Figure - uploaded by Nancy A Mcnamara
Content may be subject to copyright.
Macrophage infiltration of the central corneal (cCo) and limbus (Lm) in Aire KO mice. A: Eye tissue sections from Aire KO or WT mice were stained with antibodies directed against F4/80, MOMA-2, or IgG isotype control. F4/80+ cells infiltrating the subepithelial (arrowheads) and stromal (arrow) compartments of the cCo are indicated. The limbal region was identified using the iris insertion (Is) as an anatomical landmark. Results shown are representative of at least four independent tests. B: Quantitative analysis of MOMA-2+ and F4/80+ macrophages in the cornea and limbus of Aire KO and WT mice, shown as mean ± SE cell count. *P < 0.05 and **P < 0.01. C: CD4+ T cells from Aire KO or WT mice were adoptively transferred to immunodeficient SCID recipients. Results shown are representative of four independent tests. Mice were sacrificed at 40 days and eye sections assessed by immunofluorescence using antibodies directed against F4/80, MOMA-2, or isotype control (D). E: Quantitative analysis of corneal MOMA-2+ and F4/80+ macrophages in recipients of autoreactive CD4+ T cells (Aire KO → SCID) or WT CD4+ T cells (Aire WT → SCID), shown as mean ± SE cell number. *P < 0.05.
Source publication
Lymphocytic infiltration of the lacrimal gland and ocular surface in autoimmune diseases such as Sjögren's syndrome (SS) causes an aqueous-deficient dry eye that is associated with significant morbidity. Previous studies from our laboratory and others have established autoimmune regulator (Aire)-deficient mice as a useful model to examine exocrinop...
Contexts in source publication
Context 1
... KO mice spontaneously develop SS-like disease with associated exocrinopathy and corneal epitheliopa- thy as indicated by decreased tear secretion, severe lissamine green staining, and decreased conjunctival goblet cell density. 4,6 Using extracellular and intracellular macrophage markers, F4/80 and MOMA-2, respectively, we revealed an accumulation of macrophages in the limbus and central corneal of Aire KO mice ( Figure 1A), whereas few macrophages were noted in the eyes of Aire-sufficient, WT mice ( Figure 1A). Although F4/80 and MOMA-2 have been reported to detect different macro- phage subpopulations, 14 ...Context 2
... KO mice spontaneously develop SS-like disease with associated exocrinopathy and corneal epitheliopa- thy as indicated by decreased tear secretion, severe lissamine green staining, and decreased conjunctival goblet cell density. 4,6 Using extracellular and intracellular macrophage markers, F4/80 and MOMA-2, respectively, we revealed an accumulation of macrophages in the limbus and central corneal of Aire KO mice ( Figure 1A), whereas few macrophages were noted in the eyes of Aire-sufficient, WT mice ( Figure 1A). Although F4/80 and MOMA-2 have been reported to detect different macro- phage subpopulations, 14 ...Context 3
... Previously, we reported that adoptive transfer of autoreactive CD4 T cells from Aire KO mice to SCID recipients resulted in autoimmune dry eye and SQM in 6 weeks. 3 Consistent with dry eye development, we observed limbal and stro- mal infiltration of F4/80 macrophages after adoptive transfer of autoreactive CD4 T cells ( Figure 1C). Stain- ing with isotype control antibody produced negative re- sults ( Figure 1D). ...Context 4
... Consistent with dry eye development, we observed limbal and stro- mal infiltration of F4/80 macrophages after adoptive transfer of autoreactive CD4 T cells ( Figure 1C). Stain- ing with isotype control antibody produced negative re- sults ( Figure 1D). Quantification of MOMA-2-and F4/80- stained cells revealed that the overall infiltration of macrophages was enhanced after adoptive transfer of autoreactive CD4 T cells ( Figure 1E). ...Context 5
... ing with isotype control antibody produced negative re- sults ( Figure 1D). Quantification of MOMA-2-and F4/80- stained cells revealed that the overall infiltration of macrophages was enhanced after adoptive transfer of autoreactive CD4 T cells ( Figure 1E). MOMA-2 (38 20.41 versus 2 1.44, P 0.05) and F4/80 (32 12.66 versus 4 0.45, P 0.05) cells in the cornea were significantly increased in Aire KO ¡ SCID versus Aire WT ¡ SCID recipients. ...Similar publications
Keratinizing squamous metaplasia (SQM) of the ocular surface is a blinding consequence of systemic autoimmune disease and there is no cure. Ocular SQM is traditionally viewed as an adaptive tissue response during chronic keratoconjunctivitis sicca (KCS) that provokes pathological keratinization of the corneal epithelium and fibrosis of the corneal...
