bluuebunny/crossref_metadata_2025_split huggingface dataset:
Keratinocyte-derived TGF-β is dispensable in skin autoimmunity in mice
Authors: YANG YI , Zenke Yukari , Hirai Toshiro , Kaplan Daniel H. 
Published: January 2023

Abstract
Transforming growth factor beta 1 (TGF-β) required to suppress spontaneous autoimmunity as global deletion of TGF-β resulted in severe multi-organ autoimmunity in mice. Some TGF-β dificient mice showed skin phenotype. However, the effect of TGF-β on the autoimmunity in skin is not clear. In the epidermis of the skin, keratinocytes(KCs) are the major source of TGF-β. To test whether KCs are an obligate source of TGF-β, we generated K14-Cre-ERT2 TGF-βfl/fl mice. Topical application of tamoxifen could result in efficient deletion of TGF-β. We noted increase epidermal thickness but failed to observe evidence of cutaneous inflammation as measured by histology and flow cytometry. Notably, numbers of regulatory T cells were unaffected by the absence of KC-derived TGF-β. Since αvβ6- and αvβ8-integrins (Itgb6 and Itgb8) are principal source of activation of TGF-β in skin and then we generated Itgb6−/− Itgb8ΔKC. Similarly, we did not observe the development of spontaneous inflammation in Itgb6−/− Itgb8ΔKC mice which lack KC-derived TGFβ-activating integrins in the epidermis. From these data we concluded that KC-derived TGF-β and epidermal activation of TGF-β are not required to suppress spontaneous inflammation.
Regulation of FOXP3 Expression by c-Rel O-GlcNAcylation
Authors: de Jesus Tristan J , Tomalka Jeffery , Shukla Sudhanshu , Staback Franklin , Agarwal Ruchira , Centore Joshua , Ramakrishnan Parameswaran 
Published: January 2023

Abstract
The NF-kappaB protein c-Rel plays a critical role in controlling autoimmunity through regulating T cell function and T regulatory cell development. Type 1 diabetes is a T cell-mediated autoimmune disease associated with hyperglycemia and chronic inflammatory complications. Previously, we discovered that hyperglycemia induces hyper O-GlcNAcylation of c-Rel at serine residue 350 and enhances the transcription of c-Rel-dependent pro-autoimmune cytokines interleukin-2 (IL-2), interferon gamma (IFNG) and granulocyte macrophage colony-stimulating factor (GM-CSF). In contrast, our recent results show that c-Rel O-GlcNAcylation decreases forkhead boxP3 (FOXP3) expression in T cells. Chemically enhancing cellular O-GlcNAcylation decreases the DNA binding ability of c-Rel at the FOXP3 promoter as demonstrated both by oligonucleotide pulldown and chromatin immunoprecipitation assays. Moreover, mutation of serine 350 in c-Rel to alanine augments T cell receptor-induced FOXP3 expression. O-GlcNAcylation-dependent suppression of FOXP3 was also found in vivo in streptozotocin-induced diabetic mouse model. This study reveals a novel molecular mechanism that regulates hyperglycemia-dependent c-Rel function in autoimmune diabetes, with potential to develop novel therapeutics targeting c-Rel O-GlcNAcylation.
Activation of naive CD4+CD25+FOXP3+Treg by specific autoantigen and Th2 cytokines IL-4 and IL-5 induces autoantigen specific Treg that inhibit EAE
Authors: Hall Bruce Milne , Tran Giang , Verma Nirupama Darshan , Robinson Catherine Margaret , Wilcox Paul , Hodgkinson Suzanne Jean 
Published: January 2023

