bluuebunny/crossref_metadata_2025_split huggingface dataset:
Prohibitin-1 and -2 have diverse cell-autonomous effects on inflammatory signaling
Authors: Psaltis Christine E. , Aloor Jim , Reece Sky W. , Kilburg-Basnyat Brita , Robidoux Jacques , Fessler Michael , Anderson Ethan , Gowdy Kymberly 
Published: January 2023

Abstract
Prohibitins (PHB1 and PHB2) are proteins that assemble in hetero-oligomeric complexes within the mitochondrial inner membrane and in the plasma membrane lipid rafts where they are at the nexus of metabolic and pro-survival decisions including inflammation. We have recently observed that mice intraperitoneal (i.p.) injected with lipopolysaccharide (LPS) have increased serum prohibitin (PHB), and injection of recombinant PHB1 (rPHB1) post LPS rescues cardiac function. However, the role of PHB in the innate immune response to LPS is currently unknown. Using an in vivo model of sepsis, we injected C57Bl/6J mice i.p. with LPS and 3 subsequent doses of rPHB1. Blood was analyzed for immune cell characterization by flow cytometry, and liver, kidney and lung tissue were harvested for pro-inflammatory cytokine expression. Systemic LPS increased number of blood neutrophils (PMNs), PMN expression of the adhesion molecule CD11b, and tissue expression of TNF-α, IL-6, and IL-1β. Treatment with rPHB1 decreased blood PMN CD11b expression and expression of IL-6 in liver, kidney and lung tissues. To better characterize the role of PHB in immune cells, we knocked down PHB1 or PHB2 in RAW 264.7 macrophages. PHB1- or PHB2-deficient macrophages were then stimulated with LPS, and cell supernatant and lysate were collected for cytokine production. Cell lines deficient in PHB2 had decreased TNF-α compared to macrophages sufficient in PHBs. These data suggest PHB1 and -2 have differential and complex effects on the innate immune response. Cell-associated PHB is required for acute pro-inflammatory signaling responses to LPS that initiate the sepsis response, but extracellular PHB, released in the later phase of sepsis, may feedback to dampen the response.
Structural, functional, and evolutionary differences between PD-L1 and PD-L2
Authors: Philips Elliot A. , Techova Anna S. , Mor Adam , Kong Xiangpeng 
Published: January 2023

Abstract
The PD-1/PD-L1/PD-L2 axis is a critical immune checkpoint that tips immune responses towards tolerance. This axis has been heavily targeted in cancer immunotherapy with monoclonal antibodies, which block receptor-ligand interactions. PD-L1 is constitutively expressed at low levels and is induced on nearly all tissues upon IFN-gamma signaling. In contrast, PD-L2 expression is restricted mainly to antigen presenting cells (APC)s, such as dendritic cells and macrophages. PD-L2 has a 4-fold stronger affinity for PD-1 than does PD-L1. It has been proposed that W110 of PD-L2 accounts for this affinity difference; the corresponding PD-L1 residue is A121. To test this hypothesis, we produced, in a mammalian expression system, a set of PD-L1 and PD-L2 proteins with the W and A swapped, and studied their binding to PD-1 using surface plasmon resonance. Surprisingly, rather than decreasing affinity as expected, a W110A substitution improved affinity by decreasing the off-rate. We propose a novel structural mechanism for PD-L2’s affinity advantage, one that interestingly emerged upon the evolution of placental mammals. We examined the functional effect that these structural alterations have on the PD-Ligands ability to inhibit T cell activation and found a correlation. Additionally, we discovered that PD-Ligands homo- and hetero-dimerize in a manner similar to other B7-family members, with PD-L2 displaying the strongest homo-dimerization. Collectively, these data support our hypothesis that PD-L2 expression on APCs provides an inhibitory advantage, one that may heighten the tolerogenic hurdle that an immune response must overcome during the priming phase.
Leptin-stress response axis promotes allergic airway inflammation
Authors: Yang Xuexian O. , Zheng Handong , Zhang Xing , Wu Dandan , Luo Yan , Yang Xin , Chock Cameron , Liu Meilian 
Published: January 2023

