bluuebunny/crossref_metadata_2025_split huggingface dataset:
β-1,3-glucan induces natural killer cell memory-like immunity
Authors: Li ShuShun , Mody Christopher H 
Published: January 2023

Abstract
Natural killer (NK) cells are considered innate lymphocytes that are able to kill pathogenic microorganisms, including bacteria, parasites and fungi as well as virus-infected cells and tumor cells without prior stimulation. Recent studies suggest that they possess a memory-like property to previously encountered pathogens (e.g., cytomegalovirus). We showed that NK cells directly kill Cryptococcus and Candida, which are fatal opportunistic fungi often infecting immunocompromised individuals. We determined that NKp30 is the activating pattern recognition receptor that mediates NK cell killing of these fungi, and that β-1,3-glucan is the ligand for NKp30. Interestingly, we found that soluble β-1,3-glucan enhanced NK cell killing of C. neoformans and C. albicans. Since humans and animals are exposed to β-1,3-glucan through inhalation of components of the cell wall of microbes or consumption of dietary grains, we asked whether isolated β-1,3-glucan plays a role in shaping NK cell immunity. To determine whether β-1,3-glucan induces a training or memory effect, YT cells, an NK cell line, were treated for 24 hours with laminarin or Saccharomyces β-1,3-glucan and cultured for various times. After an initial period of enhanced C. neoformans killing, killing was reduced to the untreated level on the second week after the initial treatment. However, when cryptococcal killing was assessed on the fourth week, a much greater killing was observed compared to the enhanced killing. This suggests that a recall immune response was induced. We conclude that β-1,3-glucan may induce NK cell memory- like immunity.
Evidence for serial antibody-dependent cell-mediated cytotoxicity (ADCC) by primary unstimulated human natural killer (NK) lymphocytes
Authors: Sung Alexander P , Navarrete-Galvan Lydia T , Guglielmo Michael , Smith-Gagen Julie , Redelman Douglas D , Merica Rebecca , Hudig Dorothy 
Published: January 2023

Abstract
Serial killers are more desirable than single-hit attackers when it comes to treating cancer. Antibody-dependent cell-mediated cytotoxicity (ADCC) mediated by NK lymphocytes is a significant component of many monoclonal antibody anti-cancer immunotherapies. However, there is a significant problem associated with ADCC: the NK cells lose their CD16A Fc-receptors and can no longer recognize target cells coated with antibodies, due to the action of NK cell metalloproteases. In this report, we searched for evidence for serial ADCC before the effector cells lose their receptors. Human ADCC measured by 51Cr-release at multiple NK effector to tumor cell ratios (E:Ts) was largely finished after 4 hours when the targets were Daudi B leukemia cells coated with non-fucosylated obinutuzumab anti-CD20. We reasoned that serial killing is evident when the number of dead targets exceeds the number of initial CD16A NK effectors. We assessed the dead Daudi per NK effector (D/E) at an E:T of 1:4 to detect serial killers given an excess of targets. For 28 healthy human subjects, the total ratio of dead targets per CD16Apos NK effector cell ranged from 0.6 to 2.2, with a median of 1.5 and average of 1.5 +/− 0.4. Four donors (14%) had D/Es &gt;2. The D/E values were similar for CD16A AA158 F/F donors compared to F/V & V/V donors, even though the V-positive donors had greater initial NK cellular levels of CD16A. We conclude that this gross measurement (D/E) can detect serial ADCC even without the information provided by time-lapse cinematography to indicate the actual fraction of effectors that are serial killers. This simple measurement may be useful to screen for promotion of serial killing by reagents that reduce proteolytic loss of NK cell CD16A.
A rho GDP dissociation inhibitor produced by a subset of T-regulatory cells enhances mitophagy in Mycobacterium tuberculosis infected human macrophages
Authors: Paidipally Padmaja , Thandi Ramya Sivangala , Radhakrishnan Rajesh Kumar , McAllister Madeline K , Samten Buka , Vankayalapati Ramakrishna , Tripathi Deepak 
Published: January 2023

