bluuebunny/crossref_metadata_2025_split huggingface dataset:
Targeting intracellular check points to unleash natural killer cell cytotoxicity
Authors: Barda-Saad Mira , Ben-Shmuel Aviad , Biber Guy , Raiff Anat , Sabag Batel , Matalon Omri 
Published: January 2023

Abstract
Natural killer (NK) cells are a potent weapon of the immune system against viral infections and tumor growth. The actomyosin network generates forces through the activity of actin filaments and myosin motors. This machinery is responsible for the conversion of mechanical forces into biochemical signals in a process termed mechanotransduction. However, the mechanism by which mechanotransduction controls the immune response, and specifically lymphocyte activity, is poorly understood. Here, we demonstrate that actomyosin retrograde flow (ARF) controls NK cell response through a novel interaction between beta-actin with the SH2-domain containing protein tyrosine phosphatase-1 (SHP-1), converting its conformation state, thereby regulating NK cell cytotoxicity. Actin dynamics govern SHP-1 conformational structure dictating its catalytic activity. Indeed, blocking actin dynamics results in reduced SHP-1 activity, by confining SHP-1 to its inactivated “closed” conformation. This reduced enzymatic activity of SHP-1 leads to increased phosphorylation of SHP-1 substrates, an elevation of intracellular calcium flux, and NK cell cytotoxicity. Using multidisciplinary approaches including functional and advanced molecular imaging, we followed the signaling events in individual NK cells from the moment of target cell encounter until the final outcome – target cell killing or NK cell inhibition. Our data suggest that SHP-1 plays a major role as a sensor of ARF-generated forces in the process of mechanotransduction, and reveal a novel mechanism by which regulation of SHP-1 by ARF dictates NK cell killing decisions. Our data identify ARF as a master regulator of the lymphocyte response.
Phosphorylation of MAVS/VISA by Nemo-like kinase (NLK) for degradation regulates the antiviral innate immune response
Authors: Zhong Bo 
Published: January 2023

Abstract
MAVS is essential for antiviral immunity, but the molecular mechanisms responsible for its tight regulation remain poorly understood. Here, we show that NLK inhibits the antiviral immune response during viral infection by targeting MAVS for degradation. NLK depletion promotes virus-induced antiviral cytokine production and decreases viral replication, which is potently rescued by the reintroduction of NLK. Moreover, the depletion of NLK promotes antiviral effects and increases the survival times of mice after infection with VSV. NLK interacts with and phosphorylates MAVS at multiple sites on mitochondria or peroxisomes, thereby inducing the degradation of MAVS and subsequent inactivation of IRF3. Most importantly, a peptide derived from MAVS promotes viral-induced IFN-β production and antagonizes viral replication in vitro and in vivo. These findings provide direct insights into the molecular mechanisms by which phosphorylation of MAVS regulates its degradation and influences its activation and identify an important peptide target for propagating antiviral responses.
Glycan Remodelling Controls Germinal Centre B cell Responses
Authors: De La Pena Enterina Jhon Ralph , Sarkar Susmita , Macauley Matthew 
Published: January 2023

Abstract
Germinal centre (GC) B cells are routinely identified through distinct changes on their surface carbohydrates, called glycans; yet, whether these modifications play an essential role in GC reaction remain unknown. One striking change relates to the monosaccharide sialic acid. In mice, this change is mediated through downregulation of an enzyme called Cmah. As a consequence, GC B cells lose the preferred ligand for CD22, a member of the sialic acid-binding immunoglobulin-type lectins (Siglecs) and a co-inhibitory receptor of the B cell antigen receptor. Since CD22 regulates B cell function, we hypothesize that this specific glycan remodelling on B cells control the GC reaction by modulating the activity of CD22.
To test our hypothesis, we developed a mouse model that constitutively expresses Cmah in B cells. Following immunization, we find that sustained expression of Cmah in B cells results in impaired generation of antigen-specific GC B cells. More significantly, this observed defect is dependent on CD22, highlighting that coordinated loss of preferred ligands for CD22 on B cells plays a critical function in the GC. Furthermore, antibody affinity maturation is significantly impaired in immunized mice harbouring B cells which lack either CD22 or its cis glycan ligands, further supporting the role of CD22 and its glycan ligands in the GC. Ongoing efforts are focused on defining the GC-specific nature of these observations and examining potential mechanism related to BCR signalling, GC B cell survival and exit to plasma cells, and B-T cell interactions.
Role of peptide-MHCII complexes in germinal center B cell selection and plasma cell differentiation
Authors: Jing Zhixin , McCarron Mark , Fooksman David R 
Published: January 2023

