Kidney resident macrophages have distinct subsets and multifunctional roles.

BACKGROUND: The kidney contains distinct glomerular and tubulointerstitial compartments with diverse cell types and extracellular matrix components. The role of immune cells in glomerular environment is crucial for dampening inflammation and maintaining homeostasis. Macrophages are innate immune cells that are influenced by their tissue microenvironment. However, the multifunctional role of kidney macrophages remains unclear. METHODS: Flow and imaging cytometry were used to determine the relative expression of CD81 and CX3CR1 (C-X3-C motif chemokine receptor 1) in kidney macrophages. Monocyte replenishment was assessed in Cx3cr1CreER X R26-yfp-reporter and shielded chimeric mice. Bulk RNA-sequencing and mass spectrometry-based proteomics were performed on isolated kidney macrophages from wild type and Col4a5-/- (Alport) mice. RNAscope was used to visualize transcripts and macrophage purity in bulk RNA assessed by CIBERSORTx analyses. RESULTS: In wild type mice we identified three distinct kidney macrophage subsets using CD81 and CX3CR1 and these subsets showed dependence on monocyte replenishment. In addition to their immune function, bulk RNA-sequencing of macrophages showed enrichment of biological processes associated with extracellular matrix. Proteomics identified collagen IV and laminins in kidney macrophages from wild type mice whilst other extracellular matrix proteins including cathepsins, ANXA2 and LAMP2 were enriched in Col4a5-/- (Alport) mice. A subset of kidney macrophages co-expressed matrix and macrophage transcripts. CONCLUSIONS: We identified CD81 and CX3CR1 positive kidney macrophage subsets with distinct dependence for monocyte replenishment. Multiomic analysis demonstrated that these cells have diverse functions that underscore the importance of macrophages in kidney health and disease.

Age-Related Alterations in Macrophage Distribution and Function Are Associated With Delayed Cutaneous Wound Healing.

Ageing-related delays and dysregulated inflammation in wound healing are well-documented in both human and animal models. However, cellular and molecular changes underlying this impairment in healing progression are not fully understood. In this study, we characterised ageing-associated changes to macrophages in wounds of young and aged mice and investigated transcriptomic differences that may impact the progression of wound healing. Full-thickness wounds created on the dorsum of C57BL/6J young and aged mice were excised on Days 3 and 7 post-wounding for analysis by immunohistochemistry, flow cytometry, and RNA sequencing. Our data revealed that macrophages were significantly reduced in aged wounds in comparison to young. Functional transcriptomic analyses showed that macrophages from aged wounds exhibited significantly reduced expression of cell cycle, DNA replication, and repair pathway genes. Furthermore, we uncovered an elevated pro-inflammatory gene expression program in the aged macrophages correlated with poor inflammation resolution and excessive tissue damage observed in aged wounds. Altogether, our work provides insights into how poorly healing aged wounds are phenotypically defined by the presence of macrophages with reduced proliferative capacity and an exacerbated inflammatory response, both of which are pathways that can be targeted to improve healing in the elderly.

Hematopoietic stem and progenitor cells are present in healthy gingiva tissue.

Hematopoietic stem cells reside in the bone marrow, where they generate the effector cells that drive immune responses. However, in response to inflammation, some hematopoietic stem and progenitor cells (HSPCs) are recruited to tissue sites and undergo extramedullary hematopoiesis. Contrasting with this paradigm, here we show residence and differentiation of HSPCs in healthy gingiva, a key oral barrier in the absence of overt inflammation. We initially defined a population of gingiva monocytes that could be locally maintained; we subsequently identified not only monocyte progenitors but also diverse HSPCs within the gingiva that could give rise to multiple myeloid lineages. Gingiva HSPCs possessed similar differentiation potentials, reconstitution capabilities, and heterogeneity to bone marrow HSPCs. However, gingival HSPCs responded differently to inflammatory insults, responding to oral but not systemic inflammation. Combined, we highlight a novel pathway of myeloid cell development at a healthy barrier, defining a gingiva-specific HSPC network that supports generation of a proportion of the innate immune cells that police this barrier.

