bluuebunny/crossref_metadata_2025_split huggingface dataset:
Activation of the stimulator of interferon genes (STING) adaptor attenuates experimental autoimmune encephalitis (THER6P.848)
Authors: Mohamed Eslam , Lemos Henrique , Huang Lei , Chandler Phillip , Souza Guilherme , Li Lingqian , Pacholczyk Gabriela , Barber Glen , Hayakawa Yoshihiro , Munn David , Mellor Andrew 
Published: January 2023

Abstract
Activation of Stimulator of Interferon genes (STING) adaptor causes the release of IFN αβ in most of the cells, an activity that is tightly controlled to avoid continuous immune activation that could lead to autoimmunity. Here we show that STING activators can be used systemically to suppress experimental autoimmune encephalitis following myelin oligodendrocyte (MOG) immunization through STING-IFN αβ pathway. DNA nanoparticles (DNPs) attenuated MOG-specific effector responses in CNS and spleen. Therapeutic responses to DNPs are dependent on STING adaptor and IFN αβ receptors as they were abolished in mice lacking those genes. Additionally, indoleamine 2,3 dioxygenase enzyme (IDO) is crucial for therapeutic responses to DNPs as the loss of IDO1 gene in hematopoietic cells and IDO inhibition by 1-methyl-D-tryptophan had ablated those responses. Cyclic diguanylate monophosphate (c-diGMP), a direct STING activator also attenuated EAE progression and disease severity. Thus, Systemic administration of DNPs and c-diGMP can attenuate EAE through STING-IFN αβ-IDO regulatory pathway that is activated in certain immune cells subsets to overcome CNS specific autoimmunity. These findings emphasize a previously unappreciated role of STING activators to attenuate autoimmunity.
Cell-based kidney transplantation in adult Danio rerio (TRAN3P.879)
Authors: Vahey Delia , Delaney Tetyana 
Published: January 2023

Abstract
With the increasing incidence of kidney disease and the growing list of patients in line for kidney transplantation, new methods to restore renal function are critical. An alternative method to whole-organ transplant involves the transplantation of individual cells. Here we show a new approach to the transplantation of kidney cells in zebrafish, Danio rerio. This method involves the transplantation of kidney cells from the donor fish by direct injection into the recipient kidney. The donor zebrafish were transgenic, their cells labeled with green fluorescent protein. Both renal and hematopoetic progenitor cells were isolated from the donor kidney and prepared for transplantation in an injection solution. The recipient fish were treated with gentamicin and immunosuppressed with 10 μg/ml dexamethasone for 2 days. Upon injection of 0.1-0.5 million allogeneic cells via the dorsum of the fish into the head kidney, anesthetized Danio rerio regained complete swimming abilities within 5 min. Seven out of nine fish survived the procedure. The fish were allowed to recover for 10 days, leaving time for the transplant cells to implant. Then the fish were euthanized and a renal biopsy was performed and visualized using a fluorescent microscope. The formation of new fluorescent renal tubules was observed and analyzed. Although further research must be done to refine this technique, we demonstrated that cell-based kidney transplant is a promising alternative to whole-organ transplant.
Cross dressing by donor dendritic cells maximizes T cell responses to indirectly presented alloantigen after BMT by promoting immune synapse formation (TRAN3P.903)
Authors: Markey Kate , Koyama Motoko , Gartlan Kate , Leveque Lucie , Kuns Rachel , Lineburg Katie , Teal Bianca , MacDonald Kelli , Hill Geoff 
Published: January 2023

