bluuebunny/crossref_metadata_2025_split huggingface dataset:
FVIII tolerance induction in hemophilia A mice utilizing low dose IL2 treatment
Authors: Lyle Meghan J , Fu Richard Y. , Miao Carol H. 
Published: January 2023

Abstract
Hemophilia A patients lack or possess low levels of functional FVIII protein, which results in an inability for the blood to clot when injury occurs. The current treatment for hemophilia A patients is FVIII protein replacement, but this treatment is expensive and often results in anti-FVIII immune responses, neutralizing the clotting effect.
Regulatory T (Treg) cells help balance T effector (Teff) cells through their suppressive function during an immune response and keep autoimmunity in check. Treg cells possess a high affinity epitope of the IL2 receptor. With a low dose of IL2, the Treg population outcompetes the Teff cells, leading to an increase in activation and number. The increased suppressive activity may induce tolerance in hemophilia A mice treated with FVIII. Hemophilia A mice were hydrodynamically injected with IL2 and FVIII plasmids. Cell staining data showed a marked increase in Treg cell population and activation. In addition, the Treg/Teff ratio was significantly increased in the first 3 weeks and maintained at increased levels over several weeks afterwards. On Day 28-post FVIII injection, control mice had started to develop inhibitors associated with significant decrease of FVIII expression, while mice treated with IL2 showed no inhibitors with persistent FVIII expression. The treated mice are monitored to determine the potential long term tolerance induced by low dose IL2. Combined gene transfer of FVIII and IL2 plasmids can produce therapeutic FVIII and simultaneously prevent inhibitory antibody formation, thus providing a potentially effective treatment of hemophilia A.
PSMP, a novel chemotactic cytokine, triggers inflammatory Ly6ChiCCR2+ monocyte migration and promotes colitis in a CCR2-dependent manner
Authors: She Shaoping , Pei Xiaolei , Zheng Danfeng , Ma Jing , Guo Changyuan , Zhang Yan , Peng Xinjian , Chen Dixin , Wang Pingzhang , Mo Xiaoning , Zhang Yingmei , Song Quansheng , Ma Dalong , Wang Ying 
Published: January 2023

Abstract
Chemokines plays a crucial role in the migration of leukocytes, which are involved in immune and inflammatory reactions with their receptors. To find novel chemotactic cytokines, we carried out genome-wide bioinformatics analysis and PBMC chemoattractant platform screen. Seven novel potential chemotactic cytokines were picked up, one of them was PC3-secreted microprotein (PSMP). The mature secreted PSMP was able to chemoattract human peripheral blood monocytes, lymphocytes and THP-1 cells. CCR2 was identified as a high-affinity receptor for PSMP by in vitro chemotaxis, calcium flux, receptor internalization, and radioligand-binding assays. Furthermore, we found that PSMP was extensively expressed at low levels in normal human tissues, but was significantly up-regulated in intestinal biopsies from patients with inflammatory bowel disease and mouse DSS-induced colitic tissue in the early stage prior to IL-6, TNF-α and CCL2 up-regulation. Moreover, the results showed that PSMP triggered inflammatory Ly6ChiCCR2+ monocyte migration from the circulation into the lamina propria in a CCR2-dependent manner and promoted the progression of inflammation. A PSMP neutralizing monoclonal antibody significantly mollified inflammation in a mouse DSS colitis model. Taken together, our study provides the first evidence that PSMP is a novel chemotactic cytokines whose receptor is CCR2 and PSMP plays a vital and irreplaceable role in triggering and promoting DSS colitis. Our finding not only reveals the function and regulating mechanism of PSMP, but also provides a promising therapeutic strategy for treating inflammatory bowel disease.
IL-33-Driven Innate Tissue-Protective Function of ST2+ Treg Cells
Authors: Liu Quan , Lott Jeremy M. , Mathews Lisa R. , Li Huihua , Zhu Qiang , Matta Benjamin M. , Delgoffe Greg M. , Fan Ming-Hui , Turnquist Heth R. 
Published: January 2023

