bluuebunny/crossref_metadata_2025_split huggingface dataset:
Monocyte and NK cell dysfunctional responses to Mycobacteria in the context of chronic HIV-1 infection (P4377)
Authors: Gasper Melanie , Sherman David , Sodora Donald 
Published: January 2023

Abstract
Coinfection with M. tuberculosis (Mtb) and other Mycobacteria is a leading cause of morbity and mortality in HIV+ patients. While neither monocytes nor NK cells are directly infected with HIV, several of their phenotypic and functional properties become altered during chronic infection, and both cell types are important to the control of Mtb. We hypothesized that HIV-related NK cell dysfunction may result from impaired monocyte dysfunction and impaired crosstalk with NK cells, which we will evaluate in the context of Mycobacteria infection. In fact, we have observed a decrease in IL-12, an important cytokine in NK cell activation, by blood monocytes from HIV+ individuals following a 6h stimulation with M. Bovis BCG (opportunistic pathogen), compared to uninfected donors (p=0.017). In addition, while developing a flow-based functional assay to measure NK cell killing of autologous Mtb-infected monocytes, we have established that monocytes from HIV+ patients exhibit decreased phagocytosis of fluorescently-labeled BCG (p=0.016) but similar phagocytosis of Mtb compared to uninfected donors. Further, we found that NK cells from HIV+ donors exhibit decreased killing of target cells (p=0.04). Our data demonstrate a reduction in both monocyte and NK cell functionality in the same HIV+ patients and is suggestive of synergism in the dysfunction of two innate cell types with a potential for identifying novel therapeutic targets to enhance innate cellular function in HIV+ patients.
γδ T cells' unique WC1 PPRs direct cellular responses (P4337)
Authors: Baldwin Cynthia , Yokota Payal , Chen Chuang , Flores Carlo , Hsu Hao-Ting , Telfer Janice 
Published: January 2023

Abstract
IFNγ-producing bovine γδ T cells are the major cell population responding in recall responses to Leptospira during the first month following vaccination. The responsive cells bear a subset of the co-receptors known as WC1 which are PRR in the SRCR CD163 superfamily. Annotation of the bovine genome showed there were 13 WC1 molecules coded for by individual genes which are distributed among cells to form a number of γδ T cell subsets. RNA silencing has shown that WC1 co-receptors contribute to the γδ T cell response to leptospira and that the responsive are found within the serologically-defined WC1.1+ γδ T cell subpopulation. Since our data indicate that the WC1 molecules expressed act as pattern recognition receptors interacting directly with bacterial components, we have now evaluated the variegated gene expression of the individual WC1 molecules by specific subsets of gamma delta T cells. We have found that individual cells express more than one WC1 gene product but only a restricted subset of the available 13, generally expressing only 2 or 3. Thus the expression of the WC1 molecules is likely to direct the response by the γδ T cells to pathogens in conjunction with the TCR which is particularly interesting given that the various subsets defined by their WC1 expression also differ functionally with some being pro-inflammatory while others express cytokines consistent with regulatory functions.
CpG ODN-induced suppressor of cytokine signaling-3 expression is required for the activation of MAP kinase and NF-κB via toll-like receptors-9. (P6303)
Authors: Lee Chuhee , Kim Kyung-Chan , Kim Nam-Yi , Lee Hwa-Young 
Published: January 2023

Abstract
Toll-like receptors (TLRs) recognize pathogen-derived structures which can activate the innate immune system and play an important role in the recognition of various microbial-derived molecules; lipopolysaccharide (LPS), lipoteichoic acid (LTA), peptidoglycan (PGN), and bacterial DNA/synthetic CpG oligodeoxynucleotides (CpG ODN). In this study, we demonstrated that unmethylated CpG ODNs induces suppressor of cytokine signaling (SOCS-3) expression from murine macrophage RAW 264.7 cells. CpG ODN-mediated SOCS-3 expression is regulated at transcriptional level and requires de novo protein synthesis. Expression of SOCS-3 mRNA and protein was blocked by inhibitors of TLR-9, MAP kinases, and NF-κB, indicating that TLR-9 dependent MAP kinases and the subsequent NF-κB activation is required for the CpG ODN-induced SOCS-3 expression.
Kinetics of lymphocyte activation in cytokine expression studies (P6317)
Authors: Tran Kimvan , Clor Julie , Tyagarajan Kamala 
Published: January 2023