Abstract
Allergic asthma and obesity are both leading health problems in the world. Recent Meta-analysis studies implicated a positive relationship between serum leptin, which is elevated in obese individuals, and the risk of asthma. However, it is not well understood how obesity-associated elevation of leptin increases the risk of asthma. In the current study, we have found that leptin induces the stress response factor XBP1s in pro-allergic TH2 cells, which is required for leptin-mediated pro-allergic TH2 cell survival and cytokine production. In vivo, mice fed with high fat diet had increased serum leptin as observed in human obese population and exacerbated asthmatic symptoms, associated with increased stress responses in CD4+ T cells. Our results reveal a previously unappreciated insight that obesity-associated hyperleptinemia contributes to enhanced pro-allergic lymphocyte responses through targeting the stress response pathway, leading to exacerbation of allergic asthma.
Immunization elicits antigen-specific antibody sequestration in sensory neurons
Authors: Chavan Sangeeta S. , Gunasekaran Manojkumar , Chatterjee Prodyot K. , Shih Andrew , Kumar Gopal , Lee Annette , Graf John , Meyer Dan , Marino Michael , Puleo Christopher , Ashe Jeffrey , Metz Christine N. , Tracey Kevin J. 
Published: January 2023

Abstract
The immune and nervous systems coordinate physiological responses to infection and injury. Immunological memory to pathogens is retained in lymphocytes, but the role of neurons in this process is unknown. Here, we report the surprising discovery that peripheral sensory neurons of dorsal root ganglia (DRG) of immunized mice sequester antigen-specific antibodies. A significant increase in the number of total IgG releasing neurons was observed in DRGs isolated from immunized mice as compared to controls (p&lt;0.001). Around 20% of the neurons in DRGs are positive for antibodies. To determine whether sensory neurons synthesize antibodies, DRG neurons from mice immunized with KLH were analyzed by RNA-seq and qPCR. DRG sensory neurons lack enzymes (i.e., RAG1, RAG2, AID, or UNG) required for generating antibody diversity. Transgenic mice that express a reporter fluorescent protein under the control of IgG1 constant region fail to express Ighg1 transcripts in DRG sensory neurons. Neural sequestration of antibodies is observed in mice rendered deficient in neuronal Rag2, but not observed in sensory neurons isolated from mice that lack mature B cells (e.g., Rag1 knock out (KO) or μMT mice). Adoptive transfer of Rag1-deficient bone marrow into wild-type mice or wild-type bone marrow into Rag1 KO mice revealed that antibody sequestration is observed in sensory neurons of chimeric mice with wild-type bone marrow but not with Rag1-deficient bone marrow. Our studies demonstrate that DRGs do not synthesize antigen-specific antibodies, but rather sequester primarily IgG1 subtype antibodies. These studies reveal that antigen-specific antibodies and sensory neurons may collaborate in bidirectional communication.
A Non-Canonical Role for Interferon Regulatory Factor 3 Downstream of the Interferon Gamma Receptor
Authors: Guinn Zachary P. , Petro Thomas M. 
Published: January 2023

Abstract
IFN-γ promotes the expression of anti-viral and anti-tumoral genes which act intrinsically to the cell (e.g. ISG54, ISG49), as well as genes which express secreted factors (e.g. IP10, IL-15). The canonical pathway of the IFN-γ receptor (IFNGR) revolves around the phosphorylation of the transcription factor, STAT1, promoting its dimerization and localization to the nucleus where it induces the expression of target genes, like Interferon Regulatory Factor 1 (IRF1). Our lab has shown impaired IFNGR signaling in cells which do not express IRF3. IRF3 is a transcription factor canonically activated by Pathogen Associated Molecular Patterns (PAMPs) through Pattern Recognition Receptors (PRRs) like Toll-Like Receptor 3 (TLR3). TLR3 recognizes dsRNA and analogs like Polyinosinic-polycytidylic acid (Poly I:C). WT and IRF3KO RAW 264.7 cell lines expressing secreted luciferase (Lucia) under control of an ISG54 promoter were stimulated with IFN-γ, Poly I:C, or both. IRF3KO RAW-Lucia cells had significantly reduced ISG54 promoter activity in response to all treatments compared with WT RAW-Lucia. This was confirmed via western blot and qPCR analysis. Using qPCR, expression of ISG49 and IL-15 was also IRF3 dependent in response to all treatments. However, expression of IP10 was only IRF3 dependent in response to Poly I:C and not IFN-γ. Thioglycollate elicited peritoneal macrophages (PECs) generated from WT and IRF3KO mice also showed an IRF3 dependency for the expression of ISG54 in response to IFN-γ. These PECs did not show altered STAT1 phosphorylation or IRF1 expression. These data imply that IRF3 contributes to the non-canonical pathway of IFNGR signaling.
MARVELD1 promotes survival during septic shock in mice
Authors: Shi Ming , ZHANG XIAOJUN , FU KAI , CHEN XI , ZHANG SHUAI , TANG YU , MA ZHENGXIONG , WU QIONG 
Published: January 2023

