bluuebunny/crossref_metadata_2025_split huggingface dataset:
Targeted depletion of macrophages using an anti-human DR5 antibody prevented arthritis in a novel human/mouse chimeric DR5 Tg mouse (135.29)
Authors: Li Jun , Yang Ping-Ar , Wu Qi , Hsu Hui-Chen , Mountz John 
Published: January 2023

Abstract
The severity of rheumatoid arthritis (RA) correlates with the activation and abundance of macrophages in the inflamed tissues and circulation. The purpose of this study is to generate a humanized mouse (hu/mo) DR5 transgenic (Tg) mouse in which the hu/mo DR5 is targeted to the monocytes/macrophages and promote apoptosis by an anti-human DR5 antibody, TRA-8. The DR5 Tg consists of a human extracellular domain, a mouse transmembrane and intracellular domain under the regulation of the 3kb mo DR5 promoter. A Floxed STOP was inserted between the promoter and hu/mo DR5. Targeted expression of the DR5 Tg in monocytes/macrophages was generated by crossing the DR5 Tg with the lysozyme-Cre (LysMCre) mice. Arthritis was induced by intradermal injection of chicken type II collagen (CII) in CFA/IFA on day 0 and boost on day 21. There was a high percentage of CD11b+ macrophage in the spleen of CII immunized non-Tg mice (10.7%) compared to naïve mice (2.5%). TRA-8 (0.2 mg) was administrated I.V. every 3 days for 3 doses starting at day 14 after boost. After TRA-8 treatment, there was a significant decrease (55.6%) of CD11bhighGr-1+ macrophages. TRA-8 treatment also suppressed the development of arthritis in LysMCre+ hu/mo DR5+ Tg mice. This study demonstrated that selectively reducing the numbers of activated macrophage by using an anti-human DR5 antibody is a novel therapeutic strategy applicable to human RA, which might lead to prevention of joint inflammation and arthritis.
CD11b+Ly6C+ cells, myeloid derived suppressor cells, affect T cell responses during demyelinating disease (135.38)
Authors: Olson Julie , Glazer Alex 
Published: January 2023

Abstract
Theiler’s murine encephalomyelitis virus (TMEV) is a mouse model of multiple sclerosis in which mice develop a chronic progressive demyelinating disease associated with a myelin- specific CD4+ T cell response. Our previous studies have demonstrated that the innate immune response directly affects the initiation and progression of demyelinating disease. We recently determined that the predominant innate immune cells infiltrating into the central nervous system (CNS) following TMEV infection are CD11b+Ly6C+ cells. CD11b+Ly6C+ cells are immature myeloid cells that exit the bone marrow without maturing. Thus, we wanted to determine the role of these cells in the immune response during demyelinating disease. CD11b+Ly6C+ cells isolated from the CNS during TMEV infection expressed high levels of IL-10 and iNOS and low levels of IL-12, and expressed low levels of costimulatory molecules and MHC class II. Most interestingly, CD11b+Ly6C+ cells isolated during TMEV- induced demyelinating disease suppressed CD4+ T cell responses against virus and myelin antigens. Furthermore, depletion of Ly6C+ cells during TMEV infection lead to increased expression of IFNγ, IL-12, and IL-17 in the CNS associated with increased T cell responses. These results suggest that these CD11b+Ly6C+ cells are myeloid derived suppressor cells that suppress T cell responses and affect the progression of demyelinating disease. This work was supported by the National Multiple Sclerosis Society Grant RG3625.
Lipoteichoic acid from Staphylococcus aureus attenuates differentiation of pre-osteoclast into mature osteoclast (136.26)
Authors: Yang Jihyun , Kim Jiseon , Ryu Young Hee , Yun Cheol-Heui , Han Seung Hyun 
Published: January 2023