Abstract
Germinal center (GC) selection is critical for generating affinity-matured antibodies in T-cell-dependent immune responses. Despite the essential help follicular T cells provide to GC B cells, it is unclear how GC B cell fate decisions are made. Although cognate peptide-MHCII (pMHCII) is critical for T cell help, how pMHCII density directly contributes to B cell selection and cell fate remain unclear. The a-DEC205-OVA (dec) model system has been widely used to interrogate GC B selection, by enhancing selection of WT (Ly75+/+) B cells over untargeted Ly75−/− (KO) B cells. However, under these conditions, it is unclear what the fate of B cells presenting intermediate levels of pMHCII is and whether T cell help is indeed limited.
To investigate these questions, we used Ly75+/− (DEC-het) B cells, which expressed intermediate levels (~50%) of surface DEC205 protein compared to WT B cells, and presented proportional amount of OVA peptide after dec treatment. Using competitive transfers, we found that WT B cells expanded two-fold more than DEC-het B cells. This 2-fold difference in expansion was maintained at a wide range of dec administration, indicating that in vivo, T cells can distinguish different densities of pMHCII on multiple B cells in the same GC and respond proportionally. Surprisingly, we found that upon dec stimulation, both WT and DEC-het GC B cells were equally capable of differentiating into plasma cells, suggesting that pMHCII density does not control B cell fate directly. Blocking of CD40 signaling removed the two-fold advantage of WT over DEC-het B cells during selection after dec treatment, suggesting CD40 signaling is proportional to pMHCII density. These results provide new interpretations of how GC selection occurs in vivo.
In vivo mechanisms by which Irf5 regulates BCR-, TCR- and TLR-induced plasma cell generation and antibody secretion
Authors: Brune Zarina , Rice Matthew , Matta Bharati , Lu Ailing , Barnes Betsy J 
Published: January 2023

Abstract
Generation of antibody secreting cells (ASCs) requires multiple B cell activation pathways, including B cell receptor (BCR), T cell receptor (TCR), and Toll-like receptors (TLRs). In human naïve B cells, we reported that knockdown of interferon regulatory factor 5 (IRF5) resulted in IgD retention, reduced proliferation and plasma cell differentiation, and reduced IgG secretion. Defects were due to early impairments in B cell activation and clonal expansion. Conversely, murine Irf5 was reported to regulate antibody production through direct control of class switch recombination (CSR) at the γ2a locus. To further elucidate distinct and overlapping roles between human and murine IRF5 in the regulation of plasma cell differentiation and antibody production, we performed in vivo analysis of B and T cell differentiation and function in wild-type (wt) and Irf5 knockout (ko) littermate mice after immunization. B and T cell subsets were analyzed by multi-color flow cytometry. Naïve B cells were in vitro differentiated to ASCs with CpG-B, anti-IgM, anti-CD40 and IL-21. Kinetics of B cell activation, AID expression and IgG production were determined. BCR and TCR signaling were examined by phospho-flow. A combination of T-dependent/independent and TLR-dependent/independent immunizations were used to study ASC differentiation. RAG−/− mice were used to determine cell type-specific effects. Results indicate distinct roles for human and murine IRF5 at the early stages of BCR signaling and redundant roles at the later stages of CSR, plasma cell differentiation and antibody secretion via regulation of AID.
Antigen valency dictates the affinity of B cells recruited to germinal centers and the magnitude of extrafollicular B cell responses
Authors: Kato Yu , Groschel Bettina , Abbott Robert K. , Menis Sergey , Irvine Darrell J. , Schief William R. , Crotty Shane 
Published: January 2023

