Developing mammary terminal duct lobular units have a dynamic mucosal and stromal immune microenvironment

AbstractThe human breast and ovine mammary gland undergo a striking degree of postnatal development, leading to formation of terminal duct lobular units (TDLUs). In this study we interrogated aspects of sheep TDLU growth to increase understanding of ovine mammogenesis and as a model for the study of breast development. Mammary epithelial proliferation is significantly higher in lambs less than two months old than in peri-pubertal animals. Ki67 expression is polarized to the leading edge of the developing TDLUs. Intraepithelial ductal macrophages exhibit striking periodicity and significantly increased density in lambs approaching puberty. Stromal macrophages are more abundant centrally than peripherally. The developing ovine mammary gland is infiltrated by intraepithelial and stromal T lymphocytes that are significantly more numerous in older lambs. In the stroma, hotspots of Ki67 expression colocalize with large aggregates of lymphocytes and macrophages. Multifocally these aggregates exhibit distinct organization consistent with tertiary lymphoid structures. The lamb mammary gland thus exhibits a dynamic mucosal and stromal immune microenvironment and, as such, constitutes a valuable model system that provides new insights into postnatal breast development.Summary statementDevelopment of terminal duct lobular units in the sheep mammary gland involves distinct growth phases and macrophage and lymphocyte fluxes. Tertiary lymphoid structures are present subjacent to the mucosal epithelium.

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Developing ovine mammary terminal duct lobular units have a dynamic mucosal and stromal immune microenvironment

Communications Biology ◽

10.1038/s42003-021-02502-6 ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

Dorottya Nagy ◽

Clare M. C. Gillis ◽

Katie Davies ◽

Abigail L. Fowden ◽

Paul Rees ◽

...

Keyword(s):

Mammary Gland ◽

Immune Cell ◽

Leading Edge ◽

Breast Development ◽

Cell Aggregates ◽

Human Breast ◽

Immune Microenvironment ◽

Ki67 Expression ◽

Tertiary Lymphoid Structures ◽

Lymphoid Structures

AbstractThe human breast and ovine mammary gland undergo striking levels of postnatal development, leading to formation of terminal duct lobular units (TDLUs). Here we interrogate aspects of sheep TDLU growth as a model of breast development and to increase understanding of ovine mammogenesis. The distributions of epithelial nuclear Ki67 positivity differ significantly between younger and older lambs. Ki67 expression is polarised to the leading edge of the developing TDLUs. Intraepithelial ductal macrophages exhibit periodicity and considerably increased density in lambs approaching puberty. Stromal macrophages are more abundant centrally than peripherally. Intraepithelial T lymphocytes are more numerous in older lambs. Stromal hotspots of Ki67 expression colocalize with immune cell aggregates that exhibit distinct organisation consistent with tertiary lymphoid structures. The lamb mammary gland thus exhibits a dynamic mucosal and stromal immune microenvironment and constitutes a valuable model system that provides new insights into postnatal breast development.

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Fluorescent Multiplex Immunohistochemistry Coupled With Other State-Of-The-Art Techniques to Systematically Characterize the Tumor Immune Microenvironment

Frontiers in Molecular Biosciences ◽

10.3389/fmolb.2021.673042 ◽

2021 ◽

Vol 8 ◽

Author(s):

Anaïs Boisson ◽

Grégory Noël ◽

Manuel Saiselet ◽

Joël Rodrigues-Vitória ◽

Noémie Thomas ◽

...

Keyword(s):

Flow Cytometry ◽

Immune Microenvironment ◽

Cancer Treatments ◽

Luminal B ◽

Her2 Breast Cancer ◽

Technological Advances ◽

Tumor Immune Microenvironment ◽

Tertiary Lymphoid Structures ◽

Treatment Responses ◽

Lymphoid Structures

Our expanding knowledge of the interactions between tumor cells and their microenvironment has helped to revolutionize cancer treatments, including the more recent development of immunotherapies. Immune cells are an important component of the tumor microenvironment that influence progression and treatment responses, particularly to the new immunotherapies. Technological advances that help to decipher the complexity and diversity of the tumor immune microenvironment (TIME) are increasingly used in translational research and biomarker studies. Current techniques that facilitate TIME evaluation include flow cytometry, multiplex bead-based immunoassays, chromogenic immunohistochemistry (IHC), fluorescent multiplex IHC, immunofluorescence, and spatial transcriptomics. This article offers an overview of our representative data, discusses the application of each approach to studies of the TIME, including their advantages and challenges, and reviews the potential clinical applications. Flow cytometry and chromogenic and fluorescent multiplex IHC were used to immune profile a HER2+ breast cancer, illustrating some points. Spatial transcriptomic analysis of a luminal B breast tumor demonstrated that important additional insight can be gained from this new technique. Finally, the development of a multiplex panel to identify proliferating B cells, Tfh, and Tfr cells on the same tissue section demonstrates their co-localization in tertiary lymphoid structures.