Abstract
Bacterial virulence factors regulate differentiation and functions of osteoclasts, which mediate the bone-resorptive process. Here, we investigated the effect of a major cell wall virulence factor of Gram-positive bacteria, lipoteichoic acid (LTA), on the osteoclast differentiation. Highly-pure and structurally-intact LTA was prepared from Staphylococcus aureus through sequential application of organic solvent extraction and hydrophobic-interaction column chromatography followed by ion-exchange column chromatography. Osteoclast precursors were prepared from C57BL/6 mouse bone marrow cells using M-CSF and RANKL. Staphylococcal LTA suppressed the differentiation of osteoclast precursors to mature osteoclasts concordantly with an inhibition of the bone-resorptive capacity. Meanwhile, LTA augmented the phagocytic and inflammatory potential of the pre-osteoclasts. The LTA inhibition of osteoclast differentiation appears to be mediated through Toll-like receptor 2 (TLR2), and interestingly both MyD88-dependent and -independent signaling pathways. Furthermore, LTA reduced phosphorylation of ERK and JNK during the osteoclast differentiation process followed by a decreased DNA-binding activity of AP-1. Conclusively, our results suggest that LTA from S. aureus attenuates the differentiation of pre-osteoclast to mature osteoclast through TLR2 with partial involvement of MyD88 signaling pathways.
RIG-I mediates rhabdovirus-induced inflammatory immune responses of primary human astrocytes (136.3)
Authors: Furr Samantha , Moerdyk-Schauwecker Megan , Grdzelishvili Valery , Marriott Ian 
Published: January 2023

Abstract
Astrocytes are ideally positioned to respond to invading pathogens of the central nervous system (CNS) and produce key inflammatory mediators following exposure to neurotropic rhabdoviruses such as rabies virus and vesicular stomatitis virus (VSV). However, the mechanisms by which resident CNS cells perceive such viral challenges have not been defined. Recently, several cytosolic DExD/H box RNA helicases including retinoic acid-inducible gene I (RIG-I) have been described that function as intracellular sensors of replicative RNA viruses. Here, we demonstrate that primary human astrocytes constitutively express RIG-I and show that such expression is elevated following exposure to VSV. Evidence for the functional nature of RIG-I expression in these cells comes from the observation that this molecule associates with its downstream effector molecule, interferon promoter stimulator-1, following VSV infection and from the finding that a specific ligand for RIG-I elicits astrocyte immune responses. Importantly, RIG-I knockdown significantly reduces inflammatory cytokine production by VSV-infected astrocytes and inhibits the production of soluble neurotoxic mediator(s) by these cells. These findings directly implicate RIG-I in the initiation of inflammatory immune responses by human glial cells and provide a potential mechanism underlying the inflammation and neuronal cell damage associated with acute rhabdoviral infections of the CNS.
Accelerated wound healing in mice by local administration of sustained-release CpG oligodeoxynucleotide (136.4)
Authors: Sato Takashi , Shimosato Takeshi , Yamamoto Masaki , Kobayashi Nobuaki , Miyazawa Naoki , Ishigatsubo Yoshiaki , Klinman Dennis 
Published: January 2023

Abstract
Recent reports show that wounds in mice lacking MyD88 heal at a delayed rate, suggesting that the innate immune pathway can affect the healing process. Preliminary studies revealed that the kinetics of wound healing in TLR9 KO mice were also delayed when compared to congenic controls, suggesting that this activation pathway may contribute physiologically to wound healing. Synthetic oligodeoxynucleotides (ODN) expressing unmethylated CpG motifs mimic the ability of bacterial DNA to stimulate an innate immune response via TLR9. We incorporated CpG ODN into a vehicle (basement membrane extract) that slowly released bioactive ODN to wound sites. The effect of the ODN was then examined using a murine excisional skin biopsy model. Results show that CpG ODN treatment significantly accelerates wound repair in normal adult and aged mice (p &lt; 0.0001 and p &lt; 0.01, respectively). Consistent with these effects being mediated by CpG signaling via TLR9, no effect was observed in TLR9 KO mice. Histologic studies indicate that CpG treatment promotes local accumulation of macrophages, increases production of VEGF, and accelerates neovascularization, resulting in accelerated granulation and wound closure in wildtype (but not TLR9 KO mice, p &lt; 0.01). Current findings suggest that immunostimulatory oligonucleotides may represent a novel, inexpensive, safe and effective means of improving wound repair.
Mindbomb proteins are E3 ubiquitin ligases essential for TBK1-mediated antiviral activity (136.6)
Authors: Wang Lingyan , Li Shitao , Kong Young-Yun , Dorf Martin 
Published: January 2023

Abstract
First line host defense against virus attack includes the innate immune system which recognizes and mounts rapid antiviral responses. An important aspect in defense against viral infections is the production of type 1 interferon which provides a protective state to surrounding cells. TBK1 is a critical kinase involved in cellular transcriptional responses to infection and production of type I interferon. We found that TBK1 undergoes Lys63 ubiquitination following viral infection. The E3 ligases Mindbomb 1 (MIB1) and Mindbomb 2 (MIB2) associate with the TBK1 complex and confer K63-linked ubiquitin to TBK1, resulting in activation of downstream IRF signaling. Two lysine residues on TBK1 are ubiquitin acceptor sites and critical for innate antiviral defense. Ectopic expression of MIB1 or MIB2 augments TBK1-induced antiviral responses. Deficiency of MIB1 and MIB2 demonstrates that these genes are critical for TBK1 ubiquitination, type 1 interferon production, and TBK1-mediated antiviral activity. These results identify MIB1 and MIB2 as positive regulators of antiviral responses through polyubiquitination of TBK1.
Molecular mechanism underlying lipopolysaccharide-mediated expression of LCN2 in mesangial cells (136.9)
Authors: Glaros Trevor , Li Liwu 
Published: January 2023