Abstract
MARVELD1 is integral to the inflammatory response occurring during septic shock, although its precise function has yet to be determined. Here we show that compared with wildtype mice, listeria monocytogenes (LM)-induced septic shock in MARVELD1 knockout mice resulted in a heightened pro-inflammatory response and increased mortality. This observation was associated with impaired bacterial clearance related to marked inhibition of the macrophage phagocytic acitivity in MARVELD1 knockout mice. Consistently, the MARVELD1 knockout mice were more sensitive to a lethal endotoxin or LM challenge, while wildtype mice in the bacterial peritonitis model produced a significant survival benefit. These data highlight the crucial role of the MARVELD1 pathway in inducing an adequate macrophage response to polymicrobial sepsis, and both the therapeutic promise and potential risks of its modulation.
Inhibition of IFN-I signaling redirects STAT1 through the IL27R to enhance the generation of CXCR5+ follicular cytotoxic T cells
Authors: Huang Zhe , Zak Jaroslov , Pratumchai Isaraphorn , Teijaro John R. 
Published: January 2023

Abstract
Checkpoint blockade treatment has shown great potential in both chronic viral infection and cancer patients, however the efficacy relies on the number of responsive CD8 T cells. Recently a population of CXCR5+ follicular CD8 T cells (TFC) has been identified in chronic LCMV infection and cancer models as the cells provide the proliferation burst and effector function following anti-PD-1/PD-L1 therapy. Understanding the cellular and molecular mechanisms regulate TFC differentiation and expansion will be crucial for improving the efficacy of immunotherapy. Here, we demonstrate that blockade of IFN-I signaling promotes a cell-intrinsic and sustained generation of TFC following persistent virus infection. Paradoxically, we observed a subset of ISG in TFC were induced by the cytokine IL-27 via STAT1 transcriptional program in the absence of IFN-I signaling. Moreover, blockade of IFN-I signaling either with antibody or pharmacologically through JAK inhibition, promoted the expansion of TFC during viral infection through an IL-27 and STAT1-dependent mechanism. Our results uncover a contrasting role of IL-27 and IFN type I in regulating the expansion of TFC despite their highly overlapping transcriptional signatures. Moreover, we demonstrate that pharmacological inhibition of JAK1/2 signaling induces optimal TFC generation without compromise effector CD8 T cell proliferation and function.
Aryl-hydrocarbon receptor-driven responses of innate lymphocytes
Authors: Stojanovic Ana , Correia Margareta P. , Klein Klara , Angel Peter , Platten Michael , Müller-Decker Karin , Cerwenka Adelheid 
Published: January 2023

Abstract
Innate Lymphoid Cells (ILCs) play significant roles during immune responses to infection at barrier sites and in the control of tissue damage and repair. In addition, ILCs have been involved in several pathologies, including exacerbated inflammation and carcinogenesis. However, understanding the contribution of distinct ILC subsets and modules of their regulation in both homeostasis and disease, is still incomplete. The Aryl-hydrocarbon Receptor (AhR) is a ligand-induced transcription factor that binds various exo- and endogenous polycyclic hydrocarbons and induces transcription of numerous target genes involved in development, cell differentiation and immune response. Here, we show that subsets of NKp46-expressing ILCs, including natural killer (NK) cells and helper ILC1, express AhR and respond to stimulation with AhR ligands. Exposure of NK cells to the tryptophan degradation product kynurenine, an endogenous AhR ligand, induced changes in global gene expression and primed NK cells for enhanced effector responses, including cytokine secretion and migration. In the context of diet-induced chronic liver damage, NK cells accumulated in the liver in an AhR-dependent manner. Conditional AhR deletion abolished NK cell accumulation, which correlated with reduced liver damage. In contrast to NK cells, liver resident helper ILC1 required AhR for the development and/or maintenance. In conclusion, the AhR is expressed by ILC1 subsets and differentially affects their development and effector responses. This might impact ILC1-driven responses in the context of chronic liver damage and could be of significance for liver tissue regeneration.
Global phosphoproteomic analysis of PD-1 signaling reveals T cell subset specific PD-1 functions
Authors: Tocheva Anna S. , Peled Michael , Nayak Shruti , Philips Elliot A. , Ueberheide Beatrix , Mor Adam 
Published: January 2023