Abstract
CD4+CD25+FOXP3+Treg (tTreg) protect against induction of autoimmunity, but fresh naïve or ex vivo expanded tTreg have limited ability to control established autoimmunity. We have examined pathways of naïve tTreg activation by antigen and Th2 cytokines IL-4 and IL-5. First, IL-4 and antigen induce Ts2 cells that express IL-5Ralpha. In a second step, IL-5, not IL-4, and antigen induce Ts2 cells to Th2-like Treg.
EAE was induced in Lewis rats by immunization with myelin basic protein (MBP) in Freund’s complete adjuvant. Naïve CD4+CD25+Treg cultured with MBP and IL-4 for 4d produced Ts2 cells. To produce Th2-like Treg, Ts2 cells were cultured with IL-5 and MBP for 4d. Cells were assessed by FACS, RT-PCR and ability to suppress EAE.
5×106 Ts2 or 1×106 Th2-like Treg were given ivi 9d post-immunization. Ts2 cells activated by MBP, but not those activated by renal tubular antigen, suppressed EAE between 11 and 16d post-immunization. Clinical score: 0.1±0.5(n=6) in treated vs 1.5±2.7(n=13) in controls at 12d (p&lt;0.01); 1.3±0.7 vs 2.5±0.8 at 13d (p&lt;0.001). Onset of EAE was delayed, peak disease score reduced, as was weight loss. Naïve tTreg had no effect on EAE.
Th2-like Treg had no effect on the onset until 14d when there was rapid recovery with no weakness from 15–21d. Controls did not recover until 22d.
The Ts2 and Th2-like Treg were 99% CD4+, 98% CD25+ and 70–80% FOXP3+. Naïve tTreg expressed no il5ra whereas Ts2 and Th2-like Treg expressed Il5ra. Th2-like Treg had higher Foxp3, irf4 and gata3.
Ex vivo culture of naïve tTreg with IL-4 and autoantigen induces antigen-specific Treg that can control established autoimmune responses. These Ts2 cells further activated by culture with IL-5 and MBP also control autoimmunity. Both cells may have therapeutic potential.
sCD83 alleviates experimental autoimmune uveitis through disrupting the regulation of Rab1a/LRRK2 on F-actin rearrangements
Authors: Lin Wei 
Published: January 2023

Abstract
A soluble form of CD83, generated from membrane CD83 by splice variants or by shedding, inhibites T-cell proliferation to regulatory the immune response in many autoimmune disease and organ-rejection treatment. However, the role of sCD83 in the autoimmune uveitis is still not clear. In our study, we show that sCD83 alleviates experimental autoimmune uveitis (EAU) through a novel mechanism. During onset and recovery of EAU, the level of sCD83 rises in the serum and aqueous humor. Systemic or topical application of sCD83 application alleviates the symptom of EAU and decreases the infiltrated inflammatory cells and cytokines. Mechanistically, sCD83 induces tolerogenic DCs by decreasing the synaptic expression of co-stimulatory molecules and hampering the cell contact between DCs and T cells. sCD83 disruptes the cytoskeletal rearrangements at the DC-T cell contact zone, leading to alter localization of co-stimulatory molecules and suppress T-cell activation. Furthermore, sCD83 is proved to enter into DCs through caveolin-dependent endocytic path-way. And then, sCD83 can bind with Rab1a to inhibit Rab1a-GTP activation, which further influence the regulation of LRRK2 on F-actin rearrangements. This pathway is different from the sCD83/TLR4/MD2 path-way. Thus, the ability of sCD83 to modulate DC-mediated inflammation in the eye could be harnessed to develop new immunosuppressive therapeutics for autoimmune uveitis.
The cellular basis of the PD-1/PD-L1 regulatory pathway in diabetic autoimmunity in the NOD mouse
Authors: Hu Hao , Wan Xiaoxiao , Zakharov Pavel , Unanue Emil R 
Published: January 2023

Abstract
We report our studies on the effects of checkpoint blockade on the development of diabetic autoimmunity in the NOD mouse. One of the complications of checkpoint blockade therapy in cancer patients is the development of acute type 1 diabetes (T1D). We confirm that the PD-1/PD-L1 regulatory pathway is highly involved in T1D in the NOD mouse. In NOD autoimmunity there is an entry of CD4+ T cells specific to insulin peptides shortly after weaning followed by CD103+ dendritic cells (DC), CD4+ T cells and CD8+ T cells recognizing broader islet antigens into the pancreatic islets. The islet resident macrophages and infiltrated DCs, as well as the endothelium and the beta cells, express PD-L1. Diabetes doesn’t develop in NOD mice until about 20 weeks of age or later. Administration of anti-PD-1 antibody rapidly induces diabetes in the young mice showing a control phase established very early in the autoimmune process. Anti-PD-1 treatment induces a high level of islet leukocytes infiltration including CD4+ T cells and CD8+ T cells. The regulatory effect of PD-1 is involved in both CD4+ T cells which recognize self-peptides via H2g7MHC haplotype, as well as in CD8+ T cells. The islet resident macrophage is an essential component for the PD-1/PD-L1 regulatory pathway to take effect. In their absence, no diabetes develops even after anti-PD-1 treatment. The same observation is found in the NOD.Batf-3−/− mice. We suggest that the control mechanism takes place once T cells enter islets. To establish PD-1 regulation, IFNγ signaling is required probably via up-regulating PD-L1 expression on various types of cells in islets. Together, our study has explored the cellular basis of PD-1 in regulating diabetic autoimmunity in the NOD mouse.
Essential role of IL-17A in Tregs induction in autoimmune uveitis
Authors: Chong Wai Po , Zhong Yajie , Mattapallil Mary , Chen Jun , Caspi Rachel R 
Published: January 2023

