bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD4+ helper T cells and immunity to therapeutic proteins in gene therapy (P4431)
Authors: Bremer William , Campbell Katherine , Rodino-Klapac Louise , Goodchild Laurie , Griffin Danielle , Montgomery Chrystal , Walker Christopher 
Published: January 2023

Abstract
Adeno-associated (AAV) vectors are being assessed for gene replacement therapy in humans. Proteins encoded by the vector transgene are potentially foreign. Strategies to temper destructive immune responses may be required to achieve a therapeutic effect, particularly in diseases like Duchenne Muscular Dystrophy where the defective gene to be replaced has a large frame-shifting deletion. Here we studied the impact of immunity on expression of enhanced green fluorescent protein (eGFP) in AAV-transduced skeletal muscle of rhesus macaques. Antibodies to eGFP developed rapidly after AAV vector delivery to the tibialis anterior muscle. Cellular immunity was variable. In some animals, strong CD4+ helper and CD8+ cytotoxic T cells were detected in blood 2 weeks after vector administration. In others, low frequency responses were not detected until week 5-6. Regardless of the pace of the response, eGFP was cleared or substantially diminished in muscle after week 6. Antibody-mediated depletion of CD4+ T cells before AAV vector treatment delayed antibody and CD8+ T cell immunity and facilitated robust expression of eGFP persisted through day 42. Some positive muscle fibers were still visible at month 10, an unexpected finding that suggested an inability to sustain destructive cellular immune responses initially primed without CD4+ T cell help. The capacity for recall of eGFP-CD8+ T cell responses upon administration of a serologically distinct AAV vector is now being assessed.
Reciprocal interaction between NK cells and DCs regulates the Th17 response by controlling the innate IFN-γ/IL-27 axis (P4067)
Authors: Caspi Rachel , Chen Jun , Silver Phyllis , Horai R. , Mattapallil Mary , Zhou Ru , Chong WaiPo 
Published: January 2023

Abstract
Experimental autoimmune uveitis (EAU) is a model for human uveitis. IFN-γ deficient mice develop enhanced EAU with elevated Th17 responses. This protective effect of IFN-γ appears at odds with the ability of IFN-γ-producing Th1 cells to induce EAU. We previously showed that it is innate IFN-γ that is protective, whereas adaptive IFN-γ is pathogenic. We now demonstrate that NK cells, a major cellular source of innate IFN-γ, interact with DCs in draining lymph nodes of EAU-challenged mice to ameliorate disease by controlling the innate IFN-γ/IL-27 axis. After immunization, both NK cells and DCs were recruited to the draining lymph nodes, with recruitment of NK cells being at least partially dependent on DCs and CXCR3. Infusion of IFN-γ-sufficient WT NK cells into IFN-γ-/- recipients reduced their EAU scores and enhanced production of IL 27, a suppressor of the Th17 response, from DCs of these mice. In vivo neutralization of IL-27 abolished the protective effect of WT NK cells on EAU. In vitro experiments revealed that mature DCs induced NK cells to produce IFN-γ, which in turn caused DCs to produce IL-27. IL-27 then stimulated NK cells to produce more IFN-γ, suggesting the presence of a positive feedback loop. IL-27 also promoted generation of IL-10-producing Tr1-like cells. Our results suggest that the magnitude of the Th17 response, and consequently the severity of disease, is controlled by the interaction between NK cells and DCs through the IFN-γ/IL-27 axis.
FAS-associated death domain and tripartite motif-containing protein 21 negatively regulate virus-induced interferon production (P6126)
Authors: Nandu Suruchi , Chandra Neha , Winoto Astar 
Published: January 2023