Abstract
While it is widely agreed that antigen valency is a factor in immunogenicity, how antigen valency contributes to B cell selection during antigen-specific responses is poorly understood. Here, using HIV immunogens with a defined valency ranging from 1 to 60, we investigated the role of antigen valency during different phases of the antigen-specific B cell response. We show that highly multimerized immunogens rapidly activated cognate B cells in vivo with little affinity discrimination, initially. This subsequently led to strong extrafollicular and germinal center responses where B cells within a broad affinity range were efficiently recruited. Low-valency (1–8 copy) antigens induced less diverse effector B cell responses, with limited recruitment of low-affinity B cells. The data reveal roles of antigen valency in dictating affinity thresholds required for recruitment of B cells to germinal centers or rapid extrafollicular plasma cell responses, with important implications for vaccine design.
Liver-resident CD8 T cells do not need Levy walks to be efficient killers
Authors: Ganusov Vitaly V , Rajakaruna Harshana , O’Connor James , Cockburn Ian A 
Published: January 2023

Abstract
Movement strategies by which antigen-specific CD8 T cells search for sites of infection are not fully understood. It was recently suggested that activated CD8 T cells, specific for Toxoplasma gondii, perform generalized Levy walks in murine brains; that is cells perform a local search and then displace randomly to distant locations. Several theoretical studies found that Levy walkers are an order of magnitude more efficient at finding rare targets than Brownian walkers. We investigated movement patterns of Plasmodium-specific liver-resident CD8 T cells using intravital microscopy. Based on the distribution of movement lengths Trm cells performed Brownian walks and yet displayed superdiffusive (Levy-like) behavior, i.e., on average they displaced further from the initial position than that predicted by the standard diffusion theory. Mapping out structure of liver sinusoids revealed that superdiffiusive nature of T cell walks in the liver is likely due to small branching angles of the sinusoids, instructing T cells to move directly. By simulating T cells moving as Brownian or Levy walkers in free space or physiologically constrained liver sinusoids we showed that structure may override the intrinsic pattern of cell movement. Interestingly, while Levy walkers indeed required less time than Brownian walkers to find rare targets when search was done in space with no constrains, on average, this advantage disappeared when search was done in the liver. Our results, thus, suggest that observed patterns of movement of T cells in the liver are likely to arise as a combination of cell-intrinsic search patterns and environmental constrains, and that evolutionary benefits of Levy walks may be limited in biologically-relevant environments.
Mechanistic mathematical representation of CD8+ T cell mediated drug-induced liver injury (DILI) Part 1: initial development using experimental antigen, ovalbumin (OVA)
Authors: Battista Christina , Kenz Zackary , Shoda Lisl 
Published: January 2023

AbstractIdiosyncratic DILI (iDILI), rare liver injury not dependent on drug dose, is poorly understood and costly to patients and drug developers. Some iDILI events appear immune-mediated. This work builds on the established hepatocyte life cycle and intrinsic injury models in DILIsym, utilizing quantitative systems toxicology methods to incorporate direct activation of liver CD8+ T cells by OVA-expressing hepatocytes.Simulated hepatocyte OVA expression leads to activation of naïve OVA-specific CD8+ T cells. Activated T cells differentiate to effector, memory, or exhausted T cells. T cell cytotoxicity induces apoptosis in antigen-presenting hepatocytes, with resultant ALT release. By design, the model reproduces a differential CD8+ T cell response to low vs. high levels of hepatocyte OVA expression consistent with data 1,2. Mechanistically, the differential response is driven by effector T cell mediated clearance of low antigen vs. persistent high antigen and differentiation of exhausted T cells3. Reducing T cell functional avidity by 70x, as described for OVA altered peptide ligands4, reduced IFN-γ production and T cell cytotoxicity consistent with data5.By reproducing these results, DILIsym establishes proof-of-concept in the quantitative relationships among antigen-expressing hepatocytes, antigen-specific T cell number, cytotoxicity, and ALT release. These quantitative relationships set the stage for translation to drug-mediated CD8+ T cell cytotoxicity6.
AR blockade promotes immunosuppression and pro-tumorigenic activity of myeloid cells
Authors: Udartseva Olga , Consiglio Camila Rosat , Ramsey Kimberley , Gollnick Sandra O. 
Published: January 2023