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Prognostic Significance of Gene Signature of Tertiary Lymphoid Structures in Patients With Lung Adenocarcinoma

Frontiers in Oncology ◽

10.3389/fonc.2021.693234 ◽

2021 ◽

Vol 11 ◽

Author(s):

Hong Feng ◽

Fujun Yang ◽

Lihong Qiao ◽

Kai Zhou ◽

Junfei Wang ◽

...

Keyword(s):

Overall Survival ◽

Lung Adenocarcinoma ◽

Immune Cell ◽

Prognostic Significance ◽

Gene Signature ◽

Driver Gene ◽

Immune Microenvironment ◽

High Group ◽

Tertiary Lymphoid Structures ◽

Lymphoid Structures

BackgroundLung adenocarcinoma (LUAD) is a highly mortal cancer. Tertiary lymphoid structures (TLS) are ectopic lymphoid organs with similar morphological and molecular characters to secondary lymphoid organ. The aim of this study is to investigate the prognostic effect of a gene signature associated with TLSs, including B-cell-specific genes.MethodsClinical data of 515 LUAD patients in the TGCA cohort were used to examine the relationship of TLS signature with immune microenvironment, tumor mutational burden (TMB), and driver gene mutations. Patients were divided into the TLS signature high group and TLS signature low group, and comparative analysis of survival and its influencing factors between the two groups was performed. The resulting data were then validated in the GSE37745 cohort.ResultsTLS signature high group had significantly better overall survival (OS) and progression-free interval (PFI) as well as significantly higher infiltration of immune cell subsets, cancer immune cycle (CIC) signature except for immunogram score2 (IGS2), and expression of major checkpoint genes than the TLS signature low group. Notably, while TLS signature was not markedly associated with TMB and mutation frequencies of driver genes, there were significant differences in overall survival of patients with given mutation status of EGFR, KRAS, BRAF and TP53 genes between the TLS signature high and low groups.ConclusionThis study provided evidence that LUAD patients with high TLS signature had a favorable immune microenvironment and better prognosis, suggesting that TLS signature is an independent positive prognostic factor for LUAD patients.

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Androgen Receptor Inactivation Resulted in Acceleration in Pubertal Mammary Gland Growth, Upregulation of ERα Expression, and Wnt/β-Catenin Signaling in Female Mice

Endocrinology ◽

10.1210/en.2014-1226 ◽

2014 ◽

Vol 155 (12) ◽

pp. 4951-4963 ◽

Cited By ~ 13

Author(s):

Yan Ru (Ellen) Gao ◽

Kirsty A. Walters ◽

Reena Desai ◽

Hong Zhou ◽

David J. Handelsman ◽

...

Keyword(s):

Mammary Gland ◽

Androgen Receptor ◽

Physiological Characteristic ◽

Mammary Epithelial ◽

Breast Development ◽

Female Mice ◽

Estrogen Exposure ◽

Terminal End Buds ◽

Receptor Inactivation

The androgen receptor (AR) is widely expressed in mammary cells of female mammals including humans and mice, indicating a possible role for AR-mediated androgen actions in breast development, function, and pathology, although the specific mechanisms remain unclear. To elucidate the mechanisms of androgen action in mammary gland physiology and development, we used AR-knockout (ARΔex3KO) female mice with a universally expressed, transcriptionally inactive AR protein harboring an in-frame deletion of its second zinc finger. Although in sexually mature wild-type (WT) and ARex3ΔKO females, the mammary epithelial growth was fully extended to the edge of the fat pad, during puberty, ARex3ΔKO females exhibit significantly accelerated mammary ductal growth and an increased number of terminal end buds compared with WT females. Accelerated ARex3ΔKO female mammary growth was associated with significantly increased mammary epithelial ERα expression and activated Wnt/β-catenin signaling as shown by increased Wnt4 expression and accumulation of nuclear β-catenin. These findings are consistent with increased mammary estrogen exposure although ovarian estradiol content was unchanged compared with WT females. Furthermore, treatment with the potent pure androgen DHT markedly reduced ductal extension and terminal end bud numbers in WT but not in ARΔex3KO females, further supporting the concept that AR-mediated, androgen-induced suppression of murine mammary growth is a physiological characteristic of puberty. In summary, our findings reveal an inhibitory role of AR-mediated androgen actions in pubertal mammary gland development by reducing epithelial cell proliferation and could be mediated by regulation of Wnt/β-catenin signaling.