Abstract
PD-1 is a cell surface receptor with immunoregulatory properties expressed in activated T cells. PD-1 binds to either PD-L1 or PD-L2 and mediates downstream tyrosine dephosphorylation of key proteins critical for TCR signaling. This prevents excessive inflammation as well as T cell mediated anti-tumor immunity. Anti-PD-1 therapy is effective in ~30% of cancer patients, of which up to 25% develop off-target immune inflammation. Little is known about downstream signaling triggered by PD-1 and further analysis will provide better rationales for safer and more effective PD-1 targeting therapies. We performed tandem mass tag spectrometry and examined protein phosphorylation in Jurkat T cells following PD-1 ligation by PD-L2. Computational analysis revealed the PD-1/PD-L2 phosphoproteomic landscape. Our results confirm previous reports that PD-1 ligation dephosphorylates key proteins in the TCR signaling cascade. In addition, compared to unstimulated cells and to cells stimulated via the TCR alone, αCD3 stimulation in the presence of PD-1 ligation led to significant dephosphorylation of proteins involved in cytoskeletal organization and cellular adhesion. Surprisingly, we observed increased S/T phosphorylation of proteins associated with cell cycle regulation, chromosomal organization and negative regulation of gene expression triggered by PD-1 ligation. Further analysis in primary human cells revealed that these PD-1 triggered pathways were T cell subset specific, which is key for developing T cell subset specific targeted therapies. Our study provides a comprehensive system wide view of the signaling cascade downstream of PD-1 and may serve as a springboard for future therapeutic approaches targeting the PD-1 pathway.
Notch signaling is protective in splenic T cells against apoptosis during endotoxemia via TLR4/Myd88/TRIF/iNOS/NO/TACE pathway.
Authors: Yang Chenxuan , Deng Meihong , Scott Melanie , Billiar Timothy R. 
Published: January 2023

Abstract

Introduction
Notch signaling regulates cell communication and fate decisions, though recently implicated in inflammatory responses, its regulation and roles in sepsis are unknown. We aim to study the biology of Notch signaling in endotoxemia.

Methods
WT (C57BL/6), Myd88−/− , TRIF−/−, TLR4−/−, iNOS−/−mice were injected saline or 5mg/kg LPS i.p, spleen and serum were harvested after 0, 4, 8, 12, 24h. The activation of Notch signaling was assessed as notch intracellular domain (NICD), and apoptosis was measured as cleaved-caspase 3 level by western blot.

Results
The NICD level in spleen peaked at 8h, decreasing to baseline level at 24 h after LPS injection. Cleaved-caspase 3 level in spleen was inversely correlated to the NICD level. Inhibition of Notch activation using DAPT (5 mg/kg) resulted in an increase of cleaved-caspase-3 at 8h after LPS stimulation, suggesting a critical role of Notch signaling for splenocyte survival after LPS stimulation. The sheddase, TNFα-converting enzyme (TACE/ADAM17) participates in activation of Notch and we have previously shown its activation via TLR4/iNOS/NO pathway in hepatocytes, thus we tested whether the pathway is conserved in spleen. LPS-induced Notch activation was significantly lower in TRIF−/−, Myd88−/− , TLR4−/− and iNOS−/− mice. In vitro treatment of SNAP (200 uM) and cGMP analog 8-pCPT-cGMP(400 uM) increased NICD level in primary splenocytes. Treatment with 1400W, an iNOS inhibitor or TAPI-II (800 nM), a TACE inhibitor, reduced LPS-induced Notch activation in splenocytes while increasing cleaved-caspase3.