Abstract
Activation of lymphocytes leads to expression of cytokine receptors, cytokine secretion and the expression of specific cell surface molecules resulting in divergent immune responses. Correlation of cell surface activation with cytokine secretion levels can provide valuable information to evaluate secretion levels. In this study, we demonstrate the kinetics of CD69 and CD25 expression on human peripheral blood lymphocytes from multiple donors following activation with polyclonal activator, phytohaemagglutinin or phorbol myristate acetate and Ionomycin. The studies were performed using novel no-wash assays on the Muse™ Cell analyzer to assess CD69 and CD25 levels on total lymphocytes. Our data shows that the kinetics of CD69 and CD25 expression demonstrates that a high level of lymphocytes depict CD69 expression as early as 6 hours of stimulation, peaked at 24 hours and down regulated after 48 hours. The observed CD25 expression on lymphocytes was significantly less than that of CD69 at 6 hours, increased at 24 hours and peaked at 48 hours with differences between PHA and PMA-ionomycin activation with some variability in extent of activation between donors. Comparison of the level of cytokines secreted and lymphocyte activation will be presented. The combined measurements of activation markers at the cellular level and the secreted cytokines can provide a comprehensive analysis in monitoring lymphocyte response to polyclonal stimuli.
Cytokine/chemokine secretion profiles in differentiated mouse muscle cells (P6341)
Authors: Xiao Qiang , Sun Jingyu , Rose Tina , Samovski Dmitri , Stahl Phillip 
Published: January 2023

Abstract
Cytokines and Chemokines are a group of small proteins that are secreted primarily from immune cells and play essential roles in inflammation and infectious disease. Over the past decade, a great number of studies have demonstrated that some cytokines/chemokines are also secreted from adipocytes (adipokines), hepatocytes (hepatokines), and myocytes (myokines). These non-immune-cell secreted cytokines/chemokines are involved in the regulation of metabolic homeostasis as well as the development of metabolic diseases such as diabetes, obesity, and cardiovascular diseases. To further study muscle cell cytokines/chemokines secretion systematically, we collected mouse muscle cell (C2C12) culture medium samples at Day 0, 1, 3, 5 after cell differentiation and measured cytokine/chemokine concentration in these samples using MILLIPLEX® map Mouse Cytokine/Chemokine Panels I (32 analytes), II (12 analytes), and III (6 analytes) multiplex assays using the Luminex® xMAP® technology. Among the 50 cytokines/chemokines measured we found 9 cytokines/chemokines were easily detectable in C2C12 culture medium samples. Six of them, including IL-6, Eotaxin, IP-10, VEGF, Fractalkine, and TARC, showed a gradual concentration increase after cell differentiation. These data demonstrated that MILLIPLEX® map Mouse Cytokine/Chemokine panels are useful tools for cytokine/chemokine secretion profiling and indicate that muscle cells may secrete more myokines than have been previously reported.
A single population of tolerogenic APC induce two types of CD4+ Treg cells by two different mechanisms. (P1054)
Authors: Stein-Streilein Joan , Mathew Rose , Wu-Freeman Ying , Gordon Siamon 
Published: January 2023

Abstract
Anterior chamber associated immune deviation,ACAID, is an animal model developed to study immune privileged mechanisms of the eye. The in vitro correlate of ACAID is traditionally used to study the cellular and molecular mechanism of the Treg cell development. Tolerogenic antigen-presenting cells (TolAPC) are required to express F4/80 for induction of efferent CD8+ Treg cells. We wondered if F480 is required for the development of afferent CD4+ Treg cells subpopulations. CD4+ Treg cells from cultures containing F4/80 deficient TolAPC cells partially inhibited an in vivo proliferative response to antigen. The depletion of CD25+ T cells after (but not before) exposure to TolAPC in vitro removed all inhibitory activity of the T cells in vivo. Thus, CD4+CD25+ Treg cells generated in vitro are not dependent on F4/80 but do require PDL-1 expression by the TolAPC . Since the F4/80 dependent CD4+ CD25- Treg cells express LAP-1 and PD-1, but not FoxP3, and the F4/80 independent CD4+ CD25+ Treg cells express FoxP3, PD-1, PD-L1, but not LAP-1, they resemble the mucosally-defined Th3 and inducible (i)Treg cells, respectively. Flow cytometry analyses showed that rather than two subpopulations of TolAPC, a single population of F4/80+PD-L1+TolAPC was generated by TGFβ and antigen pretreatment. Together, these data suggest that two subpopulations of afferent CD4+ Treg cells are induced by a single population of TolAPC in vitro.
A novel mechanism of B-cell mediated immune suppression through adenosine production (P1084)
Authors: Kaku Hiroaki , Cheng Kai Fan , Al Abed Yousef , Rothstein Thomas 
Published: January 2023