Photorefractive keratectomy (PRK) represents a therapeutic option to remodel corneal stroma and to compensate refractive errors, which involves inflammatory and/or regenerative processes. In this context, the modulation of cytokines/chemokines in the conjunctival sac fluid and their role in the maintenance of the corneal microenvironment during the...
Citations
... As a result, establishing strategies for early diagnosis and effective therapy is critical for treating such diseases.Currently, many researchers are trying to investigate the potential genes involved in the pathogenesis of SS to develop clinical indicators and drug targets that contribute to implementing effective early intervention strategies. Previous studies documented the involvement of pro-in ammatory cytokines, ampli cation of T cells, and cellular super mutation of B cells in SS pathogenesis [6][7][8].The secretion of cytokines related to innate immunity induced the in ammatory hyperactivity observed during SS [9].Thus, the dysregulation of cytokines associated with abnormal immune activation is one of the main causes of SS [10][11][12].Recent studies showed that serum salivary glycochain antigen (MUC1) could be used as a biomarker of CTD-ILD, and its level can re ect the damage to alveolar epithelium and stroma, which is of great signi cance in evaluating the disease and improving the prognosis [13].Numerous studies showed the high expression of MUC1 in the tissue of patients with SS [14][15], implying that MUC1 may be potentially involved in SS-ILD pathogenesis. However, the detailed role of MUC1 in patients with SS-ILD is unclear. ...
Objectives
To clarify if the mechanism of Sanliangsan in improving Sjogren’s syndrome complicated with interstitial lung disease (SS-ILD) involves MUC1 suppression, which is involved in SS-ILD pathogenesis.
Methods
Fifty-six patients were randomly divided into two groups receiving Sanliangsan prescription therapy and conventional therapy. In-depth transcriptome profiles collected and analyzed to identify candidate genes involved in SS pathogenesis. Clinical symptom scores, metabolic compositions, lung HRCT scores, and serum MUC1 levels were compared between the two groups before and after treatment. Metabolome analyzed the metabolic composition of serum with SS-ILD before and after SP treatment.
Results
Transcriptome results identified the involvement of abnormal expression of genes relevant to the immune system, inflammatory responses, and signaling pathways. MUC1 was involved in SS pathogenesis and could be used to diagnose SS-ILD early. The SP therapy improved SS-ILD more effectively than conventional therapy. Moreover, Sanliangsan prescription therapy reduced serum MUC1 levels and restored the abnormal metabolisms, improving the abnormal inflammatory and immune responses of patients. Eugenol directly interacted with MUC1, and suppressed related genes, and was the bioactive compound of SP.
Conclusion
Modified Sanliangsan can improve the clinical symptoms, signs, and lung function of patients; the mechanism may be due to eugenol and related to MUC1 regulation
... Macrophage-mediated inflammatory response, which can result in epithelial cell injuries and/or degeneration, is believed to play a crucial role in DED. 32 Previous studies have shown that mExo can modulate the macrophage immunophenotype by promoting the differentiation of macrophages into particular phenotypes, 11,33 and the effects of AA on the immune response in ocular diseases have also been investigated in a few studies. 34,35 In the current study, LPS-induced mouse peritoneal macrophages were cultured under the different treatments to evaluate the effects of mExo@AA on macrophages. ...
