bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of follicular helper T cells as a novel cell population potentially involved in the pathogenesis of Rheumatoid Arthritis (HUM3P.251)
Authors: Zhang Xin , Singla Shikha , Pound Jerry , Zakem Jerald , Collin Kismet , Webb-Detiege Tamika , Davis William , Quinet Robert 
Published: January 2023

Abstract
Background. RA is an autoimmune disease characterized by chronic inflammation of the synovium, causing progressive joint destruction. The treatment for RA is largely based on the clinic manifestations, which are remote from the fundamental pathophysiologic defects. Identification of cellular subsets or biomarkers is a necessary first step toward more accurate prognosis and better therapeutic decisions. Here, we identified a novel T cell subset, follicular helper T cells (Tfh) in RA patients, and examined the hypothesis that Tfh cells may account for RA pathogenesis by providing a fostering environment for self-reactive B cells, resulting in autoantibody production. Methods. Peripheral blood was collected from 26 RA patients and healthy donors. Synovial fluid from actively inflamed joints of RA patients was also collected. Clinical disease activity (DAS-28) and serum laboratory results were obtained. Tfh cells were defined by their signature surface markers via flow cytometry. IL-21 expression and plasmablasts were measured by intracellular staining. Results. A novel T cell subset with Tfh cell signature molecule and cytokine was expanded in the peripheral blood of active RA patients. The percentage of Tfh cells correlated positively with the level of pathogenic auto-antibody (anti-CCP) and disease activity. Conclusions. Circulating Tfh cells may serve as a biomarker of pathogenesis in RA patients. Targeting RA-Tfh cells may provide earlier and precise treatment strategies.
Antigen-specific regulatory T cells from pluripotent stem cells ameliorates arthritis in mice (HUM3P.252)
Authors: Haque Mohammad , Fino Kristin , Song Jianxun (Jim) 
Published: January 2023

Abstract
Embryonic stem cells have the ability to grow indefinitely while maintaining pluripotency. Under the right circumstance, pluripotent stem cells (PSC) can produce almost all of the cells in the body including regulatory T cell (Tregs). Tregs are essential for normal immune surveillance systems, and their dysfunction leads to the development of diseases, such as autoimmune disorders. However, the right circumstances for the development of antigen (Ag)-specific Tregs from PSC (i.e., PSC-Tregs) has not been well defined. So the purpose of this study is to develop antigen-specific Tregs from pluripotent stem cells for treatment of Ag-induced arthritis (AIA) in mice. In this study mouse iPS cells were transduced with FoxP3 and ovalbumin-specific TCR (OTII). iPS cells were driven toward T lymphocyte lineage by co-culture on OP9 cells expressing delta-like (DL1), a Notch ligand. The combination of Notch signaling, FoxP3, and TCR drove iPS cell to differentiation into Ag-specific Tregs. In vitro generated Tregs were adoptively transferred into AIA mice. Our results showed that TCR transduced iPS cells differentiated into Tregs and express CD3, TCRβ, CD4, CD8, CD25, CTLA4 in vitro. Adoptive transfer of such Tregs dramatically suppressed autoimmunity in a well-established AIA model, including the inflammation, joint destruction, cartilage prostaglandin depletion, osteoclast activity, and Th17 production.
A commensal bacteria-derived lipid, significantly decreased in the serum of multiple sclerosis patients, causes attenuation of experimental autoimmune encephalomyelitis (HUM3P.269)
Authors: Anstadt Emily , Fujiwara Mai , Nemati Reza , Yao Xudong , Nichols Frank , Clark Robert 
Published: January 2023

