bluuebunny/crossref_metadata_2025_split huggingface dataset:
A host cell-intrinsic innate regulatory circuit limits inflammasome activity and promotes immune escape of Salmonella inside macrophages
Authors: Akhade Ajay Suresh , Atif Shaikh M , Lakshmi Bhavana S , Dikshit Neha , Qadri Ayub , Subramanian Naeha 
Published: January 2023

Abstract
Inflammasomes have been implicated in the detection and clearance of a variety of bacterial pathogens, but little is known about whether there is active cross-talk between the host sensing mechanism and the expression of stimulatory ligands by the pathogen. Here we show that inflammasome activation regulates expression of the NLRC4 and TLR5 ligand, flagellin, by Salmonella. Host lysophospholipids released upon NLRC4-mediated pyroptosis increase flagellin expression by extracellular bacteria that enhances pyroptosis upon internalization, establishing a positive feedback loop that potentiates Salmonella detection and clearance. A TLR-dependent host negative feedback response later inhibits inflammasome activation and lysophospholipid biosynthesis within cells, prompting the pathogen to switch to a flagellin-low phenotype and establish an intracellular survival niche inside macrophages. Ablation of this host regulatory circuit prevents downregulation of flagellin by intracellular Salmonella and promotes bacterial clearance in vivo. Our data identify modulation of expression of a bacterium-derived inflammasome ligand by the very process of inflammasome activation as a novel mode of host-pathogen cross-talk and reveal a host mechanism that is adapted by Salmonella for flagellin downregulation and immune escape within macrophages.
PtdIns4P on dispersed trans-Golgi network mediates NLRP3 inflammasome activation
Authors: Chen Jueqi , Chen Zhijian J 
Published: January 2023

Abstract
The NLRP3 inflammasome, which has been linked to human inflammatory diseases, is activated by diverse stimuli. How these stimuli activate NLRP3 is unknown. Here we show that different NLRP3 stimuli lead to disassembly of the trans-Golgi network (TGN). NLRP3 is recruited to the dispersed TGN (dTGN) through ionic bonding between its conserved polybasic region and negatively charged phosphatidylinositol-4-phosphate (PtdIns4P) on the dTGN. The dTGN then serves as a scaffold for NLRP3 aggregation into multiple puncta, leading to polymerization of the adaptor protein ASC, thereby activating the downstream signalling cascade. Disruption of the interaction between NLRP3 and PtdIns4P on the dTGN blocked NLRP3 aggregation and downstream signalling. These results indicate that recruitment of NLRP3 to dTGN is an early and common cellular event that leads to NLRP3 aggregation and activation in response to diverse stimuli.
Nucleic acid sensing in a 3D hydrogel model of B16F10 mouse melanoma
Authors: Heller Loree , Bulysheva Anna , Arpag-Mcintosh Sezgi , Semenova Nina , Cemazar Maja 
Published: January 2023

Abstract
Pattern recognition receptors (PRRs) are nearly ubiquitous in immune and non-immune mammalian cells. PRRs detect molecular pattern molecules associated with pathogen invasion as well as internal damage markers. A subset of pattern-recognition receptors, found endosomally or cytosolically, detect nucleic acids including DNA. Recently, a number of cytosolic DNA-specific PRRs expressed in non-immune cells have been described. Binding and activation of these sensors induces the production of Type I interferons and pro-inflammatory cytokines and chemokines.
Many cell types, including B16F10 mouse melanoma cells, respond to DNA electroporation with the upregulation of specific PRRs on the mRNA and protein levels. We developed B16F10 melanoma cell hydrogels to test whether cells in these constructs (“3D cells”) responded similarly to cells in suspension. The cells formed small spheroids within the hydrogels, similar to in vivo tumor morphology, cell-cell, and cell-matrix microenvironment. 3D cells maintained a nearly 73% proliferation rate compared to a 94% rate for cells cultured in 2D monolayers. The relative metabolism kinetics of 3D cells were similar to monolayers in culture, as was transgene expression after delivery by electroporation. However, while similar PRR mRNAs were upregulated, including DAI/ZBP1, DDX60, and p204, the relative levels of upregulation differed significantly between suspended B16F10 and 3D cells. In addition, p204 was upregulated in control constructs but not in suspension cells receiving pulses without DNA. Supporting these observations, the induced cytokine profile also differed. These data emphasize the importance of testing multiple models before generalizing to tissue responses.
Outcomes of ERK Signaling Differ in Macrophages and Dendritic Cells
Authors: Groft Sarah Grace , Nagy Nancy , Shukla Supriya , Sweet David , Boom W Henry , Harding Clifford V 
Published: January 2023

