bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bifidobacterium longum strain change CD4 and CD8 T cells profile in inflammatory bowel disease induced in mice (MPF6P.663)
Authors: Wasilewska Ewa , Zlotkowska Dagmara 
Published: January 2023

Abstract
Probiotics are defined as a live microorganisms which, when given to the host, beneficially affect the properties of the natural microflora. In our earlier study, we found reduction of the amount of CD8+ and CD4+ T cells in PP in mice fed with different commercial probiotic preparations. Nevertheless, some of them significantly induced CD8+CD4+ cells, most effectively enhanced CD4+ and lowered CD8+ cells in splenocytes, but later, this activating effect was reduced and changed. The presented study compared the effect of potentially probiotic Bifidobacterium longum strain on DSS - induced colitis in older mice. Colitis was induced for 7 days via administration of 2.5% DSS in drinking water. BALB/c mice were gavaged 200 µl of the probiotic preparation (5x109 CFU/mL) or physiological saline, for 14 days. Feeding mice with B. longum significantly increased CD4+ and decreased CD4+CD8+ T cells in MLNs. Significant reduction of CD4+CD8+ T cells was also observed in PPs, however, with no changes in the other populations tested. Splenocytes responded to bacterial feeding by significant increasing both CD4+ and CD8+ T cells, whereas, lymphocytes from peripheral lymph nodes respond by further increase of CD4+, but decrease of CD8+ cells. No changes in lymphocytes phenotyping in the two tested groups were observed in the serum. The experiment showed that the strain tested changed T lymphocyte profile, and further studies are necessary to evaluate its safety and effectiveness.
Protective IL-10-producing Th1 cells also make IL-21, which is regulated by the T-bet/Bcl6 ratio, in a mouse model of chronic malaria (MPF7P.709)
Authors: Carpio Victor , Opata Michael , Montañez Marelle , Dent Alexander , Banerjee Pinaki , Stephens Robin 
Published: January 2023

Abstract
Chronic exposure to the malaria parasite is required for full clinical protection, possibly due to improved survival versus maintenance of effector function of CD4 memory T cells (Tmem). IFN-γ is required for protection; therefore, we investigated the development of Th1 memory in chronic Plasmodium chabaudi malaria infection using ifng-reporter mice. We observed that the expression of Th1 markers (T-bet, CXCR3) peaked well before T cell contraction, suggesting poor long-term commitment to IFN- γ production. In addition, markers of T follicular helper cells (Tfh), such as CXCR5 and Bcl6, were on most Th1 cells, and some co-produced IL-21 and IL-10. While Th1 effector cells differentiate into CXCR3+ Tmem in malaria, they are Tbetlo and do not maintain ifng expression, unless they are dividing, though they do express Bcl6 and IL-21. Interestingly, Bcl6-deficient Th1 cells produce more IL-21, while infection of IL-10-deficient mice increased T cell expression of both T-bet and Bcl6, generating more mixed phenotype cells. P. chabaudi infected IL-10-/- mice die of T cell-mediated pathological inflammation, suggesting that the dominant Th1/Tfh phenotype is pathogenic if not regulated. Expression of Bcl6 and IL-21 in Th1 cells has been described as an early and transient event before full commitment; however, these results suggest that this intermediate stage can be maintained in chronic infection, potentially to prevent both exhaustion and pathology, via IL-10.
Role of interleukin-33 during Schistosoma mansoni infection (MPF7P.713)
Authors: Kalenda Yombo , Masuda Kyoko , Sayed Eman , Nakamura Risa , Kato Kentaro , Uematsu Satoshi , Akira Shizuo , Hamano Shinjiro 
Published: January 2023