Abstract
The role of the microbiome in multiple sclerosis (MS) remains unknown. We have identified a novel commensal bacteria-produced lipodipeptide, Lipid 654 (L654), which functions as a toll-like receptor 2 agonist. We reported that L654 can be recovered in healthy human serum and that levels are significantly lower in patients with MS. The purpose of this study was to understand the functional relevance of the decreased serum L654 level in MS by determining: 1) if L654 attenuates clinical disease in a transfer model of EAE, and 2) the mechanism by which L654 mediates this attenuation. L654 administration significantly attenuated clinical disease severity in an adoptive transfer model of EAE in SJL/J mice (p &lt; 0.0001). Cellular analysis of mononuclear cells in the spinal cord at day 18 showed a significant decrease in total cell number (p = 0.02) as well as a decrease in percentage of IL-17 producing, IFNγ producing and IL-17/IFNγ double-producing CD4+ T cells in mice administered L654. At day 11, we found a decrease in total mononuclear cell number and an increase in percentage of CD39+ Tregs and IL-10 producing CD11b+ cells in mice administered L654. These results suggest that L654, normally present in human serum but at significantly lower levels in MS serum, has the ability to mediate an immunoregulatory and disease inhibitory effect on CNS autoimmunity. L654 may thus have a role both in the abnormal immunoregulation in MS and as a potential therapeutic in this disease.
CD160 expression defines a functional CD8 T cell subset during HIV-1 infection (HUM4P.270)
Authors: Zhang Anli 
Published: January 2023

Abstract
T cell exhaustion, indicated by elevated expression of inhibitory receptors such as PD-1 and Tim-3 and impaired T cell function, is a hallmark of HIV-1 chronic infection. Glycosylphosphatidylinositol-anchored molecule CD160 expression on CD8+T cells is significantly increased in HIV-1 infected individuals. We show here, CD160 expression is higher in slow progressors than in typical progressors. Levels of CD160 expression on CD8+T cells correlated positively with CD4 T cell count and inversely with HIV-1 plasma viral load. CD160 co-expressed with HLA-DR but not CD38. Functionally, CD160 expressing T cells produced more IFN-γ and proliferate more vigorously in response to HIV-1 specific peptides stimulation. Together, our current study suggests CD160 defines a functional instead of a typical exhausted CD8 T cell subset, targeting CD160 pathway might help in controlling HIV-1 infection.
Immunodeficiency in a hypomorphic mouse model of propianic acidemia (HUM4P.300)
Authors: Castillejo Becerra Clara , Guenzel Adam , Barry Michael 
Published: January 2023

Abstract
Propionic Acidemia (PA) is a recessive disorder resulting from mutations in the PCCA or PCCB genes encoding the two subunits of propionyl-CoA carboxylase. Patients with PA generate propionate and potentially toxic metabolites after consumption of certain amino acids and odd-chain fatty acids leading to systemic health complications. In addition, PA patients are exceptionally sensitive to infections due to a compromised immune system. We developed a hypomorph mouse model of PA (Pcca-/-(A138T) that recapitulates many phenotypes associated with the human disease. In this study, we tested A138T mouse immune function and their susceptibility to infections. A138T mice had an overall reduction in white blood cells, lymphocytes, monocytes, granulocytes, and immunoglobulins when compared to control mice. When wild type and A138T mice were infected i.v. with adenovirus 5 (Ad5) expressing GFP-Luciferase, we observed higher luciferase expression in the livers of the Pcca-/-(A138T) suggesting that the mutant mice were more susceptible to the virus due to defects in the reticuloendothelial system. The A138T mice also generated lower levels of anti-GFP antibodies than control mice 7 weeks after injection suggesting weaker adaptive immune responses. Overall, our results show that the Pcca-/-(A138T) mouse model recapitulates the immunodefiency seen in humans and can be used to further explore their susceptibility to infections as well as the benefits or dangers of vaccination to PA patients.
Common and differential roles of inducible NO synthase and poly (ADP-ribose)polymerase in allergen-induced inflammation and airway hyperresponsiveness: a potential connection to NO levels (HYP5P.320)
Authors: Ibba Salome' , Ghonim Mohamed , Pyakurel Kusma , Naura Amarjit , Mishra Anil , Boulares Hamid 
Published: January 2023