Abstract
The neutrophil gelatinase-associated lipocalin 2 (LCN2) is a critical inflammatory mediator highly expressed in both acute and chronic inflammatory conditions. In particular, LCN2 produced in kidney mesangial cells contributes to tubular damage and kidney failure. In this report, we demonstrated that the interleukin 1 receptor associated kinase 1 (IRAK-1) is essential for lipopolysaccharide (LPS)-induced expression of LCN2 in kidney mesangial cells in vtro and kidney tissues in vivo. Mechanistically, we demonstrated that C/EBPδ is critically involved in the expression of LCN2. Through chromatin immunoprecipitation assays, we showed that LPS recruits C/EBPδ to the LCN2 promoter from WT, but not IRAK-1-/- cells. Furthermore, we revealed that both de novo synthesis and activation of C/EBPδ mediated by IRAK-1 are required for LCN2 expression. Blockage of new protein synthesis using cyclohexamide significantly reduces C/EBPδ and LCN2 expression in WT, but not IRAK-1-/- cells. The expression of C/EBPδ and LCN2 can also be effectively blocked by an IKKϵ inhibitor in WT, but not IRAK-1-/- cells, suggesting the IRAK-1 contributes to C/EBPδ expression though an IKKϵ-dependent mechanism. Taken together, this study has identified an integrated intracellular network involved in LPS-induced expression of LCN2. In addition, this research provides evidence that IRAK-1 is an important target with therapeutic potential in treating acute and chronic inflammatory diseases.
An Imbalance in sVEGFR1 and VEGF-A mediates corneal neovascularization after HSV-1 ocular infection (137.23)
Authors: Mulik Sachin , Suryawanshi Amol , Sharma Shalini , Rouse Barry 
Published: January 2023

Abstract
Ocular transparency is essential for optimal vision. The cornea maintains avascularity by expressesing soluble VEGFR1 (sVR) which binds resident VEGFA and constrain its pro-angiogenic activity. Here we report that early on, herpes simplex virus 1 (HSV-1) infection leads to an increase in VEGFA expression while low sVR levels in vitro as well as in vivo in infected mice corneas. In addition, we found degradation of sVR protein which was subsequently attributed to matrix metallopeptidase 7 and 9 as observed by in-vitro and ex-vivo degradation assays. Furthermore, mice receiving an inhibitor of MMPs showed decreased degradation of sVR and diminished HSV induced corneal angiogenesis. The second peak of VEGFA observed during clinical phase of herpes stromal keratitis (HSK) was attributed to infiltrating neutrophils and their depletion during this stage resulted in low VEGFA load and diminished corneal angiogenesis. Finally, the provision of recombinant sVR to HSV infected mice diminished angiogenic response supporting the notion that the activity of sVR has a constraining effect on the angiogenic activity of VEGFA. Taken together our results support the notion that the binding of sVR to VEGFA acts to limit the extent of ocular angiogenesis in pathological situations such as observed in HSK. Thus therapies designed to increase the presence, or diminish the breakdown, of sVR may be a useful therapeutic approach to manage the severity of HSK, an important cause of human blindness.
Dual roles of CD4+CD25+Foxp3+ Tregs in regulating Th17 cells and promoting host resistance to Candida albicans (137.7)
Authors: Pandiyan Pushpa , Conti Heather , Gaffen Sarah , Lenardo Michael 
Published: January 2023

Abstract
Th17 cells and CD4+CD25+Foxp3+ regulatory T cells (Tregs) are thought to promote and suppress inflammatory responses, respectively. However, their direct interaction in the context of an inflammation is unclear. Here we show that under Th17 conditions, Tregs suppress late but not early Th17 responses. Moreover, Tregs boost IL-17A, IL-17F and IL-22 in Th17 cells at early time-points. Transient up-regulation of Th17 cytokines by Tregs plays a protective role in clearing Candida albicans infection in mice. However, Tregs regain their suppressive functions over time and suppress Th17 cell mediated inflammation in Candida infected mice. Thus, Tregs play dual roles by initially promoting protective Th17 cells, but subsequently reducing the pathology of Th17 driven inflammation.
Control of RSV-induced lung injury by alternatively activated macrophages is IL-4Rα-, TLR4-, and IFN-β-dependent (137.8)
Authors: Shirey Kari Ann , Pletneva Lioubov , Puche Adam , Keegan Achsah , Prince Gregory , Blanco Jorge , Vogel Stefanie 
Published: January 2023