Abstract
Stimulation of dendritic cells and macrophages by pathogen-related products activates a variety of immune responses. Pathogen-induced cell signaling and associated outcome research is abundant in macrophages, but minimal in dendritic cells. We investigated the role of ERK signaling in the induction of cytokines such as IL-12p40 and IL-10 following activation by Pam3Cys or Mycobacterium tuberculosis (Mtb) H37Ra. ERK ablation was accomplished pharmacologically with MERK1/2 inhibitor U0126 or genetically by using cells from Tpl2−/− mice (TPL2 connects TLR to ERK). We utilized western blotting to examine ERK activation, ELISA to assess cytokine production, and qRT-PCR to investigate mRNA levels. In macrophages, blockade of ERK signaling inhibited IL-10 production and increased IL-12p40 production (6–10 fold increase in IL-12p40 mRNA and 3–4 fold increase in protein). In dendritic cells, ERK blockade similarly inhibited IL-10 production, but produced a very different change in IL-12p40 expression: ERK blockade decreased IL-12p40 production (2-fold decrease in IL-12p40 mRNA and protein). This result suggests that the impact of ERK signaling in response to these stimuli differs between macrophages and dendritic cells for a subset of ERK-regulated genes. ERK-mediated suppression of IL-12p40 in macrophages may prevent excess inflammation and associated tissue damage, while ERK-mediated induction of IL-12p40 in dendritic cells may promote priming of Th1 responses. Future studies will examine other genes and proteins involved in infection responses. Greater understanding of the role that ERK signaling plays in different cell types may inform the development of host-directed therapy for infections such as tuberculosis.
Analysis of health and lung injury markers in mice treated with TLR4-interacting SPA4 peptide
Authors: Awasthi Shanjana , Rahman Negar , Hedrick Andria , Awasthi Vibhudutta , Breshears Melanie , Kosanke Stanley 
Published: January 2023

Abstract
The SPA4 peptide (GDFRYSDGTPVNYTNWYRGE) interacts with TLR4, affects intracellular signaling, improves host defense, and alleviates tissue injury in mouse models of bacterial lung infection and lipopolysaccharide-induced lung inflammation. In this study, we investigated the effect of different doses of SPA4 peptide on health, and systemic and lung injury parameters in outbred CD-1 mice. Mice were intratracheally administered with 0.625, 1.25, 2.5, 5, and 10 μg SPA4 peptide per g body weight every day for three days. Mice left untreated or those treated with equivalent volume of vehicle were included as controls. Signs of sickness and body weights were monitored every 24 h; food and water were provided ad libitum. After 24 h of last intratracheal instillation, mice were euthanized, and blood and major organs were collected. Blood gas and electrolytes (pCO2, total CO2, pO2, pH, anion gap, base excess, total hemoglobin, Na+, K+, Cl−, HCO3−) were analyzed, and markers of lung injury (lung wet/dry weight ratio, and levels of albumin, lactate, and total protein) were determined in lung tissue homogenates. Major organs (stomach, intestines, lung, liver, spleen, kidney, heart, and thymus) were fixed, sectioned, stained with hematoxylin and eosin, and examined for tissue damage or inflammatory response. No signs of sickness or changes in body weight were noted. Blood gas and electrolyte parameters were not significantly altered in SPA4 peptide-treated mice. The lung injury parameters were also not significantly different in SPA4 peptide-treated mice. Histological examination revealed no sign of tissue injury or inflammation in any of the major organs. Together, these results provide an initial safety assessment of the SPA4 peptide.
NHR-49/PPARα regulates HLH-30/TFEB-mediated innate immune response via a flavincontaining monooxygenase in C. elegans
Authors: Wani Khursheed , Taubert Stefan , Irazoqui Javier 
Published: January 2023