Abstract
While induction of type I Interferon (IFN) during the host antiviral response is imperative for efficient viral clearance, regulation of this response is crucial for immune system homeostasis. We have reported that FADD, a well characterized apoptotic protein, is involved in negatively regulating RNA virus-induced IFN-α production through a novel interaction with the E3 ubiquitin ligase, Tripartite Motif-Containing Protein 21 (TRIM21). Interaction between FADD and TRIM21 cooperatively represses IFN-α activation in Sendai virus infected cells. This occurs through TRIM21 targeted ubiquitin-mediated degradation of transcription factor IRF7. Recently, it was also determined that FADD and TRIM21 can be found in 2 different complexes within cells: one containing FADD/TRIM21/IRF7 and a second containing FADD/TRIM21/RIP1 (receptor-interacting protein 1). We hypothesized that TRIM21 may have different functions dependent on complex composition and that its activity could be regulated via its autoubiquitination status. Interaction of TRIM21 with FADD yields increased autoubiquition while interaction with RIP1 yields decreased autoubiqtuitination suggesting a regulation mechanism. However, studies involving mutant forms of TRIM21 which lack autoubiquitination demonstrate that autoubiquitination status does not regulate TRIM21’s ability to target IRF7 for degradation. We are currently investigating the function of TRIM21 in other FADD/RIP1 dependent processes like cell death.
Bovine natural killer cells are infected by BVDV and the infection results in changes in phenotype and activation markers (P6128)
Authors: Darweesh Mahmoud Darweesh , Rajput Mrigendra , Lebedev Max , Young Alan , Braun Lyle , Chase Christopher 
Published: January 2023

Abstract
Bovine viral diarrhea virus infection results in immunosuppression and persistence infection of fetuses. Understanding the interaction between BVDV and NK cells is important to understand the ability of the virus to establish its infection especially through placental barrier to generate persistently infected animals. In this study, we demonstrated that BVDV is able to infect bovine NK cells by detecting the presence of BVDV RNA level up to 24 hours post infection without any increase in RNA levels over time. We also confirmed that although BVDV viral RNA was present, there was neither infectious virus release at any time post infection nor decrease in cell viability due to infection. Using immunofluorescence staining of NK cells with a mAb specific for BVDV E2 protein, we demonstrated the presence of BVDV E2 proteins in NK cells. The effect of BVDV strains that differ in its virulence or biotype on NK activation marker CD25 and phenotype markers CD2, NKp46 and the granule protein granulysin was determined. Highly virulent BVDV strains significantly decreased surface expression of each of these markers with a slight increase in the total number of cells expressing the markers 24 hour post infection. On the other hand less virulent strains increased significantly the percent of expression but with a slight decrease in the number of cells expressing the markers.
Prenatally acquired vitamin A deficiency alters innate immune responses to human rotavirus in a gnotobiotic pig model (P6145)
Authors: Saif Linda , Chattha Kuldeep , Kandasamy Sukumar , Siegismund Christine , Vlasova Anastasia 
Published: January 2023

Abstract
We examined how prenatally acquired vitamin A deficiency (VAD) modulates innate immune responses and human rotavirus (HRV) vaccine efficacy in gnotobiotic (Gn) pigs. The VAD and vitamin A sufficient (VAS) Gn pigs were vaccinated with attenuated HRV (AttHRV) with or without concurrent oral vitamin A supplementation (100,000IU) and challenged with virulent HRV (VirHRV). Regardless of vaccination status, the numbers of conventional and plasmacytoid dendritic cells (cDCs and pDCs) were higher in VAD vs. VAS piglets pre-challenge, but decreased substantially post-challenge. We observed significantly higher frequency of CD103 (integrin αEβ7) expressing DCs in VAS vs. VAD piglets post-challenge, indicating that VAD may interfere with homing (including intestinal) phenotype acquisition. Post VirHRV challenge, we observed longer/more pronounced diarrhea and higher VirHRV fecal titers in non-vaccinated VAD piglets. Consistent with higher VirHRV shedding titers, higher IFNα levels were induced in control VAD vs. VAS piglet sera at post-challenge day (PCD)2. HRV-stimulated mononuclear cells (MNCs) isolated from spleen and blood of VAD pigs pre-challenge also produced more IFNα. In contrast at PCD10, we observed reduced IFNα levels in VAD pigs that coincided with decreased TLR3+ MNC frequencies. Thus, prenatal VAD caused an imbalance in innate immune responses and exacerbated VirHRV infection, whereas vitamin A supplementation failed to compensate for these VAD effects.
Corneal-derived dendritic cells are critical in T cell expansion after HSV-1 ocular infection (P6102)
Authors: Buela Kristine-Ann , Hendricks Robert 
Published: January 2023