Abstract
Severe RSV-induced bronchiolitis has been associated with a mixed “Th1” and “Th2” cytokine storm. We hypothesized that differentiation of “alternatively activated” macrophages (AA-Mϕ) would mediate resolution of RSV-induced lung injury. RSV infection of murine lung and peritoneal macrophages resulted in IL-4 and IL-13 production, IL-4Rα/STAT6-dependent AA-Mϕ differentiation, and led to significantly enhanced inflammation in lungs of IL-4Rα-/- mice. Adoptive transfer of wild-type macrophages to IL-4Rα-/- mice restored RSV-inducible AA-Mϕ phenotype and diminished lung pathology. RSV-infected TLR4-/- and IFN-β-/- macrophages and mice also failed to express AA-Mϕ markers, but exhibited sustained proinflammatory cytokine production (e.g., IL-12) in vitro and in vivo and epithelial damage in vivo. TLR4 signaling was required for PPARγ expression, a DNA-binding protein that induces AA-Mϕ genes, while IFN-β regulates IL-4, IL-13, IL-4Rα, and IL-10 expression in response to RSV. RSV-infected cotton rats treated with a COX-2 inhibitor increased expression of lung AA-Mϕ. These data suggest novel treatment approaches for RSV that promote AA-Mϕ differentiation.
ITAM-containing adaptors regulate NKG2D-mediated cytotoxicity in human NK cells (138.15)
Authors: Neff Jadee , Bida Anya , Schoon Renee , Dick Christopher , Leibson Paul , Billadeau Daniel 
Published: January 2023

Abstract
NKG2D is expressed on cytotoxic lymphocytes and is an important immunosurveillance receptor that recognizes stress ligands such as MICA on tumor cells or virally infected cells. Unlike traditional immune activating receptors, NKG2D couples to the non-ITAM-containing transmembrane adaptor DAP10, which recruits PI3K and Grb2 to initiate target cell lysis. Current understanding predicts an ITAM-independent mechanism of NKG2D-mediated cytotoxicity. However, we show that in primary human NK cells, the ITAM-containing adaptors DAP12, FcϵR1γ, and CD3ζ selectively regulate the Grb2-Vav1 branch of NKG2D-mediated signal transduction and cytotoxicity. Furthermore, this ITAM-adaptor regulation of NKG2D is independent of ITAM activation. Thus our data reveal a more complex interplay between NKG2D and other activating receptors than was previously imagined, and we have uncovered a novel mechanism of ITAM-adaptor regulation of cell-mediated cytotoxicity. A more thorough understanding of immunoreceptor signaling networks will lead to more precise therapies when fighting diseases such as cancer, autoimmunity, and infection.
Signaling through kainate receptors enhances murine B cell proliferation and Ig production. (138.17)
Authors: Chaimowitz Natalia , Sturgill Jamie , Conrad Daniel 
Published: January 2023

Abstract
Abundant evidence indicates an interplay between the immune and nervous systems. Altered plasma glutamate levels, for example, have been strongly correlated with immunodeficiency states, such as AIDS and malignancies, suggesting that glutamate is capable of acting as an immunomodulator. While glutamate receptor signaling has been examined in some immune cells (e.g. T cells), the effects of glutamatergic stimuli on B lymphocytes remain unknown. To determine if glutamate is capable of modulating B cell function, we first examined mouse B cells for glutamate receptor expression. Furthermore, mouse splenocytes were cultured in glutamate-enriched media; proliferation and antibody production were determined. FACS analysis shows that splenic B2 B cells express the receptor subunits that comprise the kainate receptor (KAR), a specific subtype of ionotropic glutamate receptor. KAR surface expression was found to be upregulated upon B cell stimulation. Moreover, when splenocytes and B cells were stimulated with IL-4 and CD40 ligand in glutamate enriched media, there was a significant increased in cellular proliferation and antibody (IgG1, IgE) production. The KAR specific antagonist NS-102 is being used to confirm that these effects are mediated by KARs. The data also suggests that ADAM10 is required for the proliferative enhancing activity of glutamate. These results suggest that glutamate is capable of enhancing B cell function.
Human peripheral CD4+ T-cells expressing complement receptor 1 (CR1, CD35) (138.3)
Authors: Reilly Brian , Pfeiffer Anne , Petney Katrina , Atkinson John 
Published: January 2023