Abstract
Non-lymphoid tissue-resident CD4+ Foxp3+ regulatory T (Treg) cells with the capacity to modulate non-immunological processes including organismal metabolism and tissue repair have been recently described. Notably, a large fraction of non-lymphoid tissue-resident Treg cells express ST2, the receptor for the tissue-derived cytokine and alarmin, IL-33. However, the relationship between IL-33 and ST2+ Treg cells in quiescent and pathogenic states is only starting to be understood. Using FACS and Foxp3 reporter mice, we demonstrate that ST2+ Treg cells from naïve animals are phenotypically and functionally unique relative to ST2− Treg isolated from the same location. Yet, our studies establish that peripheral and lymphoid tissue ST2+ Treg cells are highly comparable. Interestingly, ST2+ Treg exhibit a high level of aerobic glycolysis, which supports their augmented potential for cytokine secretion, especially that of IL-5, IL-10, and IL-13. IL-5 and IL-13 secretion in response to IL-33 is an innate function of ST2+ Treg cells as it does not require TCR-stimulation. Conversely, ST2+ Treg release of other cytokines, including IL-10, is antigen-restricted and amplified by IL-33. Among Treg subsets, ST2+ Treg cells are the exclusive source of Type 2 cytokines, especially IL-13, which we reveal supports Treg reparative responses. Using mice deficient in IL-33 or Treg cells, we establish that Treg cells and IL-33 are both required to protect mice from lethal lung injury. In line with these data, delivery of IL-13, but not IL-33, at the time of lung injury rescues Treg-depleted mice. This study establishes that ST2+ Treg cells possess an innate capacity for cytokine production that is crucial to tissue repair and inflammation control.
Down-regulation of E protein activity augments an ILC2 differentiation program in the thymus
Authors: Sun Xiao-Hong , Wang Hong-Cheng , Qian Miranda Liangyue , Zhao Ying , Zhuang Yuan , Urban Joseph F. , Fung Kar-Ming 
Published: January 2023

Abstract
Innate lymphoid cells (ILCs) have gained recognition as important regulators in various immune responses. How these cells are generated is just beginning to be understood. Current paradigm states that all three subsets of ILCs originate from common lymphoid progenitors (CLP) in the bone marrow through an array of downstream ILC progenitors. Id2, a member of the Id family that inhibits the function E protein transcription factors, is expressed in these ILC progenitors and is indispensable for ILC differentiation. Unexpectedly, we found that specifically down-regulating E protein function in the thymus by either Id1 expression or conditional deletion of two E protein genes resulted in massive expansion of ILC2s in the thymus as well as other organs where ILC2 normally reside. Consequently, the mutant mice exhibit augmented spontaneous infiltration of eosinophils and heightened responses to allergen, papain, in the lung. Furthermore, these mice also display increased ability in the expulsion of helminth parasites, N. Brasiliensis. These results raise an interesting question as to whether the thymus is a natural site for ILC2 production and the phenotypes of our mutant mice simply highlight such capacity. Indeed, lineage tracing experiments using lck-Cre/ROSA26-stop-tdTomato mice demonstrate that tdTomoto specifically marks a fraction of ILC2s in the thymus as well as in the lung, suggesting that thymus-derived ILC2s preferentially home to the lung. We also found that thymic progenitors were able to differentiate into ILC2 cells in vitro. Together, our data reveal a new ILC2 differentiation program present in the thymus, which enriches our knowledge about ILC2 ontogeny and has potential biological significance.
Differential role of ID2 in the group 3 innate lymphoid cells for the development of different gut associated lymphoid tissues.
Authors: Guo Xiaohuan , Wang Wenyang , Fu Yang-Xin 
Published: January 2023

Abstract
Gut associated lymphoid tissues (GALTs), including the mesentery lymph nodes (mLNs), Peyer’s patches (PPs), and isolated lymphoid follicles (ILFs), are important locations for the initiation of adaptive immune responses in the intestine. Group 3 Innate lymphoid cells (ILC3s) are defined by their expression of RORγt and include lymphoid tissue inducer (LTi) cells, which play a critical role in the development of GALT. However, the function of ID2 in these well-differentiated ILCs is still unclear. In our study, RorccreId2fl/fl mice showed normal LNs, but completely lacked PPs and ILFs compared with WT mice. And ILC3s were significantly reduced in the gut of RorccreId2fl/fl mice compared with WT mice, but were actually increased in mLNs, indicating that the development or maintenance of ILC3s in the gut, but not LNs, requires ID2 regulation. Moreover, ID2 deficient ILC3s exhibited significantly reduction of the lymphotoxin expression in the gut, which is important for the formation of GALT. We also found IL-7 stimulation could induce lymphotoxin expression in WT ILC3s, while the IL-7Rα expression was dramatically reduced in ID2 deficient ILC3s, suggesting that ID2 may regulate the lymphotoxin expression through IL-7 signaling. On the other hand, we found that the expression of Rank was upregulated in ID2 deficient ILC3s, which indicates that the elevated Rank may rescue ILC3s’ survival and lymphotoxin expression through NF-κB pathway. Taken together, our results suggest that ID2 could regulate the function and maintenance of ILC3s through IL-7 and Rank signaling to further determine the differential development of GALTs.
The synthesis of VEGF in allergen-stimulated mast cells is through a leukotriene B4 receptor-2-dependent signaling pathway
Authors: Lee A-Jin , Ro MyungJa , Park JaeIn , Jang Jae-Hyun , Kim Jae-Hong 
Published: January 2023