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Ultrastructural Changes in the Mammary Epithelial Cell Population During Neoplastic Development Induced by A Chemical Carcinogen.

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100091238 ◽

1976 ◽

Vol 34 ◽

pp. 250-251 ◽

Cited By ~ 2

Author(s):

J. Russo ◽

W. Isenberg ◽

M. Ireland ◽

I.H. Russo

Keyword(s):

Mammary Gland ◽

Cell Population ◽

Virgin Female ◽

Histological Change ◽

Mammary Epithelial ◽

Female Rats ◽

Ultrastructural Changes ◽

Post Inoculation ◽

Chemical Carcinogen ◽

Sprague Dawley

The induction of rat mammary carcinoma by the chemical carcinogen DMBA is used as a model for the study of the human disease (1). We previously described the histochemical changes that occur in the mammary gland of DMBA treated animals before the earliest manifested histological change, the intraductal proliferation (IDP), was observed (2). In the present work, we demonstrate that a change in the stable cell population found in the resting mammary gland occurs after carcinogen administration.Fifty-five day old Sprague-Dawley virgin female rats were inoculated intragastrically with 20mg of 7,12-dimethylbenz(a)anthracene (DMBA) in 1ml sesame oil. Non-inoculated, age-matched females were used as controls. Mammary glands from control and inoculated rats were removed weekly from the time of inoculation until 60 days post-inoculation. For electron microscopy, the glands were immersed in Karnovsky's fixative, post-fixed in 1% OsO4, dehydrated, and embedded in an Epon-Araldite mixture. Thick (lμ) sections were stained with 1% toluidine blue and were used for selecting areas for ultrastructural study.

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Faculty Opinions recommendation of Induction of secondary and tertiary lymphoid structures in the skin.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1022294.253695 ◽

2004 ◽

Author(s):

David Chaplin

Keyword(s):

Tertiary Lymphoid Structures ◽

Lymphoid Structures

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Faculty Opinions recommendation of Stromal fibroblasts in tertiary lymphoid structures: A novel target in chronic inflammation.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.727013457.793528869 ◽

2017 ◽

Author(s):

Theresa Lu ◽

William Shipman ◽

Dragos Dasoveanu

Keyword(s):

Chronic Inflammation ◽

Stromal Fibroblasts ◽

Tertiary Lymphoid Structures ◽

Lymphoid Structures ◽

Novel Target

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Mature tertiary lymphoid structures predict immune checkpoint inhibitor efficacy in solid tumors independently of PD-L1 expression

Nature Cancer ◽

10.1038/s43018-021-00232-6 ◽

2021 ◽

Author(s):

Lucile Vanhersecke ◽

Maxime Brunet ◽

Jean-Philippe Guégan ◽

Christophe Rey ◽

Antoine Bougouin ◽

...

Keyword(s):

Solid Tumors ◽

Immune Checkpoint ◽

Immune Checkpoint Inhibitor ◽

Checkpoint Inhibitor ◽

Tertiary Lymphoid Structures ◽

Lymphoid Structures

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Agonistic CD40 therapy induces tertiary lymphoid structures but impairs responses to checkpoint blockade in glioma

Nature Communications ◽

10.1038/s41467-021-24347-7 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Luuk van Hooren ◽

Alessandra Vaccaro ◽

Mohanraj Ramachandran ◽

Konstantinos Vazaios ◽

Sylwia Libard ◽

...

Keyword(s):

Checkpoint Inhibitors ◽

Systemic Delivery ◽

Systemic Induction ◽

T Cell Infiltration ◽

Treatment Naïve ◽

Tertiary Lymphoid Structures ◽

Lymphoid Structures ◽

Immunosuppressive Microenvironment ◽

The Brain ◽

Opening Up

AbstractGliomas are brain tumors characterized by an immunosuppressive microenvironment. Immunostimulatory agonistic CD40 antibodies (αCD40) are in clinical development for solid tumors, but are yet to be evaluated for glioma. Here, we demonstrate that systemic delivery of αCD40 in preclinical glioma models induces the formation of tertiary lymphoid structures (TLS) in proximity of meningeal tissue. In treatment-naïve glioma patients, the presence of TLS correlates with increased T cell infiltration. However, systemic delivery of αCD40 induces hypofunctional T cells and impairs the response to immune checkpoint inhibitors in pre-clinical glioma models. This is associated with a systemic induction of suppressive CD11b+ B cells post-αCD40 treatment, which accumulate in the tumor microenvironment. Our work unveils the pleiotropic effects of αCD40 therapy in glioma and reveals that immunotherapies can modulate TLS formation in the brain, opening up for future opportunities to regulate the immune response.

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