Dry eye disease (DED), a complex ocular surface disease with a high prevalence rate, is associated with corneal injury, excess oxidative stress and inflammation. Current therapeutic strategies, including artificial tears and anti-inflammatory agents, are unable to address all the deleterious factors or to achieve a clinical cure due to their temporary or side effects. Here, we prepared a multiple-functional eyedrop based on the deposition of gold nanoparticles (AuNPs) reduced by ascorbic acid (AA) onto the exosomal phospholipid membrane of mesenchymal stem cell (mExo)-derived exosomes in situ (mExo@AA). The therapeutic value of mExo@AA for DED was demonstrated in a mouse DED model. Combining the benefits of mExo and AA, mExo@AA effectively improves corneal epithelium recovery and anti-inflammation capacity, decreases corneal reactive oxygen species, and restores tear secretion without adverse effects. Thus, this study suggests that mExo@AA is effective and safe as a therapeutic agent for the treatment of DED.
... Interestingly, IL-20 is also detected on the infiltrated F4/80 + macrophage population in the DED corneas, suggesting that IL-20 could be an autocrine or paracrine factor to activate macrophages and result in forming a vicious cycle of the inflammatory responses in DED. Macrophages are crucial immune mediators in inflammatory dry eye disease [14,47,60] and that macrophage depletion was found to protect against DED in an experimental model by reducing proinflammatory cytokine release [16]. Further study to investigate whether IL-20-mediated macrophage activation is crucial during the DED progression is required. ...
Background
Dry eye disease (DED) is a common disease in ophthalmology, affecting millions of people worldwide. Recent studies have shown that inflammation is the core mechanism of DED. IL-20 is a proinflammatory cytokine involved in various inflammatory diseases. Therefore, we aimed to explore the role of this cytokine in the pathogenesis of DED and evaluate the therapeutic potential of the anti-IL-20 monoclonal antibody (mAb) 7E for DED treatment.
Methods
Clinical tear samples from patients with DED and non-DED controls were collected and their IL-20 protein levels were determined. We established three DED animal models to explore the role of IL-20 and the efficacy of IL-20 antibody in DED. Benzalkonium chloride (BAC)-induced over-evaporative DED, extra-orbital lacrimal gland excision (LGE)-induced aqueous tear-deficient DED, and desiccating stress (DS)-induced combined over-evaporative and aqueous tear-deficient DED animal models were established to investigate the role of IL-20. The anti-IL-20 antibody 7E was established to neutralize IL-20 activity. The effects of IL-20 or 7E on human corneal epithelial cells and macrophages under hyperosmotic stress were analyzed. 7E was topically applied to eyes to evaluate the therapeutic effects in the DED animal models.
Results
IL-20 was significantly upregulated in the tears of patients with DED and in the tears and corneas of DED animal models. Under hyperosmotic stress, IL-20 expression was induced via NFAT5 activation in corneal epithelial cells. 7E suppressed hyperosmotic stress-induced activation of macrophages. IL-20 induced cell death in corneal epithelial cells and 7E protected cells from hyperosmotic stress-induced cell death. Blocking IL-20 signaling with 7E protected mice from BAC-induced, LGE-induced, and DS-induced DED by reducing DED symptoms and inhibiting inflammatory responses, macrophage infiltration, apoptosis, and Th17 populations in the conjunctiva and draining lymph nodes.
Conclusions
Our results demonstrated the functions of IL-20 in DED and presented a potential therapeutic option for this condition.
Graphical Abstract
... To localize and eventually identify the cellular compartments contributing to these changes, we calculated the score corresponding to the expression of genes involved in these pathways for each spot (Figure 8, Supplemental Figure 9). The most significant pathway enriched in upregulated DEGs was the "TYROBP Causal Network", for which we observed a higher score in the NOD.H-2b LG samples ( Figures 8A, B), particularly around and within infiltration foci ( Figure 8B, black arrows) where the majority of activated macrophages are present (65,66). The Violin plots ( Figures 8B', B'') confirms the results of bulk RNAseq and shows that the "TYROBP Causal Network" pathway is highly enriched in the cluster 4 (B and T cells). ...