Abstract
CD4+ T-cells display an array of functions that can be traced to distinct lineages or to a variety of effector fates open to a naïve T cell. The path taken is dependent not only on the antigen-specific signal received through the TCR, but also on individual signals received through other cellular receptors. Earlier studies have shown the existence of CD35 on human CD4+ T-cells. To date, no further characterization of CD35 expression on other CD4+ subsets has been described. Here, using purified CD4+ T-cells, the molecular basis of CD35 expression is further explained. Known subsets were characterized by flow cytometry as naïve, central memory (CM), effector memory (EM), or T follicular helper cells (Tfh). All subsets contained small numbers of CD35+ cells with preferential expression on naïve and CM cells. If CFSE-loaded T-cells were activated in vitro by TCR cross-linking, the division index of CD35+ cells was 2-fold greater than that of CD35neg cells. This suggests that expression of CD35 defines a functional subset of CD4+ T-cells. Further analysis using cytokine microarrays demonstrated several cytokines to be elevated following co-engagement of CD3 and CD35 compared to TCR triggering alone. Employing cytokine secretion assays at the single-cell level, a bystander effect was observed with strong enhancement of IFN-γ production in CD35neg T-cells following co-engagement of CD3 and CD35. These results point to a regulatory role for CD35+ cells in T-cell activation.
Specific deletion of TRAF3 in B lymphocytes leads to B lymphoma development in mice (138.6)
Authors: Moore Carissa , Liu Yan , Shao Changshun , Bishop Gail , Covey Lori , Morse Herbert , Xie Ping 
Published: January 2023

Abstract
TRAF3 is a cytoplasmic adaptor protein utilized by the tumor necrosis factor receptor superfamily and Toll-like receptors for signaling. To explore the in vivo functions of TRAF3 in B cells, we recently generated B cell-specific TRAF3-deficient (B-TRAF3-/-) mice, and reported that TRAF3 is a critical regulator of peripheral B cell survival. In this study, we examined the hypothesis that the prolonged survival of TRAF3-/- B cells may predispose them to transformation. We found that approximately 50% of B-TRAF3-/- mice spontaneously developed B lymphomas by the age of 18 months. Our histopathological and immunophenotyping data revealed that B1 lymphomas, marginal zone B lymphomas, and immunoblastic B lymphomas developed in different individual B-TRAF3-/- mice. TRAF3-/- B lymphoma cells were malignant, as demonstrated by the identification of clonal B lymphoma cells in multiple organs in diseased mice, including spleen, ascites, bone marrow, cervical and mesenteric lymph nodes, and occasionally, kidney and lung. Corroborating our findings, recent evidence demonstrates that deletions and mutations of the TRAF3 gene frequently occur in human patients with B-cell lymphomas and multiple myeloma. Thus, our results, together with the evidence from human patients, indicate that TRAF3 is a tumor suppressor gene in B lymphocytes. Our findings suggest that TRAF3 downstream signaling components may serve as important therapeutic targets for the treatment of B lymphomas.
PKC-θ mediates negative feedback on regulatory T cell function (139.1)
Authors: Zanin-Zhorov Alexandra , Ding Yi , Kumari Sudha , Attur Mukundan , Hippen Keli , Brown Maryanne , Blazar Bruce , Abramson Steven , Lafaille Juan , Dustin Michael 
Published: January 2023

Abstract
T cell receptor (TCR) dependent regulatory T cell (Treg) activity controls effector T cell (Teff) function and is acutely inhibited by tumor necrosis factor (TNF). Protein kinase C-θ (PKC-θ) recruitment to the immunological synapse is required in effector cells to recruit Carma-1 in the pathway to NF-κB activation. Surprisingly, PKC-θ was sequestered away from the Treg immunological synapse. Furthermore, PKC-θ inhibition or suppression by RNAi enhanced Treg function, demonstrating PKC-θ mediated negative feedback. Treg integrate TCR and TNF signals through control of PKC-θ location such that inhibition of PKC-θ protected Treg from inactivation by TNF, restored activity of defective Treg from rheumatoid arthritis patients, and enhanced protection of mice from inflammatory colitis. Treg freed of PKC-θ mediated negative feedback can function in the presence of TNF and thus have therapeutic potential in control of inflammatory diseases.