bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pathogen clearance by engineering novel host innate immunity (P1251)
Authors: Gupta Goutam , Dandekar Abhaya , Gouran Hossein , Aguero Cecilia , Bruening George , A. Feldstein Paul , Nascimento Rafael , Goulart Luiz , Norvell Meghan , Pardington Paige , Chaudhary Anu , Civerolo Edwin 
Published: January 2023

Abstract
A few years ago, we introduced a concept that a protein chimera of pathogen recognition and lysis domains would be able to rapidly clear a broad-spectrum of pathogens [Crit Rev Immunol 2007;27(3):233-245]. For a wide variety of viral, bacterial, and fungal pathogens, appropriate recognition and lysis domains can be chosen from the host innate immune repertoire. A chimera of the recognition and lysis domains would be designed with the aid of a flexible linker to ensure synergy of the two functions and therefore, the rapid clearance of the targeted pathogen. In this work, we demonstrate the design of such a chimera and the efficacy of this chimera in clearing a plant pathogen Xylella fastidiosa (Xf) that causes diseases in multiple plants of economic importance. The most notable ones are Pierce’s disease (PD) in grape and variegated chlorosis (CVC) in citrus. Specifically, we show the construction of the transgenic grapevines expressing a protein chimera of recognition and lysis domains specific for Xf. This chimera clears Xf from the xylem (the site of colonization) and blocks the development of PD [Proc Natl Acad Sci U S A. 2012;109(10):3721-3725]. The same chimera can be applied to block CVC. Finally, we indicate how such chimeras of recognition and lysis domains can be developed to target multiple human pathogens.
Memory CD8 T cells are programmed to rapidly synthesize core 2 O-glycans for tissue-specific trafficking in response to inflammatory cues (P5112)
Authors: Nolz Jeffrey , Harty John 
Published: January 2023

Abstract
Immune cell trafficking from the circulation into peripheral tissues is a highly orchestrated process involving a number of receptor-ligand interactions. Previously, it has been demonstrated that memory CD8 T cells localize to areas of infection/inflammation in an antigen-independent manner and that these recruited CD8 T cells are highly cytolytic, thus providing rapid protection against pathogens expressing cognate antigen. However, the molecular mechanisms regulating memory T cell recruitment to inflamed tissue remain ill defined. Herein, we demonstrate that memory CD8 T cells utilize core 2 O-glycans, which mediate interactions with P- and E-selectin, for trafficking to inflamed tissues. Following infection, antigen-specific effector CD8 T-cells strongly express core 2 O-glycans, but this glycosylation pattern is lost by almost all memory CD8 T cells. However, following an inflammatory challenge, de novo generation of core 2 O-glycans occurs on memory, but importantly, not naïve CD8 T cells. Mechanistically, memory and naive CD8 T-cells exhibit the opposite pattern of histone H3 modifications at the Gcnt1 locus, a critical enzyme for core 2 O-glycan synthesis and its expression is regulated by IL-15. Thus, initial antigen encounter epigenetically programs memory CD8 T cells to rapidly generate functional selectin ligands in response to inflammatory cues, a previously unknown mechanism controlling differential localization of naïve and memory CD8 T cells.
Sumoylation is crucial for the stability of Leukocyte-specific protein 1 (P5118)
Authors: Hossain Md Mokarram , Huang Jiannan , Cayabyab Francisco , Liu Lixin 
Published: January 2023

Abstract
Leukocyte-specific protein 1 (LSP1) is an intracellular protein expressed in leukocytes and endothelial cells and regulates several leukocyte and endothelial cell functions including transmigration and chemotaxis of leukocytes in inflammation. There is a discrepancy between its predicted molecular weight (≈37KDa) and that detected in SDS-PAGE (52KDa). To solve this discrepancy, we tested the hypothesis that LSP1 is sumoylated immediately after its translation. Here we demonstrate that, in transfected cells, SUMO1 is conjugated to LSP1 and Ubc9 acts as a SUMO1 ligase for LSP1. Moreover, the sumoylated LSP1 can be desumoylated by the over-expression of SENP1, a SUMO-specific protease. Single lysine to alanine mutation at lysine residue 270 or 318 of recombinant mouse LSP1 prevented its sumoylation in vitro. These LSP1 mutants were degraded rapidly compared to their wild type counterpart. Our results show that sumoylation enhances LSP1 stability, which may ensure its functions in leukocytes and endothelial cells.
Adipose-derived mesenchymal stem cells restore impaired mucosal immune responses in aged mice (P3162)
Authors: Aso Kazuyoshi , Kimura Koh , Tokuhara Daisuke , Kobayashi Taira , Takagaki Kentaro , Sugimoto Shinya , Oki Katsuyuki , Tanemura Hideki , Urushihara Naoki , Lu Shen , Mcghee Jerry , Fujihashi Kohtaro 
Published: January 2023

