bluuebunny/crossref_metadata_2025_split huggingface dataset:
Innate CD4+ T cells associated with development of virus- induced demyelinating disease (INC1P.362)
Authors: Olson Julie , Penm Steven 
Published: January 2023

Abstract
Theiler’s murine encephalomyelitis virus (TMEV) infection of susceptible mice results in a persistent infection in the central nervous system (CNS) that leads to the development of a chronic progressive demyelinating disease associated with a pro-inflammatory immune response. TMEV- induced demyelinating disease serves as a mouse model for multiple sclerosis. TMEV infection of resistant mice leads to clearance of the virus from the CNS without development of demyelinating disease. We have previously shown that the innate immune response to TMEV directly affects development of demyelinating disease in susceptible mice. We recently identified a population of CD4+ T cells that infiltrate into the CNS during the innate immune response to TMEV in susceptible mice. Most interestingly, the resistant mice infected with TMEV do not have innate immune infiltration of CD4+ T cells into the CNS. The CNS infiltrating innate immune CD4+ T cells in the TMEV- infected susceptible mice were NK1.1-, αβ+ and CD3+. The innate immune CD4+ T cells expressed high levels of IL-17A and IFNγ but low levels of IL-22. Furthermore, susceptible mice expressed high levels of IL-23, IL-1β, and IL-6 during the innate immune response to TMEV which promotes IL-17A expression in CD4+ T cells. Therefore, the innate immune response in susceptible mice promotes the infiltration of innate CD4+ T cells into the CNS which express pro-inflammatory cytokines associated with development of demyelinating disease.
Basal forebrain cholinergic signaling regulates peripheral cytokine responses (INC1P.349)
Authors: Pavlov Valentin , Silverman Harold , Dancho Meghan , Regnier-Golanov Angelique , Ochani Mahendar , Hanes William , Olofsson Peder , Chavan Sangeeta , Golanov Eugene , Prado Vania , Prado Marco , Tracey Kevin 
Published: January 2023

Abstract
Brain cholinergic signaling regulates peripheral cytokine release and inflammation (Nat Rev Endocrinol, 2012, 8:743). We have previously shown that the centrally-acting acetylcholinesterase inhibitor galantamine suppresses peripheral pro-inflammatory cytokine release through a brain muscarinic receptor-mediated and vagus nerve-dependent mechanism. Here, we provide insight into the brain mechanisms of this regulation. VAChTNkx21-Cre-flox/flox mice with selective, basal forebrain cholinergic neuron genetic ablation of the vesicular acetylcholine transporter (an important determinant of acetylcholine release), and control VAChTflox/flox mice, were treated with galantamine or saline and subjected to endotoxemia. Galantamine (3mg/kg) i.p. administration suppressed serum TNF levels in control mice only. In addition, serum TNF and IL-10 in VAChTNkx2 1-Cre-flox/flox mice were significantly higher (2.9-fold, P&lt;0.05) and lower (57%, P&lt;0.05) respectively, as compared to control mice. Furthermore, electrical stimulation of the medial septum, an important basal forebrain cholinergic region, significantly lowered serum TNF (79%, P&lt;0.01) and other pro-inflammatory cytokine levels in endotoxemic mice, as compared to sham-stimulation. These results demonstrate a previously unrecognized role of basal forebrain cholinergic signaling in the regulation of peripheral inflammation.
Gene array analysis of PD-1H overexpressing monocytes reveals a pro-inflammatory profile (INC2P.417)
Authors: Shankar Premlata , bharaj preeti , Yi Guohua , Swamy Manjunath 
Published: January 2023