Abstract
The signature cytokine of Th17 cells, IL-17A, is a pro-inflammatory cytokine associated with various autoimmune diseases in patients and in animal models. However, clinical trials targeting IL-17A in autoimmune disease have been disappointing. We found that Foxp3+ T regulatory cells (Tregs), express both IL17A receptor subunits, i.e. IL17RA and IL17RC., We therefore hypothesized that IL-17A may regulate the induction and/or function of Tregs. Recombinant IL-17A significantly increased the in-vitro polarization of T cells into Tregs. We next investigated the number of Tregs in WT and IL-17A−/− mice that were immunized for experimental autoimmune uveitis. Although there was no significant difference in Treg frequency between WT and IL-17A−/− mice before disease onset, IL-17A− /− mice failed to induce Tregs as efficiently as WT mice when disease progressed. Interestingly, IL-17A−/− mice developed similar disease severity when compared to WT mice, which may in part be due to an impairment in Treg induction. Our data suggest that IL-17A is involved in Tregs induction and/or maintenance, and that this has a role in controlling the (auto)immune response.
Type-1 interferons inhibition of Interleukin-10 signaling in T cells in Type 1 Diabetes development
Authors: Lozano Marcos Iglesias , Chicco Maria , Bibicheff Darrel , Brandacher Gerald , Raimondi Giorgio 
Published: January 2023

Abstract
Type 1 Diabetes (T1D) is caused by T cell-mediated destruction of pancreatic beta cells. The “fertile field” hypothesis of T1D proposes that, in a genetically predisposed individual, an inflamed environment prompts the organ towards autoimmunity. In this context, the protective role of the cytokine IL-10 remains unclear and somewhat controversial. Here we tested whether chronic and localized accumulation of type-I interferons (TI-IFNs) creates a “fertile field” that releases diabetogenic T cells from IL-10 mediated regulation and prompts T1D development. In vitro incubation of mouse T cells with IFN-b induces a dramatic defect in the production of phospho-STAT3 in response to IL-10 (but not in response to IL-6). This alteration was NOT caused by reduction of IL-10 receptor expression, nor induction of SOCS-1 or 3. Instead, microarray analysis showed that IFN-b increases STAT1 levels causing a reversal of the STAT1/STAT3 protein ratio in T cells, suggesting a novel role for this transcription factor. In agreement, STAT1-KO T cells exposed to IFN-b did not show any defect in IL-10 signaling. In diabetes-prone NOD mice we discovered a persistent reduction of IL-10 signaling in T cells of pancreatic and mesenteric lymph nodes (but not in T cells of other lymphoid tissues), correlating with the reported aberrant and localized accumulation of TI-IFNs in these mice. Early treatment of NOD mice with an IFNRA blocking antibody (that delays the onset and reduces the incidence of T1D) showed a clear restoration of IL-10 signaling.
Overall, these data reveal a previously unknown molecular interplay between IL-10 and TI-IFNs that, if thoroughly elucidated, could reveal a novel target of intervention to enhance the efficacy of T1D immunotherapies.
Protein kinase C activates multiple pathways to induce dendritic cell differentiation
Authors: Chavel Colin A , Lee Kelvin P 
Published: January 2023

Abstract
Dendritic cell (DC) differentiation is a complex process that requires multiple signaling pathways to induce this response. Tumor derived factors can block this process, and one way in which this occurs is through repression of the protein kinase C (PKC) isoform PKCβ at the gene level. However, the role PKCβ plays in DC differentiation has yet to be fully characterized. To address this question, we used the K562 cell line, which our lab and others has shown to be capable of differentiating into DCs with PMA. Inhibition of either the ERK or NFκB pathways in the presence of PMA led to DCs that were unable to activate T-cells. Upon further examination of either pathway, we uncovered novel interactions for PKC signaling and ERK or NFκB during the early stages of DC differentiation. Multiple PKCs induced C-Raf activation, in part through RKIP repression. PKC signaling also induced changes in multiple ERK targets, including degradation of Foxo3a, an interaction not shown previously in DCs. PKCβ was the sole PKC isoform required for canonical NFκB signaling through a mechanism involving the CARD11 complex and PKCβ was required for CARD11 complex formation. However, the IKK kinase TAK1 was not required for downstream NFκB signaling, as opposed to its established role in B-cell and T-cell receptor induced NFκB signaling, indicating an alternative mechanism by which PKCβ induces NFκB signaling. PKCβ was also required for non-canonical NFκB signaling outside of the established TRAF and cIAP cascade. We also found crosstalk between the ERK and NFκB pathways at both protein-protein interaction and protein expression levels. Taken together, these results indicate PKC as an important signaling node for DC differentiation.
IL-9-secreting tissue-resident memory CD4+ T cells contribute to allergic airway recall responses
Authors: Ulrich Benjamin Joseph , Fu Yongyao , Koh Byunghee , Kharwadkar Rakshin , Kaplan Mark H 
Published: January 2023