Abstract
It has been shown that adipose-derived mesenchymal stem cells (AMSCs) can differentiate into red blood cells and epithelial cells as well as mesenchymal cells such as adipocytes, chondrocytes and osteoblasts. Various clinical trials have shown the ability of AMSCs to regenerate these cell types. Age-associated dysregulation of the gastrointestinal (GI) immune system has been well documented. Our previous studies showed that impaired mucosal immunity in the GI tract occurs earlier than is seen in the systemic compartment. In this study, we examined the potential of AMSCs to restore the GI mucosal immune system in aged mice. Aged (&gt;2yr) mice were adoptively transferred with AMSCs. Two weeks later, mice were orally immunized with ovalbumin (OVA) plus cholera toxin three times at weekly intervals. Seven days after the final immunization, when fecal extract samples and plasma were subjected to OVA-specific ELISA, elevated levels of mucosal secretory IgA (SIgA) and plasma IgG antibody (Ab) responses were noted in aged mouse recipients. When cytokine production was examined, OVA-stimulated Peyer’s patch CD4+ T cells produced increased levels of IL-4. Further, CD4+ T cells from the lamina propria revealed elevated levels of IL-4 and IFN-γ production. In contrast, aged mice without AMSC transfer showed essentially no OVA-specific mucosal SIgA or plasma IgG Ab or cytokine responses. These results suggest that AMSCs can restore impaired mucosal immunity in the GI tract of aged mice.
Food allergenic proteins in lactose as the pharmaceutical excipient (P6222)
Authors: Sakai Shinobu , Nakamaura Ryosuke , Nakamaura Rika , Adachi Reiko , Teshima Reiko 
Published: January 2023

Abstract
Most drugs contain pharmaceutical excipients which are pharmacologically inactive substances used as vehicles to deliver the active ingredients of a medication. Some of these pharmaceutical excipients are produced from foods containing allergenic proteins (e.g., milk, egg, peanut, soybean, and sesame) and removing these completely from the excipients is difficult. Therefore, if individuals with food allergies consume drugs containing allergenic food-derived excipients, they are at risk for developing specific allergic symptoms. We determined the levels of total protein in lactose found in pharmaceutical excipients and ethical drugs known to contain lactose by spectrophotometric analyses. The level of total protein in lactose was approximately 1 mg/g. We also determined the levels of allergenic proteins in lactose using commercial enzyme-linked immunosorbent assay systems. The milk proteins in lactose were detected in the range of 1.4-13.1 μg/g. Furthermore, we performed a luciferase-based in vitro elicitation test (EXiLE test) in addition to western-blot analysis using the sera from patients with milk allergies in order to examine the allergenicity of protein residues in lactose. The result of the EXiLE test indicated that the protein residues in lactose have the ability to crosslink human FcϵRI on mast cells.
Complement protein C3 exacerbates disease in a mouse model of chronic wasting disease. (P1343)
Authors: Michel Brady , Zabel Mark , Ferguson Adam 
Published: January 2023

Abstract
Accumulating evidence shows a critical role of the complement system in facilitating attachment of prions to both B cells and FDCs and assisting in prion replication. Complement activation intensifies disease in prion-infected animals, and elimination of complement components inhibits prion accumulation, replication, and pathogenesis. Chronic wasting disease is a highly infectious prion disease of captive and fee-ranging cervid populations that has utilized the complement system for efficient peripheral prion replication and most likely efficient horizontal transmission. Here we show that complete genetic or transient pharmacological depletion of C3 prolongs incubation times and significantly delays splenic accumulation in a CWD transgenic mouse model. Using a semiquantitative prion amplification scoring system we show that C3 impacts disease progression in the early stages of disease by slowing the kinetic rate of accumulation and/or replication of PrPRES. The delayed kinetics in PrPRES replication correlates with delayed disease kinetics in mice deficient in C3. Taken together, these data support a critical role of C3 in peripheral CWD prion pathogenesis.
NICIP, an inhibitory molecule for lamprey variable lymphocyte receptor to signal transduction (P1291)
Authors: Wu Fenfang 
Published: January 2023