Abstract
The nematode C. elegans mounts an innate immune response against Gram-positive bacteria S. aureus by the transcriptional induction of multiple host defense genes. The expression of a majority of these genes is under the control of HLH-30, C. elegans homolog of TFEB (transcription factor EB) that also functions in the innate immune response in mammals. In addition to its role in host defense against S. aureus, HLH-30 acts as a nutritionally-controlled stress response factor. We wanted to deconvolve the response of C. elegans to nutritional challenge from the antibacterial response against S. aureus. To this end, we performed RNAseq profiling in wildtype and hlh-30 mutants that were either infected with S. aureus or were starved. Our RNAseq analysis showed that multiple HLH-30-dependent or-independent genes are specifically induced by S. aureus infection, and not by starvation. One of the genes that is highly induced by infection and not starvation is fmo-2, which encodes a member of the flavin-containing monooxygenase (FMO) family in C. elegans. We decided to characterize the role of FMO-2 in host defense in C. elegans by employing genetic, transgenic, and molecular approaches. We found that FMO-2 is important for host defense against S. aureus infection, and its induction during S. aureus infection is controlled by two transcription factors, HLH-30 and NHR-49 (C. elegans ortholog of the mammalian nuclear-hormone receptor, PPARα). Our results propose a model in which NHR-49 functions upstream of HLH-30 to bind to fmo-2 promoter and to induce its upregulation by functioning specifically in C. elegans intestine (the site of innate immune response), thereby providing host defense against S. aureus infection.
Induction of COX-2 gene transcription and prostaglandin E2 (PGE2) production in primary human monocytes by MDP adjuvant: role of MAPK and p300 HAT
Authors: Liu Fengjie , Romantseva Tatiana , Golding Hana , Zaitseva Marina 
Published: January 2023

Abstract
Muramyl dipeptide (MDP) is the minimal structure responsible for adjuvant activity of peptidoglycans. Early clinical trials of vaccines formulated with MDP revealed adjuvant-induced pyrogenic response. Here we studied signaling pathway leading to production of mediator of fever, PGE2, in human monocytes activated with MDP.
MDP alone induced nuclear translocation of NF-kB but only minimal levels of COX-2 mRNA expression and PGE2 production in monocytes. T cell conditioned medium (TCM) prepared from CD3 bead-purified T cells dramatically upregulated COX-2 mRNA and PGE2 in MDP-activated monocytes but did not increase nuclear NF-kB. COX-2 mRNA expression and PGE2 production in monocytes activated with MDP and TCM were sensitive to the inhibitors of ERK1/2, p38, and JNK MAPK. Western blotting confirmed that MDP alone and TCM alone induced phosphorylation of p38 and SAPK/JNK and of ERK1/2, respectively. In addition to MAPK, the role of p300 histone acetyltransferase (HAT) was studied by ChIP assay. TCM induced binding of p300 to a COX-2 promoter. Inhibition of p300 blocked binding of p300 to COX-2 promoter, acetylation of H3 and H4 histones, upregulation of COX-2 mRNA and of PGE2 in monocytes activated with MDP/TCM. Importantly, inhibitors of either p38 or ERK1/2 prevented recruitment of p300 to COX-2 promoter. Taken together these results suggest cooperation between two signaling pathways in monocytes: nuclear translocation of NF-kB and activation of SAPK/JNK and p38 MAPK are induced by MDP, while activation of ERK1/2 and recruitment of p300 to COX-2 promoter are induced by TCM. These two pathways are equally important for p300-facilitated acetylation of H3 and H4 histones at the COX-2 promoter and for up-regulated COX-2/PGE2 production.
Lung-resident memory CD4 T cells do not divide extensively during protective responses against Influenza virus
Authors: McKinstry Karl Kai , Finn Caroline M 
Published: January 2023

Abstract
Tissue-resident memory (TRM) CD4 T cells are emerging as critical orchestrators of immunity but little is known about their response kinetics during secondary challenges. We recently developed an adoptive transfer model in which virus-specific CD4 TRM isolated from lungs of Influenza A virus (IAV)-primed mice are transferred intranasally to new congenic hosts. Here, we track these donor TRM cells after IAV challenge of the adoptive hosts to gain insight into unique aspects of CD4 TRM responses to infection. Our findings demonstrate that the TRM rapidly activate elements of innate defense in the lung, including dendritic cells and NK cells. The TRM response is limited to the lung as no donor cells are detected in the spleen or mediastinal draining lymph nodes through the first week of infection. Although the CD4 TRM are highly protective and can clear even lethal IAV challenge in unprimed hosts, they undergo only limited division. To ask if the TRM-mediated protection requires influx of systemic T cells to the lung, we treated adoptive hosts with FTY720 to block the migration of host T cells during infection. TRM-mediated protection was not impacted by FTY720 treatment, indicating that the local memory CD4 cells can protect independently of T cell responses initiated in secondary lymphoid organs. Finally, we could detect the donor TRM in lungs for at least 21 days after infection and they were rapidly activated by antigen given intranasally. Our results thus show that lung CD4 TRM initiate highly protective responses in the lungs of otherwise naïve mice and have the capacity to respond against multiple rounds of infection. As such their generation through vaccination is a powerful way to improve immunity against seasonal and pandemic IAV.
An Intranasally Administered Antibody Cocktail Prevents Ricin Toxin-Induced Lung Inflammation and Alveolar Macrophage Death
Authors: Mantis Nicholas J , Rong Yinghui , Torres-Velez Fernando J 
Published: January 2023