Abstract
Mast cells are among the principal effector cells in the pathogenesis of allergic asthma. In allergic reactions, allergen(Ag)-induced cross-linking of IgE bound to FcɛRI on mast cells results in the production of vascular endothelial growth factor(VEGF), which is essential for the initiation and development of the allergic response. Despite the central role of VEGF in allergic asthma, the signaling events responsible for the production of VEGF remain unclear, particularly in Ag-stimulated mast cells. In the present study, we observed that blocking leukotriene B4 receptor-2(BLT2) completely abrogated the production of VEGF in Ag-stimulated bone marrow-derived mast cells(BMMCs). The synthesis of BLT2 ligands (leukotriene B4 [LTB4] and 12(S)-hydroxyeicosatetraenoic acid [12(S)-HETE]) was also required for VEGF production, suggesting a mediating role of an autocrine BLT2 ligands-BLT2 axis in the production of VEGF in mast cells. The Nox1-reactive oxygen species(ROS)-NF-kB cascade is downstream of BLT2 during Ag signaling to VEGF synthesis in mast cells. Furthermore, the level of VEGF synthesis in genetically mast cell-deficient Kit(W/Wv) mice was significantly lower than that in wild-type(WT) mice in the ovalbumin(OVA)-induced asthma model, suggesting that mast cells play a critical role in the synthesis of VEGF in OVA-induced allergic asthma. Importantly, VEGF production was restored to the levels observed in WT mice after adoptive transfer of normal BMMCs into Kit(W/Wv) mice but was not restored in BLT2−/− BMMC-reconstituted Kit(W/Wv) mice in the OVA-induced asthma model. Taken together, our results suggest that BLT2 expression in mast cells is essential for the production of VEGF in OVA-induced allergic asthma.
Epitope spreading in experimental autoimmune encephalomyelitis by the infection of periodontitis pathogen porphyromonas gingivalis
Authors: Guan Hongbing , Zheng Zhichao , Li Zhengmao 
Published: January 2023

Abstract
Deep cervical lymph nodes (dcLNs) were reported to be the first location for the initiation of encephalitogenic CD4+ T cells in experimental autoimmune encephalomyelitis (EAE). However, the precise mechanisms are not crystal clear. Here, we explored the epitope spreading in the two anatomically adjacent diseases, periodontitis and EAE. C57bl/6 mice were immunized with porphyromonas gingivalis (p.gingivalis), a virulent peridontopathogen, in the buccal submucosal tissue. Activation of T cells and MOG-specific CD4+ T cells in dcLNs and superficial cervical lymph nodes were detected with respect to the expression of CD69, CD44, and MOG-tetramer. A clinical symptom was observed in p.gingivalis-immunzed mice similar to that in MOG35-55 peptide-immunized mice. Dendritic cells in the buccal submucosal tissue and dcLNs were analyzed for the spreading of MOG epitope. Our results demonstrated the existence of the epitope spreading to the endogenous myelin in the inflammatory reponse of periodontitis caused by infection of p.gingivalis.
Visualization of IFN-β mRNA in Chlamydia-infected and adjacent uninfected cells and the contribution of CX43 in intercellular transfer of cGAMP
Authors: Tripathy Manoj Kumar , Zhang Yugen , Rodriguez-Nieves Jennifer , Bodnar Wanda M , Nagarajan Uma 
Published: January 2023

Abstract

Background
IFNβ has been implicated a key effector of oviduct pathology after genital chlamydial infection in mice. We reported that the host DNA sensor cGAS (cGAMP synthase) is essential for IFNβ expression during Chlamydia trachomatis infection. Sensing during infection leads to generation of the second messenger, 2′3-cGAMP, which can migrate to uninfected adjacent cells via gap-junction mediated transfer. cGAMP binding to central adaptor molecule, STING drives IFNβ induction. The goals of this study were to visualize the mechanism of c-GAMP transfer and to identify the DNA source triggering this response.

Methods
cGAMP production was detected by mass spectrometry. The contribution of gap junction proteins, connexins CX43 and CX45 in intercellular transfer of cGAMP was assayed by siRNA depletion and subsequent co-culture experiments. view RNATM methodology was used to detect IFNβ mRNA transcripts in Hela cells. Mitochondrial DNA as IFNβ inducer was assessed in mtDNA depleted cell lines.