Conclusion
Our data suggests that Notch signaling is activated during endotoxemia under the regulation of iNOS/NO/TACE and plays a critical role in regulating splenocytes survival.
The incorporation of active learner centered strategies to support improved memory recall and classroom engagement within postgraduate education blended learning education frameworks
Authors: Sika-Paotonu Dianne 
Published: January 2023

Abstract
Immunology education is a vital component for the clinical health sciences context. The inclusion of undergraduate Immunology teaching content within nursing, midwifery and health related courses however remains inconsistent. Blended learning environments provide the opportunity for learner centered classroom activities to support more traditional learning methods. To promote effective teaching and learning of Immunological related content at postgraduate level for classes with students with mixed experiences of Immunology education, integration of active learning and teaching strategies into postgraduate clinical sciences education are needed. The purpose of this work was to explore and assess the impact of Immunological based active memory recall exercises within a blended learning environment for postgraduate clinical health sciences students. Face-to-face Immunology teaching sessions for the Postgraduate Pathophysiology course were delivered with the incorporation of short memory recall sessions throughout in-class time where students recalled their understanding of recent learning content. Students demonstrated improved understanding, memory recall of Immunological concepts in addition to improved in-class engagement and ability to answer verbal in-class questions. In addition to enjoying the session time in class, improved confidence regarding Immunological content covered in class was also noted. Enhanced student knowledge and understanding of Immunology was achieved using integration of active learning and teaching strategies such as Immunology based memory recall exercises within blended learning postgraduate clinical sciences education frameworks.
The antagonist/agonist exam question: thinking like a scientist.
Authors: Parkin Kirstin 
Published: January 2023

Abstract
The goal of my undergraduate immunology course is to teach students how to think like a scientist, specifically how to think like an immunologist. The course has 150 students enrolled each year and I use a variety of team-based, active learning strategies in the classroom and four exams with a similar format that reflect on the class activities. Each exam consists of 4 short answer questions. One question involves students designing their own graphs to represent data from a primary research article covered during the class, and then using labeled drawings to demonstrate how the data is connected to cellular/molecular interactions in an organism. The second question requires students to propose methods for identifying two test tubes containing different contents, such as IL-10 and IL-12, when the test tubes are unlabeled. The third question assesses the ability of students to use labeled drawings to demonstrate cellular interactions, such as the receptor/ligand interactions that would likely lead to a successful vs. unsuccessful pregnancy. Finally, the students are asked to create treatments for conditions such as allergies, transplant rejection, specific autoimmune diseases, and immunodeficiencies. One of the treatments must involve an agonist and the other must involve an antagonist. The student use graphs to demonstrate expected results when the treatments are effective or ineffective. When students were polled on which of the four questions addressed the goal of teaching them how to think like an immunologist, over ninety percent of the students chose the antagonist/agonist question. The use of the agonist/antagonist question promotes deeper understanding of the activating and inhibitory signals that control immune responses.
Immunology Education for English Second Language learners within a Postgraduate Nursing Science program in New Zealand
Authors: Sika-Paotonu Dianne , Tait Carolyn 
Published: January 2023

Abstract
Immunology education is fundamental to support in-depth understanding of many pathological conditions. It is imperative to ensure all students are supported in their learning efforts and that curriculum and teaching content is relevant and appropriately delivered for all students, and in particular English Second Language (ESL) learners. Postgraduate Clinical Health Science ESL learners who have undertaken undergraduate Nursing studies in alternative settings with limited Immunology content may require support at postgraduate level. The purpose of this work was to ensure incorporation of Immunological components within a postgraduate Clinical Health Sciences course was appropriate for all, including ESL learners. In-class Immunology teaching sessions incorporated within a postgraduate Pathophysiology course were delivered to ensure cultural appropriateness for all students, including ESL learners. Active learning approaches employing the use of scientific on-line animations, interactive websites, Youtube scientific content and selected readings were adopted to ensure content delivery was culturally appropriate, stimulating, supported student engagement and participation for in-class learning and teaching activities. Active student engagement and in-class participation was observed with positive student satisfaction scores in response to the Immunological course content from formal student evaluation by all students recorded. Favorable assessment outcomes for Immunological components were noted for all students including ESL learners. These findings support the need to ensure Immunology education for postgraduate Clinical Health Sciences students is appropriate for all, including ESL learners.