Abstract
The potential role of iNOS as a viable therapeutic target for the treatment of asthma was severely hampered by the negative clinical trial results showing that a selective iNOS inhibitor did not affect airway hyperreactivity (AHR) or airway inflammation after allergen challenge in human subjects with asthma. Our laboratory has shown that iNOS inhibition is protective against airway inflammation upon acute, but not chronic, exposures to ovalbumin. More importantly, iNOS inhibition protected against lung fibrosis suggesting that iNOS inhibition may be protective against some aspects of asthma. Here, we show that iNOS inhibition, pharmacologically by L-Nil or by gene knockout, provided an excellent protection against AHR upon an acute, but not chronic, exposure to ovalbumin. Our laboratory also established a reciprocal relationship between iNOS and PARP-1. However, PARP inhibition, protected against inflammation and AHR upon acute and chronic exposure to ovalbumin. It is interesting that PARP-1 inhibition does not completely abrogate expression of iNOS leaving the possibility that the protective effect of PARP inhibition against inflammation and AHR may be associated with reduction but not the complete inhibition of iNOS and associated production of moderate levels of NO. Studies are being conducted to verify this hypothesis and clarify the intricate roles of iNOS in asthma pathogenesis allowing the possibility of being a viable target for the treatment of the disease.
Chimeric antigen receptors based on low affinity mutants of FcεRI re-direct T cell specificity to cells expressing membrane IgE (HYP5P.321)
Authors: Ma Zhengyu , Ward Dana , Han Huihui 
Published: January 2023

Abstract
IgE is the key mediator of allergic immune responses. Targeting the source of IgE, the IgE-expressing B cells, represents an attractive therapeutic approach for atopic diseases. An adoptive T cell therapy (ACT) approach that redirects T cell killing to IgE-expressing B cells is becoming increasingly feasible due to the recent advance of ACT in efficacy and safety. Here, we designed chimeric antigen receptors (CARs) based on FcεRIα and tested their abilities to recognize cells expressing the transmembrane form of IgE (mIgE), a marker for IgE-expressing B cells. The ectodomains of the wild type (WT) and six low affinity mutants of FcεRIα were fused with CD3ζ to form WT and mutant CARs. When expressed on Jurkat cells, four of the six mutant CARs mediated robust T cell responses to U266 myeloma cells expressing mIgE, and the WT CAR mediated lower T cell responses. The presence of free IgE significantly inhibited the activity of WT CAR but did not affect mutant CARs. CAR+ Jurkat cells did not respond to cells with IgE bound through FcεRI or FcεRII (CD23). CAR+ Jurkat cells did not lead to degranulation of LAD2 mast cells with IgE bound through FcεRI. Taken together, CARs based on low affinity FcεRIα mediated robust and specific T cell responses to mIgE+ cells, even in the presence of free IgE, but not to cells with IgE bound through FcεRII or FcεRI. These CARs therefore have the potency and specificity required for targeting IgE-expressing B cells in ACT for atopic diseases.
Mitigation of bacterial translocation and subsequent sepsis in γ-irradiated mice treated with CCL1 antisense ODN (INC4P.331)
Authors: Kobayashi Makiko , Ito Ichiaki , Nishiguchi Tomoki , Suzuki Sumihiro , Loucas Bradford , Suzuki Fujio 
Published: January 2023