Abstract
The role of trogocytosis in generating naïve T cell responses is controversial and highly relevant to transplantation. We transplanted B6.CD11cGFP donor bone marrow (BM) grafts (H2Db, IAb, GFP off the CD11c promoter) into irradiated B6D2F1 (H2Db/d, IAb/d,) and tracked donor DC. Using standard and imaging flow cytometry, we noted that all donor cells become “cross-dressed” in high levels of recipient hematopoietic cell-derived MHC early after BMT. To assess the functional consequences of cross-dressed MHC, we transplanted B6.MHCII-/- BM into WT B6 recipients to generate DC cross-dressed with IAb. WT DC induced proliferation in BALB/c CD4 T cells transferred into B6.MHCII-/- recipients but no responses were seen to B6.MHCII-/- DC bearing cross-dressed Ag. Next, DC were generated that were either cross-dressed (B6.MHCII-/-→B6D2F1), bore cross-dressed and indirectly presented Ag (B6→B6D2F1), or could present indirect Ag only (B6→B6.MHCII-/-xD2F1) to the allospecific TEa Tg T cell. T cell proliferation was maximal in response to processed recipient alloAg presented via the indirect pathway in the presence of cross-dressed Ag, markedly reduced to indirect Ag when cross-dressing was absent, and absent to cross-dressed Ag alone. Imaging flow cytometry confirmed that cross-dressing promotes formation of the immunologic synapse. Thus, cross-dressing serves to maximize naïve T cell responses to indirectly presented alloAg and exerts this effect after BMT via the immunologic synapse.
Tools for optimization of adoptive cellular therapy (TRAN3P.908)
Authors: Brunstedt Katja , Wallace Paul , Pan Dalin , Halgreen Charlotte , Pedersen Henrik , Brix Liselotte 
Published: January 2023

Abstract
The transfusion of T lymphocytes is a promising treatment for viral infections in immune-compromised patients, and has induced regression of cancer in early-stage clinical trials. Adoptive T cell therapy can be optimized in several ways: -Patient stratification - only those that benefit from therapy are treated -Measurement of T-cell immunity pre and post treatment - has the desired immune response been induced and is additional therapy needed -Quantitative quality control of the cellular product prior to infusion Multiple adoptive cell therapy trials are investigating the benefit of transferring CMV-specific T cells to transplant patients to avoid reactivation of CMV. We used the MHC multimer based Dextramer CMV assay, to quantitate CMV-specific T cells in whole blood and processed cell samples. The reconstitution of CMV immunity was followed in &gt;90 transplant patients and it was shown that induction of CMV-specific immunity upon adoptive transfer of CMV-specific T cells could be reliably measured. We showed that CMV Dextramers can be used to characterize cellular products with respect to CMV-specific T-cell composition upon in vitro expansion. The CMV Dextramer assay is a valuable tool that may improve CMV-specific adoptive cellular therapy by allowing: -Stratification - Identification of patients with low CMV-specific immunity -Immune monitoring - Determination of the induced cellular response in patients -Quality control - QC of manufactured cellular products
Age-associated increase of IL-6 dampens anti-tumor immune responses through attenuating Th1 differentiation of tumor-specific CD4 T cells (TUM7P.928)
Authors: Tsukamoto Hirotake , Senju Satoru , Swain Susan , Nishimura Yasuharu 
Published: January 2023

Abstract
With age immune function progressively decreases and pro-inflammatory status is concomitantly developed. We focused on the impact of this inflammatory environment on tumor-specific CD4 T cells. We demonstrated that in contrast to tumor elimination in young hosts, adoptive transfer of OVA-specific young CD4 T cells and vaccination with OVA peptide-pulsed DC failed to induce efficient regression of OVA-expressing tumor cells in aged hosts. This was consistent with the result that in aged mice generation of tumor-specific Th1 cells was significantly attenuated. Interestingly, a blockade of IL-6 signaling partially restored Th1 differentiation of donor CD4 T cells and elicited tumor regression in aged hosts, but did not alter their expansion, IL-2 or IL-17A production. Furthermore, in vitro analyses revealed that IL-6 directly acted on CD4 T cells to attenuate Th1 differentiation. IL-6 blockade also restored the defect in an ability of tumor-specific CD4 T cells to promote anti-tumor CD8 T cells induction in aged mice. These results were confirmed by aged IL-6-deficient mice. However, the Th1-mediated help for tumor-specific CD8 T cells was not observed in aged mice when IFN-γ-deficient CD4 T cells were transferred into anti-IL-6 Ab-treated mice. These results imply that systemic increase of IL-6 is an extrinsic factor counteracting tumor-specific Th1 differentiation, and is responsible for the age-associated defect in mounting protective anti-tumor immunity in aged individuals.
Combination Immunotherapy of Melanoma by inhibiting HSP90 and targeting its client proteins. (TUM7P.934)
Authors: Raveendran Subhara , Rao Aparna , Storkus Walter 
Published: January 2023