Abstract
We have previously reported that overexpression of Programmed Death -1 Homolog (PD-1H) in human monocytes leads to activation and spontaneous secretion of multiple pro inflammatory cytokines. Here we evaluated changes in monocytes gene expression after enforced PD-1H expression by gene array. Total RNA isolated from PD-1H-expressing and control monocytes were analyzed with human Illumina HT-12 v4 Expression BeadChip. Fold change of genes were confirmed by real time PCR. Data revealed alterations in 51 potential candidate genes that relate to immune response, cell adhesion and metabolism. Genes corresponding to pro-inflammatory cytokines showed the highest upregulation. Compared to vector control, PD-1H induced a 7, 3.2, 3.0, 5.8, 4.4 and 3.1 fold upregulation of TNF-α, IL-1beta, IFN-α, γ, λ and IL-27 respectively. Further, the difference was abrogated in presence of specific but not scrambled siRNA. The data are in agreement with our cytometric bead array analysis showing induction of proinflammatory cytokines, IL-6, IL-1β and TNF-α by PD-1H. Other genes related to inflammation, including transglutaminase 2, the transcription factor NF-kB (p65 and p50) and toll like receptors 3 and 4 were also upregulated 5, 4.5 and 2.5 fold respectively. The analysis also revealed a novel set of previously unreported genes that need to be further mined and validated. We conclude that PD-1H functions to enhance monocyte activation.
A Rho GDP dissociation inhibitor produced by apoptotic T cells inhibits growth of Mycobacterium tuberculosis (INC2P.415)
Authors: Venkatasubramanian Sambasivan , Dhiman Rohan , Paidipally Padmaja , Cheekatla Satyanarayana , Tripathi Deepak , Welch Elwyn , Tvinnereim Amy , Jones Brenda , Theodorescu Dan , Barnes Peter , Vankayalapati Ramakrishna 
Published: January 2023

Abstract
In the current study, we found that a subpopulation of apoptotic T-cells (CD4+CD25+ and 85% Foxp3+) from persons with latent tuberculosis (TB) infection, inhibits growth of M. tuberculosis (M. tb) in human monocyte-derived macrophages (MDMs) from 16.5 ± 1.2 x 106 vs 2.2 ± 1.1 x 106 CFU (p=0.001). Analysis of culture supernatants from T-cells by 2D gel electrophoresis and LC MS/MS indicated that a soluble factor, Rho GDP dissociation inhibitor (D4GDI), produced by apoptotic T-cells is responsible for this inhibition of M.tb growth in MDMs and in mice. M. tb-expanded CD4+CD25+Foxp3+D4GDI+ cells do not produce IL-10, TGF-β and IFN-γ. D4GDI inhibited growth of M. tb from 10.4 ± 1.1 x 106 vs 2.8 ± 0.6 x 106 CFU (p&lt;0.001) and D4GDI siRNA reversed T-cells-dependent M. tb growth inhibition in MDMs from 1.8 ± 0.3 x 106 to 5.5 ± 1 x 106 CFU (p=0.001). D4GDI enhanced production of IL-1β (43 ± 33 to 193 ± 104 pg/ml, p=0.02), TNF-α (88 ± 126 to 129 ± 111 pg/ml, p=0.03) and ROS production and enhanced apoptosis of M. tb-infected MDMs to inhibit M.tb growth. D4GDI was concentrated at the site of disease in TB patients, with higher levels detected in pleural fluid than in serum. However, in response to M. tb, PBMC from TB patients produced less D4GDI than PBMC from persons with LTBI (13.7 ± 9.3 vs. 40.6 ± 3.2, p=0.02). Our study provides the first evidence that a subpopulation of T-cells enhances immunity to M. tb, and that production of D4GDI by this subpopulation inhibits M. tb growth.
Macrophage production of innate immune protein C1q is associated with M2 polarization (INM1P.434)
Authors: Fraser Deborah , Melzer Emily , Camacho Alexis , Gomez Mariam 
Published: January 2023