Abstract
Previously, we found that a Rho GDP dissociation inhibitor (D4GDI), produced by apoptotic T regulatory cells inhibit Mycobacterium tuberculosis (Mtb) growth in human macrophages through reactive oxygen species (ROS) mediated IL-1β production. Mitochondria is an important source of ROS and critical for antibacterial properties. In the current study, we evaluated the effects of D4GDI on mitochondrial homeostasis and anti-microbial scheme during Mtb infection. Recombinant D4GDI (rD4GDI) treatment of Mtb infected human macrophages enhanced the accumulation of LC3B protein on the mitochondria and triggered mitophagy. rD4GDI treatment significantly enhanced the expression of a mitophagy receptor BCL2 and adenovirus E1B 19-kDa-interacting protein 3-like (BNIP3L). BNIP3L siRNA inhibited rD4GDI dependent accumulation of LC3B protein on the mitochondria and enhanced Mtb growth in human macrophages. Further we found that rD4GDI treatment of Mtb infected macrophages induces the expression of antimicrobial peptide phospholipase family protein phospholipase A2 group VII (PLA2G7). Further studies are underway to determine the interactions between BNIP3L, PLA2G7 and mitophagy in rD4GDI treated macrophages to inhibit Mtb growth in human macrophages.
Activation of TREM1 Signaling in Kupffer Cells Following HIV and HCV Stimulation Augments Hepatic Inflammatory Responses
Authors: Thomas Emmanuel , Hyun Jinhee 
Published: January 2023

Abstract
In this study, we sought to examine inflammatory responses following HIV or HCV stimulation of macrophages or Kupffer cells (KCs), that may contribute to virus mediated inflammation and subsequent liver disease. KCs are liver-resident macrophages and reports have provided evidence that HIV can stimulate and infect them. In order to characterize HIV-intrinsic innate immune responses that may occur in the liver, we performed microarray analyses on KCs following HIV stimulation. Our data demonstrate that KCs upregulate several innate immune signaling pathways involved in inflammation, myeloid cell maturation, stellate cell activation, and Triggering Receptor Expressed on Myeloid cells 1 (TREM1) signaling. TREM1 is a member of the immunoglobulin superfamily of receptors and it is reported to be involved in systemic inflammatory responses due to its ability to amplify activation of host defense signaling pathways. Our data demonstrate that stimulation of KCs with HIV or HCV induces the upregulation of TREM1. Additionally, HIV viral proteins can upregulate expression of TREM1 mRNA through NF-κB signaling. Furthermore, activation of the TREM1 signaling pathway, with a targeted agonist, increased HIV or HCV-mediated inflammatory responses in macrophages due to enhanced activation of the ERK1/2 signaling cascade. Silencing TREM1 dampened inflammatory immune responses elicited by HIV or HCV stimulation. Finally, HIV and HCV infected patients exhibit higher expression and frequency of TREM1 and CD68 positive cells. Taken together, TREM1 induction by HIV contributes to chronic inflammation in the liver and targeting TREM1 signaling may be a therapeutic option to minimize HIV induced chronic inflammation.
Nerve associated lung resident interstitial macrophage subset exhibits distinct localization and polarization
Authors: Yeung Stephen T , Ural Basak Burcu , Damani-Yokota Payal , Devlin Joseph C. , De Vries Maren , Samji Tasleem , Jang Geunhyo , Loke P’ng , Dittmann Meike , Reizis Boris , Khanna Kamal M 
Published: January 2023

Abstract
Tissue resident macrophages are a diverse population of cells that perform specialized functions: sustaining tissue homeostasis and tissue surveillance. Here we report an interstitial subset of CD169+ lung resident macrophages that are transcriptionally and developmentally distinct from alveolar macrophages (AM). They are primarily localized around the airways and in close contact with the innervating pulmonary nerves. These nerve and airway associated macrophages (NAMs) are tissue resident, yolk sac derived, self-renewing, and do not require CCR2+ monocytes for development or maintenance. We show that unlike AMs, CSF-1, but not CSF-2 is required for their development. Bulk population and single cell transcriptome analysis indicates that NAMs are distinct from other lung resident macrophage subsets and highly expressed immune regulatory genes under steady state and inflammatory conditions. NAMs proliferated robustly following TLR activation and influenza infection, and in their absence, the inflammatory response was remarkably augmented resulting in excessive production of inflammatory cytokines and innate immune cell infiltration. We show that AMs contribute to viral clearance while NAMs, contribute to regulating virus-induced inflammation in the lung. Overall, our study relates structure to function at a fundamental level, and provides essential insights into a distinct subset of resident macrophages in the lung that function to maintain immune and tissue homeostasis.
IL-4R signaling mediated polarization of neutrophils promoted monocytes derived alveolar macrophages function
Authors: Chen Fei , El-Naccache Darine W. , Wu Wenhui , Inclan-Rico Juan M. , Ponessa John J. , Espinosa Vanessa , Lothstein Katherine E , rivera-Medina Ameriliz , Siracusa Mark C , Gause William 
Published: January 2023