Abstract
Corneas possess resident DC (cornea-resident DC), and infiltrating DC migrate to the draining lymph nodes (DLN) following HSV-1 corneal infection (cornea-derived DC). The relative contribution of these two DC populations and DLN-resident DC to CD4+ and CD8+ T cell expansion in DLN following HSV-1 corneal infection is unknown. Local depletion of cornea-resident DC in CD11c-DTR mice did not affect T cell expansion in the DLN. Depletion of cornea-derived DC through 3 days post infection (dpi) abrogated early (3 dpi) CD8 T cell expansion (BrdU incorporation), but only reduced CD4 T cell expansion by 50%. Systemic DC depletion through 3 dpi completely abrogated expansion of both CD4 and CD8 T cells at 3 dpi. Depletion of cornea-derived DC through 7 dpi abrogated expansion of both CD4 and CD8 T cell at 7 dpi. Cornea-derived DC showed greater expression of CD70, CD80, and CD86 relative to DLN-resident DC. Blocking CD70 co-stimulation significantly reduced CD8 T cell survival and numbers, but did not affect their proliferation. We conclude that: (i) DC are required for expansion of both HSV-specific CD4+ and CD8+ T cells; (ii) up to 3 dpi CD8 T cells absolutely require HSV-1 presentation by cornea-derived DC possibly due to their enhanced costimulation, whereas DLN-resident DC can stimulate CD4 T cells (presumably with free virus that reaches the DLN from the cornea); (iii) after 3 dpi expansion of both CD4+ and CD8+ T cells requires transport of viral antigens by cornea-derived DC.
IL-33 induces nuocytes and modulates liver injury in viral hepatitis (P6112)
Authors: Liang Yuejin , Jie Zuliang , Hou Lifei , Aguilar-Valenzuela Renan , Vu David , Soong Lynn , Sun Jiaren 
Published: January 2023

Abstract
Our aim was to investigate the role of IL-33, the damage-associated molecular pattern (DAMP) molecule, in acute viral hepatitis. C57BL/6 mice were i.v. infected with a recombinant adenovirus. IL-33 and its receptor ST2 expression increased steadily in the liver in the first week of the infection. Treatment of exogenous IL-33 resulted in a great decrease in the serum alanine aminotransferase (ALT) levels and the number of Councilman bodies. Attenuated liver injury by IL-33 correlated with an increase in T regulatory (Treg) cells but with a decrease in macrophages, dendritic cells and NK cells in the liver. IL-33 enhanced both type 1 (IL-2, IFN-γ) and type 2 (IL-5, IL-13) immune responses in infected mice. However, IL-33 inhibited TNF-α in hepatic T cells and macrophages, and significantly reduced TNF-α levels in the liver. In addition to direct effects of IL-33, we found that IL-33 strongly induced nuocytes in the liver and spleen of infected mice. When co-cultured with nuocytes, hepatic T cells and macrophages expressed lower levels of TNF-α. In addition, the adoptive transfer of nuocytes led to lower serum ALT levels in the Ad-infected recipient mice. In conclusion, this study reveals that IL-33 acts as a potent immune stimulator and a hepatoprotective cytokine in acute viral hepatitis. Its direct immunoregulatory functions and ability to induce nuocytes further suggest to us that it may be a potentially promising therapeutic candidate for the management of viral hepatitis.
Common activation mechanism of beta 2 integrins (P5073)
Authors: Yuki Koichi , Sen Mehmet , Springer Timothy 
Published: January 2023

Abstract
Beta 2 integrins are α/β heterodimeric adhesion molecules on leukocytes consisting of αLβ2, αMβ2, αXβ2 and αDβ2. They undergo significant conformational changes to bind to their ligands at the domain called the α I domain of the α subunit. It has been postulated that the domain called the β I domain of the β subunit undergoes conformational changes and crosstalks with the α I domain in favor for ligand binding. However, the regions important for the β I domain activation have not been determined yet. Our unpublished and published αXβ2 structures suggested that the α1 helix of the β I domain was important in the activation of the β I domain. To examine the importance of the α1 helix, alanine scanning mutagenesis was performed on this region and transient transfectants in 293T cells were used for the experiments. Our β2 V124A and L132A mutations were constitutively active in all αL, αM, and αX integrins as demonstrated by ligands’ binding (αLβ2 to ICAM-1, αMβ2 to fibrinogen, ICAM-1, and iC3b, αXβ2 to iC3b) and activation-sensitive epitope exposures (KIM127, m24 and MEM148) even without activation. On the other hand, β2 L127A and L135A mutations were deactivating in all αL, αM and αX integrins as shown by diminished ligands’ binding and activation-sensitive epitope exposures even in an activating condition. Our data suggested that b2 integrins shared the common activation pathway of the β I domain, which would subsequently relay to the αI domain.
Permission of pathogen expansion inside a specialized DC by a co-inhibitory pathway dictates T cell response (P4249)
Authors: Fu Yang-Xin , Yang Xuanming 
Published: January 2023

