Lymph Node Subcapsular Sinus Microenvironment-On-A-Chip Modeling Shear Flow Relevant to Lymphatic Metastasis and Immune Cell Homing

Objectives: The aim of this study was to assess the number of tumour-infiltrating lymphocytes (TILs) and the expression of Programmed Cell Death 1 Ligand 1 (PD-L1) in Acinic Cell Carcinoma (AciCC) of the salivary glands, to enable a correlation with clinico-pathological features and to analyse their prognostic impact. Methods: This single centre retrospective study represents a cohort of 36 primary AciCCs with long-term clinical follow-up. Immunohistochemically defined immune cell subtypes, i.e., those expressing T-cell markers (CD3, CD4 and CD8) or a B-cell marker (CD20) were characterized on tumour tissue sections. The number of TILs was quantitatively evaluated using software for digital bioimage analysis (QuPath). PD-L1 expression on the tumour cells and on immune cells was assessed immunohistochemically employing established scoring criteria: tumour proportion score (TPS), Ventana immune cell score (IC-Score) and combined positive score (CPS). Results: Higher numbers of tumour-infiltrating T- and B- lymphocytes were significantly associated with high-grade transformation. Furthermore, higher counts of T-lymphocytes correlated with node-positive disease. There was a significant correlation between higher levels of PD-L1 expression and lymph node metastases as well as the occurrence of high-grade transformation. Moreover, PD-L1 CPS was associated with poor prognosis regarding metastasis-free survival (p = 0.049). Conclusions: The current study is the first to demonstrate an association between PD-L1 expression and lymph node metastases as well as grading in AciCCs. In conclusion, increased immune cell infiltration of T and B cells as well as higher levels of PD-L1 expression in AciCC in association with high-grade transformation, lymph node metastasis and unfavourable prognosis suggests a relevant interaction between tumour cells and immune cell infiltrates in a subset of AciCCs, and might represent a rationale for immune checkpoint inhibition.

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Identification of lymph node metastasis-related microRNAs in lung adenocarcinoma and analysis of the underlying mechanisms using a bioinformatics approach

Experimental Biology and Medicine ◽

10.1177/1535370216677353 ◽

2016 ◽

Vol 242 (7) ◽

pp. 709-717 ◽

Cited By ~ 16

Author(s):

Li Yan ◽

Demin Jiao ◽

Huizhen Hu ◽

Jian Wang ◽

Xiali Tang ◽

...

Keyword(s):

Transcription Factors ◽

Lymph Node ◽

Lymph Node Metastasis ◽

Lung Adenocarcinoma ◽

Lymphatic Metastasis ◽

Target Genes ◽

Bioinformatics Analyses ◽

Node Metastasis ◽

Primary Lung Adenocarcinoma ◽

Underlying Mechanisms

This study aimed to screen lymphatic metastasis-related microRNAs (miRNAs) in lung adenocarcinoma and explore their underlying mechanisms using bioinformatics. The miRNA expression in primary lung adenocarcinoma, matched adjacent non-tumorigenic and lymph node metastasis tissues of patients were profiled via microarray. The screened metastasis-related miRNAs were then validated using quantitative real-time PCR in a second cohort of lung adenocarcinoma patients with lymphatic metastasis. Significance was determined using a paired t-test. Target genes of the metastasis-related miRNAs were predicted using TargetScan, and transcription factors (TFs) were predicted based on the TRANSFAC and ENCODE databases. Furthermore, the related long non-coding RNAs (lncRNAs) were screened with starBase v2.0. The miRNA-TF-mRNA and lncRNA-miRNA-mRNA networks were constructed to determine the key interactions associated with lung adenocarcinoma metastasis. According to the miRNA microarray results, there were 10 miRNAs that were differentially expressed in metastatic tissues compared with primary tumor and adjacent non-tumorigenic tissues. Among them were increased levels of miR-146a-5p, miR-342-3p, and miR-150-5p, which were validated in the second cohort. Based on the miRNA-TF-mRNA network, vascular endothelial growth factor A and transcription factors (TFs) including TP53, SMAD4, and EP300 were recognized as critical targets of the three miRNAs. Interactions involving SNHG16–miR-146a-5p–SMAD4 and RP6-24A23.7–miR-342-3p/miR-150-5p–EP300 were highlighted according to the lncRNA-miRNA-mRNA network. miR-146a-5p, miR-342-3p, and miR-150-5p are lymphatic metastasis-related miRNAs in lung adenocarcinoma. Bioinformatics analyses demonstrated that SNHG16 might inhibit the interaction between miR-146a-5p and SMAD4, while RP6-24A23.7 might weaken miR-342-3p–EP300 and miR-150-5p–EP300 interactions in metastasis.