Abstract
We have reported that alternatively activated neutrophils (N2) enhance lung anti-helminth trained macrophage effector function in response to infection with N. brasiliensis(Fei Chen, et al., NI, 2014). However, it is not clear how these neutrophils develop and what lung macrophage subsets actually mediate resistance. We report here that IL-4R signaling is required for the polarization of neutrophils within 2 days after N. brasiliensisinfection and that this characteristic N2 phenotype can develop in Rag−/− but not Rag2cg−/− mice, suggesting a role for innate lymphoid cells in initiating N2 cell differentiation. N2 neutrophils further showed significant proliferative capacity and oxidative phosphorylation at day 2 after N. brasiliensisinfection. Using CD45 congenic transfers and fate mapping Cx3cr1creER-YFP/+R26tdTomato/+ mice, we further show that monocyte-derived alveolar macrophages are the major effector macrophage subset mediating parasite killing through Arginase 1 dependent mechanisms. In vivo depletion of neutrophils with anti-Ly6G Ab blocked the development of this anti-helminth pulmonary macrophage subset and RNAseq analyses show marked differences from tissue resident alveolar macrophages. These studies demonstrate heterogeneity in lung macrophage and neutrophil subsets during helminth infection resulting in distinct activation pathways required for helminth resistance.
Simplified size-based immune cell monitoring using a hand-held particle counting device
Authors: Smith Janet L , Cappione Amedeo 
Published: January 2023

Abstract
Biological samples are often heterogeneous cell mixtures that differ by type and functional or responsive capacity. Beyond differences in protein expression, many cell types and physiological states are also distinguishable by size (cell diameter). The ability to distinguish population subsets is critical to many aspects of research. Such variation in cellular attributes is most commonly determined by multicolor fluorescent antibody detection of cell type-specific surface markers using flow cytometry. While flow cytometers are complex, trained operator-dependent platforms of core facilities, the availability of simplified methods for identifying activation status and measuring differences in the relative cell frequencies of more than one cell type in samples or co-cultures is somewhat limited. Moreover, despite the need for speed and accuracy, hemocytometry, the most widely used technique for cell counting, is quite tedious and fraught with user-driven variability. To address these needs, we present application of a hand-held cell counter that operates on the basic Coulter Principle of measuring changes in electrical impedance produced by particles (cells or beads) as they pass through a defined aperture. Specifically, instrumentation has been collapsed to the size of a pipette; the precision-made, microfluidic sensor contains the cell-sensing zone that enables size discrimination at the submicron with high counting accuracy. To demonstrate performance, the platform was used to assess cell distribution in human PBMC isolates and determine immune cell activation in response to CD3/CD28 co-stimulation or PHA exposure. All experiments were conducted in comparison to flow cytometry and a standard Coulter device.
Functional responses of salivary gland resident T cells are suppressed by extracellular adenosine
Authors: Springer Lauren E , Smith Corinne , Snyder Christopher M 
Published: January 2023

Abstract
Tissue-resident memory T cells (TRM) survive and persist within non-lymphoid tissues throughout the body, and are therefore subjected to an array of tissue-specific environments. Previous work showed that T cells extracted from the salivary gland were markedly dysfunctional for cytokine secretion upon ex vivo stimulation. This finding is important since the salivary gland is a site of shedding for several viruses. Cytomegalovirus (CMV) is a herpesvirus that persists in the salivary gland and is shed in saliva. Using the natural mouse pathogen murine (M)CMV, we previously showed that most MCMV-specific T cells in the salivary gland readily become TRM. Interestingly, CD8+ TRM in the salivary gland, but not T cells in the spleen, expressed both CD39 and CD73 ectoenzymes that break down extracellular adenosine triphosphate (ATP) to the immunosuppressive molecule adenosine. Expression of CD73 was dependent on TGF-β and neither CD39 nor CD73 expression was dependent on MCMV infection. Importantly, blocking the adenosine receptor A2AR during stimulation restored the function of CD8+TRM recovered from the salivary gland, resulting in IFN-γ and TNF-α production by salivary gland TRM that was comparable to T cells from the spleen. These data suggest that extracellular adenosine inhibits the function of salivary gland CD8+ TRM and that any ATP released during infection may be converted to adenosine by the TRM cells themselves, an avenue we are currently pursuing by use of mass spectrometry with and without MCMV infection.
CD6-ALCAM signaling regulates multiple effector/memory T cell functions
Authors: Ampudia Jeanette , Young-Greenwald William W. , Badrani Jana , Gatto Sole , Pavlicek Adam , Doherty Taylor , Connelly Stephen , Ng Cherie T 
Published: January 2023