Abstract
B and T lymphocyte attenuator (BTLA), a co-inhibitory receptor, is required to suppress T cell mediated immunity. We demonstrated that cross-linking BTLA by herpesvirus entry mediator (HVEM) suppresses T cell responses against Listeria, while blocking BTLA enhances the T cell response at not only primary but also memory phase. Unexpectedly, BTLA-/- mice have significantly lower CD8+ T cell effector and memory cells while paradoxically being more resistant to Listeria monocytogenes infection. Although there is no reduction in listerial entry into BTLA-/- CD8+ DCs, intriguingly, Listeria fails to expand within these DCs. We show that BTLA signaling is essential for limiting Fas/FasL-mediated suppression of Listeria expansion within host CD8+ DCs, which more effectively alert adaptive immune cells. This study provides new evidence that a co-inhibitory pathway for T cells can increase overall T cell responses by permitting pathogen proliferation within specialized DCs.
A role for IL-10 in efferocytosis of apoptotic neutrophils and resolution of lung inflammation produced by STAT1-regulated lung MDSC-like cells (P4255)
Authors: Arora Meenakshi , Poe Stephanie , Ray Anuradha , Ray Prabir 
Published: January 2023

Abstract
Bacterial pneumonia remains a significant burden worldwide. Little is known about cellular and molecular mechanisms in the lung tissue that remove apoptotic neutrophils to prevent collateral tissue damage during bacterial infection. In a mouse model of bacterial infection with Klebsiella pneumoniae, we observed a role for IL-10, produced in a highly orchestrated fashion by tissue-resident cells, in resolution of pulmonary inflammation and recovery of mice post-infection. While IL-10-/- mice cleared bacteria, they displayed increased morbidity with progressive weight loss and persistent lung inflammation. The major source of tissue IL-10 was CD11b+Gr1intF4/80+ cells resembling myeloid-derived suppressor cells (MDSCs) that accumulated with a delayed kinetics in the lung tissue after infection and efficiently efferocytosed apoptotic neutrophils. However, increased bacterial burden handicapped this function by decreasing the ratio of the efferocytic MDSC-like cells:inflammatory neutrophiils in the lung. To overcome this undesirable outcome, we took advantage of functional antagonism between STAT1 and STAT3. In STAT1-/- mice, the excessive neutrophil infiltration in the lungs elicited by a high dose of K. pneumoniae was attenuated with concomitant increase in the frequency of the MDSC-like cells that expressed increased pSTAT3 levels. Thus, inhibiting STAT1 may be a novel therapeutic strategy to address inefficient resolution of bacterial pneumonia.
Critical role of TRIF in modulating innate and adaptive responses during Citrobacter rodentium infection. (P4037)
Authors: Gurung Prajwal , Kanneganti Thirumala 
Published: January 2023

Abstract
Citrobacter sp. is a gram negative coliform facultative bacteria that is mostly associated with the inflammation of the gut and is widely studied as the mouse model organism for studying colitis. We previously showed that Citrobacter rodentium induces non-canonical inflammasome activation through a TRIF-dependent mechanism, however the exact role of TRIF during C. rodentium induced colitis is still unclear. Herein, we have used C. rodentium induced colitis model to study the role of TRIF in vivo and showed that TRIF plays a critical role in protection during C. rodentium induced colitis. Absence of TRIF results in increased bacterial burden and morbidity, which is dramatically increased when TRIF is absent in a Myd88-deficient background. TRIF-deficiency resulted in increased neutrophil numbers in the spleen. Similarly, dendritic cells frequency and numbers were dramatically increased and displayed activated phenotype. T cell frequencies and numbers were similar, however TNF-alpha producing CD4+ and CD8+ T cells were dramatically reduced. Further studies are being carried out in the lab to understand how TRIF regulates these various populations and functions of the innate and adaptive cells.
Innate defence regulator peptide 1018 protects mice in the Pseudomonas aeruginosa acute lung infection model (P4045)
Authors: Hilchie Ashley , Magrangeas-Janot Laure , Wuerth Kelli , Hancock Robert 
Published: January 2023