The lacrimal gland (LG) is an exocrine gland that produces the watery part of the tear film that lubricates the ocular surface. Chronic inflammation, such as Sjögren’s syndrome (SS), is one of the leading causes of aqueous-deficiency dry eye (ADDE) disease worldwide. In this study we analyzed the chronic inflammation in the LGs of the NOD.B10Sn-H2b/J ( NOD.H-2b ) mice, a mouse model of SS, utilizing bulk RNAseq and Visium spatial gene expression. With Seurat we performed unsupervised clustering and analyzed the spatial cell distribution and gene expression changes in all cell clusters within the LG sections. Moreover, for the first time, we analyzed and validated specific pathways defined by bulk RNAseq using Visium technology to determine activation of these pathways within the LG sections. This analysis suggests that altered metabolism and the hallmarks of inflammatory responses from both epithelial and immune cells drive inflammation. The most significant pathway enriched in upregulated DEGs was the “TYROBP Causal Network”, that has not been described previously in SS. We also noted a significant decrease in lipid metabolism in the LG of the NOD.H-2b mice. Our data suggests that modulation of these pathways can provide a therapeutic strategy to treat ADDE.
... some immune cells in T1DM inflammation were preliminarily studied. During the development of T1DM, many organs were infiltrated by immune cells, and CD4+ T cells promote the activation and infiltration of macrophages aggravating the inflammatory state [26][27][28] , so the spleen, pancreas and kidney of mice were stained by immunohistochemistry, and the degree of infiltration of CD4 + T cells and macrophages was observed in the DM Group and normal group. CD4 + T cells play an important role in the occurrence and development of T1DM, and can mediate the destruction of pancreatic β cells, thus inducing the occurrence of type 1 diabetes 29,30 . ...
Immune cells play an important role in the development of inflammation in type 1 diabetes mellitus, so we want to explore the changes of CD4 ⁺ T cells and macrophages in vivo, which can provide an experimental basis for immunotherapy based on CD4 ⁺ T cells and macrophages. The intraperitoneal injection of streptozocin was used to induce a type 1 diabetes mellitus mouse model; the blood glucose, body weight, and the expression of inflammatory factors in the kidney were measured. Immunohistochemistry was applied to determine and analyze the infiltration of CD4 ⁺ T cells and macrophages in the spleen, pancreas, and kidney. The subtypes of macrophages in the kidney and CD4 ⁺ T cells in the spleen were analyzed by flow cytometry. Our study suggests that CD4 ⁺ T cells and macrophages increase, while the inflammatory immune response system is activated in the development of T1DM. CD4 ⁺ T cells positively correlated with macrophages in the pancreas and kidney of T1DM. CD4 ⁺ T cells turn to pro-inflammatory subtypes in the spleen of T1DM, while macrophages turn to pro-inflammatory subtypes in the kidney of T1DM. Therefore, regulation of CD4 ⁺ T cells and macrophages may be a potential target for T1DM and kidney complications.
... Activation of APCs, migration to eye-draining lymph nodes, priming of Th1 and Th17 cells that migrate to the ocular surface epithelium and secretion of IFN-γ and IL-17, causing further insult to the ocular surface, breaking the corneal barrier, and activating the epithelium, reinitiating the vicious circle and becoming self-perpetuating (Fig. 4) 229,230 . Interruption of APC migration, achieved by subconjunctival administration of liposome-encapsulated clodronate (which depletes APCs), or lymphadenectomy of eye-draining nodes will decrease the generation of eye-pathogenic T cells 231,232 . The subject has recently been extensively reviewed-the interested reader is referred to reference 233 for a comprehensive review of dry eye pathophysiology. ...
... There is a complex relationship between APCs and conjunctival goblet cells 264 . Goblet cells and their products not only act to mechanically lubricate the eye, but also actively contribute to immune homeostasis at the ocular surface by secreting TGF-β and retinoic acid, as mentioned before 29,30,58,62,[231][232][233]240,243,265,266 . Studies have shown that experimental dry eye changes the polarization of conjunctival macrophages from M2 to M1 267,268 . ...