Abstract
It has been recently recognized that sex hormones can regulate hematopoiesis and modulate immune phenotypes that underlie sex disparities in immune response. Androgen receptor (AR) antagonism is known to compromise T cell response; however, the role of AR signaling in myeloid cell biology is poorly understood. To address this question, we analyzed the effect of AR blockage on functional activity of myeloid cells in MARKO (Myeloid AR KnockOut) mice and after pharmacological inhibition of AR. We demonstrated that AR antagonism promoted VEGFa, Arg1 and PD-L1 expression in MDSCs, and enhanced their immunosuppressive activity. AR blockage induced metabolic switch in myeloid cells by suppression of mitochondrial respiration and upregulation of glycolysis. AR-mediated metabolic reprogramming was regulated in a MPC/AMPK-dependent manner. To understand the role of AR antagonism in anti-tumor immunity, we utilized the AR-independent models of MC-38 colon and TRAMP C2 prostate carcinomas. Neither AR inhibition nor AR deletion in myeloid cells affected tumor infiltration with TAMs, MDSCs or T cells. However, in both models AR blockade and genetic deletion promoted tumor growth in immunocompetent but not immunodeficient mice, suggesting an important role of AR-mediated regulation of immunosuppression in anti-tumor immunity. Taken together, our findings reveal that AR signaling represents an important regulator of myeloid cell functional activity, and suggest a novel tumor-extrinsic mechanism of prostate cancer relapse.
Endoplasmic reticulum-targeting delivery of curcumin alters reactive oxygen species production and phenotypes in activated macrophages
Authors: Yen Chia Liang , Chen Ru-Fen , Shieh Dar-Bin , Jiang Si-Tse , Shieh Chi-Chang K 
Published: January 2023

Abstract
Chronic granulomatous disease (CGD) is a primary immunodeficiency disease caused by defects in the leukocyte NADP oxidase. We previously reported that organelle-targeted delivery of sarcoplasmic/endoplasmic reticulum calcium pump (SERCA) inhibitor curcumin with the endoplasmic reticulum (ER)-targeting peptide decorated PLGA nanoparticles (NPs) rescues mutant H338Y-gp91phox protein maturation of a particular type of CGD with a CybbC1024T mutation, leading to ER retention of the mutant protein. However, how the rescuing of mutant H338Y-gp91 phox protein with ER-targeting curcumin-PLGA NPs on macrophages functions and differentiation are still unknown. In this study, we analyzed gp91phox maturation and macrophage M1/M2 polarization in bone marrow-derived macrophages (BMDMs) from wild-type (WT), Cybb−/−, and CybbC1024T transgenic Cybb−/−mice. First, we found that the levels of CD86 and CD206 expression with lipopolysaccharide+interferon-γ (LPS+IFN-γ) stimulation were similar in WT and CybbC1024T transgenic Cybb−/−BMDMs and CD206 expression was lowered in Cybb−/−mice when compared with WT and CybbC1024T transgenic Cybb−/−BMDMs. Second, we found that CD206 levels after LPS+IFN-γ stimulation were enhanced by free curcumin, curcumin-PLGA NPs and ER-targeting curcumin-PLGA NPs treatment in CybbC1024T transgenic Cybb−/−BMDMs, but not in WT and Cybb−/− BMDMs. Meanwhile, we found that ER-targeting curcumin-PLGA NPs enhanced gp91phox glycosylation in CybbC1024T transgenic Cybb−/−BMDMs. These findings may indicate that both the expression and maturation of gp91phox are crucial for macrophage M1/M2 polarization. Gp91phox maturation may be important for M2 phenotypes in activated macrophages.
Metabolic characteristics of Th2 and Th17 cells drive airway inflammation
Authors: Contreras Diana C , Cephus Jacqueline Y , Dahunsi Debolanle O , Newcomb Dawn C , Rathmell Jeffrey C 
Published: January 2023