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The C-terminal peptide of CCL21 drastically augments CCL21 activity through the dendritic cell lymph node homing receptor CCR7 by interaction with the receptor N-terminus

Cellular and Molecular Life Sciences ◽

10.1007/s00018-021-03930-7 ◽

2021 ◽

Author(s):

Astrid Sissel Jørgensen ◽

Emma Probst Brandum ◽

Jeppe Malthe Mikkelsen ◽

Klaudia A. Orfin ◽

Ditte Rahbæk Boilesen ◽

...

Keyword(s):

Lymph Node ◽

Electrostatic Interactions ◽

Immune Cell ◽

Direct Interaction ◽

Receptor Interaction ◽

Binding Model ◽

Lymph Node Homing ◽

Gating Mechanism ◽

N Terminus ◽

Positively Charged

AbstractThe endogenous chemokines CCL19 and CCL21 signal via their common receptor CCR7. CCL21 is the main lymph node homing chemokine, but a weak chemo-attractant compared to CCL19. Here we show that the 41-amino acid positively charged peptide, released through C-terminal cleavage of CCL21, C21TP, boosts the immune cell recruiting activity of CCL21 by up to 25-fold and the signaling activity via CCR7 by ~ 100-fold. Such boosting is unprecedented. Despite the presence of multiple basic glycosaminoglycan (GAG) binding motifs, C21TP boosting of CCL21 signaling does not involve interference with GAG mediated cell-surface retention. Instead, boosting is directly dependent on O-glycosylations in the CCR7 N-terminus. As dictated by the two-step binding model, the initial chemokine binding involves interaction of the chemokine fold with the receptor N-terminus, followed by insertion of the chemokine N-terminus deep into the receptor binding pocket. Our data suggest that apart from a role in initial chemokine binding, the receptor N-terminus also partakes in a gating mechanism, which could give rise to a reduced ligand activity, presumably through affecting the ligand positioning. Based on experiments that support a direct interaction of C21TP with the glycosylated CCR7 N-terminus, we propose that electrostatic interactions between the positively charged peptide and sialylated O-glycans in CCR7 N-terminus may create a more accessible version of the receptor and thus guide chemokine docking to generate a more favorable chemokine-receptor interaction, giving rise to the peptide boosting effect.

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Adaptive and innate immune cell responses in tendons and lymph nodes after tendon injury and repair

Journal of Applied Physiology ◽

10.1152/japplphysiol.00682.2019 ◽

2020 ◽

Vol 128 (3) ◽

pp. 473-482 ◽

Cited By ~ 6

Author(s):

Andrew C. Noah ◽

Thomas M. Li ◽

Leandro M. Martinez ◽

Susumu Wada ◽

Jacob B. Swanson ◽

...

Keyword(s):

