Provide a reproducible method for culturing confluent monolayers of hfRPE cells that exhibit morphology, physiology, polarity, and protein expression patterns similar to native tissue.Human fetal eyes were dissected on arrival, RPE cell sheets mechanically separated from the choroid cultured in specifically designed medium comprised entirely commercially available components. Physiology experiments performed with previously described techniques. Standard techniques used immunohistochemistry,...

Abstract Th1 and Th17 cells are characterized by their expression of IFN-γ or IL-17, respectively. The finding Th producing both IL-17 suggested, however, that certain may modify selective cytokine expression. In this study, we examined changes in an experimental system which polarized specific against hen egg lysozyme induce ocular inflammation recipient mice expressing eyes. Whereas only was expressed eyes mice, substantial proportions donor possibility nonpolarized preparations were...

Recently reported lines of Th9 cells, producing IL-9 and IL-10, were generated by polarization with IL-4 TGF-β activation Abs against CD3 CD28. In this paper, we analyzed features similarly polarized but activated the "natural mode" (i.e., exposure CD4 cells to their target Ag, hen egg lysozyme [HEL] APCs). Main observations are following: 1) both IL-10 expressed line strikingly different kinetics, being produced rapidly, reaching a peak on day 3 in culture declining sharply thereafter,...

purpose. Proinflammatory cytokines in degenerative diseases can lead to the loss of normal physiology and destruction surrounding tissues. In present study, physiological responses human fetal retinal pigment epithelia (hfRPE) were examined vitro after polarized activation proinflammatory cytokine receptors. methods. Primary cultures hfRPE stimulated with an inflammatory mixture (ICM): interleukin (IL)-1β, tumor necrosis factor (TNF)-α, interferon (IFN)-γ. Western blot analysis...

Inflammation and oxidative stress are involved in age-related macular degeneration (AMD) possibly associated with an activation of neuronal apoptosis inhibitor protein/class II transcription activator the Major Histocompatibility Complex (MHC)/heterokaryon incompatibility/telomerase-associated protein 1, leucine-rich repeat or nucleotide-binding domain, repeat-containing family, pyrin domain-containing 3 (NLRP3) inflammasome. In present study, we used a translational approach to address this...

Regulatory B cells (Breg cells) that secrete IL-10 or IL-35 (i35-Breg) play key roles in regulating immunity tumor microenvironment during autoimmune and infectious diseases. Thus, loss of Breg function is implicated development diseases while aberrant elevation prevents sterilizing immunity, exacerbates diseases, promotes cancer metastasis. identified thus far are largely antigen-specific derive mainly from B2-lymphocyte lineage. Here, we describe an innate-like IL-27–producing natural...

Abstract Müller cell is the most common type of glial in human and mouse retina, playing a crucial role maintaining retinal homeostasis. In addition to providing structural support cells can also supply trophic substances neurons, remove metabolic waste, mitigate oxidative stress, promote synaptic activities. However, many roles remain largely unknown, particularly for those inflamed retina. this article, we reanalyzed single RNA-seq (scRNA-seq) dataset from Aire-/- mice, which exhibits...

IAN is an R package that addresses the challenge of integrating, analyzing and interpreting high-throughput "omics" data, using a multi-agent artificial intelligence (AI) system. leverages popular pathway regulatory datasets (KEGG, WikiPathways, Reactome, GO, ChEA) STRING database for protein-protein interactions to perform standard enrichment analysis. The individual results are then used generate insightful summaries, each datasets, large language model (LLM) through architecture. These...

The role of Th17 lymphocytes in immunopathogenic processes has been well established, but little is known about their basic cell features. In this study, we compared polarized Th1 and for key biological activities related to pathogenicity trafficking. lineages were derived from TCR-transgenic CD4 murine cells specific against hen egg lysozyme. When adoptively transferred into mice expressing lysozyme eyes, both induced ocular inflammation with slight differences histological pathology. PCR...

Both Th1 and Th17 T cell subsets can mediate inflammation, but the kinetics of pathogenic processes mediated by these two have not been investigated. Using an experimental system in which TCR-transgenic or cells specific for hen egg lysozyme induce ocular inflammation recipient mice expressing eye-restricted lysozyme, we found important differences vivo behavior subsets. initially proliferated considerably faster invaded eye more quickly than their counterparts, then disappeared rapidly. By...