Abstract
The host immune response to Schistosoma infection is characterized by two main phases, Th1 and Th2 consecutively, the later one being related to the excretion of mature eggs by the adult female worm. Interleukin-33 (IL-33), a Th2 inducing cytokine, has been reported to be associated with schistosomiasis. Here we assessed the role of this epithelial related cytokine on the time course of the experimental S. mansoni infection in mouse. IL-33 knockout (KO) and wild type (WT) 6-week-old BALB/c female mice were infected with 50 cercariae and sacrificed at 0, 6, 8 and 10 weeks post infection. We monitored symptoms of mice, the worms and eggs burden, kinetics of immune cells in the liver and spleen, and cytokine levels. The size of granuloma induced in the liver and intestine were also measured. There was no significant difference between KO and WT mice in the survival rate, worms and eggs burden; nevertheless some sparse differences related to the increase of eosinophils were detected at 8 weeks post-infection. A very few number of KO mice displayed a severe anemia and colonic bleeding at the end of the experiment, suggesting some intestinal effects of IL-33 as epithelial alarmin. Our results suggest that although IL-33, the Th2 inducing cytokine, was reported to be crucial in other helminthic diseases, it may have little impact on the course of S. mansoni infection.
Inhibiting the mammalian target of rapamycin as an adjunctive therapy for cerebral malaria (MPF7P.714)
Authors: Hart Geoffrey , Gordon Emile , Waisberg Michael , Tran Tuan , Akkaya Munir , Skinner Jeffrey , Zinöcker Severin , Pena Mirna , Yazew Takele , QI chenfeng , Miller Louis , Pierce Susan 
Published: January 2023

Abstract
Malaria is an infectious disease caused by parasites of the Plasmodium spp. In endemic areas, in approximately 1% of cases, almost exclusively in young children, malaria becomes severe resulting in nearly seven hundred thousand deaths each year in Africa alone. Cerebral malaria (CM) is a common form of severe malaria and one for which we have no effective adjunctive therapy. The kinase, mammalian target of rapamycin (mTOR), through its regulation of cellular metabolism, is a central regulator of immune responses and drugs that inhibit the mTOR pathway have been shown to be antiparasitic, raising the possibility that mTOR inhibitors could be effective adjunctive therapies for CM. Here we show in a mouse model of CM, experimental CM (ECM), that the mTOR inhibitor rapamycin protects against ECM when administered within the first four days of infection. Treatment with rapamycin increased survival, blocked breakdown of the blood brain barrier and brain hemorrhaging, decreased influx of both CD4+ and CD8+ T cells into the brain and the accumulation of parasitized red blood cells in the brain. Remarkably, animals were protected against ECM even though rapamycin treatment significantly increased the inflammatory response induced by infection in both the brain and spleen and elevated the levels of peripheral parasitemia. These results open a new avenue for the development of highly selective adjunctive therapies for CM, by targeting pathways that regulate host and parasite metabolism.
IL-27 signaling is crucial for survival of mice infected with African trypanosomes via preventing lethal effects of CD4+ T cells and IFN-γ (MPF7P.719)
Authors: Liu Gongguan , Xu Jinjun , Sun Donglei , Shi Meiqing 
Published: January 2023

Abstract
African trypanosomes are extracellular protozoan parasites causing a chronic debilitating disease with a persistent inflammatory response. IL-10 signaling is essential to maintain the balance of the inflammatory response. Here we demonstrate that IL-27R-deficient (WSX-1-/-) mice are highly susceptible to infection with Trypanosoma congolense and T. brucei, developing exacerbated Th1-mediated immune responses with highly activation of CD4+ T cells and strongly enhanced production of inflammatory cytokines including IFN-γ, which result in severe liver immunopathology and dramatically reduced survival. IL-10 signaling was not impaired in infected WSX-1-/- mice. Either depletion of CD4+ T cells, but not CD8+ T cells, accompanied by a significantly reduced production of IFN-γ, or neutralization of IFN-γ, prevented the early mortality of infected WSX-1-/- mice with a significantly reduced inflammatory response and a major amelioration of liver pathology, providing direct evidence that CD4+ T cells mediated the early mortality of infected WSX-1-/- mice directly via excessive secretion of IFN-γ. Thus, our data identify IL-27 signaling, in addition to known IL-10 signaling for limiting M1-type myeloid cells, as a novel pathway to prevent early mortality via inhibiting hyperactivation of Th1 cells during infection with African trypanosomes. These data are the first to demonstrate the essential role of IL-27 signaling in regulating immune responses to extracellular protozoan infections.
Metalloprotease NleC suppresses host NF-κB/inflammatory responses by cleaving p65 and interfering with the p65/RPS3 interaction (MPF7P.703)
Authors: Hodgson Andrea , Wier Eric , Fu Kai , Sun Xin , Zheng Wenxin , Sham Ho Pan , Yu Hongbing , Johnson Kaitlin , Bailey Scott , Vallance Bruce , Wan Fengyi 
Published: January 2023