T Cells ◽

Lymph Node ◽

Immune System ◽

Innate Immune System ◽

Immune Cell ◽

Tendon Healing ◽

Tendon Injury ◽

Innate Immune ◽

Adaptive Immune ◽

Immune Cell Response

Tendon injuries are a common clinical condition with limited treatment options. The cellular components of the innate immune system, such as neutrophils and macrophages, have been studied in tendon injuries. However, the adaptive immune system, comprising specialized lymphocytes, plays an important role in orchestrating the healing of numerous tissues, but less is known about these cells in tendon healing. To gain a greater understanding of the biological processes that regulate tendon healing, we determined how the cellular components of the adaptive and innate immune system respond to a tendon injury using two-month-old male mice. We observed that lymphatic vasculature is present in the epitenon and superficial regions of Achilles tendons, and that the lymphatics drain into the popliteal lymph node. We then created an acute Achilles tenotomy followed by repair, and collected tendons and popliteal lymph nodes 1, 2, and 4 wk after injury. Tendon injury resulted in a robust adaptive immune cell response that followed an initial innate immune cell response in tendons and lymph nodes. Monocytes, neutrophils, and macrophages initially accumulated at 1 wk after injury in tendons, while dendritic cells and CD4+ T cells peaked at 2 wk after injury. B cells and CD8+ T cells progressively increased over time. In parallel, immune cells of the popliteal lymph node demonstrated a similarly coordinated response to the injury. These results suggest that there is an adaptive immune response to tendon injury, and adaptive immune cells may play a role in regulating tendon healing. NEW & NOTEWORTHY While the innate immune system, consisting of macrophages and related hematopoietic cells, has been studied in tendon injury, less is known about the adaptive immune system. Using a mouse model of Achilles tendon tenotomy and repair, we observed an adaptive immune cell response, consisting of CD4+ and CD8+ T cells, and B cells, which occur through 4 wk after tendon injury. This response appeared to be coordinated by the draining popliteal lymph node.

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A Mushroom Extract Piwep fromPhellinus igniariusAmeliorates Experimental Autoimmune Encephalomyelitis by Inhibiting Immune Cell Infiltration in the Spinal Cord

BioMed Research International ◽

10.1155/2014/218274 ◽

2014 ◽

Vol 2014 ◽

pp. 1-11 ◽

Cited By ~ 1

Author(s):

Lan Li ◽

Guang Wu ◽

Bo Young Choi ◽

Bong Geom Jang ◽

Jin Hee Kim ◽

...

Keyword(s):

Multiple Sclerosis ◽

Spinal Cord ◽

Lymph Node ◽

Immune Cell ◽

Therapeutic Potential ◽

Immune Cell Infiltration ◽

Autoimmune Encephalomyelitis ◽

Phellinus Igniarius ◽

Mushroom Extract ◽

Experimental Autoimmune

The present study aimed to evaluate the therapeutic potential of a mushroom extract fromPhellinus igniariusin an animal model of multiple sclerosis. The medicinal mushroom,Phellinus igniarius, contains biologically active compounds that modulate the human immune system. Experimental autoimmune encephalomyelitis (EAE) was induced by immunization with myelin oligodendrocyte glycoprotein (MOG 35–55) in C57BL/6 female mice. A water-ethanol extract ofPhellinus igniarius(Piwep) was delivered intraperitoneally every other day for the entire experimental course. Three weeks after the initial immunization, demyelination and immune cell infiltration in the spinal cord were examined. Piwep injection profoundly decreased the daily incidence rate and clinical score of EAE. The Piwep-mediated inhibition of the clinical course of EAE was accompanied by suppression of demyelination and infiltration of encephalitogenic immune cells including CD4+ T cells, CD8+ T cells, macrophages, and B cells in the spinal cord. Piwep reduced expression of vascular cell adhesion molecule-1 (VCAM-1) in the spinal cord and integrin-α4in the lymph node of EAE mice. Piwep also inhibited proliferation of lymphocytes and secretion of interferon-γin the lymph node of EAE mice. The results suggest that a mushroom extract, Piwep, may have a high therapeutic potential for ameliorating multiple sclerosis progression.

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Immunofluorescent characterization of non-myelinating Schwann cells and their interactions with immune cells in mouse mesenteric lymph node

European Journal of Histochemistry ◽

10.4081/ejh.2017.2827 ◽

2017 ◽

Vol 61 (3) ◽

Cited By ~ 4

Author(s):

Zhongli Shi ◽

Wayne K. Greene ◽

Philip K. Nicholls ◽

Dailun Hu ◽

Janina E.E. Tirnitz-Parker ◽

...