Abstract
Asthma is a chronic intermittent inflammatory lung disease for which there is no cure. Allergen-specific CD4 T helper (Th) cells that secrete cytokines including IL-4, IL-13, and IL-9 mediate asthma pathogenesis. In a six week chronic model of allergic airway disease, we were able to detect a robust IL-9+CD4+ T cell population in the lungs. Interestingly, the in vivo derived IL-9+CD4+ T cells had a multi-cytokine profile including the production of IL-5, IL-13, and IL-10. IL-9-reporter-positive cells demonstrated high gene expression levels of innate cytokine receptors Il1rl1, Il17rb, and Crlf2 and the transcription factor Gata3. IL-9+CD4+ T cells were found to be tissue-resident through intravenous labeling and had CD4+ tissue-resident memory (Trm) cell markers CD69 and CD11a. To determine the Trm phenotype of lung resident IL-9+ T cells, we developed a memory model using a five week period of rest after six weeks of chronic challenge, before a recall allergen challenge. In the memory model, IL-9+ T cells maintained tissue residency and demonstrated increased cell frequency and IL-9 production, characteristic of a secondary response. Antibody blockade of IL-9 immediately prior to the recall challenge significantly reduced overall allergic lung inflammation, suggesting that IL-9 plays an obligate role in the allergic memory response following pulmonary allergen challenge. These observations further suggest that IL-9 from Trm cell populations plays a novel role in allergen recall responses and is a potential therapeutic target for patients suffering from chronic intermittent asthma.
The complex allergen house dust mite (HDM) acts directly on macrophages to stimulate noncanonical autophagy
Authors: Gao Hongjuan , Qi Xiulan , Jackson William , Keegan Achsah 
Published: January 2023

Abstract
Asthma is a chronic inflammatory disease that results in narrowing of the airways. Inhalation of house dust mites (HDM) is a main cause of allergic asthma. Macrophages (Mf) are abundant in the lung and are one of the first cells exposed to HDM, however little is known about the effects of HDM on Mf. Autophagy has been linked to Mf phenotypes and asthma but the mechanism by which autophagy contributes to these processes is unclear. Autophagy is a homeostatic process involving degradation of intracellular components, including proteins, organelles, and foreign bodies. To determine whether HDM stimulates autophagy, we treated bone marrow-derived Mf (BMM) with HDM and evaluated autophagosome foci formation. HDM induced foci formation that was similar to treatment with the autophagy inducer Torin 1. Further, electron microscopy revealed an increase in double membraned vesicles in response to HDM and Torin 1, which could be due to an increase in autophagosome formation or inhibition of their fusion with lysosomes. To differentiate these hypotheses, we investigated markers of autophagy. During canonical autophagy, LC3-I is converted to membrane-associated LC3-II by lipidation, while the cargo adapter protein SQSTM1/p62 is degraded. Surprisingly, treatment with HDM caused a general increase in LC3-I and LC3-II without a substantial change in the LC3-II to LC3-I ratio. As expected, Torin 1 induced autophagic degradation demonstrated by reduction of SQSTM1/p62. However, SQSTM1/p62 increased dramatically in response to HDM, indicating a block in autophagic degradation. We observed a similar trend in THP-1 and human CD14+ Mf. These results suggest that HDM engages autophagy machinery but does not stimulate degradative autophagy.
Impaired tumor-necrosis-factor-α-driven dendritic cell activation limits lipopolysaccharide-induced protection from allergic inflammation in infants
Authors: Kaur Kamaljeet , Bachus Holly , Papillion Amber M. , Marquez-Lago Tatiana T. , Yu Zhihong , Ballesteros-Tato Andre , Matalon Sadis , Leon-Ruiz Beatriz 
Published: January 2023

