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

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.

Download Full-text

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

10.1101/2020.11.05.369843 ◽

2020 ◽

Cited By ~ 1

Author(s):

Dorottya Nagy ◽

Clare M. C. Gillis ◽

Katie Davies ◽

Abigail L. Fowden ◽

Paul Rees ◽

...

Keyword(s):

Mammary Gland ◽

Leading Edge ◽

Mammary Epithelial ◽

Breast Development ◽

Growth Phases ◽

Immune Microenvironment ◽

Ki67 Expression ◽

Tertiary Lymphoid Structures ◽

Summary Statement ◽

Lymphoid Structures

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.

Download Full-text

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.

Download Full-text

Abstract A40: The balance between activated follicular helper T cells and follicular regulatory T cells within tertiary lymphoid structures guides antitumor immune responses in human breast cancer

10.1158/2326-6074.tumimm18-a40 ◽

2020 ◽

Author(s):

Grégory Noël ◽

Mireille Langouo ◽

Soizic Garaud ◽

Anaïs Boisson ◽

Hugues Duvillier ◽

...

Keyword(s):

Breast Cancer ◽

T Cells ◽

Regulatory T Cells ◽

Immune Responses ◽

Human Breast Cancer ◽

Helper T Cells ◽

Human Breast ◽

Follicular Helper ◽

Tertiary Lymphoid Structures ◽

Lymphoid Structures

Download Full-text

Multiplex Immunofluorescence Histology for Immune Cell Infiltrates in Melanoma-Associated Tertiary Lymphoid Structures

Methods in Molecular Biology - Melanoma ◽

10.1007/978-1-0716-1205-7_40 ◽

2021 ◽

pp. 573-587

Author(s):

Ileana S. Mauldin ◽

Adela Mahmutovic ◽

Samuel J. Young ◽

Craig L. Slingluff

Keyword(s):

Immune Cell ◽

Tertiary Lymphoid Structures ◽

Lymphoid Structures ◽

Immune Cell Infiltrates

Download Full-text

The Immune System of Mesothelioma Patients: A Window of Opportunity for Novel Immunotherapies

10.5772/intechopen.98617 ◽

2021 ◽

Author(s):

Fabio Nicolini ◽

Massimiliano Mazza

Keyword(s):

Immune System ◽

Immune Cells ◽

Immune Cell ◽

Current Knowledge ◽

Cell Types ◽

Screening Strategies ◽

Tertiary Lymphoid Structures ◽

Lymphoid Structures ◽

Review Current

The interplay between the immune system and the pleural mesothelium is crucial both for the development of malignant pleural mesothelioma (MPM) and for the response of MPM patients to therapy. MPM is heavily infiltrated by several immune cell types which affect the progression of the disease. The presence of organized tertiary lymphoid structures (TLSs) witness the attempt to fight the disease in situ by adaptive immunity which is often suppressed by tumor expressed factors. In rare patients physiological, pharmacological or vaccine-induced immune response is efficient, rendering their plasma a valuable resource of anti-tumor immune cells and molecules. Of particular interest are human antibodies targeting antigens at the tumor cell surface. Here we review current knowledge regarding MPM immune infiltration, MPM immunotherapy and the harnessing of this response to identify novel biologics as biomarkers and therapeutics through innovative screening strategies.

Download Full-text

Metagenomic and single-cell RNA-seq survey of the H. pylori-infected stomach in asymptomatic individuals

10.1101/2021.12.04.21267139 ◽

2021 ◽

Author(s):

Chiara Sorini ◽

Kumar P. Tripathi ◽

Shengru Wu ◽

Shawn M. Higdon ◽

Jing Wang ◽

...

Keyword(s):