Keyword(s):

Nervous System ◽

Dendritic Cells ◽

Lymph Node ◽

Immune System ◽

Mesenteric Lymph Node ◽

Immune Cell ◽

Cell Interactions ◽

Immunofluorescent Staining ◽

Local Interactions ◽

Mesenteric Lymph

The central nervous system (CNS) influences the immune system in a general fashion by regulating the systemic concentration of humoral substances, whereas the autonomic nervous system communicates specifically with the immune system according to local interactions. Data concerning the mechanisms of this bidirectional crosstalk of the peripheral nervous system (PNS) and immune system remain limited. To gain a better understanding of local interactions of the PNS and immune system, we have used immunofluorescent staining of glial fibrillary acidic protein (GFAP), coupled with confocal microscopy, to investigate the non-myelinating Schwann cell (NMSC)-immune cell interactions in mouse mesenteric lymph nodes. Our results demonstrate i) the presence of extensive NMSC processes and even of cell bodies in each compartment of the mouse mesenteric lymph node; ii) close associations/interactions of NMSC processes with blood vessels (including high endothelial venules) and the lymphatic vessel/sinus; iii) close contacts/associations of NMSC processes with various subsets of dendritic cells (such as CD4+CD11c+, CD8+CD11c+ dendritic cells), macrophages (F4/80+ and CD11b+ macrophages), and lymphocytes. Our novel findings concerning the distribution of NMSCs and NMSC-immune cell interactions inside the mouse lymph node should help to elucidate the mechanisms through which the PNS affects cellular- and humoral-mediated immune responses or vice versa in health and disease.

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Fc chimeric protein containing the cysteine-rich domain of the murine mannose receptor binds to macrophages from splenic marginal zone and lymph node subcapsular sinus and to germinal centers.

Journal of Experimental Medicine ◽

10.1084/jem.184.5.1927 ◽

1996 ◽

Vol 184 (5) ◽

pp. 1927-1937 ◽

Cited By ~ 125

Author(s):

L Martínez-Pomares ◽

M Kosco-Vilbois ◽

E Darley ◽

P Tree ◽

S Herren ◽

...

Keyword(s):

Lymph Node ◽

Lymph Nodes ◽

B Cell ◽

Marginal Zone ◽

Chimeric Protein ◽

Mannose Receptor ◽

Germinal Centers ◽

White Pulp ◽

Subcapsular Sinus ◽

Splenic White Pulp

Ligands for the cysteine-rich (CR) domain of the mannose receptor (MR) were detected by incubating murine tissues with a chimeric protein containing CR fused to the Fc region of human IgG1 (CR-Fc). In naive mice, CR-Fc bound to sialoadhesin+, F4/80low/-, macrosialin+ macrophages (M phi) in spleen marginal zone (metallophilic M phi) and lymph node subcapsular sinus. Labeling was also observed in B cell areas of splenic white pulp. Western blotting analysis of spleen and lymph nodes lysates revealed a restricted number of molecules that interacted specifically with CR-Fc. In immunized mice, labeling was upregulated on germinal centers in splenic white pulp and follicular areas of lymph nodes. Kinetic analysis of the pattern of CR-Fc labeling in lymph nodes during a secondary immune response to ovalbumin showed that CR ligand expression migrated towards B cell areas, associated with cells displaying distinctive dendritic morphology, and accumulated in developing germinal centers. These studies suggest that MR+ cells or MR-carbohydrate-containing antigen complexes could be directed towards areas where humoral immune responses take place, through the interaction of the MR CR domain with molecules expressed in specialized macrophage populations and antigen transporting cells.

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Lymph-Borne Chemokines and Other Low Molecular Weight Molecules Reach High Endothelial Venules via Specialized Conduits While a Functional Barrier Limits Access to the Lymphocyte Microenvironments in Lymph Node Cortex

Journal of Experimental Medicine ◽

10.1084/jem.192.10.1425 ◽

2000 ◽

Vol 192 (10) ◽

pp. 1425-1440 ◽

Cited By ~ 408

Author(s):

J. Elizabeth Gretz ◽

Christopher C. Norbury ◽

Arthur O. Anderson ◽

Amanda E.I. Proudfoot ◽

Stephen Shaw

Keyword(s):