Abstract
HSP90 plays a major role in human melanoma progression, by facilitating the over expression of its client proteins, many of which provide growth, survival and migratory advantages to tumor cells and support a pro-angiogenic tumor microenvironment. HSP90-mediated stabilization also limits the proteasome-dependent processing of client proteins and the subsequent presentation of derivative peptides in tumor cell-associated MHC class I complexes which is required for effective host CD8+ T cell recognition of melanoma cells. When considered in the combination of active vaccination, inhibiting HSP90 function might serve as a highly effective anti-tumor therapy. As a consequence, we analyzed the anti-tumor impact of a range of clinically-relevant HSP90 inhibitors (HSP90i; i.e. NVP-AUY922, STA9090, 17-DMAG, Novobiocin) on client protein expression in vitro (human and murine melanoma cell lines) and in vivo (s.c. B16 tumors established in syngenic C57BL/6 mice). We observed that HSP90i promote the proteasome-dependent (i.e. blocked by inhibitors PS-341 and MG-132) degradation of a range of client proteins. We have also identified H-2Kb/Db-presented peptide epitopes derived from these client proteins that may be recognized by Type-1 CD8+ T cells after specific vaccination. We are currently investigating the capacity of NVP-AUY922 and STA9090 to therapeutically sensitize established melanomas to client protein-specific CD8+ T cells in combination therapy approaches.
Mitochondrial-mediated inflammasome activation controls production of Hepatoma-derived IL-1β (TUM7P.942)
Authors: Baseler Walter , O'Connor Geraldine , McVicar Daniel 
Published: January 2023

Abstract
Obesity is emerging as a critical driver of chronic inflammation and inflammation-associated cancers, such as hepatocellular carcinoma. The obesity-associated proinflammatory cytokine IL-1β is upregulated in multiple solid tumors and correlates with poor overall prognosis. Recently, fatty acid-induced inflammasome activation in macrophages has been shown to be dependent on mitochondrial reactive oxygen species (ROS) production. Here we asked whether in addition to traditional sources of IL-1β, the tumor itself might participate in maintenance of the inflammatory state. Stimulation of murine hepatoma line Hep1-6 cells with LPS substantially increased the mRNA level of IL-1β. Addition of LPS and palmitate together significantly elevated Hep1-6 IL-1β mRNA and mature IL-1β abundance. Inflammasome constituents were detectable in Hep1-6 cells with Nlrp3 mRNA content increased in LPS/palmitate treated cells. Cellular ROS levels were significantly increased in the LPS/palmitate treated group. AICAR, a potent activator of AMPK and mitophagy, decreased cellular ROS and mature IL-1β production by LPS/palmitate treated Hep1-6 cells, but did not affect IL-1β mRNA levels. Collectively, our data indicates that fatty acid-induced mitochondrial ROS results in inflammasome activation and cooperates with LPS-driven IL-1β mRNA resulting in tumor cell production of mature IL-1β. Through this mechanism, plasma fatty acids may promote tumor-associated inflammation.
Estrogen receptor α signaling inhibits T cell responses against gynecologic tumors (TUM7P.943)
Authors: Svoronos Nikolaos , Allegrezza Michael , Nguyen Jenny , Perales-Puchalt Alfredo , Rutkowski Melanie , Stephen Tom , Tesone Amelia , Conejo-Garcia Jose 
Published: January 2023