Ligands for aryl hydrocarbon receptor (AHR), such as dioxins, are highly toxic. One ligand, TCDD, was found to exert potent immunosuppressive capacities in mice developing pathogenic autoimmune processes, including EAU, but its toxicity makes it unusable humans. A recently identified endogenous AHR ITE, is also immunosuppressive, nontoxic and could therefore be useful therapy Here, we tested ITE capacity inhibit EAU related immune responses.

The human facilitative transporter Glut1 is the major glucose present in all cells, has a central role metabolism, and an archetype of superfamily protein facilitators. Here we describe three-dimensional structure based on helical packing schemes proposed for lactose permease predictions secondary structure, refined using energy minimization, molecular dynamics simulations, quality environmental scores. Ramachandran scores stereochemical obtained were as good those known structures KcsA...

Microbial products are assumed to play a major role in triggering pathogenic autoimmunity. Recently accumulated data have shown that these stimulate the immune system by interacting with TLRs, expressed on APCs. To examine capacity of various TLR ligands trigger autoimmunity, we used which naive CD4 cells, specific against hen egg lysozyme (HEL), injected into recipient mice expressing HEL their eyes. Only when stimulated, cells acquire and induce ocular inflammation. Seven were tested this...

The interaction between TLRs and their cognate ligands triggers both the innate adaptive immune systems, thus can play a pivotal role in defense against pathogen invasion. This work investigates differentiation of naive CD4 cells into Th1 or Th17 phenotypes mice treated with different TLR ligands. We use model system which transgenic specific to hen egg lysozyme are adoptively transferred recipients that express lens eye. T induce ocular inflammation only Treatment LPS preferentially...

Background Mutations in LRRK2 are related to certain forms of Parkinson's disease and, possibly, the pathogenesis Crohn's disease. In both these diseases inflammatory processes participate pathogenic process. is expressed lymphoid cells interestingly, Lrrk2 (-/-) mice were reported develop more severe experimental colitis than their wild type (WT) controls. Here, we examined possible involvement autoimmune uveitis (EAU), an animal model for human uveitis, by testing capacity this eye and...

Summary Synthetic oligodeoxynucleotides (ODN) expressing ‘suppressive’ TTAGGG motifs down-regulate a variety of proinflammatory and T helper type 1 (Th1)-mediated pathological immune responses. The ability the archetypal suppressive ODN A151 to inhibit ocular inflammation was examined in two murine models: experimental autoimmune uveitis, induced by immunization with retinal antigen, interphotoreceptor retinoid-binding protein (IRBP) adoptively transferred inflammation, transferring Th1...

Purpose: Digoxin, a major medication for heart disease, was recently reported to have immunosuppressive capacity. Here, we determined the capacity of digoxin on development experimental autoimmune uveitis (EAU) and related immune responses. Methods: The B10.A mice were immunized with interphotoreceptor retinoid-binding protein (IRBP) treated daily or vehicle control. On postimmunization day 14, mouse eyes examined histologically, while spleen cells tested cytokine production in response IRBP...

Abstract Experimental autoimmune uveitis (EAU), an animal model for severe intraocular inflammatory eye diseases, is mediated by both Th1 and Th17 cells. Here, we examined the capacity of TMP778, a selective inhibitor RORγt, to inhibit development EAU, as well related immune responses. EAU was induced in B10.A mice immunization with interphotoreceptor retinoid‐binding protein (IRBP). Treatment TMP778 significantly inhibited determined histological examination. In addition, treatment...

Subpopulations of pathogenic or nonpathogenic Th17 cells were reported to develop when presensitized CD4 activated with their target Ag during polarization by either IL-23 IL-6 and TGF-β, respectively. In this study, we generated two subpopulations using a system in which naive from TCR transgenic mice specific hen egg lysozyme (HEL) are polarized IL-6/TGF-β and, concurrently, HEL presented APCs, anti-CD3/CD28 Abs. Only the former pathogenic, inducing inflammation eyes expressing HEL. Naive...

<i>Purpose:</i> Microbial products stimulate the immune system by interacting with Toll-like receptors (TLR) on antigen-presenting cells. This study examined hypothesis that microbial products, which function as TLR ligands, are playing a major role in triggering pathogenic autoimmunity. <i>Methods:</i> An experimental was developed ligands were tested vivo for their capacity to naïve CD4 cells specific against hen egg lysozyme (HEL) become effector capable of...