Lymph Node ◽

Basement Membranes ◽

High Endothelial Venules ◽

Reticular Fibers ◽

Lymph Node Enlargement ◽

Subcapsular Sinus ◽

Sinus Floor ◽

Draining Lymph Node ◽

Reticular Cells ◽

Draining Lymph Nodes

Lymph-borne, soluble factors (e.g., chemokines and others) influence lymphocyte recirculation and endothelial phenotype at high endothelial venules (HEVs) in lymph node cortex. Yet the route lymph-borne soluble molecules travel from the subcapsular sinus to the HEVs is unclear. Therefore, we injected subcutaneously into mice and rats a wide variety of fluorophore-labeled, soluble molecules and examined their distribution in the draining lymph nodes. Rather than percolating throughout the draining lymph node, all molecules, including microbial lipopolysaccharide, were very visible in the subcapsular and medullary sinuses but were largely excluded from the cortical lymphocyte microenvironments. Exclusion prevailed even during the acute lymph node enlargement accompanying viral infection. However, low molecular mass (MW) molecules, including chemokines, did gain entry into the cortex, but in a very defined manner. Low MW, fluorophore-labeled molecules highlighted the subcapsular sinus, the reticular fibers, and the abluminal and luminal surfaces of the associated HEVs. These low MW molecules were in the fibers of the reticular network, a meshwork of collagen fibers ensheathed by fibroblastic reticular cells that connects the subcapsular sinus floor and the HEVs by intertwining with their basement membranes. Thus, low MW, lymph-borne molecules, including chemokines, traveled rapidly from the subcapsular sinus to the HEVs using the reticular network as a conduit.

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Reconstitution of immune cell in liver and lymph node of adult- and newborn-engrafted humanized mice

BMC Immunology ◽

10.1186/s12865-016-0157-9 ◽

2016 ◽

Vol 17 (1) ◽

Cited By ~ 5

Author(s):

Crystal Dykstra ◽

Amanda J. Lee ◽

Evan J. Lusty ◽

Mira M. Shenouda ◽

Mahsa Shafai ◽

...

Keyword(s):

Lymph Node ◽

Immune Cell ◽

Humanized Mice

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Hyperion Image Analysis Depicts a Preliminary Landscape of Tumor Immune Microenvironment in OSCC with Lymph Node Metastasis

Journal of Immunology Research ◽

10.1155/2021/9975423 ◽

2021 ◽

Vol 2021 ◽

pp. 1-7

Author(s):

Shang Xie ◽

Xin-Yuan Zhang ◽

Xiao-Feng Shan ◽

Vicky Yau ◽

Jian-Yun Zhang ◽

...

Keyword(s):

Lymph Node ◽

Lymph Node Metastasis ◽

Treatment Planning ◽

Immune Cell ◽

Imaging System ◽

Individual Treatment ◽

Immune Microenvironment ◽

Node Metastasis ◽

Tumor Immune Microenvironment ◽

Formalin Fixed Paraffin Embedded

Background. Oral squamous cell carcinoma (OSCC) constitutes the most common types of oral cancer. Because its prognosis varies significantly, identification of a tumor immune microenvironment could be a critical tool for treatment planning and predicting a more accurate prognosis. This study is aimed at utilizing the Hyperion imaging system to depict a preliminary landscape of the tumor immune microenvironment in OSCC with lymph node metastasis. Methods. We collected neoplasm samples from OSCC patients. Their formalin-fixed, paraffin-embedded (FFPE) tissue sections were obtained and stained utilizing a panel of 26 clinically relevant metal-conjugated antibodies. Detection and analysis were performed for these stained cells with the Hyperion imaging system. Results. Four patients met our inclusion criteria. We depicted a preliminary landscape of their tumor immune microenvironment and identified 25 distinct immune cell subsets from these OSCC patients based on phenotypic similarity. All these patients had decreased expression of CD8+ T cells in tumor specimens. Variety in cell subsets was seen, and more immune activated cells were found in patient A and patient B than those in patient C and patient D. Such differences in tumor immune microenvironments can contribute to forecasting of individual prognoses. Conclusion. The Hyperion imaging system helped to delineate a preliminary and multidimensional landscape of the tumor immune microenvironment in OSCC with lymph node metastasis and provided insights into the influence of the immune microenvironment in determination of prognoses. These results reveal possible contributory factors behind different prognoses of OSCC patients with lymph node metastasis and provide reference for individual treatment planning.

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