Abstract
Estradiol is a pleiotropic steroid hormone that regulates homeostasis, cell growth, and the immune system. In estrogen receptor positive breast cancer, anti-estrogen therapies are used to inhibit tumor cell growth. However, estrogen receptor α is also found on non-tumor cells within the tumor microenvironment. Therefore, we hypothesized that estradiol is able to affect tumor progression independent of its direct effect on tumor cells by regulating the tumor microenvironment, specifically immune cells. Using an orthotopic mouse model for ovarian cancer, we show that even in an estrogen receptor negative tumor, estradiol is able to drive tumor progression. Conversely, dependent upon an adaptive immune response, depletion of estrogen impedes tumor growth, as WT but not recombinase activating gene KO (RAG KO) estrogen-depleted mice survive longer than control littermates. This effect is dependent on estrogen receptor α expression in T cells. Mice reconstituted with a 1:1 mixture of WT and estrogen receptor α KO bone marrow reveal that estrogen receptor α KO T cells exhibit greater antitumor effector functions and pro-inflammatory cytokine production in a cell-intrinsic manner. These findings suggest anti-estrogens as a novel adjuvant therapy to augment T cell-dependent immunotherapies, such as adoptive cell transfer and programmed cell death protein 1 (PD-1) inhibition.
Chitinase-3 like-protein-1 (CHI3L1) expression associated with pulmonary inflammation accelerates breast cancer metastasis (TUM7P.960)
Authors: Libreros Stephania , Garcia-Areas Ramon , Iragavarapu Vijaya 
Published: January 2023

Abstract
Disseminated metastasis accounts for a majority of breast cancer deaths. Recently, elevated serum levels of a glycoprotein known as chitinase-3 like-protein-1 (CHI3L1) has been correlated with poor prognosis in both breast cancer and asthmatic patients. We have combined mouse models of breast cancer and pulmonary inflammation to determine if CHI3L1 associated pulmonary inflammation accelerates metastasis. We found that allergic pulmonary inflammation significantly enhances primary tumor growth in 4T1, 4T07 and 67NR mammary tumors by 10-fold, while decreasing survival. 4T1 tumor bearers with allergic pulmonary inflammation showed a 100-fold increase in metastatic tumor formation. We also assessed CHI3L1 levels and myeloid cells in the lungs of wild type and CHI3L1 knockout mice with allergic pulmonary inflammation and 4T1 mammary tumors. CHI3L1 levels were higher in the lungs of mammary tumor bearers with allergic pulmonary inflammation and correlated with increased metastasis. Wild type mammary tumor bearers with allergic inflammation had higher numbers of myeloid cells in the lungs in comparison to CHI3L1 knockout tumor bearers with allergic pulmonary inflammation. CHI3L1 knockout mice tumor bearers had significantly fewer myeloid cells in the lungs, decreased tumor growth and metastasis, along with increased survival. We propose that increased CHI3L1 in the lungs attracts myeloid cells that promote tumor growth and breast cancer metastasis.
TREM2 differential expression is associated with BAL cells that express high levels of ARG1 (VET2P.1044)
Authors: Hiremath Jagadish , Binjawadagi Basavaraj , Ouyang Kang , Dhakal Santosh , Manickam Cordelia , Dwivedi Varun , Gourapura Renukaradhya 
Published: January 2023