Abstract
Gut bacteria-associated sepsis is a serious concern in persons who have radiation-induced injuries in the gastrointestinal system. Such infection does not normally occur in healthy hosts with M1Mϕ. However, M1Mϕ are not generated in γ-irradiated mice (WBI-mice), even though Mϕ are still present. WBI-mice are carriers of M2bMϕ, which inhibit Mϕ conversion from resident Mϕ to M1Mϕ. We have previously reported that sepsis stemming from Enterococcus faecalis oral infection is markedly mitigated in 5 Gy WBI-mice after M2bMϕ polarization using CCL1 antisense ODN. In this study, we examined the effect of CCL1 antisense ODN on spontaneous bacterial translocation and subsequent sepsis in 8 Gy WBI-mice. The ODN was administered (s.c., twice daily) to 8 Gy WBI-mice for 1 week starting 3 days after γ-irradiation. The efficacy of the treatment was evaluated by an increase in the percentage of survival in test groups, as compared with that of control groups. All control mice died within 22 days of γ-irradiation. However, 70% of the WBI-mice treated with 25 to 125 μg/mouse of the ODN survived more than 30 days after γ-irradiation. Severe bacteremia and bacterial growth in MLNs were not seen in WBI-mice treated with the ODN. M1Mϕ were isolated from MLNs of these mice, while M2bMϕ were not. These results indicate that the Mϕ polarization therapy using CCL1 antisense ODN is effective to mitigate spontaneous bacterial translocation and subsequent sepsis in mice exposed to 8 Gy of γ-irradiation.
Possible roles of LC3-independent autophagy in elimination of Shigella in human colon cells (INC4P.343)
Authors: Ra Eun , Lee Sungwook , Park Boyoun 
Published: January 2023

Abstract
Autophagy is essential for the defense against bacterial invasion. The intestinal tract is quite specialized in order to co-exist with extracellular bacteria, and thus, this system efficiently exploits autophagy to regulate the number of bacteria and to control the immune response to preventing chronic inflammation. Bacterial elimination is mediated by microtubule-associated protein 1A/1B-light chain 3 (LC3), which is critical for the activation of the autophagic canonical pathway in mammals. Although both human colon HT-29 cell line and the human cervical HeLa cell line display normal LC3 expression, HeLa cells are very susceptible to Shigella infection, whereas HT-29 cells efficiently restrict the number of intracellular Shigella. Our results suggest that colon cells may utilize LC3-independent noncanonical autophagy to regulate bacterial growth and survival.
Caspase-11 regulates HMGB1 release from liver during polymicrobial sepsis (INM2P.348)
Authors: Deng Meihong , Scott Melanie , Billiar Timothy 
Published: January 2023

Abstract
Caspase-11 was recently identified as the cytosolic LPS receptor, although little is known about its role in a polymicrobial sepsis. We subjected WT and caspase-11-/-(casp-11-/-) mice to polymicrobial sepsis in the form of cecal ligation and puncture(CLP). By 18h caspase-11 was upregulated and activated in multiple tissues, including liver. Casp-11-/- mice had significantly lower bacterial levels in peritoneum (3.9E+5±2.4E+5vs 8.3E+6±4.8E+6CFU/mL;p&lt;0.05), lower systemic cytokine levels (IL1b;IL6) and reduced peritoneal neutrophil death (3.1±0.3vs6.7±0.5%;p&lt;0.05) vs WT. HMGB1 has been associated with late mortality during sepsis and hepatocytes (HC) are major producers of HMGB1 after CLP, as shown by significantly decreased plasma HMGB1 in HC-specific HMGB1-/- mice at 18h after CLP vs WT(11±3vs53±9ng/mL;p&lt;0.05). Similarly, plasma HMGB1 levels in casp-11-/- mice were significantly lower after CLP vs WT(14±2vs39±8ng/mL;p&lt;0.05). To confirm a role for caspase-11 in HMGB1 release, we stimulated primary isolated HC with LPS(100ng/mL) for 24h in vitro. Media HMGB1 level increased significantly in WT HC after LPS stimulation, but not in casp-11-/- HC. Furthermore, HC cell death was not different between WT and casp-11-/- HC +/- LPS suggesting the release of HC-HMGB1 regulated by caspase-11 is not passive via induction cell death. Our data suggest novel roles for caspase-11 in regulation of immune responses during sepsis including the regulation of HMGB1 release by epithelial cells.
The STING of inflammation (INM2P.356)
Authors: Packard Thomas , Mohning Michael , Barthel Lea , Henson Peter , Janssen William , Cambier John 
Published: January 2023