Comparación de las estructuras de difusión de información errónea y verídica en las redes sociales durante una emergencia de salud pública / Contrasting Misinformation and Real-Information Dissemination Network Structures on Social Media During a Health Emergency

RESUMEN Objetivos. Elaborar un esquema operativo integral para detectar la información errónea principal sobre el zika distribuida en Twitter® en el 2016; reconstruir las redes por las que se difunde información mediante retuiteo; contrastar la información verídica frente a la errónea con diversos parámetros; e investigar cómo se difundió en las redes sociales la información errónea sobre el zika durante la epidemia. Métodos. Revisamos sistemáticamente los 5 000 tuits más retuiteados con información sobre el zika en inglés, definimos "información errónea" a partir de la evidencia, buscamos tuits que tuvieran información errónea y conformamos un grupo equiparable de tuits con información verídica. Elaboramos un algoritmo para reconstruir las redes de retuiteo de 266 tuits con información errónea y 458 tuits equiparables con información verídica. Calculamos y comparamos nueve parámetros para caracterizar la estructura de las redes a varios niveles, entre los dos grupos. Resultados. En los nueve parámetros se aprecian diferencias estadísticamente significativas entre el grupo de información verídica y el de información errónea. La información errónea en general se difunde mediante estructuras más sofisticadas que la información verídica. También hay una considerable variabilidad intragrupal. Conclusiones. Las redes de difusión de la información errónea sobre el zika en Twitter fueron sustancialmente diferentes que las de información verídica, lo cual indica que la información errónea se sirve de mecanismos de difusión distintos. Nuestro estudio permitirá formar una comprensión más holística de los desafíos que plantea la información errónea sobre salud en las redes sociales.

ABSTRACT Objectives. To provide a comprehensive workflow to identify top influential health misinformation about Zika on Twitter in 2016, reconstruct information dissemination networks of retweeting, contrast mis- from real information on various metrics, and investigate how Zika misinformation proliferated on social media during the Zika epidemic. Methods. We systematically reviewed the top 5000 English-language Zika tweets, established an evidence-based definition of "misinformation," identified misinformation tweets, and matched a comparable group of real-information tweets. We developed an algorithm to reconstruct retweeting networks for 266 misinformation and 458 comparable real-information tweets. We computed and compared 9 network metrics characterizing network structure across various levels between the 2 groups. Results. There were statistically significant differences in all 9 network metrics between real and misinformation groups. Misinformation network structures were generally more sophisticated than those in the real-information group. There was substantial within-group variability, too. Conclusions. Dissemination networks of Zika misinformation differed substantially from real information on Twitter, indicating that misinformation utilized distinct dissemination mechanisms from real information. Our study will lead to a more holistic understanding of health misinformation challenges on social media.

Amphiregulin-producing γδ T cells are vital for safeguarding oral barrier immune homeostasis.

γδ T cells are enriched at barrier sites such as the gut, skin, and lung, where their roles in maintaining barrier integrity are well established. However, how these cells contribute to homeostasis at the gingiva, a key oral barrier and site of the common chronic inflammatory disease periodontitis, has not been explored. Here we demonstrate that the gingiva is policed by γδ T cells with a T cell receptor (TCR) repertoire that diversifies during development. Gingival γδ T cells accumulated rapidly after birth in response to barrier damage, and strikingly, their absence resulted in enhanced pathology in murine models of the oral inflammatory disease periodontitis. Alterations in bacterial communities could not account for the increased disease severity seen in γδ T cell-deficient mice. Instead, gingival γδ T cells produced the wound healing associated cytokine amphiregulin, administration of which rescued the elevated oral pathology of tcrδ-/- mice. Collectively, our results identify γδ T cells as critical constituents of the immuno-surveillance network that safeguard gingival tissue homeostasis.

Direct effects of conjugated linoleic acid isomers on P815 mast cells in vitro.

Conjugated linoleic acid (CLA) is a dietary fatty acid which causes extensive remodeling and mast cell recruitment in the mouse mammary gland. Two CLA isomers, 9,11- and 10,12-CLA, have differing effects in vivo, with only 10,12-CLA increasing mast cell number. The purpose of this project is to test the hypothesis that CLA acts directly on the mast cell. The P815 mastocytoma cell line was assayed for the effects of CLA on mast cell number, proliferation, apoptosis, and differentiation. Both CLA isomers decreased viable mast cell number, with no effect on membrane integrity, or cell cycle distribution. 10,12-CLA induced an increase in apoptosis, assessed by Annexin-FITC binding. Both isomers increased mast cell granularity, and secretion of MMP-9. The complex effects of CLA isomers on mast cells in the mammary gland are distinct from direct effects on mast cells in vitro, and may require interactions between multiple cell types present in vivo.