The eye is a sensory organ exposed to the environment and protected by a mucosal tissue barrier. While it shares a number of features with other mucosal tissues, the ocular mucosal system, composed of the conjunctiva, Meibomian glands, and lacrimal glands, is specialized to address the unique needs of (a) lubrication and (b) host defense of the ocular surface. Not surprisingly, most challenges, physical and immunological, to the homeostasis of the eye fall into those two categories. Dry eye, a dysfunction of the lacrimal glands and/or Meibomian glands, which can both cause, or arise from, sensory defects, including those caused by corneal herpes virus infection, serve as examples of these perturbations and will be discussed ahead. To preserve vision, dense neuronal and immune networks sense various stimuli and orchestrate responses, which must be tightly controlled to provide protection, while simultaneously minimizing collateral damage. All this happens against the backdrop of, and can be modified by, the microorganisms that colonize the ocular mucosa long term, or that are simply transient passengers introduced from the environment. This review will attempt to synthesize the existing knowledge and develop trends in the study of the unique mucosal and immune elements of the ocular surface.
... cells and cellular super mutation of B cells, facilitating salivary gland inflammation. [5][6][7][8] Innate immune-inflammatory responses relevant are activated by microbial components and toxins that promote the secretion of IL-1 and IL-18 which induce the inflammatory hyperactivity observed during pSS. 9 The dysregulation of cytokines secreted by different cell subsets is the main cause of pSS. ...
Objective:
The objective of this study is to investigate the expression levels of Th9 CD4+ T cells and IL-9 secretion in peripheral blood mononuclear cells of patients with primary Sjogren's syndrome. Further, this study aimed to investigate the role of Th9 cells in the occurrence and development of pSS.
Methods:
A total of 20 pSS patients and 20 healthy people, matched with age and gender, were selected as the experimental and control group, respectively. Flow cytometry and ELISA were used to detect the expression of Th9 cytokines in peripheral blood mononuclear cells and IL-9 in serum, respectively. These factors were then correlated to other clinical indicators.
Results:
The levels of Th9 CD4+ T cells and IL-9 of pSS patients were significantly higher than those of the control group. Th9 CD4+ T cells and IL-9 levels in peripheral blood of pSS patients were negatively correlated with salivary flow rate, while IL-9 level was positively correlated with globulin. The transcription levels of IL-9 and immune-related genes including IL-4, IL-7, IL-17, SMAD3, STAT5 and JAK3 were dramatically increased in serum of pSS patients.
Conclusion:
The expression levels of Th9 in peripheral blood and serum IL-9 of patients with pSS were significantly increased, which were correlated with clinical immunological indexes. Together, these data suggest that Th9 cells and IL-9 may be involved in the pathogenesis of pSS.
... We next tested whether Lacripep restored basal cell identity and barrier function by reducing immune cell infiltration. Prolonged desiccating stress from reduced tear production elicits an immune response in which activated CD4+ T cells home to the corneal epithelium and stroma (primarily accumulating at the limbal region) to modulate epithelial differentiation (Pflugfelder et al., 2008) and direct the development of autoimmunemediated dry eye (Zhou et al., 2012). Intriguingly, at day 15 we found all Aire KO corneas ( Figures 1I and 1J), as well as their corresponding lacrimal glands (Figures S1A and S1B), regardless of treatment, to be extensively infiltrated by CD4+ T cells. ...
Corneal architecture is essential for vision and is greatly perturbed by the absence of tears due to the highly prevalent disorder dry eye. With no regenerative therapies available, pathological alterations of the ocular surface in response to dryness, including persistent epithelial defects and poor wound healing, result in lifelong morbidity. Here, using a mouse model of aqueous-deficient dry eye, we reveal that topical application of the synthetic tear protein Lacripep reverses the pathological outcomes of dry eye through restoring the extensive network of corneal nerves that are essential for tear secretion, barrier function, epithelial homeostasis, and wound healing. Intriguingly, the restorative effects of Lacripep occur despite extensive immune cell infiltration, suggesting tissue reinnervation and regeneration can be achieved under chronic inflammatory conditions. In summary, our data highlight Lacripep as a first-in-class regenerative therapy for returning the cornea to a near homeostatic state in individuals who suffer from dry eye.
... Macrophage tissue infiltration is a known phenomenon in most autoimmune diseases including SSc [16][17][18][19][20][21][22][23][24]. Accumulating evidence has revealed a crucial role of innate immune reactions in driving not only disease flare-ups (causing progression) but also contributing to the ignition processes in such diseases [25,26]. ...