Abstract
Ricin toxin, a plant-derived, mannosylated glycoprotein, elicits an incapacitating and potentially lethal, pulmonary inflammatory response in the airways following inhalation. Uptake by alveolar macrophages (AM) and other pulmonary cell types occurs via ricin’s B subunit (RTB), a galactose-specific lectin, and the mannose receptor (MR; CD206). Ricin’s A subunit (RTA) is a ribosome-inactivating protein capable of inducing apoptosis in all mammalian cell types. It was recently reported that a single monoclonal antibody (MAb), PB10, directed against an immunodominant epitope on RTA, is able rescue Rhesus macaques against lethal dose of ricin administered by aerosol. In this study, we now demonstrate in mice that the effectiveness PB10 is significantly improved when combined with a second MAb, SylH3, against RTB. Mice treated with PB10 alone survived lethal-dose intranasal ricin challenge, but experienced significant weight loss, moderate pulmonary inflammation (e.g., elevated IL-1 and IL-6 levels, PMN influx), and apoptosis of alveolar and tissue macrophages. In contrast, mice treated with the PB10/SylH3 cocktail were essentially impervious to pulmonary ricin toxin exposure, as evidenced by no weight loss, no change in local IL-1 and IL-6 levels, retention of alveolar and tissue macrophage numbers, and a significant dampening of PMN recruitment to the bronchoalveolar lavage (BAL) fluids. The PB10/SylH3 cocktail only marginally reduced ricin binding to target cells in the BAL, suggesting that the antibody mixture neutralizes ricin by interfering with one or more steps in the RTB- and MR-dependent uptake pathways.
Inducible thymic epithelial cell-based rejuvenation in thymic aging-associated inflammaging (Supported by NIH/NIAID R01AI121147)
Authors: Oh Jiyoung , Thomas Rachel , Wang Weikan , Su Dongming 
Published: January 2023

Abstract
Age-related thymic involution primarily results from a decline in the transcription factor forkhead box N1 (FOXN1), while input of exogenous FOXN1 is able to significantly rescue thymic aging. Given that FOXN1 over-expressing fibroblasts (inducible thymic epithelial cells, iTECs) can generate an ectopic de novo thymus and engrafted young TECs can lead to middle-aged thymus regrowth, we asked whether engrafting iTECs directly into the aged thymus and perithymus can be a strategy to rescue aged thymic function. The re-establishment of central tolerance is crucial to attenuate age-associated chronic inflammation in the elderly (termed inflammaging). This will reduce increased self(auto)-reactivity, an important contributor to inflammaging. Intrathymic transplantation of iTECs into the aged murine thymus revealed that iTECs drove partial regrowth of the aged thymus, equally well in both male and female mice. The iTECs also rejuvenated aged thymic architecture, including regeneration of well-organized thymic medulla, and improvement of aged thymic function, including re-enforcement of thymocyte negative selection-related Aire gene expression. This led to reduction of self-reactivity-resulted inflammaging, including pro-inflammatory cytokines (IL-6 and IL-1b) and lymphoinfiltration to non-lymphoid organs, in old mice. Our findings support this potential novel strategy for enforcing the declined thymocyte negative selection to partially restore central tolerance in the elderly and reduce inflammaging.
T regulatory antigen-specific multi-peptide immunotherapy induce clinical responses in autoimmune diseases
Authors: Marquez-Manriquez Juan P , MATUTE-BRISENO JOSE ANTONIO , Alejandro Lucero-Diaz PEDRO ALEJANDRO , CAMACHO-HERNANDEZ ALEJANDRO 
Published: January 2023

Abstract

Background
The current treatment for several fatal autoimmune diseases is mainly palliative with minimal clinical benefit. By other hand the biological and clinically relevant potential targets to stop the auto immune response are mainly unknown, including the innate and adaptive auto-immune micro-environment. We performed a pilot trial with (n=5) progressive patients with myasthenia gravis (MG), multiple sclerosis (MS) and amyotrophic sclerosis lateral (ASL) using a multi-peptide T regulatory immunotherapy.

Methods
We performed a systematic review to found potential proteins involved in the auto-immune response in these three autoimmune diseases using the terms “misfolded proteins and autoimmune disease and prognosis and multivariate analysis”. initially N=25 repository samples from autoimmune disease patients were analyzed to test the predicted T regulatory peptides immunogenicity for ELISA, IL-10 ELISPOT and T –cell expansion.