Abstract
Immune suppression is a critical mechanism to limit excess inflammation and autoimmunity. Treg and Breg (also called B10) are both involved in this process. So far, IL-10 is believed to be a major factor of B cell mediated immune suppression. Here, we report a novel mechanism that B-1 cells suppress inflammation through adenosine generation by CD73. CD73 is an ecto-enzyme that catalyzes the extracellular dephosphorylation of AMP to adenosine. We identified a novel B-1 cell subset, CD73-high B-1 cells. CD73-high B-1 cells exist in both of C57 BL/6 (50-60 %) and BALB/c mice (30-40 %) whereas B-2 cells don’t express CD73. All of B cells express CD39 but its expression levels in B-1 cells are 2-3 fold higher than those in B-2 cells. B-1 cells produced adenosine in the presence of the substrate of these enzymes whereas B-2 cells didn’t, suggesting that coordinated expression of CD39 and CD73 on B-1 cells is important for adenosine generation. CD73 KO mice were more susceptible to dextran sulfate sodium salt (DSS)-induced colitis than WT, and transfer of WT B-1 cells ameliorated the severity of colitis judging from body weight loss, suggesting that CD73 on B-1 cells has a regulatory role on DSS-induced colitis. Thus, our findings suggest a novel regulatory role of B-1 cells on colitis through adenosine generation. We will discuss which cytokines are regulated by B-1 cell-derived adenosine and how adenosine works in cooperation with IL-10 for suppression.
Apoptotic cell-induced tolerance implicates pdc and shapes adaptive immunity (P1100)
Authors: Perruche Sylvain , Bonnefoy Francis , Kaminski Sandra , Saas Philippe 
Published: January 2023

Abstract
Plasmacytoid dendritic cells (pDC) are involved in innate immune response to viral infection. They can mount a strong anti-viral response with the release of high amount of type I IFN after recognition of pathogen-derived material. Since pDC demonstrate potent antigen (Ag) presentation capacity, they have been involved in adaptive immunity. Effector T cell polarization is highly dictates by the microenvironment created by the Ag presenting cell, itself affected by the microenvironment shaped during pathogen elimination. We demonstrated that pDC are critical cells in the tolerance mechanism to apoptotic cells. The microenvironment created by apoptotic cell favors pDC to induce Treg polarization. However, we demonstrated that sole TGF-β polarizes pDC to induce Th17 commitment. Thus we recently identified factors in the secretome of phagocytes eliminating apoptotic cells that prevent pDC to promote Th17 polarization but favor pDC to drive Treg commitment. Such factors implicate chemokine released by phagocytes eliminating apoptotic cells and chemokine receptor signaling in the pDC rendering pDC no longer sensitive to inflammatory stimulus. A lack in this pathway strongly induced inflammatory pDC and inhibited apoptotic cell-induced tolerance through pDC. Thus we identified a chemokine/chemokine receptor critical pathway that prevents inflammation to apoptotic bodies and therefore favor tolerance induction to apoptotic cells though the specific involvement of pDC.
Classifying malaria using multidimensional space maps (P3108)
Authors: Torres Brenda , Oliveira Jose , Schneider David 
Published: January 2023

Abstract
If the path to sickness differs from the path to recovery, we should treat each route differently. Current treatments that inhibit pathogen growth might not be useful when pathogens are already being cleared. To determine the trajectory of both disease and recovery we created a multidimensional map of a malaria model infection in mice, tracing the route from initial infection through recovery. We did this using physiological parameters like red blood cell counts, temperature, weight and blood sugar and included molecular markers from blood transcriptome that was serially collected daily. We then plotted transcript levels versus parasite load and anemia to create a map for the full course of the infection. We can now use this map, constructed from longitudinal data, to scaffold data obtained in human cross sectional studies where we do not know the point of the patient in the infection timeline. This reveals previously unseen structure to these data and should provoke a re-examination of past conclusions. As we add data from non-recovering mice we will build maps that predict outcomes. Our goal is to not simply use these maps as a way of identifying biomarkers but to measure how treatments warp this multidimensional disease space.
The role of the central region of YopD in Yersinia pseudotuberculosis virulence (P3144)
Authors: Adams Walter , Kwuan Laura , Duncan Miles , Auerbuch Victoria 
Published: January 2023