Results
Mass spectroscopy confirmed cGAMP in infected host cytosol. Connexin depletion studies revealed that CX43 but not CX45 was essential for optimal IFNβ expression during infection. IFNβ mRNA transcripts were visualized in infected and adjacent, uninfected cells. In contrast, distal uninfected cells lacking cell-cell contact were negative. IFNβ induction after chlamydia infection was unchanged in mtDNA depleted cells.

Conclusion
Overall, our results confirm the production of cGAMP in chlamydia-infected cells and its transfer via CX43 to adjacent cells resulting in IFNβ production. Our studies exclude mtDNA as a potential source initiating this response. We are currently investigating the role of Chlamydial-DNA as an initiator.
Protein kinase C dependent development of dendritic cells.
Authors: Chavel Colin A , Lee Kelvin P 
Published: January 2023

Abstract
Dendritic cells (DC) are professional antigen presenting cells that are important activators of adaptive immune responses and play a role in the activation of CD8+ T cells that target tumor cells. Our lab has also previously shown that protein kinase C (PKC) βII is essential for DC differentiation and that tumor derived factors, particularly IL-6, inhibit DC differentiation by repressing the expression of PKCβII through a Stat3 dependent mechanism. However, the pathways activated downstream of PKCβII during DC differentiation have yet to be fully characterized. To study this, we used the K562 cell line, which differentiates into DCs with addition of PMA, a known chemical activator of conventional PKC isoenzymes. We have also shown previously that PMA specifically activates PKCβII in K562 cells. Addition of PMA to K562 cells showed the activation of the ERK1/2 and NFκB pathways, including both the canonical and non-canonical NFκB pathways. Chemical inhibition of either the ERK1/2 or NFκB pathway yielded immature DCs with a reduced capacity to stimulate T-cell proliferation when incubated with PMA. Inhibition of ERK1/2 also showed a change in levels of RelB, a NFκB transcription factor, with PMA treatment and during PMA induced DC differentiation. Our lab has also shown that matured DCs reduce Foxo3a protein levels with the stimulation of CD80/CD86 through an Akt dependent mechanism. Inhibition of either the ERK1/2 or NFκB pathways showed an increase in Foxo3a levels, indicating a role for the ERK1/2 and NFκB pathways in Foxo3a regulation during differentiation into mature DCs outside of Akt. Taken together, these results show an integral role for both the ERK and NFκB pathways downstream of PKCβII activation during DC differentiation.
ItpkB regulates CCR7 expression on dendritic cells prior to lymph node trafficking
Authors: Vella Jennifer L , Wang Xinxin , Huang Yina H 
Published: January 2023

Abstract
Dendritic cells (DC) are fundamental in the initiation of the adaptive immune response. Following antigen uptake, tissue-resident DCs undergo maturation in preparation for their migration to lymph nodes where they present processed antigen to T cells. Down-regulation of tissue-homing receptors and upregulation of lymphoid-homing receptors, including CCR7 is required for dendritic cell entry into afferent lymphatic vessels. CCR7 binding to its ligands, CCL19 and CCL21, provides the appropriate directional cues for DCs to traffic to lymph nodes. Although both maturation and migration of DCs have been extensively studied, it still remains unclear what signaling pathways are responsible for inducing CCR7 expression. We found that DCs deficient in inositol-trisphosphate 3-kinase B (ItpkB) express higher levels of CCR7. ItpkB-deficient BMDCs migrate to draining lymph nodes more efficiently in both competitive and noncompetitive environments compared to their wild-type counterparts. We have previously shown that ItpkB deficiency leads to enhanced Akt activation in NK cells. Western blot analysis showed that ItpkB-deficiency in DCs also leads to increased basal and LPS-induced Akt1 activation. To determine whether hyper-Akt activity is sufficient to trigger enhanced lymph node homing, we expressed two forms of constitutively active Akt1, Myr-Akt1 and Akt1 T308E, S473D, in wildtype DCs. Akt1 hyperactivity was sufficient to promote CCR7 expression but did not affect DC maturation. Our findings suggest that ItpkB regulates CCR7 expression on dendritic cells as they prepare to traffic to draining lymph nodes for the purpose of primary T cell responses.
Transcriptional regulation of basophil development and function in Th2 immunity
Authors: Zhang Sai , Henry Everett , Siracusa Mark C , Sant’Angelo Derek B 
Published: January 2023