Results
We found that predicted T regulatory peptides from Bcl-2, peptide C (p=0.001), survivi, peptide A (0.001) and VCP, peptide 8 = (P=0.005) were able to expand antigen-specific T regulatory cells and there was a clinical correlation in symptoms improvement in all the three diseases (mOS=28 months).

Discussion
The multi-peptide T regulatory peptide immunotherapy is feasible therapeutic approach to treat incurable autoimmune diseases and this data could be used as proof of principle to prepare a large clinical study in order to potentially improve clinical outcomes in combination with other approaches that may impact the clinical scenario in autoimmune diseases with limited or nor therapeutic options. No clinical toxicity was observed.
Lipid Nanoparticle-Mediated Delivery of Enhanced Costimulation Blockade to Prevent Type 1 Diabetes
Authors: ZHANG YICHUAN , Wang Jialu , Calderon-Colon Xiomara , Tiburzi Olivia N. , Lozano Marcos Iglesias , Patrone Julia B. , Raimondi Giorgio 
Published: January 2023

Abstract
Type 1 diabetes (T1D) remains an untreatable autoimmune disease caused by the destruction of pancreatic beta cells by autoreactive T cells. Costimulation blockade, blocking the CD28 pathway via CTLA4-Ig, is one promising immunotherapeutic, but recent observations suggest its efficacy is antagonized by inflammatory factors. As antigen presenting cells (APCs) malfunctions and aberrant accumulation of type 1 interferons are associated with T1D, we pose that inhibiting the signaling of inflammatory cytokines via Tofacitinib (Tofa), a JAK inhibitor, would enhance the efficacy of CTLA4-Ig in preventing T1D development. The objective of this study was to design a controlled and localized delivery of Tofa via biocompatible lipid nanoparticles, Nanostructured Lipid Carrier (NLC), and assess the immunomodulatory impact of this strategy.
We identified a specific composition of NLC that had negligible toxicity, could be readily taken up by multiple immune cells, and had a favorable Tofa encapsulation efficiency. Live animal imaging confirmed these particles have unique property of accumulating in lymphoid tissues, particularly in spleen, pancreatic and mesenteric lymph nodes when administrated via oral gavage. Ex-vivo, Tofa-NLC rapidly delivered Tofa to mouse APCs preventing their maturation and the release of inflammatory cytokines. Ongoing experiments show that short-term administration of Tofa-NLC via oral gavage at early (3-week-old) or late (10-week-old) stage in disease prone NOD mice promotes a significant reduction of T1D onset.
Overall, Tofa-NLC represent a promising strategy to complement CTLA4-Ig and we envision this combined strategy could enhance the efficacy of antigen specific immunotherapy.
Foxp3 cooperates with Ikaros to control the suppressive function of regulatory T cells
Authors: Thomas Rajan M , Wang Liqing , Chen Chunxia , Hancock Wayne W , Wells Andrew D 
Published: January 2023

Abstract
Ikaros (encoded by Ikzf1) is a lymphocyte-specific transcription factor that controls a wide spectrum of T cell functions, including T helper differentiation, cytokine production, and tolerance in response to antigenic stimuli. Using heterozygous and dominant-negative mouse models, we have shown previously that Ikaros controls IL-2 production by conventional T cells, T helper differentiation, CD8+ effector differentiation, and anergy induction. While the role of Ikaros in many T cell subsets has been established, its role in Treg-mediated peripheral tolerance has not been examined. To examine the Treg-specific role of Ikaros, we generated Ikzf1-fl/fl-Foxp3-YFP-Cre+ mouse with conditional deletion of Ikaros in Tregs. Ikzf1-fl/fl-Foxp3-YFP-Cre+ mouse exhibited enlarged secondary lymphoid tissues, with increased frequencies of effector memory Tconv and Treg. We show that Ikaros expression by Tregs is required for immune tolerance in an allogenic cardiac transplant model. While CD28- and CD40L-blockade induced long-term cardiac allograft tolerance in control recipients, Ikzf1-fl/fl-Foxp3-YFP-Cre+ recipients rejected the allografts in a short duration (p&lt;0.01). We show that Ikaros forms a complex with Foxp3, which is required for Foxp3 to bind to and repress inflammatory cytokine genes. Ikaros is also required for normal iTreg development in vitro, as TGF-B/IL-2-induced iTreg development is impaired in conventional CD4+ T cells with reduced Ikaros activity. These studies suggest that Ikaros interacts with Foxp3 and directs Foxp3 to exert its suppressive function in Tregs.