, and shows potential for comprehensive biopsy evaluation in oncoimmunology.