Abstract
C1q is an important recognition molecule of the innate immune response. It plays a dual role in vivo, triggering inflammation via activation of the classical complement pathway, to clear pathogens. However, it can also directly opsonize targets including apoptotic cells and damaged-self molecules, leading to increased macrophage phagocytosis and M2-like anti-inflammatory cytokine production. C1q is primarily synthesized by macrophages. Therefore we investigated C1q production by macrophages under various conditions. Raw 264.7, a murine macrophage cell line, were incubated with different targets or under conditions associated with M1 or M2 polarization. C1q gene expression levels were measured by quantitative RT-PCR from isolated mRNA. C1q protein expression and secretion was measured in cell media by hemolytic titer. The results showed a decrease in C1q production at the gene and protein level when cells were treated with M1-associated inflammatory stimuli such as high levels of LPS, and/or IFNγ. Interestingly, treating the cells with M2-associated stimuli such as IL-4, or interaction of macrophages with “self” targets such as damaged self molecules oxidized or acetylated LDL yielded an increase in C1q production. These data support a non-complement associated role for C1q in resolving inflammation and prevention of autoimmune disease, and suggest it may act as a novel biomarker for M2-like inflammation resolving macrophage polarization.
Gata3 regulates the maturation program required for bone marrow exit and proliferation in Natural Killer cells (INM1P.435)
Authors: Lee Seung-Hwan , Ali Alaa 
Published: January 2023

Abstract
Natural killer (NK) cells are innate lymphocytes capable of killing virally infected cells or transformed cells. NK cells differentiate in the bone marrow before they migrate out to peripheral organs. The developmental program leading to functionally mature NK cells has been studied in the context of several transcription factors. However, the role of GATA-3 in NK cell development has not been completely understood. Using NK-cell specific Gata3 knockout mice (NK-Gata3-/-), we demonstrated that GATA-3 is required for the NK cell maturation beyond the CD27 single positive stage and for efficient IFN-g production. The frequencies of NK cells from NK-Gata3-/- mice were found higher in the bone marrow but lower in peripheral organs compared to control littermates, indicating that GATA-3 controls the maturation program required for bone marrow egress. Despite this, upon MCMV infection, NK cells from NK-Gata3-/- mice were able to expand vigorously, achieving NK cell frequencies surpassing those in controls, and therefore provided comparable protection. The enhanced proliferation of GATA-3-deficient NK cells was associated with enhanced upregulation of CD25 expression. Thus, the NK-Gata3-/- mice serve as a model to study the NK cell developmental process linked to cell migration, effector functions and proliferation.
p47phox, a subunit of NADPH oxidase, enhances the function of Nrf2 (INM1P.441)
Authors: Joo Myungsoo , Kim Kyun Ha 
Published: January 2023

Abstract
Although an essential component to clear invading bacteria in innate immunity, reactive oxygen species (ROS) can adversely affect tissue. However, ROS activate Nrf2 and induce the expression of anti-oxidant enzymes including NQO1, HO-1 and GCLC, which in turn reduce the adverse effects of ROS. Given a high reactivity of ROS, we postulate that NADPH oxidase, a major producer of ROS in macrophages, exists proximal to Nrf2-Keap1 complex to effectively activate Nrf2. Here, we show that p47phox, a cytosolic subunit of NADPH oxidase, physically interacted with Nrf2 and facilitated the protective function of Nrf2 in inflammation. Biochemical and functional analyses show that p47phox bound to Nrf2 and enhanced the expression of Nrf2-dependent genes such as NQO1, HO-1 and GCLC. Immunoprecipitation assays indicate that p47phox did not interfere with the binding between Nrf2 and Keap1 but suppressed the ubiquitination of Nrf2, suggesting that p47phox enhances Nrf2 transcriptional activity by disturbing the ubiquitination of Nrf2 by Keap1. Consistent with these results, bone marrow-derived macrophage (BMDM) from p47phox -/- mice showed reduced expression levels of Nrf2-dependent genes, compared with BMDM from wild type littermates. In addition, overexpression of p47phox in the mouse lung was sufficient to suppress LPS-induced neutrophilic lung inflammation. Together, our results suggest that p47phox binds to Nrf2, enhancing the protective function of Nrf2 in inflammation.
The heightened inflammatory response makes females are more susceptible to F. tularensis-mediated death (INM1P.444)
Authors: Spencer Charles , Martinez Danielle , Setzu Nicole , Ramos Mireya 
Published: January 2023