Abstract
Lung immunopathology is the major cause of influenza induced morbidity and mortality. DAP12 (DNAX-Activating Protein of 12kDa) is a membrane adaptor protein associated with varied surface receptors, and it is known to regulate influenza induced lung immunopathology. DAP12 associated receptor expression and their association with phenotype of bronchoalveolar lavage fluid (BAL) cells during swine influenza virus (SIV) infection in pigs is unknown. Since pig is a suitable large animal model for influenza research, our aim was to understand the effects of zoonotic SIV H1N1 infection on expression of DAP12 associated MDL-1, TREM-1 and TREM-2 receptors in pig BAL cells through qRT-PCR profiling. Results indicated that DAP12, MDL-1 and TREM-1 were constitutively expressed, but TREM-2 was upregulated. Stimulation of uninfected pig BAL cells in vitro using the cytokines IFNgamma and IL4 revealed that upregulation of TREM-2 was associated with enhanced expression of ARG1, but not with high levels of expression of TNFalpha and iNOS. ARG1 is a phenotype of alternatively activated alveolar macrophages that are known to protect lung tissue; hence TREM2 upregulation in pig BAL cells with similar phenotype suggests the possible beneficial role of these molecules in preventing the lung immunopathology. In conclusion, DAP12 and associated receptors are differentially expressed in pig BAL cells and TREM2 appears to have a beneficial role in regulating the lung immunopathology in SIV infection.
Polyfunctional CD4 T cells in the response to bovine tuberculosis. (VET2P.1034)
Authors: Fernanda Maggioli Mayara , Palmer Mitchell , Vordermeier H. Martin , Whelan Adam , Waters Wade Ray 
Published: January 2023

Abstract
Polyfunctional CD4 T cells simultaneously produce interferon-gamma (IFN-γ), interleukin-2 (IL-2) and tumor necrosis factor-alpha (TNF-α) and play relevant roles in several chronic infections, including human TB and HIV. However, the assessment of this response in bovine infections was not feasible due to the lack of monoclonal antibodies that recognize bovine IL-2. Recently, an antibody specific for bovine IL-2 enabled the evaluation of polyfunctional T cells in cattle. The objective of the present study was to access antigen-specific polyfunctional ex vivo responses after aerosol Mycobacterium bovis infection of cattle. Peripheral blood mononuclear cells (PBMCs) were collected from infected cattle and stimulated with rESAT-6:CFP-10 (E:C), media or pokeweed (PWM) for 16 hours. After stimulation, the expression of CD4, CD45RO, CCR7, IL-2, IFN-γ, TNF-α and cell viability were evaluated. The ex vivo response to E:C consisted of 63% effectors (CD4+ CD45RO- CCR7-), 32% effector memory (CD4+ CD45RO+ CCR7-), and 9% central memory (CD4+ CD45RO+ CCR7+) phenotypes. In regard to the cytokine profile, 70% of cells producing cytokines expressed both IFN-γ and TNF-α, 31% expressed all three cytokines, 6% expressed both TNF-α and IL-2, and 3% expressed IL-2 and IFN-γ. Cells producing only IFN-γ, IL-2, or TNF-α represented, 5%, 8% and 1%, respectively. These findings demonstrate that the polyfunctional response consists mainly of effector cells co-producing IFN-γ and TNF-α.
Serologic markers of Epstein-Barr virus (EBV) reactivation in the conditions of viral encephalitis in young patients (VIR7P.1063)
Authors: Goleva Olga , Murina Elena 
Published: January 2023

Abstract
Viral encephalitis is one of the most severe disorders that is accompanied by the processes of tissue demyelization. It can be connected with both significantly causative viruses and a potential activation of latently persisting viruses of herpes group. The impact of persisting viruses on the activation of viral encephalitis is unclear. We analyzed the blood samples from 1-14 year-old patients diagnosed with acute viral encephalitis who had a history of Epstein-Barr virus (EBV) infection. Interestingly, there were low levels of IgM to early antigen (EA), nuclear antigen (NA) and capsid antigen (VCA) but high levels of IgG as determined by ELISA. Next, we used specific stripes with the immonobilized extract of EBV antigens (Euroimmun, Germany). The analysis of IgG antibodies in all blood samples demonstrated the presence of the late phase infection markers: VCA and NA. Importantly, the early antigen EA (areas: p43, p45, p93) that is induced within 3 weeks after the initial infection, was clearly present as well. These results suggest that EBV reactivation could be one of the favorable factors that accompanies the processes of tissue demyelization of central nervous system.
Escherichia coli Nissle 1917 colonization ameliorates human rotavirus diarrhea and modulates B cell responses in a neonatal gnotobiotic pig disease model (VIR7P.1064)
Authors: Kandasamy Sukumar , Vlasova Anastasia , Chattha Kuldeep , Fisher David , Shao Lulu , Kumar Anand , Rajashekara Gireesh , Saif Linda 
Published: January 2023