Abstract
Basophils are the most rare granulocyte population, accounting for less than 1% of circulating leukocytes in mice and human. Recent studies have shown that basophils are associated with multiple immunological processes, such as allergic diseases, autoimmunity, and protective immunity against parasites. It has been show that the rapid secretion of Th2 cytokines by basophils in response to various stimuli plays key roles in immunity against parasitic infections. Basophils also release histamine and, therefore, are key contributors to allergy. In addition, basophil responses are associated with multiple human diseases such as asthma, atopic dermatitis and cancer. Given their important roles in inflammatory signaling and human diseases, remarkably little is known about the molecular regulation of basophil effector functions. Therefore, we have been exploring transcription factors that may play a direct role in controlling basophil development and effector functions. In this study, we have developed multiple in vitro and in vivo systems to study the influence of the loss of a potentially key transcription factor on the basophil response to allergen-induced Th2 immunity. We are in the process of performing gene expression arrays to understand the molecular mechanisms underlying the transcriptional regulation. Collectively, our studies provide new insight into the transcriptional network controlling basophil effector functions in Th2 immune response and lead to potential therapies that modulate basophil activities during the treatment of allergic diseases.
The advancing age affects the frequency, functions and antibody repertoire of human B-1 cells
Authors: Hernandez Ana Maria , Rodríguez-Zhurbenko Nely , Quach Tam , Hopkins Thomas J , Rothstein Thomas 
Published: January 2023

Abstract
Aging decreases the efficiency of several physiological processes, including immune function, which has been associated with an increased incidence of infections, autoimmune diseases and cancer. Human B cell populations, like naïve and memory B cells, suffer significant quantitative and qualitative changes in the elderly. The frequency and function of human B-1 cells, which play critical housekeeping and anti-microbial defensive roles, have not been studied. In the present work we analyzed how the frequency and function of human B-1 cells (CD20+CD27+CD43+CD38low) change with age. Our results show that not only the percentage of B-1 cells but also their capacity to spontaneously secrete IgM decreased with age. In fact, expression levels of Xbp1 and Blimp1, associated with a secreting B cell phenotype, were significantly lower, while those of Pax5, characteristic of non-secreting B cells, were significantly higher in elderly donors. Since aging affects the humoral immune response both quantitatively and qualitatively, next we studied the antibody repertoire of B-1 cells of healthy young and elderly donors. Elderly donors showed a higher number of repeated clones and therefore, a reduction of the antibody repertoire variability in comparison with young individuals. Furthermore it was interesting to observe in aged individuals an increased frequency of VH3-23 and VH4-34 usage, genes which have been linked to autoimmune diseases and hematopoietic malignancies. Moreover, there is a significant reduction in older individuals of clones expressing the sequence VH3-33, frequently found on antibodies against PC, oxidized LDL, apoptotic cells and induced after vaccination with Pneumavax23.
Neutrophil extracellular traps (NETs) promote disseminated intravascular coagulation in sepsis.
Authors: McDonald Braedon , Davis Rachelle , Jenne Craig N 
Published: January 2023

Abstract
Neutrophil extracellular traps (NETs; webs of DNA coated in anti-microbial proteins) are released into the vasculature during sepsis and contribute to organ damage. Various components of NETs have been shown to modulate the coagulation cascade in vitro. We hypothesized that NETs released into the vasculature during sepsis activate intravascular coagulation, leading to microvascular hypoperfusion and end-organ damage. Multi-color confocal intravital microscopy was used in mouse models of sepsis (endotoxemia and E. coli peritonitis) to visualize and quantify neutrophil and platelet trafficking, NETs production, real-time perfusion analysis, and intravascular thrombin activity (using novel in vivo zymography). In vivo imaging of the liver microcirculation in septic mice revealed spatial co-localization between NETs and intravascular coagulation (thrombin activity, fibrin deposition, and platelet aggregation). Inhibition of NETs using i.v. DNase infusion (to digest NETs), or PAD4-deficient mice (that have impaired NETs production) resulted in significantly lower quantities of intravascular thrombin activity, and improved microvascular perfusion. In a model of Gram-negative sepsis (E. coli peritonitis), PAD4-deficient mice had reduced markers of disseminated intravascular coagulation (intravascular thrombin, PAI-1), and end-organ damage (serum lactate, ALT) compared to wild-type controls. Together, these data demonstrate that NETs activate coagulation in vivo, and promote disseminated intravascular coagulation in sepsis. Inhibition of NETs in septic animals reduces intravascular coagulation, improves microvascular perfusion, and attenuates end-organ damage.