Gastric Mucosa ◽

Single Cell ◽

Immune Cell ◽

Human Stomach ◽

List Type ◽

Whole Genome ◽

Tertiary Lymphoid Structures ◽

Lymphoid Structures ◽

H Pylori ◽

Asymptomatic Individuals

ABSTRACTObjectiveHelicobacter pylori colonization of the gastric niche can persist for years in asymptomatic individuals. Although latent H. pylori infection can progress to cancer, a detailed survey of the microbiome and immune composition in the chronically infected stomach is still lacking.DesignWe collected human gastric tissues and performed metagenomic sequencing, single-cell RNA sequencing (scRNA-seq), flow cytometry and fluorescent microscopy to deeply characterize the host-microbiota environment in H. pylori-infected (HPI) stomachs.ResultsHPI asymptomatic individuals showed dramatic changes in the composition of gastric microbiome and immune cells compared to non-infected individuals. With metagenomic data, we also demonstrated antibiotic resistant genes, enzymes and pathway alterations related to metabolism and immune response. scRNA-seq and flow cytometry data revealed that in contrast to murine stomachs, ILC2 are virtually absent in the human gastric mucosa, whereas ILC3 are the dominant population in asymptomatic HPI individuals. Specifically, NKp44+ ILC3s were highly increased in the gastric mucosa of asymptomatic HPI individuals, and their proportions correlated with the abundance of selected microbial taxa found to be enriched in the infected mucosa. In addition, CD11c+ myeloid cells, activated CD4 T cells and B cells were expanded in HPI individuals. In HPI individuals, B cells acquired an activated phenotype and progressed into a highly proliferating germinal center stage and plasmablast maturation, which correlated with the presence of tertiary lymphoid structures within the gastric lamina propria.ConclusionOur study provides a comprehensive atlas of the gastric mucosa-associated microbiome and immune cell landscape when comparing asymptomatic HPI and uninfected individuals.SIGNIFICANCE OF THE STUDYWhat is already known on this subject? Previous studies on the gastric microbiome were performed via 16S rRNA gene sequencing.Acute Helicobacter spp. infection in murine models and symptomatic H. pylori-driven pathology in humans result in remodeling of the stomach immune cell compartment.ILC2 is the dominant ILC population in the murine stomach.What are the new findings? We described the effect of chronic asymptomatic H. pylori infection on the gastric microbiome via whole-genome sequencing.Single cell census of the gastric mucosa reveals ILC3 to be the dominant ILC population in the human stomach, whereas ILC2 were virtually absent.scRNA-seq reveals the gastric immune cell programs in asymptomatic H. pylori-infected individuals, which is characterized by the formation of tertiary lymphoid structures.How might it impact on clinical practice in the foreseeable future? Whole genome sequencing of uninfected and H. pylori-infected gastric mucosa bolsters collective knowledge of stomach physiology with respect to the gastric microbiome and microbiota function.We present a comprehensive immune cellular landscape of the human stomach, which will be a valuable resource to interrogate pathology of gastric diseases.

Download Full-text

Nerve fibers in the Tumor Microenvironment are co-localized with Tertiary Lymphoid Structures

10.1101/2020.08.07.232322 ◽

2020 ◽

Author(s):

Lara R. Heij ◽

Xiuxiang Tan ◽

Jakob N. Kather ◽

Jan M. Niehues ◽

Shivan Sivakumar ◽

...

Keyword(s):

Pancreatic Ductal Adenocarcinoma ◽

Immune Cells ◽

Immune Cell ◽

Nerve Fibers ◽

Ductal Adenocarcinoma ◽

Fiber Density ◽

Tertiary Lymphoid Structures ◽

Lymphoid Structures ◽

Nerve Fiber Density ◽

Small Nerve

ABSTRACTBackgroundB cells and tertiary lymphoid structures (TLS) are reported to be important in the improvement of survival of cancer patients. These secondary lymphoid organs have been associated with the generation of an anti-tumor response. Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancer types and the stromal architecture shapes the intratumoral heterogeneity. The stroma of PDAC is a complex system in which crosstalk takes place between cancer-associated fibroblasts, immune cells, endothelial cells and the cancer cells. Besides immune cells and fibroblasts, there is some limited data about the influence of nerve fibers on cancer progression.Patients and methodsNerve Fiber Density (NFD) was analysed in our cohort of 188 patients with Pancreatic Ductal Adenocarcinoma who underwent pancreatic surgery. We used immunohistochemistry and multiplex imaging to phenotype the immune cell infiltrate. The cell detection classifier measured distance from immune cell to cancer gland and with a heat map we could count TLS. By using Machine learning we were able to define the spatial distribution and counting Tertiary Lymphoid Structures.ResultsHigh NFD is significantly associated with prolonged overall survival (HR 1.676 (95%CI 1.126,2.495) for low vs. high NFD, p-value 0.0109). The immune cells surrounding the nerve fibers were phenotyped in B cells, T cells and dendritic follicular cells, matching a TLS. Here we show that small nerve fibers are located at the TLS in Pancreatic Cancer and a high Nerve Fiber Density combined with more than 5 TLS is associated with a better survival (HR 0.388 (95%CI 0.218, 0.689).ConclusionThe co-localization of small nerve fibers with TLS is a new finding which has not been described before. However the precise roles of these TLS and nerve fibers remains unknown. These findings unravel future pathways and has the potential to reach new directions into already existing targeted therapy.

Download Full-text

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.

Download Full-text