Abstract
Human pathogenic Yersinia express a type III secretion system (T3SS) critical for virulence. The T3SS proteins YopD and YopB form pores in host membranes and mediate injection of effector proteins inside host cells to disrupt innate immune defenses such as production of reactive oxygen species (ROS). Discrete functional domains within YopD have been identified, yet the YopD central region (amino acids 150-227) remains largely uncharacterized. We assessed the importance of the YopD central region by generating Y. pseudotuberculosis YopDΔ150-170 and YopDΔ207-227 mutants and analyzing their virulence in mouse infection models. Y. pseudotuberculosis YopDΔ150-170 and YopDΔ207-227 grew normally in mesenteric lymph nodes and Peyer’s patches following oral infection, but displayed a 20-29,000-fold defect in the spleen and liver. These mutants were also highly attenuated following intraperitoneal infection. These data indicate that the YopD150-227 central region is critical for Y. pseudotuberculosis disseminated infection. To identify the underlying cause of this virulence defect, we examined the ability of the YopDΔ150-170 and YopDΔ207-227 mutants to translocate T3SS cargo and inhibit ROS production in macrophages. While only the YopDΔ150-170 mutant exhibited translocation defects, both mutants failed to fully inhibit ROS production. We hypothesize that the YopD central region facilitates optimal T3SS effector protein delivery to effectively disarm innate immune defenses.
Controlling inflammation: the neurotrophic anti-inflammatory pathway in Chagas disease (P3110)
Authors: Salvador Ryan , PereiraPerrin Mercio 
Published: January 2023

Abstract
Trypanosoma cruzi is the protozoan parasite that causes Chagas disease. The acute phase of infection is characterized by a strong inflammatory response, which subsides in the chronic indeterminate phase. Clinical manifestations emerge decades after initial infection in 30% of those infected, mainly affecting the heart, contributing to chronic chagasic cardiomyopathy. Several neurotrophic receptors of the Trk family and their ligands impact inflammation; nerve growth factor activates TrkA and induces monocyte chemotactic protein-1 (MCP-1) chemokine in cardiomyocytes. We showed that T cruzi activates TrkA and TrkC through a surface-bound neuraminidase named parasite derived neurotrophic factor (PDNF). We hypothesize T cruzi PDNF induces MCP-1 expression in the heart and modulate immune and reparative responses. In T cruzi infected mice, MCP-1 expression correlates with heart parasite burden. Recombinant sPDNF induces MCP-1 in a Trk-dependent manner in cardiac cells, in vitro. In naïve mouse hearts, intravenous sPDNF induces MCP-1 and inflammatory/anti-inflammatory cytokine expression, while multiple sPDNF injections increases MCP-1 receptor CCR2 and stem cell antigen-1 (Sca-1) marker transcripts. In chronically infected mouse hearts, sPDNF treatment yields decreased inflammatory cytokine expression, inflammation and fibrosis. T cruzi PDNF may modulate inflammatory and anti-inflammatory responses, which may reflect the pathogenesis of the parasite and be of therapeutic use.
Vitamin D receptor regulates mucosal immune responses to Clostridium difficile infection (P3076)
Authors: Viladomiu Pujol Monica , Hontecillas Raquel , Pedragosa Mireia , Michalak Pawel , Michalak Katarzyna , Lu Pinyi , Bassaganya-Riera Josep 
Published: January 2023

Abstract
Clostridium difficile is an anaerobic bacterium that has re-emerged as a facultative pathogen and can cause diarrhea, colitis or even death. Vitamin D deficiency contributes to a 5-fold increase in healthcare costs during C. difficile infection (CDI). We investigated the role of vitamin D signaling during CDI by using vitamin D receptor (VDR) -/- and wild-type (WT) mice. The loss of VDR increased disease activity, weight loss, and the severity of colonic inflammatory lesions following CDI. RNAseq analyses showed upregulation of sentrin/SUMO-specific proteases (SENPs), downregulation of phosphatidylinositol 3-kinase regulatory subunit and cAMP-specific 3’5’-cyclic phophodiesterase in colons of VDR-/- mice. SENPs mediate deSUMOylation reactions that can affect both the transcriptional activity of nuclear receptors such as VDR and peroxisome proliferator-activated receptor (PPAR) γ as well as their ability to antagonize NF-kB activity. These results provide a mechanistic basis for the upregulation of IFNγ, IL-6, IL-17 and IL-1β in colons of VDR-/- mice infected with C. difficile. In addition, miR-760-5p was down-regulated in VDR-/- mice and upregulated in WT mice after CDI. Of note, miR-760 has 128 mRNA targets, including lanthionine synthetase component C-like 2 (LANCL2), which is upstream of PPARγ. We provide novel molecular evidence in vivo suggesting novel mechanisms involving LANCL2 and PPARγ by which vitamin D regulates C. difficile-associated disease and immunopathology.
Co-infection with Nippostrongylus brasiliensis does not impair host resistance during chronic Mycobacterium tuberculosis infection: a protective role for granulomas? (P3094)
Authors: Rafi Wasiulla , Potian Julius , Bhatt Kamlesh , Gause William , Salgame Padmini 
Published: January 2023