Abstract
Asthma is an airway inflammatory disease that is mediated by T effector (Teff) cells, specifically Th2 and Th17 cells. Increases in disease severity leading to neutrophilic asthma is characterized by a shift towards a higher Th17 response. Typical treatment of asthmatic patients include the use of glucocoriticoid (GC) steroids. These drugs are effective at controlling inflammation in mild cases but as disease severity increases towards neutrophilic disease there is resistance by Th17 cells to the effects of these drugs. A key therapeutic objective is to identify either alternative treatments for asthma or targets that make Th17 cells more susceptible to GCs. Studies have shown that cells that have increased glycolysis and oxidative phosphorylation are more resistant to the effects of GCs. We hypothesized that the intrinsic metabolic phenotype of Th17 cells allow them to be more resistant to GCs. In this study we show, that in the murine model of airway inflammation there are differences in the expression of metabolic proteins between Th2 and Th17 cells that are present in the lung. Additionally, we show that metabolic inhibitors alter the functionality of the Teff cells and can be a target to restore sensitivity to GCs. We have also performed CyTOF analysis of asthmatic patients’ PBMCs, these data demonstrate a much higher metabolic phenotype of asthmatic patients when compared to healthy controls. Overall our studies suggest that metabolic inhibition may be a viable therapeautic in conjuction with GCs in order to dampen the function of Teffs in the airway.
Maternal immunometabolic adaptation in pregnancy
Authors: Rees April , Jones Nick , Jenkins Ben , Richards Oliver , Cronin James , Thornton Cathy 
Published: January 2023

Abstract
Pregnant women undergo dynamic metabolic and immunologic changes to ensure provision of nutrients to, and prevent rejection of, the fetus. The mother increases glucose production, glucose intolerance and insulin resistance to support fetal-placental development and transitions from lipid storage to lipolysis to meet her own energy demands. Maternal immunoregulation prevents rejection of the fetal semi-allograft whilst maintaining protection against pathogens.

Hypothesis
Immunometabolic adaptation underpins maternal immune function changes in pregnancy.

Aim
To determine whether monocytes undergo metabolic adaptation in pregnancy and at which gestational stage this might occur.

Results
Peripheral blood monocytes of term pregnant (&gt;37 weeks) and not pregnant women were compared using flow cytometry. Pregnant women had significantly more non-classical CD16+ monocytes, and CD16+ and CD16− subsets had more CD36 (fatty acid transporter; p = 0.0025) and less CD98 (amino acid transporter; p = 0.0043).
Bioenergetic analysis of monocytes showed oxidative phosphorylation was significantly reduced in pregnant women; glycolysis was unchanged. Also, oxidative phosphorylation was reduced significantly in the mononuclear cells of women with gestational diabetes at 28 weeks compared to healthy pregnant women at the same gestational stage.

Discussion
Ongoing work includes analysis of mitochondrial health and measuring transcriptomics, biosynthetic lipids, cytokines, and enzyme responses. Results are vital to understanding immunological adaptation during pregnancy. This could provide insight into adverse pregnancy outcomes and women’s long-term health in relation to immune-mediated diseases.
Metformin effect on human macrophage inflammatory response and phagocytosis of Mycobacterium tuberculosis
Authors: Cervantes Jorge L , Sanca Angel , Barragan Jose 
Published: January 2023

Abstract
Metformin (MTF) has a well-documented ability to cfontrol hyperglycemia, which has been shown to have effects on macrophage and lymphocyte functions that are key to controlling tuberculosis (TB) infection.
Here, we aimed to better understand the effects of MTF on the phagocytosis of Mycobacterium tuberculosis (Mtb) by human macrophages. PMA-differentiated THP-1 cells with two reporters for nuclear factor-κB (NF-κB), and interferon-regulatory factors (IRFs) were treated with 2mM of MTF for 4 hours, and then inoculated with Mtb from various lineages. Since MTF can also directly inhibit key metabolic processes of Mtb, we controlled this variable by using of gamma-irradiated mycobacteria. Phagocytosis was assessed by immunofluorescent assay.
Phagocytosis of Mtb increased in MTF-treated macrophages. NF-κB activation after Mtb stimulation was lower in MTF-treated macrophages. The effect on IRF activation was minimal.
Our results indicate that MTF improves phagocytosis of Mtb by macrophages, while it at the same time modulating their inflammatory response. Downregulation of type I IFN pathways, associated with active TB infection, could allow for improved activation of macrophages in the presence of TB infection. These results support the effects of MTF in keys steps TB infection control, and support its use as an additional treatment for TB.