Abstract
The zoonotic infectious disease, Francisella tularensis, the causative agent for human tularemia, elicits high cytokine levels as a result of rampant inflammation, a condition referred to as “cytokine storm.” This cytokine storm is responsible for the severity of symptoms and ultimately for the death of the host. Though men have a tendency to be exposed to the pathogen more frequently than women, do to its presence in wild animals, women have a significantly greater inflammatory response than do men. Therefore, we asked whether their enhanced inflammatory response might make women more susceptible to tularemia-mediated death. Male and female mice were infected with F. tularensis and their survival and inflammatory response monitored. Subsequently, the role of sex hormones was considered by infecting gonadectomized mice. On average, 15% female mice survived infection compared with 90% of male mice. In addition, the female mice displayed an increased serum cytokine storm compared with male mice despite similar levels of bacterial growth. We seek to identify the factors controlling this increased susceptibility of female mice. This line of investigation may lead to the development of novel therapeutics and/or treatment strategies specific to women exposed to F. tularensis.
Regulating role of transmembrane protein 173 (Tmem173) on RANKL-induced osteoclastogenesis (INM1P.446)
Authors: Jang Yong-Suk , Choe Chung-Hyeon , Park Jisang , Yu Kang-Yeol , Kim Ju 
Published: January 2023

Abstract
Tmem173, also known as STING, MITA and MPYS, is a potential stimulator of IFN-β production which is essential for innate immune response in monocyte/macrophage cells. Based on the known function of IFN-β as a negative feedback controller for RANKL-induced osteoclast differentiation of monocytic macrophage cells, we investigated the effect of Tmem173 on RANKL-induced osteoclastogenesis via monitoring the osteoclast-associated gene expression and protein activation in RAW264.7 cells. TRAP activity and bone resorption activity were significantly suppressed in Tmem173-overexpressed RAW264.7 cells compared to control vector-transfected RAW264.7 cells. Tmem173 overexpression also down-regulated expression of osteoclast marker genes, such as TRAP, MMP-9 and cathepsin K, in RANKL-treated RAW264.7 cells. These marker genes were up-regulated efficiently in Tmem173 knock-downed RAW264.7 cells. Tmem173 overexpression down-regulated the expression of osteoclast-specific transcription factors, such as c-Fos and NFATc1, and RANKL-induced activation of ERK mitogen-activated protein kinase. In addition, we observed the down-regulation of RANKL-induced expression of COX-2 and iNOS. Finally, we found that Tmem173 expression was up-regulated by osteoprotegerin (OPG). These results suggest that regulatory role of Tmem173 in RANKL-mediated osteoclast differentiation.
A miRNA regulatory circuitry tightly tunes macrophage inflammatory response (INM1P.426)
Authors: Mann Mati 
Published: January 2023

Abstract
Regulation of the immune system is vital to preventing many pathogenic disorders, and mammals have developed a complex system of checks and balances for immune regulation in order to allow immune responses to foreign pathogens while avoiding chronic inflammatory responses. Recently, it has become evident that micro-RNAs (miRNAs) play an important role in regulating immune response. Among these, miR-155 and miR-146a were identified as inflammatory response miRNAs that are upregulated by NF-κB signaling. Here, we set to characterize the hierarchy and genetic interaction of miR-146a and miR-155 in regulating macrophage inflammatory response during acute and chronic conditions. We show a regulatory network where miR-155 expression amplify the inflammatory response by prolonged NF-κB activity through PI3K signanling, while miR-146a attenuates the inflammatory response by inhibiting NF-κB activity and miR-155 expression. This regulatory circuitry enables strong acute inflammatory response while avoiding the development of chronic inflammation.
Sulforaphane blocks HIV infection of macrophages (IRC4P.459)
Authors: Furuya Andrea , Sharifi John , Jellinger Robert , Cristofano Paul , de Noronha Carlos 
Published: January 2023