Abstract
Lamprey and hagfish are the most ancient jawless vertebrates, which are serving as a “living fossil” of immunological origin and evolution. NICIP is a novel gene that we cloned from lamprey’s lymphocyte-like cells. Structural analysis indicates that NICIP contains two Ig-like domains, a PKD domain, a TM region and a cytoplasm immunoreceptor tyrosine-based activation motifs (ITAM) region. It belongs to IgSF family and may involve in immune response, cell signaling and other functions. Our present research showed that there are interaction between NICIP and VLR, and this interaction can effectively inhibit VLR to transduce signal to intracellular.
Validating cathepsins as selective therapeutic targets for the treatment of particulate-mediated sterile inflammatory disease. (P1373)
Authors: Orlowski Gregory , Kataoka Hiroshi , Rock Kenneth 
Published: January 2023

Abstract
IL-1β mediates a multitude of debilitating, and sometimes fatal, acute and chronic inflammatory diseases. However, existing therapies for these diseases target IL-1β directly, suppressing both its pathologic and protective activities. We propose that cathepsins are therapeutic targets specific to IL-1β-mediated inflammatory diseases caused by sterile particulates. These particulates, such as cholesterol crystals (atherosclerosis), silica (silicosis) and monosodium urate (gout), activate the NLRP3 inflammasome in immune cells, leading to active IL-1β secretion. Rather than conferring protection, this response causes disease. Here, we show that a bioavailable inhibitor of cathepsins B, L & C, called K777, specifically suppresses particulate-mediated IL-1β production by primary macrophages. Using primary and immortalized macrophages deficient in cathepsins B & L, we demonstrate these proteases are critical targets likely responsible for much of K777’s effect. Moreover, we show for the first time that, unlike other NLRP3 stimuli, particulate-mediated inflammatory cell death is inflammasome-independent and cathepsin-dependent. Finally, in a murine model of acute peritonitis, both subcutaneous and intravenous K777 treatment substantially and preferentially reduced IL-1β-dependent neutrophil influx caused by particulate stimuli. Together, these data are the first to translate cathepsin inhibition in vitro into an in vivo model, validating cathepsins as potentially selective therapeutic targets in particulate-mediated inflammatory disease.
The enigma of B cell memory: the scaffolding protein synapse-associated protein 97 is required for enhanced activation signaling through isotype-switched IgG memory B cell receptors (P1124)
Authors: Liu Wanli , Xu ChenGuang , Zhao XingWang , Chen XiangJun 
Published: January 2023

Abstract
The acquired immunological memory was probably first described by the Greek historian Thucydides who observed during a plague in Athens in 430 BCE that “the same man was never attacked twice”. Now 2442 years later, the mechanisms by which the IgG-BCRs function to enhance B cell responses are not completely understood. We showed earlier that the membrane proximal region of the mIgG1 cytoplasmic tail is necessary and sufficient to dramatically enhance the initiation of BCR signaling. We identified that the conserved cytoplasmic tail of the IgG BCR, which contains a putative PDZ-binding motif, associated with SAP97, a PDZ domain containing scaffolding molecule that is involved in controlling receptor density and signal strength at neuronal synapses. SAP97 accumulated and bound to IgG BCRs in the immunological synapses (IS) that formed in response to B cell engagement with antigen. Knocking down SAP97 in IgG+ B cells or mutating the putative PDZ-binding motif on mIgG impaired formation of the IS, initiation of IgG BCR signaling, and downstream activation of the mitogen-activated protein kinase p38. Our subsequent studies showed that SAP97 binding protein, ERM family member ezrin, actively involved in the burst-enhanced memory B cell activation through isotype-switched IgG memory B Cell Receptors. Thus, our studies propose that SAP97 serves as a scaffold protein stabilizing the antigen-driven formation of IgG BCR oligomers and facilitating assembly of the BCR signalosome.
Erk2 is a critical regulator of spontaneous osteochondromas (P1016)
Authors: Wehenkel Marie , Edwards Benjamin , Calabrese Christopher , Pagès Giles , Pouysségur Jacques , Hedrick Stephen , Corr Maripat , Vogel Peter , McGargill Maureen 
Published: January 2023