Abstract
Host defence peptides (HDPs) are small peptides with antibacterial and immunomodulatory properties that represent a novel approach to anti-infective therapy. Innate defence regulator (IDR) peptides, such as IDR-1018, are synthetic variants of HDPs that are more potent and less toxic than their naturally-occurring counterparts. IDR-1018 kills bacteria, exhibits potent chemokine activities, and decreases LPS-induced TNF-α release. The goal of the current study was to determine whether IDR-1018 protects mice in the Pseudomonas aeruginosa acute lung infection model. To this end, CD1 mice were given IDR-1018 (4 mg/kg) or the saline control intravenously 4 h prior to infection. Mice were infected intranasally with P. aeruginosa (1 x 10^6). Mice were sacrificed 18 h post infection, and blood and bronchoalveolar lavage fluid (BALF) was collected. P. aeruginosa CFUs and leukocyte counts in the BALF were determined. Cytokine concentrations (TNF-α, MCP-1, KC, and IL-6) in the blood and BALF were quantified by ELISA. Our data showed that IDR-1018-treated mice had fewer CFUs and neutrophils and more monocytes in the BALF, and lower concentrations of proinflammatory cytokines in the BALF and in the blood than did saline-treated mice. These results suggest that IDR-1018 may have potential for the treatment of acute lung infections. Future studies aim to optimize the delivery strategy by identifying a delivery vehicle that enhances the half life and ease of peptide delivery.
Celecoxib induces apoptosis, ER stress, and cyclooxgenase-2 expression on lung cancer cells (P2090)
Authors: Kim Ja-Young , Kim Hae jin , Choi Yunock , Park Ga Bin , Kim Seonghan , Hur Dae Young , Kim Yeong Seok 
Published: January 2023

Abstract
Cyclooxygenase-2 (COX-2) is an enzyme that is inducible by various stimuli such as inflammation. COX-2 expression is increased in a variety of human premalignant and malignant tumors. Celecoxib is a selective inhibitor of COX-2 that have been shown to inhibit cancer cell growth and exert anti-proliferative and pro-apoptotic effects on various cancer cell lines. In this study, we investigated that celecoxib induced apoptosis of lung cancer cell lines, A549 and H460. Increase of apoptotic cells was investigated on celecoxib treated lung cancer cell lines. Cell proliferation was decreased and sub G0 peak was increased. We also observed that celecoxib induced activation of sensor proteins involved in endoplasmic reticulum (ER) stress. Expression of CHOP was induced after treatment of celecoxib on lung cancer cells on time-dependent and dose-dependent manners using RT-PCR and western blot. However, salubrinal, specific inhibitor of eIF-2 alpha phosphatase, did not block CHOP expression and cell death after celecoxib treatment. Paradoxically, celecoxib stimulated COX-2 mRNA and protein on A549, but indomethacin, inhibitor of COX-1 and COX-2 did not induce COX-2 expression. Taken together, we suggest that celecoxib induces apoptosis and ER stress on lung cancer cell line.
α-Galactosylceramide-loaded tumor cell vaccine promotes NKT-dependent protective immunity against multiple myeloma (P2096)
Authors: Choi Inhak , Hong Sungyoul , Lee Hyeonsoo , Jeon You-Kyoung , Lee Eungchang 
Published: January 2023

Abstract
Despite progress in the treatment of multiple myeloma (MM), this disease still remains fatal in the majority of patients. Thus, new immunotherapeutic approaches are required to eradicate minimal residual myeloma cells. Tumor cells can be a source of tumor-specific antigens for immunotherapy. However, tumor cells have been weak to promote tumor-specific immune responses and maintain the tumor-specific responses with large amounts of tumor burden in vivo. In this study, we evaluated and compared the efficacy of the vaccination of autologous myeloma cells loaded with artificial ligand for T cell receptor of NKT cells, α-galactosylceramide (αGC), in the myeloma mouse model. Our results showed that αGC-loaded MOPC315BM myeloma cell vaccines efficiently retarded tumor growth, induced tumor regression against established tumor, and protected surviving mice from tumor rechallenge. The therapeutic responses were associated with an induction of strong humoral immune responses including anti-myeloma cell antibodies and cellular immune responses including myeloma-specific CD8+ cytotoxic T lymphocytes and memory T cells. In addition, we found that regulatory T cells were significantly decreased in mice receiving αGC-loaded myeloma cell vaccines. Thus, our results demonstrate that αGC-loaded myeloma vaccine efficiently promote NKT-dependent anti-tumor immunity in our study.