Abstract
Infants have a higher risk of developing allergic asthma than adults. However, the underlying mechanism remains unknown. We show here that sensitization of mice with house-dust mites (HDMs) in the presence of low-dose lipopolysaccharide (LPS) prevented T helper 2 (Th2) cell allergic responses in adult, but not infant, mice. Mechanistically, adult CD11b + migratory dendritic cells (mDCs) upregulated the transcription factor T-bet in response to tumor necrosis factor-α (TNF-α), which was rapidly induced after HDM + LPS sensitization. Consequently, adult CD11b+ mDCs produced interleukin-12 (IL-12), which prevented Th2 cell development by promoting T-bet upregulation in responding T cells. Conversely, infants failed to induce TNF-α after HDM + LPS sensitization. Therefore, CD11b+ mDCs failed to upregulate T-bet and did not secrete IL-12 and Th2 cell responses normally developed in infant mice. Thus, the availability of TNF-α dictates the ability of CD11b+ mDCs to suppress allergic Th2-cell responses upon dose-dependent endotoxin sensitization and is a key mediator governing susceptibility to allergic airway inflammation in infant mice.
[The above abstract is a summary from our recently published paper under the same title – Immunity; published: Jan 8, 2019 DOI: https://doi.org/10.1016/j.immuni.2018.11.012]
Tissue driven influences on human NK cell development, function and residence
Authors: Dogra Pranay , Rancan Chiara , Ma Wenji , Toth Marta , Senda Takashi , Carpenter Dustin J. , Thapa Puspa , Szabo Peter Anthony , Shen Yufeng , Fong Lawrence , Lanier Lewis L , Farber Donna L 
Published: January 2023

Abstract
Natural killer (NK) cells are innate immune cells with the inherent ability to kill tumor and virus infected cells without any prior priming. Recent studies have demonstrated the ability of NK cells to develop adaptive memory and tissue residence. However, our understanding of human NK cells in tissues and their site-specific properties is limited. Here we use our unique tissue resource established in collaboration with LiveOnNY to elucidate the diversity of human NK cell phenotypic subsets, functional profile and transcriptional signatures in non-mucosal and mucosal tissue sites. We identified tissue-specific patterns of NK cell maturation and show that NK cell subset (immature and mature) and memory NK cell distribution is a function of tissue site which is not affected by age. Our results also show that while NK cells from different tissue sites express similar levels of granzyme B, NK cell antibody-dependent cellular cytotoxicity response and cytokine driven activation may be influenced by tissue site. Whole transcriptome profiling of NK cell subsets revealed that mature NK cells from all the sites (blood, bone marrow, spleen, lung and LN) have very similar transcriptional profiles. Interestingly, immature NK cells from all tissue sites have very different transcriptional profiles compared to their blood counterpart and are enriched for tissue resident memory T cell transcriptional signature. Furthermore, we show that immature NK cells show site-specific expression patterns of tissue residence surface markers. Overall, our data demonstrates the effects of tissue microenvironment on the developmental and function potential of tissue NK cells and provides insight into the development of future NK cell mediated immunotherapies.
DNA origami probes for single-cell paired BCR heavy and light chain immune repertoire analysis
Authors: Chaudhary Neelam , Schoettle Louis , Dubois Courtney , Jones Kylee , Ulrich Peaches Rebecca , Yan Hao , Blattman Joseph N 
Published: January 2023

Abstract
Antibodies are a diverse set of glycoproteins produced by B-cells in response to antigen exposure, infection, or certain malignancies, making it crucial to develop a high throughput and robust technique to obtain paired heavy and light chain information from individual B-cells. We have developed DNA origami-based capture probes to isolate paired heavy and light sequence information from single B-cells. The capture probes are designed to hybridize with the constant regions of heavy and light chains. Each capture probe also incorporates sequences for hybridization with DNA origami nanostructures. The nanostructures can be electroporated into B-cells and re-isolated by cell lysis and streptavidin affinity purification. Heavy and light chains are then reverse transcribed, appending the isolated sequences to the capture probes. Following PCR-amplification, each amplicon will contain BCR heavy or light chain sequence, as well as a unique barcode. Matching barcodes from complementary heavy and light chain amplicons can then be paired via next-generation sequencing. We have used this technique to analyze the antibody repertoire from splenic tissue of unimmunized C57BL/6J mice. We designed a single IgM-Igk specific capture probe and synthesized DNA origami nanostructures. The nanostructures were transfected into naïve B-cells, purified, reverse transcribed, and PCR-amplified. The isolated IgM and Igk amplicons were confirmed by sequencing. Through this approach, we present a novel DNA origami-based technique for single cell paired BCR repertoire analysis. This is a highly versatile technique that can be used to isolate any gene pair functioning in cellular dynamics such as signaling pathways or co-occurring mutations in cancer.