Abstract
Stimulator of Interferon Genes (STING, a.k.a. MPYS, TMEM173) is a critical intracellular transducer of innate immune signals induced by cytosolic DNA. Activation of STING drives both apoptosis and production of type 1 interferon (IFN1). Thus STING is a vital defensive player on the front line of innate immunity to infectious agents. Recent studies have suggested that STING may also be activated by host-derived DNA, particularly in the context of reduced nuclease activity, and could play a role in autoimmune processes. We hypothesize that during inflammation STING can be activated by host-derived DNA acting as a damage-associated molecular pattern (DAMP). To begin to address this role of STING, we utilized multiple models of in vivo sterile inflammation. Our studies demonstrate that STING-deficient animals have reduced inflammatory responses and more rapid resolution of sterile acute lung injury (ALI) and peritonitis. Further, using in vitro and in vivo models we found that STING plays a role in responses to infiltrating neutrophils and neutrophil-derived debris. Studies to be presented support a model wherein STING is activated by host-derived cellular components, such as DNA, and drives production of type 1 interferon and cell death. Ultimately this exacerbates tissue damage, driving a vicious cycle serving to amplify and perpetuate inflammation.
Dendritic cells metabolize and activate retinol into all-trans retinoic acid which stimulates antimicrobial activity in M. tuberculosis-infected cells via paracrine interaction (INM3P.406)
Authors: Liu Philip , De Leon Avelino , Kim Elliot , Radu Roxana , Montoya Dennis 
Published: January 2023

Abstract
Understanding the innate immune response to the Mycobacterium tuberculosis, is a key step to development of effective immunotherapies for tuberculosis (TB). Epidemiological evidence correlates low serum vitamin A (retinol) levels to active TB; however, retinol is biologically inactive. Previously, we demonstrated that stimulation of M. b-infected macrophages with the active vitamin A hormone, all-trans retinoic acid (ATRA), triggers a NPC2-dependent antimicrobial activity. Therefore, understanding the mechanism by which the immune system metabolizes and activates retinol at the site of infection will solve the mystery of how retinol, despite being biologically inactive, can influence the immune response to infection. Through gene expression and functional analysis of human macrophage subsets and dendritic cells (DCs) we identified that GM-CSF-derived DCs express enzymes capable of the two step conversion of retinol into ATRA including, ALDH1A2 and DHRS9, which is subsequently released from the cell. Conditioned medium, from DCs cultured with retinol, stimulated expression of NPC2 from monocytes which was inhibited by a retinoic acid receptor inhibitor (RARi). Inclusion of DEAB, an ALDH1A2 inhibitor, during DCs and retinol incubation also abrogated this response. Expression levels of vitamin A metabolic enzymes are lower in TB lung tissues compared with normal lung. These results help us incorporate vitamin A into clinical care and therapeutic modalities for TB.
Cigarette smoke exposure augments the ability ofStaphylococcus aureus to evade host innate immune responses. (INM3P.407)
Authors: Kulkarni Ritwij , Singh Sanjay , Baral Pankaj , Jeyaseelan Samithamby 
Published: January 2023