Systemic Sclerosis (SSc) is a multiphase autoimmune disease with a well-known triad of clinical manifestations including vasculopathy, inflammation and fibrosis. Although a plethora of drugs has been suggested as potential candidates to halt SSc progression, nothing has proven clinically efficient. In SSc, both innate and adaptive immune systems are abnormally activated fuelling fibrosis of the skin and other vital organs. Macrophages have been implicated in the pathogenesis of SSc and are thought to be a major source of immune dysregulation. Due to their plasticity, macrophages can initiate and sustain chronic inflammation when classically activated while, simultaneously or parallelly, when alternatively activated they are also capable of secreting fibrotic factors. Here, we briefly explain the polarization process of macrophages. Subsequently, we link the activation of macrophages and monocytes to the molecular pathology of SSc, and illustrate the interplay between macrophages and fibroblasts. Finally, we present recent/near-future clinical trials and discuss novel targets related to macrophages/monocytes activation in SSc.
... Sjögren's syndrome (SS) dry eye is an intractable autoimmune disorder, characterized by lymphoid and myeloid cell infiltration of the lacrimal glands (LGs) and subsequent tissue damage, reflected clinically by ocular discomfort and even visual loss (1)(2)(3). Although the etiology of SS remains ill defined, available evidence have pointed out that macrophages play an important role both during the onset and resolution of inflammation in SS dry eye (4)(5)(6). ...
... Macrophages are important regulators in inflammation of LGs (5,28). To clarify its underlying mechanism of how hUC-MSC-sEVs prevented the rabbit autoimmune dacryoadenitis, we investigated the effects of hUC-MSC-sEVs on macrophages accumulated in inflamed LGs. ...
Background:
Mesenchymal stem cell-derived small extracellular vesicles (MSC-sEVs) have been increasingly proved as promising immunomodulators against some autoimmune disorders. However, the possible effect and the underlying mechanism of MSC-sEVs in autoimmune dry eye have been rarely studied.
Methods:
Small extracellular vesicles from human umbilical cord mesenchymal stem cells (hUC-MSC-sEVs) were subconjunctivally injected to rabbit dry eye model, and their preventive or therapeutical effects were assessed by recording the clinical and histological scores. Quantitative real-time PCR (Q-PCR), western blot and flow cytometry were performed to evaluate the immunomodulatory effects of hUC-MSC-sEVs on macrophages and T regulatory cells (Tregs) both in vivo and in vitro, and the in vitro T cell proliferation was detected by Bromodeoxyuridine (BrdU) assay. In addition, high expression of miR-100-5p in hUC-MSC-sEVs was identified by Q-PCR, and the functional role of sEVs-miR-100-5p on macrophages was explored by a series of co-culture experiments using sEVs derived from hUC-MSCs transfected with miR-100-5p inhibitor.
Results:
We firstly demonstrated that hUC-MSC-sEVs had the preventive and therapeutical effects on rabbit autoimmune dacryoadenitis, an animal model of Sjögren's syndrome (SS) dry eye. Further investigation revealed that hUC-MSC-sEVs administration effectively elicited macrophages into an anti-inflammatory M2 phenotype and elevated the proportion of Tregs both in vivo and in vitro, which contributed to reduced inflammation and improved tissue damage. Importantly, hUC-MSC-sEVs-educated macrophages with M2-like phenotype exhibited strong capacity to inhibit CD4+ T cell proliferation and promote Treg generation in vitro. Mechanistically, miR-100-5p was highly enriched in hUC-MSC-sEVs, and knockdown of miR-100-5p in hUC-MSC-sEVs partially blunted the promotion of hUC-MSC-sEVs on M2 macrophage polarization and even attenuated the effect of hUC-MSC-sEVs-educated macrophages on T cell suppression and Treg expansion.
Conclusion:
Our data indicated that hUC-MSC-sEVs alleviated autoimmune dacryoadenitis by promoting M2 macrophage polarization and Treg generation possibly through shuttling miR-100-5p. This study sheds new light on the application of MSC-sEVs as a promising therapeutic method for SS dry eye.