Abstract
Despite highly active anti-retroviral treatment (HAART), HIV infection still claims millions of lives annually. HAART therapy is accompanied by high financial costs, the emergence of resistant viruses and short- and long-term side-effects. For this adjunctive therapeutic based on natural products should be considered to ameliorate complications due to HIV infection. Here we show for the first time that sulforaphane (SFN), a natural product produced in cruciferous vegetables blocks HIV infection in primary macrophages but not in primary T cells or several other cell types tested by mobilizing the transcription factor nuclear erythroid 2-related factor 2 (Nrf2). SFN blocked HIV infection in macrophages and macrophage-like cell lines by triggering a block before proviral integration. Furthermore we found that other compounds known to mobilize Nrf2 in macrophages also establish an intracellular state that counters HIV infection. Additionally we found that depleting Nrf2 boosts HIV infection in primary macrophage cultures. This led us to hypothesize that SFN and other compounds that trigger Nrf2 action, in macrophages, act through this protein to establish an intracellular antiviral state that counters HIV. We propose that SFN and other modulators of Nrf2 can be used to block HIV infection and therefore SFN could serve as an auxiliary therapeutic in ART experienced patients.
Expression of cationic amino acid transporter 2 is required for myeloid derived suppressor cell-mediated control of T cell immunity (IRC4P.460)
Authors: Cimen Bozkus Cansu , Elzey Bennett , Crist Scott , Ellies Lesley , Ratliff Timothy 
Published: January 2023

Abstract
Myeloid derived suppressor cells (MDSC) are a heterogeneous population of immature cells that expand during benign and cancer-associated inflammation. MDSC regulate innate and adaptive immunity, notably by inhibiting T cell functions. Increased metabolism of L-Arginine (L-Arg), through arginase 1 (ARG1) and nitric oxide synthase 2 (NOS2), is well documented as a major MDSC suppressive mechanism. However, the mechanism by which extracellular L-Arg is transported into MDSC is not defined. Using murine models of prostate specific inflammation and cancer, we show that MDSC recruited to localized inflammation and tumor sites upregulate cationic amino acid transporter 2 (Cat2) coordinately with Arg1 and Nos2. CAT2 contributes to the transport of L-Arg in MDSC and acts as an important regulator of MDSC suppressive function. MDSC that lack CAT2 have significantly reduced suppressive ability ex vivo and display impaired capacity for regulating T cell responses in vivo as evidenced by increased T cell expansion and decreased tumor growth in Cat2-/- mice. The abrogation of suppressive function is due to lower L-Arg uptake, which leads to the impaired ability of NOS2 to catalyze L-Arg metabolic processes. These findings demonstrate that CAT2 is an important regulator of MDSC suppressive function. In the absence of CAT2 MDSC display diminished capacity for controlling T cell immunity in prostate inflammation and cancer models, where the loss of CAT2 results in enhanced antitumor activity.
Distinct features of newly found liver-draining lymph nodes in mice (IRC4P.612)
Authors: Yu Jiali , Sun Rui , Tian Zhigang 
Published: January 2023