Abstract
Osteochondromatosis is the most frequent tumor of the skeletal system. Although these bone tumors are benign, they create deformities resulting in impaired use of the affected joints, severe pain, and potential paralysis due to nerve compression. Patients often have a single tumor, although a disorder characterized by the presence of multiple osteochondromas is well established. Surprisingly, mice with a germline deficiency in Erk1 bred to mice with a conditional deletion of Erk2 in the T cell lineage (Erk1-/-.Erk2fl/fl.CD4cre) [DKO] developed multiple osteochondromas by 28 weeks of age. Gross protrusions of the spine and carpus were visible in the DKO mice, and radiological imaging revealed boney abnormalities in the spine, carpus, and sternum. Histological analysis of these protrusions showed an excessive proliferation of cartilage originating from the growth plates, but no mononuclear infiltrate indicative of inflammation. Moreover, we did not find autoantibodies in the serum of the DKO mice. Flow cytometric analysis indicated that an innate-like population of cells was more prominent in the DKO mice compared to littermate controls. These data suggest that the presence of Erk2 in a CD4-expressing, non-T cell population may play a central role in preventing the development of osteochondromas.
GPI-GM-CSF protein transferred onto H5 influenza VLPs remains stably expressed and functionally active (P6168)
Authors: Patel Jaina , He Sara , Amaram Nikhil , Yamanaka Satoshi , Vartabedian Vincent , Radhakrishnan Vijayaraghavan , Song Jae Min , Shashidharamurthy Rangaiah , Compans Richard , Kang Sang-moo , Selvaraj Periasamy 
Published: January 2023

Abstract
Pathogenic H5N1, a lipid-enveloped influenza virus, is a pandemic threat. The use of virus-like particles (VLPs) as an alternative to current influenza vaccines is highly promising. VLPs are similar in structure to their live viral counterparts but do not contain viral genome that is required for replication; hence VLPs provide for a safe and immunogenic vaccine. Although the particulate nature of VLPs allows them to be highly immunogenic, the need for protection against heterotypic viruses still remains. Therefore, inclusion of immunostimulatory molecules (ISMs) onto the VLP surface can help to induce cross-protection against strains and provide for stronger immunity. We were able to show that GPI-anchored-GM-CSF can incorporate stably onto H5 influenza VLP surfaces after a simple and quick protein transfer method. This method allows for the incorporation of the GPI-anchor onto the surface of lipid-bilayered VLPs within a matter of hours and allows for the incorporation of multiple GPI-ISMs in a concentration-dependent manner. Furthermore, protein transferred VLPs were functional in leading to bone marrow derived cell proliferation compared to unmodified VLPs, and incorporated GM-CSF was as functional as equal concentrations of commercially available recombinant soluble GM-CSF. Therefore, VLPs expressing GPI-GM-CSF could lead to enhanced immunogenicity and antiviral immune responses compared to unmodified VLPs.
CD4+ but not CD8+ memory T cells escape DN Tregs-mediated regulation via expression of Serpin Protease Inhibitor 6 (P2160)
Authors: Zhang Zhu Xu , Su Ye , Huang Xuyan , Lian Dameng , Jevnikar Anthony 
Published: January 2023

Abstract
Memory T cell (Tm) homeostasis is poorly understood and is a critical challenge to prevent transplant rejection. At present there are no effective clinical strategies to control Tm and contradictory reports exist as to their control by regulatory T cell (Treg). We previously found that TCRab+CD3+CD4-CD8-NK1.1- (double-negative, DN) Treg could effectively suppress CD4+ and CD8+ effector T cells and thus prevent transplant rejection. We utilized this model to test whether DN Tregs could similarly suppress Tm. Interestingly DN Tregs suppressed CD8+ Tm but had no effect on CD4+ Tm cells. As well DN Tregs express very high levels of granzyme B (GrB), suggesting a potential control mechanism and indeed no suppression was observed using perforin null DN Tregs. In BALB/c (H-2d) to B6-Rag1-/- mice (H-2b) skin allograft transplantation, DN Tregs significantly reduced total CD44high CD8+ Tm in recipients and significantly prolonged graft survival. In contrast, co-transfer of DN Tregs had no effect on CD4+CD44high Tm mediated rejection. There was no effect with co-transferred CD4+ Tm. CD4+ Tm cells express higher levels of GrB inhibitory Serpin Protease Inhibitor 6 (SPI-6) mRNA and protein than CD8+ Tm. Furthermore, SPI-6 null CD4+ Tm is not resistant to DN Treg-mediated regulation. We show for the first time, that DN Treg can control CD8+ Tm and suggest that DN Treg along with targeting SPI-6 may be useful to limit the expansion of both CD4+ and CD8+ Tm cells in transplantation.
Generation of secondary TCRs is critical for T cell development and the formation of an alloreactive T cell repertoire (P2132)
Authors: Ni Peggy , Allen Paul , Morris Gerald 
Published: January 2023