Abstract
The nose is the most common reservoir for Staphylococcus aureus, which is present 30-40% of normal adults. Its presence in the nasopharynx varies with age, but greater in children than adults. The oxidant chemicals in cigarette smoke (CS) induce S. aureus biofilm formation by suppressing agr (accessory gene regulator) regulon. In addition to quorum sensing, agr regulon regulates virulence gene expression of S. aureus. Therefore, we hypothesized that CS exposure augments virulence of S. aureus by evading host immunity. Using RNAseq, S. aureus transcriptome analysis revealed upregulation of gene expression which includes the induction of effectors involved in the nasal colonization (clfB), survival inside macrophages (spa) and the escape from neutrophil extracellular traps (NETs; nuc). In accordance with the RNAseq data, CS-exposed S. aureus exhibited significantly reduced phagocytosis, improved intracellular survival, increased evasion NET-mediated killing in the in vitro assays using human and murine macrophages and neutrophils. Importantly, the mouse model of acute pneumonia by intranasal inoculation resulted in significantly increased pulmonary bacterial burden in mice infected with CS-exposed S. aureus as compared to those infected with control bacteria. These results, which are clinically relevant, support a novel paradigm that the normal microflora becomes hypervirulent following exposure to environmental stressors such as CS, leading to devastating infections in humans.
Existence of LysRS-MITF/Hint1 pathway in dendritic cells (IRM12P.642)
Authors: SHU SHIN LA , Yang Chris , Razin Ehud , Kemeny David 
Published: January 2023

Abstract
Using murine bone marrow derived dendritic cell line JAWS II, we investigated the existence of LysRS-MITF/Hint1 pathway in dendritic cells (DC). Based on florescence microscopy, unstimulated cells expressed LysRS predominantly in the cytoplasm. LysRS localization was increased in the nuclei 2 h after stimulation with LPS, RANKL, GM-CSF (LRG) and was sustained over 6 h. To examine its functional role, doxycycline (Dox)-controlled plasmid overexpressing a single point mutant of LysRS (Ser207 to Asp) that mimics constitutive phosphorylation (LysRSS207D) was stably transfected into the JAWS II cell line. Dox-treated LysRSS207D cells were more dynamic, showed increased cell spreading and motility compared to wild-type cells and to untreated LysRSS207D cells. Moreover, Dox-treated LysRSS207D cells exhibited up-regulation of MHCII, but not CD11c, CD80, CD86 and CD40. Primary cultures of bone marrow derived dendritic cells (BMDCs) from C57BL/6 mice were also examined. Bone marrow cells transfected with NUDT2 siRNA inhibited development into BMDCs compared to scrambled siRNA control. Taken together, LysRS-MITF/Hint1 pathway exists in DC and appears to play an important role in DC morphology, maturation and development. Next steps include measurement of cytokine levels, primary allogeneic T cell activation, and adoptive transfer in our asthma and flu models.
Investigating the factors that regulate the rate of intracellular IL-1β degradation (IRM12P.652)
Authors: Ainscough joseph , Gerberick G , Brough David , Kimber Ian , Dearman Rebecca 
Published: January 2023

Abstract
Interleukin (IL)-1β is a key player in the initiation and orchestration of inflammation. The secretion of IL-1β requires 2 independent processes; an initial signal to induce the upregulation of the inactive precursor (pro-IL-1β) and, a second signal to drive cleavage and subsequent secretion. In previous investigations using murine dendritic cells, we have demonstrated that in lipopolysaccharide-stimulated cells, pro-IL-1β is polyubiquitinated and degraded by the proteasome, resulting in a rapid reduction in the amount of intracellular IL-1β available for secretion. The current study has employed an immortalized bone marrow derived murine macrophage cell line that stably expresses fluorescent IL-1β to track the rate of degradation. Here, proteasome inhibition using MG132 led to an increase in cell fluorescence compared with baseline expression, as measured by flow cytometry. This was coupled with a corresponding increase in IL-1β protein levels, as measured by ELISA, and an increase in polyubiquitinated IL-1β, as detected by Western blot. These data indicate that polyubiquitination and subsequent proteasomal degradation of IL-1β are also features of macrophages and independent of cell activation. Furthermore, fluorescence is a reliable readout for measuring degradation in this cell line and thus, represents a useful tool to measure factors that regulate the rate of IL-1β degradation and potentially, the potency of the inflammatory response.