Abstract
The NF-κB signaling pathway has long been recognized as a crucial activator and regulator of the innate and adaptive immune responses. More recently, the ribosomal protein, RPS3 has been identified as a non-rel component of NF-κB and functions as a “specifier” protein critical for the induction of selective proinflammatory gene transcription. Attaching/effacing pathogens including enteropathogenic Escherichia coli, enterohemorrhagic E. coli and the rodent equivalent Citrobacter rodentium are important causative agents of foodborne diseases. A/E pathogens have evolved many strategies to evade the host immune response and encode various effector proteins that target NF-κB at different stages. Our work here, demonstrates how the A/E encoded metalloprotease NleC, specifically cleaves the p65 molecule at the N-terminus generating a fragment that is capable of selectively blocking the function of RPS3. This selective blockade of RPS3 results in a dampened immune response following infection, which amplifies the effect of cleaving only a small percentage of p65 to modulate NF-κB-mediated gene expression. Thus, our results reveal a novel mechanism for A/E pathogens to specifically block NF-κB signaling and inflammatory responses by cleaving a small percentage of p65 and targeting the p65/RPS3 interaction in host cells, thus providing novel insights into the pathogenic mechanisms of foodborne diseases.
Vanilloid receptor 1 acts as a regulator of immune homeostasis in the gut (MUC8P.731)
Authors: Acharya Nandini 
Published: January 2023

Abstract
Endogenous cannabinoids (endocannabinoids) are small molecules biosynthesized from membrane glycerophospholipids. Endogenous intestinal cannabinoids such as Anandamide, control appetite and energy balance by engagement of the enteric nervous system through cannabinoid receptors. Here, we show that Anandamide and its vanilloid receptor, VR1, mediate immune homeostasis in the gut and pancreas. Capsaicin, a vanilloid, with which Anandamide shares VR1, acts similarly. Engagement of the cannabinoid/vanilloid receptor augments the number and immune suppressive function of the regulatory CX3CR1hi macrophages, which express the highest levels of such receptors among the gut immune cells. Additionally, mice that are genetically deficient in VR1 have reduced proportions of CX3CR1hi macrophages in the gut. Treatment of mice with capsaicin also leads to differentiation of a regulatory subset of CD4+ T cells called Tr1 cells in an IL27 dependent manner both in vitro and in vivo. In a functional demonstration, tolerance elicited by engagement of VR1 can be transferred to naïve NOD mice (model of type 1 diabetes) by transfer of CD4+ T cells. Our study unveils a novel role for cannabinoids in maintaining immune homeostasis in the gut/pancreas, and reveals a new conversation between the nervous and immune systems, using distinct receptors.
Chronic ethanol consumption alters intestinal microbiota composition (MUC9P.746)
Authors: Barr Tasha , George Michael , Grant Kathleen , Messaoudi Ilhem 
Published: January 2023

Abstract
It is well known that chronic alcohol consumption leads to organ damage and increased susceptibility to bacterial and viral infections. Ethanol exposure in the intestinal lumen alters tight junction proteins leading to disruption of the epithelial barrier. Increased permeability in turn leads to translocation of gut-derived endotoxins causing inflammatory responses in the liver that contribute to alcoholic liver disease. Recent studies have shown that gut mucosa homeostasis is also modulated by the microbiome. Disruptions in the gut microbiome have been described for many diseases including IBD, diabetes and colon cancer. In order to gain a better understanding of the dose-dependent effects of ethanol on the gut microbiome, we performed ribosomal RNA sequencing to determine the differentially expressed bacteria in the small and large intestine in a rhesus macaque model of voluntary ethanol self-administration. In the colon we observed a 2-fold increase in heavy drinkers compared to nondrinkers in the phylum Bacteroidetes that subsequently results in a decrease in Firmicutes, the phylum containing most beneficial lactic acid bacteria. In the duodenum, jejunum, ileum and colon we found a 3-fold decrease in the commensal species Lactobacillus intestinalis and a 2-fold increase in the potential pathogenic species Prevotella copri in heavy drinkers compared to nondrinkers. These studies are likely to reveal mechanisms of ethanol-related gut dysregulation and related inflammation.
Quantitation of activating signaling microclusters’ size and dynamics and their influence on killing kinetics by cytotoxic lymphocytes (TECH2P.915)
Authors: Sykulev Yuri , Steblyanko Maria , Anikeeva Nadia , Keen James 
Published: January 2023