Abstract
A small number of peripheral T cells in humans and mice express two TCRs. Secondary TCRs arise from a lack of allelic exclusion at the TCRα locus. A functional role for secondary TCRs has been demonstrated in several immune responses, including a particularly important contribution to alloreactivity, the molecular basis for transplant rejection. Genetic elimination of secondary TCRs in mice resulted in a 50% reduction in the naive alloreactive T cell frequency. Recently, studies have elucidated a high degree of peptide specificity in alloreactivity, but the role of secondary TCRs is unknown. We hypothesize that secondary TCRs are important for the formation of the naive T cell repertoire capable of responding to individual allopeptides. To address this, we analyzed T cells in mice heterozygous for Trac (Cα+/-), which can only generate a single functional TCRα rearrangement, to look at T cell selection and alloantigen specificity. In TCR Cα+/- mice, there is a defect in the generation of mature T cells attributable to altered thymic selection. The effect on T cell development does not impact peripheral T cell numbers. Despite this, there are functional consequences for the inability to express secondary TCRs: binding to individual allopeptide-MHC tetramers was significantly diminished in naive T cells from TCR Cα+/- mice. Our work reveals a fundamental role for secondary TCRs in thymic selection and in the formation of complete allopeptide-binding T cell populations.
Impaired selectin-dependent leukocyte trafficking induces T cell exhaustion and prevents chronic allograft vasculopathy and rejection (P2140)
Authors: Sarraj Bara , Ye Junsheng , Zhang Zheng , Miller Stephen , Kansas Geoffrey , Ansari M. Javeed 
Published: January 2023

Abstract
Selectin-selectin ligand interactions mediate the initial steps in leukocyte emigration, an integral part of immune response. Fucosyltransferase-VII (FucT-VII) enzyme, encoded by Fut7 gene, is essential for post-translational modification and function of selectin ligands. In an established model of cardiac allograft vasculopathy and chronic rejection, Fut7-/- recipients, with impaired selectin-dependent leukocyte trafficking, enjoyed long-term graft survival with minimal vasculopathy. This was associated with T cell exhaustion in the periphery characterized by impaired effector cytokine production, proliferative defect, increased expression of inhibitory receptors PD-1 and Tim-3 and low levels of IL-7Ra on CD4 T cells and reduced trafficking of polyfunctional CD4 memory T cells to the allograft. Blocking PD-1, triggered rejection only in Fut7-/- recipients, whereas depleting regulatory T cells triggered rejection in WT recipients. Adoptive transfer experiments confirmed that the exhausted phenotype is seen primarily in CD4 T cells with impaired trafficking ability. In summary, these data suggest that impaired leukocyte trafficking is a novel mechanism of CD4 T cell exhaustion and our experimental system serves as an excellent model to study CD4 T cell exhaustion as a dominant mechanism of transplant tolerance. Further, targeting FucT-VII may serve as a promising strategy to prevent chronic allograft rejection and promote tolerance.
Modulation of CD8 expression represents a novel mechanism of peripheral tolerance (BA8P.120)
Authors: Rodriguez-Rodriguez Noe , Apostolidis Sokratis , Nguyen Tran , Martin Villa Jose , Tsokos George , Crispin Jose 
Published: January 2023

Abstract
The ontogeny and raison d'être of peripheral TCR-αβ+CD3+CD4-CD8- T cells, henceforth “DN T cells”, remain controversial. They represent a small population within the bulk of peripheral T cells that expands in certain inflammatory conditions such as systemic lupus erythematosus (SLE) and autoimmune lymphoproliferative syndrome. In fate-mapping experiments, we demonstrate that DN T cells are thymic emigrants. In peripheral lymphoid organs they display an activated phenotype and their TCR repertoire is similar to that of CD8+ T cells. In unmanipulated mice, DN T cells express PD-1 and Helios, molecules induced by TCR activation. Using adoptive transfer experiments with OT-I and HY TCR T cells, we prove that CD8+ but no CD4+ T cells downregulate the expression of CD8 and become DN T cells after encountering their cognate antigen as a self-antigen. After an initial activation and expansion phase where CD8+ T cells upregulate PD-1, Sca-1, and Helios, CD8 is downregulated. Importantly, this phenomenon is not observed during productive immune responses induced by foreign cognate antigens. Notably, the phenotype of DN T cells induced by self-antigen exposure is identical to that of naturally occurring DN T cells from unmanipulated mice. In conclusion, CD8 silencing represents a mechanism that controls activation of self-reactive CD8+ T cells. The presence of small numbers of DN T cells in unmanipulated mice suggests that this mechanism is constantly active.