Abstract
Helminths are known to exert immunomodulatory effects on the host, an effect largely mediated by their induction of generalized Th2 responses. In a recent study, using the murine nematode Nippostrongylus brasiliensis (Nb), we have shown that a prior infection with Nb makes the host more susceptible to tuberculosis (TB). In this study we sought to investigate whether coinfection would exacerbate chronic infection, a phase where Mycobacterium tuberculosis (Mtb) is contained within granulomas. Mice chronically infected with Mtb were coinfected with Nb. We report herein that despite a helminth induced Th2 component, coinfected animals had a similar bacterial burden in comparison with Mtb alone infected animals. This observation could be due to the fact that the Mtb is already established in the granuloma and any effect of the helminth induced Th2 response is unable to shift this balance. In order to further understand the kinetics of helminths influencing TB, we infected mice with Mtb and these were infected 5days later with Nb. Interestingly we observed a significant increase in lung bacterial burden in coinfected animals. These results suggest that the timing of a helminth infection in an Mtb infected individual could have completely different outcomes. Studies are underway to unravel the mechanisms behind these observations. This finding is significant as a helminth infection can occur even during adulthood in endemic areas of the world, regions which are also endemic with TB
SIGIRR limits colitic and epithelial homeostatic responses, but promotes microbiota dependent colonization resistance to enteric bacterial pathogens (P3067)
Authors: Sham Ho Pan , Yu Emily , Gulen Muhammet , Chan Justin , Bhinder Ganive , Brewster Lara , Morampudi Vijay , Gibson Deanna , Hughes Michael , McNagny Kelly , Li Xiaoxia , Vallance Bruce 
Published: January 2023

Abstract
Enteric bacterial pathogens such as Enterohemorrhagic E. coli (EHEC) and Salmonella typhimurium target the intestinal epithelial cells (IEC) lining the gastrointestinal (GI) tract. Despite expressing Toll like receptors (TLRs), IEC are generally hypo-responsive to invading bacteria and their products. One reason is Single Ig IL-1 Related Receptor (SIGIRR), a negative regulator of interleukin (IL)-1R/ TLR signaling expressed by IEC. To address whether SIGIRR expression impacts on enteric host defense, Sigirr deficient (-/-) mice were infected with the EHEC related pathogen Citrobacter rodentium. Sigirr -/- mice responded with accelerated IEC proliferation and strong pro-inflammatory and antimicrobial responses that were primarily IL-1R signaling dependent. Surprisingly the Sigirr -/- mice were highly susceptible to infection, carrying 100-1000x heavier pathogen burden at Day (D) 6 and D10 post infection (pi). Sigirr -/- mice were also found to be unusually susceptible to intestinal S. typhimurium colonization, developing enterocolitis without the requirement for antibiotic based removal of commensal microbes. Strikingly, the exaggerated antimicrobial responses in the Sigirr -/- mice led to the rapid loss of competing commensal microbes (~80%) from the infected intestine, reducing colonization resistance as early as D1 pi. Thus, despite limiting IEC responses to infection, SIGIRR aids host defense by promoting commensal based resistance to pathogen colonization of the GI tract.
NFIL3 suppresses Rorγt transcription and links intestinal Th17 cell differentiation to the circadian clock network (P1223)
Authors: Yu Xiaofei , Rollins Darcy , Kashiwada Masaki , Rothman Paul , Takahashi Joseph , Hooper Lora 
Published: January 2023

Abstract
Environmental factors can profoundly impact the development and function of the mucosal immune system. The microbiota is one such factor that is known to regulate intestinal interleukin-17 (IL-17) producing CD4+ T helper (Th17) cells, but little is known about other environmental cues that influence their development. Here, we show that the transcription factor NFIL3 suppresses Th17 development by directly binding to the Rorγt promoter and repressing its activity. NFIL3 is linked to the circadian clock network through the transcription factor REV-ERBα, and accordingly we observed altered intestinal Th17 cell frequencies in mice that lacked REV-ERBα or were subjected to disturbed light cycles. Together, our findings reveal a new transcriptional regulatory network that regulates intestinal Th17 cell differentiation and unveil a molecular link between the circadian clock circuitry and Th17 cell development.