Abstract
Stimulation of cytotoxic lymphocytes via activating receptors results in aggregation of the receptors and recruitment of proximal signaling proteins at distinct location of the contact surface term signaling microclusters. The microclusters that are initially arise in the periphery of the contact surface then move centripetally to the center and signal when they are on the move. We examined the size and dynamics of CD16 activating signaling microclusters at the interface of CD16.NK-92 cells and supported lipid bilayers containing anti-CD16 antibody and ICAM-1, a ligand for integrin receptors, at different densities. We have found that variations in the ligation of integrins but not CD16 receptor significantly affect the microclusters’ size and their displacement and mobility. These and other data provided evidence that integrin signaling effectively modulate activating receptor proximal signaling and regulate the kinetics of cytolytic granule delivery and killing kinetics by cytotoxic lymphocytes.
Single cell gene expression in lupus patient monocytes reveals a differential impact of interferon signaling between monocyte subtypes (TECH2P.920)
Authors: Niewold Timothy , Jensen Mark , Dorschner Jessica , Vsetecka Danielle , Amin Shreyasee , Makol Ashima , Ernste Floranne , Osborn Thomas , Moder Kevin , Chowdhary Vaidehi , Jin Zhongbo 
Published: January 2023

Abstract
Background/Purpose Interferon (IFN) gene expression signatures have been observed in whole blood and mixed immune cell populations in systemic lupus erythematosus (SLE), but the significance of this IFN signaling in immune cell subsets is not well understood. We examined gene expression in individual SLE patient monocytes to explore the impact of increased IFN-induced transcription in single cells. Methods CD14++CD16- classical monocytes and CD14dimCD16+ non classical monocytes from SLE patients were purified by magnetic separation. Single cells were isolated, and gene expression in each individual cell was examined by rtPCR. Results Monocytes from the same SLE patient blood sample demonstrated varying levels of IFN-induced gene expression. In classical monocytes, high IFN score correlated with CD32a, IL1B, and IL8 expression. In non-classical monocytes, high IFN score was correlated with a larger number of inflammatory mediators, including cytokines such as IL12, IL23, and IL15; the immune receptors CD36, CD32a, CD80, and TLR7; and inflammatory signaling genes such as RELA, STAT2, IRAK1, IRAK4, and MyD88. Conclusion We observed striking diversity in the IFN responses of individual monocytes, demonstrating that IFN signaling has distinct effects upon classical and non-classical monocytes. Single cell studies can reveal effects of IFN on single immune cells and uncommon populations such as non-classical monocytes, which would be masked in whole blood or mixed cell populations.
Profiling a structure activity relationship library for immunological activity in human PBMCs: multiplexed measurements of cell phenotype, viability, and cytokine secretion from a single high throughput screen (TECH2P.923)
Authors: Duensing Thomas , Liu Zhaoping , Brodbeck Daniela , Parel Serge , Luu Yen 
Published: January 2023

Abstract
The success of immunomodulatory approaches for the treatment of disease has generated tremendous interest in discovery of new immunotherapy based therapeutics. Immunotherapy has the potential of affecting - positively and negatively - the myriad interactions among cells and signaling molecules regulating the immune system, and projects should include high throughput screening campaigns aimed at profiling the effects of compounds on these complex interactions. We describe a no-wash, high throughput screen that provides a multiparameter activity profile for a structure activity relationship (SAR) library of immunomodulatory compounds. Seven known immunomodulatory compounds were used as templates to generate an SAR expansion library of 1438 compounds. The selection utilized a combination of various substructure and similarity search methods combined with the generation of virtual templates. Phytohemagglutinin (PHA) activated PBMCs were treated with the compounds at two doses, 10µM and 50µM. Immunophenotyping was done by α-CD3 and α-CD8 antibodies and differential toxic effects of compounds on the viability of individual cell subsets plus effects on the secretion of three cytokines were used to generate an immunological profile for each compound tested. We demonstrate the combination of a robust SAR expansion system with high throughput, multiplex screening on primary cells to generate activity profiles that can be used for identifying viable therapeutic candidates.
Whole blood fixation and cryopreservation procedures allow cellular analysis by flow cytometry for up to 30 days after cell isolation (TECH2P.763)
Authors: Paredes R. , Dalle Lucca Jurandir , Crossland Randy , Sheppard Forest 
Published: January 2023