Abstract
CD6 is a T-cell costimulatory receptor that has been implicated in the pathogenesis of multiple autoimmune and inflammatory diseases. Primarily expressed on CD4 T cells, CD6 promotes immune synapse formation, T-cell activation and T-cell migration via interaction with its ligand activated leukocyte cell adhesion molecule (ALCAM). While the contribution of CD6 to T cell activation has been well described, less is known regarding the role of CD6 on effector and memory T cells (Teff). Thus, to better characterize this, we examined phosphorylation signaling patterns during CD6 co-stimulation together with the impact of CD6 on differentiated Teff functions. Profiling of ~100 phosphorylation targets associated with T-cell receptor signaling revealed that CD6 co-stimulation on T cells activates factors in pathways involved in actin polymerization, motility, integrin activation and T-cell activation. Comparison of Tnaive cells vs. Teff demonstrated differing levels of phosphorylation in response to CD6 stimulation. Furthermore, CD6 signaling on Teff sustained phosphorylation of these pathways at later timepoints compared to CD28 stimulation. Blockade of the CD6 pathway, using the clinically tested anti-CD6 mAb itolizumab during re-stimulation of CD4 Teff cells in the presence of ALCAM, inhibited multiple effector functions including proliferation and changes in blast size. This effect was observed exclusively in the presence of ALCAM, indicating that the effect was specific to blockade of the CD6-ALCAM pathway. These findings demonstrate that the CD6-ALCAM pathway is a key regulator of effector T-cell functions and further support targeting this pathway to directly inhibit both naïve and effector T cell populations.
Antigen presenting group 3 innate lymphoid cells control neuroinflammation
Authors: Grigg John B , Shanmugavadivu Arthi , Regen Tommy , Parkhurst Christopher , Eberl Gerard , Waisman Ari , Sonnenberg Gregory F 
Published: January 2023

Abstract
Pro-inflammatory T cells in the central nervous system (CNS) are causally associated with multiple demyelinating and neurodegenerative diseases, but the pathways controlling these responses remain unclear. Here we define a population of group 3 innate lymphoid cells (ILC3s) that are uniquely enriched in the CNS during a mouse model of multiple sclerosis. CNS-associated ILC3s exhibit distinct transcriptional profiles, localize in proximity to infiltrating T cells, and express major histocompatibility complex class II (MHCII) and co-stimulatory molecules. Critically, antigen presentation by ILC3s was required to selectively promote pro-inflammatory T cell responses in the CNS and the development of multiple sclerosis-like disease. In contrast, antigen presenting ILC3s in the periphery lack co-stimulatory molecules and did not appear to contribute to disease induction, but rather could induce tolerance in autoimmune T cells and prevent multiple sclerosis-like disease when experimentally targeted to present myelin antigen. Collectively, our data demonstrate an essential role for CNS-associated ILC3s in promoting T cell-dependent neuroinflammation and reveal the potential to harness peripheral ILC3s for the prevention of autoimmune disease.
Laquinimod arrests development of experimental autoimmune uveitis (EAU) and inhibits related immune processes, in the context of altered gut microbiota
Authors: Xu Biying , Jia Xiuzhi , Tang Jihong , Caspi Rachel R , Gery Igal 
Published: January 2023