Abstract
Microbiota regulate gut immune homeostasis, but the role of gram positive (G+)/negative (G-) commensals/probiotics in modulating human rotavirus (HRV) diarrhea and immunity is largely undefined. Gnotobiotic (Gn) piglets, the only animal model susceptible to HRV diarrhea, were colonized with G- Escherichia coli Nissle 1917 (EcN), G+ Lactobacillus rhamnosus strain GG (LGG) or LGG+EcN and challenged with virulent HRV (VirHRV) to assess probiotic effects on HRV pathogenesis and immunity. EcN+VirHRV piglets had significantly lower mean peak virus shedding titers and significantly reduced mean cumulative fecal scores and duration of diarrhea compared to LGG+VirHRV or control (uncolonized)+VirHRV piglets. LGG+VirHRV pigs had lower, but statistically similar fecal scores compared to control+VirHRV pigs. Combining LGG+EcN did not moderate VirHRV shedding or diarrhea more than EcN alone. Coinciding with reduced HRV infection, EcN+VirHRV piglets had significantly lower small intestinal (SI) HRV IgA antibody titers and HRV IgA and IgG antibody secreting cells (SC) compared to control+VirHRV piglets. Though not significantly different, SI HRV IgA antibody titers were 2-4-fold lower in the LGG+EcN+VirHRV and LGG+VirHRV piglets. Notably EcN+VirHRV piglets had higher SI total IgA levels and IgA SC compared to LGG+VirHRV piglets. Our results indicate that EcN alone was more effective than LGG or LGG+EcN in ameliorating HRV diarrhea and modulating B cell responses in a Gn piglet disease model.
Pivotal pathways that control host responses to lentivirus infection. (VIR7P.1053)
Authors: Mellor Andrew , Huang Lei , Li Lingqian , Lemos Henrique , Pacholczyk Gabriela , King William 
Published: January 2023

Abstract
Lentiviruses such as Human Immunodeficiency Virus establish persistent infections but it is unclear how lentiviruses evade host immunity. To elucidate pivotal pathways driving tolerogenic host responses to lentiviral infection we used the murine leukemia virus (LP-BM5) infection model, which causes persistent infections leading to splenomegaly, immunosuppression, and increased risk of leukemia. LP-BM5 infection induced indoleamine 2,3 dioxygenase (IDO) enzyme activity in a subset of dendritic cells expressing the B cell marker CD19 that regulate T cells. Different mouse strains exhibit variable resistance to LP-BM5 infection but all mice succumb to persistent LP-BM5 infection. In contrast, mice with genetically-induced, functional defects in Foxp3-lineage regulatory CD4 T cells (Tregs) cleared LP-BM5 infections ~4-6 weeks post infection. LP-BM5 clearance did not occur unless mice were homozygous for defective Foxp3 alleles, indicating that wild-type Tregs mediate dominant tolerogenic responses to LP-BM5 infection. Depleting CD8 T cells prevented LP-BM5 clearance in Treg defective mice, and LP-BM5 resistance correlated with diminished IDO induction, suggesting that Tregs block CD8 T cells via IDO to promote virus persistence. Thus Tregs are pivotal in promoting tolerogenic responses to LP-BM5 infection, and interfering with pathways that activate Tregs and mediate responses to activated Tregs may enhance host resistance to lentivirus infections.