Abstract
Background: Pre-clinical and clinical immunotyping of whole blood (WB) by flow cytometry (FC) is commonly used to assess immune system changes. Current recommendations require FC analysis of pre-stained samples to be completed within 24 h, imposing logistical constraints. Development of sample preservation methods for FC is crucial for extending this technology to locations not equipped with FC and high-throughput studies. Here, we tested a sample preservation method to extend the time window for FC analysis. Methods: Paired fresh WB samples from 4 Rhesus macaques were untreated or stimulated (100 µg/ml LPS, 37oC, 22 h), surface stained, fixed, permeabilized, and intracellular (IC) stained immediately or after preservation by freezing (-20oC; 5, 7, 15, 30 days). Mean fluorescence intensity (MFI) was quantified (BD FACSAria II). Statistical analysis between frozen and fresh samples was performed by the Friedman test with significance at p≤0.05. Results: There were no significant changes over 30 d in MFI from surface markers (CD3, p=0.69; CD4, p=0.32; CD8, p=0.11) or IC cytokines in either lymphoid (IL-8, p=0.98; IL-10, p=0.33; TNFα, p=0.32) or myeloid populations (IL-8, p=0.76; IL-10, p=0.23; TNFα, p=0.14). Conclusion: In frozen WB cells, FC revealed surface marker fluorescence can be maintained for up to 30 days with stable staining of IC markers. These findings have implications for the use of FC in locations with resource constraints, as well as in high-throughput analysis.
RNA fluorescence in situ hybridization and flow cytometry (FISH-Flow) detects antigen-specific T cell responses associated with latent tuberculosis infection (TECH3P.940)
Authors: Vir Pooja , Arrigucci Riccardo , Lakehal Karim , Pine Richard , Bushkin Yuri , Tyagi Sanjay , Davidow Amy , Lardizabal Alfred , Gennaro Maria 
Published: January 2023

Abstract
We have previously described fluorescence in situ hybridization for mRNA detection combined with flow cytometry (FISH-Flow) as a novel method to detect gene expression at a single-cell level. Using mRNA as analyte in flow cytometry presents multiple, biological and technical advantages. Proof-of-principle experiments have also shown that FISH-Flow can detect rare events including antigen-specific T cell responses. Whether the method is applicable to a biomedical problem remains to be determined. Here we applied FISH-Flow to detection of latent Mycobacterium tuberculosis infection (LTBI), a condition affecting approximately a third of the world population. LTBI diagnosis is based on measuring IFN-γ release from peripheral blood mononuclear cells (PBMC) stimulated ex vivo with M. tuberculosis antigen. We obtained PBMC from 60 donors equally distributed between LTBI+ and LTBI- individuals, stimulated them for 6 hrs with M. tuberculosis purified protein derivative (PPD), and measured expression of IFNG and IL2 mRNA by FISH-Flow. The proportions of antigen-induced IFNG and IL2 expression in CD3+ cells were &gt;2-fold higher in LTBI+ than in LTBI- donors (p&lt;0.005). Detection of multiple surface protein markers in combination with mRNA probes determined that CD4+ T cells are the primary cytokine-expressing subset. These results show that the FISH-Flow assay can identify a biological response and demonstrates the usefulness of this technique for basic and clinical immunological studies.
New multiplex interferon panel for simultaneous quantification of 13 human cytokines involved in anti-virus responses (TECH3P.934)
Authors: Sun Binggang , Lopes Patricia , Ji Shaoquan 
Published: January 2023

Abstract
In response to pathogens, especially viruses, cells release interferons and other cytokines to fight the infections. Interferons are typically divided into three types: I (e.g., INF-α, IFN-β), II (e.g., IFN-γ), and III (e.g., IFN-λ1, IFN-λ2). All interferons are important for fighting viral infections and for regulating the immune system. In addition, interferons are critically involved in cancer and autoimmune diseases such as psoriasis, systemic lupus erythematosus, and multiple sclerosis. Expression profiling of interferons and other related cytokines is critical in achieving a deeper understanding of the immune responses and various disease processes. We have developed a multiplex Human Interferon Panel, using fluorescence-encoded beads that are suitable for use on various flow cytometers. This panel allows simultaneous quantification of 13 cytokines involved in anti-virus responses including interferons (α, β, γ, λ1 and λ2), interleukins (1β, 6, 8, 10, 12), TNF-α, IP-10 and GM-CSF. Each antibody pair was carefully selected for assay specificity, sensitivity, accuracy and reproducibility. The panel has been validated by detecting expected changes in biological samples. Further advantages include high sensitivity, small sample volume, flexible assay configurations, and time- and cost-effectiveness. The Interferon Panel can be used for serum, plasma, cell culture supernatant and other sample types, offering a useful tool for biomedical research and drug discovery.