Abstract
EAU is an animal model for autoimmune uveitis in humans and is used to study mechanisms and potential therapeutics for this blinding disease. Here, we tested the immunosuppressive efficacy of Laquinimod (LAQ), a potent AhR (aryl hydrocarbon receptor) ligand on development of EAU.
C57BL/6J mice were challenged for EAU with 300ug of the uveitogenic antigen, IRBP 651–670 and were treated from day 0 or day 7, relative to immunization, with 25mg/Kg LAQ, or PBS, by oral gavage. EAU progression was monitored by fundus examination and confirmed by histology on day 14 post immunization (p.i.). Treatment with LAQ from day 0 p.i. completely prevented EAU development and inhibited proliferation and production of pro-inflammatory cytokines to peptide 651–670. Treatment with LAQ from day 7 p.i. also significantly inhibited the development of EAU and cellular immune responses, but to a lesser degree than in day 0 mice. In contrast to its inhibitory effect on pro-inflammatory processes, LAQ treatment increased the proportions of T-regulatory FoxP3+CD4+ T cells in treated mice, as compared to the control group.
Metagenomic analyses revealed that immunized mice treated with LAQ had altered composition of gut microbiota when compared with PBS controls, with an increased population of Bifidobacteriaceae, and decreased Clostridaceae 1, Deflubiitaleaceae and Anaeroplasmataceae bacteria.
Our results demonstrate that LAQ, a water-soluble AhR ligand, is a potent inhibitor of EAU development and exerts its effect primarily during the inductive phase of the disease. The connection of the observed changes in gut microbiota to its inhibitory effects on disease is under investigation.
T cell-intrinsic C5L2 activation protects against uncontrolled inflammatory Th responses and autoimmunity
Authors: West Erin E , Freeley Simon , Kolev Martin , Niyonzima Nathalie , Mogensen Trine Hyrup , Woodruff Trent M , Monk Peter , Lappegard Knut , Espevik Terje , Mollnes Tom Eirik , Christiansen Mette , Kemper Claudia 
Published: January 2023

Abstract
Intracellular/autocrine complement proteins have emerged as critical regulators of human Th1 induction and contraction. T cells contain both intracellular C3 and C5 activation systems, with intracellular C3aR1 and C5aR1 stimulation driving T cell homeostatic survival and normal Th1 IFN-gamma production, respectively. Here we demonstrate how the intracellular/autocrine C5 system is regulated by using T cells from the first described patient with C5aR2 deficiency. This patient suffers from an autoinflammatory syndrome, with enhanced inflammatory Th responses and a profound loss of naïve CD4 T cells (95% of CD4 T cells in the blood have a memory T cell phenotype). Thus, in contrast to intracellular C5aR1 stimulation, cell surface expressed C5aR2 is an important negative regulator of inflammatory Th induction. While both C5aR1 and C5aR2 can bind C5a, we found that the carboxypeptidase-processed form of C5a, C5a-desArg, was twice as potent as C5a in reducing Th1 induction. In addition, carboxypeptidase M (CPM) expression was highly induced upon T cell activation indicating that CPM may be mediating T cell-derived C5a-desArg generation and thus, C5aR2 stimulation. In this vein, activation of T cells in the presence of a CPM inhibitor induced Th1 hyper-induction, which was rescued by addition of a C5aR2 agonist and reduced by adding C5a-desArg, but not C5a. The in vivo importance of T cell-expressed CPM and autocrine C5aR2 activation is demonstrated by the fact that CPM−/− mice have enhanced inflammatory Th responses and CPM−/− T cells cause increased pathology in an in vivo T cell transfer colitis model. These data highlight the complex auto-regulatory functions of complement in T cell responses.
Toll like receptor 4 ligation promotes B7 family costimulatory molecules expression in a time and cytokine dependent fashion
Authors: Soto Henry Velazquez , Real Fernanda , Salazar Alberto , Jiménez Martínez Maria C 
Published: January 2023

Abstract

Introduction
The interplay between costimulatory and coinhibitory molecules allows adequate integration of instructions for T cell fate. In this study, we explored how antigen concentration and time exposition to a pathogen associated molecule pattern may modulate the expression of B7 family costimulatory molecules CD86, ICOSL, PDL1, PDL2 and the Interferon Regulatory Factor 4 in human monocyte-derived dendritic cells (MoDCs).

Methods
MoDCs were obtained from healthy donors. MoDCs were stimulated with different concentrations of E. coli lipopolysaccharide and stimulated for 12, 24 or 48 h. Supernatant from stimulated MoDCs were collected for soluble cytokines determination using cytometric bead arrays. Cells were collected and membrane stained with fluorochrome-conjugated monoclonal antibodies to evaluate PDL1, PDL2, CD86 and ICOSL trough flow cytometry. Intracellular staining was performed for IRF4 after cell permeabilization.

Results
TNF α and IL-6 were significantly increased at 12 h, while IFN-γ and IL-2 were increased at 48 h. Costimulatory molecules showed a differential expression pattern. ICOSL was preferentially incremented at 48 h post stimuli independently of concentration used. PDL1 was induced at low concentrations and short stimulation periods, maintaining their expression all along with the stimulation kinetic. PDL2 increased its expression at short time of stimulation. CD86 and IRF4 did not show statistically significant differences.

Conclusion
Our results suggest that costimulatory molecules have time-dependent expression profile, possibly influenced by the cytokines contained in the microenvironment and a lesser by the antigen concentration.