Abstract
Draining lymph nodes (LNs) are specialized sites where adaptive immune responses take place. Recently, the anatomical location of two LNs draining the mouse liver, named as portal LN (PLN) and celiac LN (CLN) respectively, has been characterized. However, the immune responses in the liver-draining LNs and their contribution to the establishment and maintenance of liver tolerance are still not fully understood. In our further study, we find that these two liver-draining LNs (PLN and CLN) have different lymphatic vessels, excluding the possibility that one LN is secondary to the other one. Interestingly, although both PLN and CLN can drain liver lymph directly, they are different in many aspects. The two liver-draining LNs have different cellular composition, with PLN being highly similar to mesenteric LN but different from periphery LNs, and CLN being special with unique composition of innate immune cells. Importantly, the development of these two liver-draining LNs might be controlled by different transcription factors. Moreover, our preliminary data shows that the immune responses of these two liver-draining LNs exhibit different features during exogenous pathogen infection. Overall, PLN and CLN are two liver-draining LNs with distinct characteristics and their contribution to liver tolerance remains to be defined.
CD14hi equine monocytes are preferential initial targets for infection with equine arteritis virus (IRC4P.615)
Authors: Chelvarajan R Lakshman , Mondal Shankar P , Cook R Frank , Go Yun , Sarkar Sanjay , Henney Pamela , Marti Franscesc , Bailey Ernest , Balasuriya Udeni 
Published: January 2023

Abstract
Equine arteritis virus (EAV), the causative agent of equine viral arteritis (EVA) in horses, is distributed worldwide with clinical signs including an influenza-like illness in adult horses, abortion in pregnant mares, interstitial pneumonia in neonatal foals and persistent infection in genetically predisposed stallions. EAV is a small,enveloped virus with a positive-sense, single-stranded RNA genome and primarily infects epithelial and endothelial cells, plus unidentified subpopulations of monocytes and macrophages. The primary objective of this study was to identify the susceptible CD14+ monocyte subpopulation using a synthetic recombinant virus expressing mCherry derived from the prototypical virulent Bucyrus stain (VBS) of EAV. The recombinant virus had identical tropism to parental VBS supporting its use as a surrogate for EAV infection. Multi-color flow cytometric analysis showed that a minor population of CD14hi cells were preferentially infected by EAV at 12 h post infection (hpi), whereas by 36 hpi (three virus replication cycles) almost all infected monocytes were CD14lo. However, this CD14lo population was also characterized by expression of CXCL16, a molecule implicated in the establishment of persistent EAV infections in stallions. CXCL16 expression was not detected in CD14hi infected monocytes at 12 hpi. Work is in progress to determine the eventual differentiation status of CD14hi/CXCL16- and CD14lo/ CXCL16+monocyte populations in terms of M1/M2 polarization.
Molecular chaperoning facilitates the inflammasome activation and an innate immune response to microbial pathogens (IRC4P.455)
Authors: Wang Hongxia , Wang Xiang-Yang 
Published: January 2023

Abstract
While certain molecular chaperones are known to regulate an adaptive immune response by shutting antigenic polypeptides and facilitating cross-presentation, its potential impact on innate immune responses to pathogens is not well understood. Assembly and activation of NLRP3 inflammasome plays a central role in sensing microbial infections and the host innate immunity. We report that extracellular glucose-regulated protein 170 (Grp170), a large molecular chaperone, profoundly enhances microbial CpG oligodeoxynucleotides (CpG-ODN) induced NLRP3 inflammasome activation, which is independent of ATP. Mechanistic studies reveal that Grp170 potentiates the endocytosis of CpG-ODN, followed by lysosomal damage and the cathepsin B release. Further, the elevation of intracellular calcium and calcium-dependent activation of calpain are crucial for Grp170-potentiated lysosomal instability and the inflammasome activation. Lastly, chaperoning-enhanced inflammasome activation results in more efficient bacterial elimination in mice compared with CpG-ODN alone. These results indicate that molecular chaperoning of pathogen-associated molecules (e.g., microbial DNA) by Grp170 facilitates the immune recognition of invading pathogens via modulation of NLRP3 inflammasome activation. Our findings underscore an important role of extracellular chaperone molecules, possibly released from tissues injury following pathogen infection, in amplifying an innate immune response.