bluuebunny/crossref_metadata_2025_split huggingface dataset:
Metabolomics reveals the impact of vitamin D on the microbiome (MUC4P.856)
Authors: Bora Stephanie , Ooi Jot Hui , Smith Philip , Patterson Andrew , Cantorna Margherita 
Published: January 2023

Abstract
Dysbiosis of the microbiome is an important factor in susceptibility to diseases such as inflammatory bowel disease (IBD). Vitamin D has known immunoregulatory roles and decreased levels of vitamin D are linked to IBD. Mice lacking the enzyme 1-α-hydroxylase (Cyp27B1 KO) cannot metabolize 25(OH)D3 into bio-active 1,25(OH)2D3. These mice are more susceptible to experimental colitis compared to WT controls. Here we wanted to determine the effect of vitamin D3 on the urinary metabolome, since microbial metabolites can be detected in urine. Urine was collected from WT, WT+20,000IU vitamin D3, and WT and Cyp27B1 KO littermates. Urine samples were profiled by ultra-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-QTOFMS) and multivariate data analysis tools. WT mice supplemented with vitamin D and WT and Cyp27B1 KO mice displayed changes in microbial metabolites compared to unsupplemented mice. Metabolites associated with the gut microbiota: p-cresol sulfate, indoxyl sulfate, xanthurenic acid, and betaine were different as a function of vitamin D3 supplementation or Cyp27B1 expression. Alterations in the urinary metabolome suggest a role for vitamin D3 in regulating the microbiota. Metabolite analysis will aid our understanding of the mechanisms by which vitamin D3 affects host health and the microbiota, and future work will address how changes in the microbiota affect susceptibility to experimental IBD.
Formyl-peptide receptor 2-expressing cells in Peyer’s patch are closely associated with the regulation of antigen-specific mucosal immune induction (MUC4P.832)
Authors: Jang Yong-Suk , Kim Sae-Hae , Lee Ha-Yan , Park Jisang , Lee Kyung-Yeol 
Published: January 2023

Abstract
It is known that formyl peptide receptor-2 (FPR-2) acts as innate immune sensor through recognition of peptides derived from pathogens such as H. pylori and HIV, mitochondrial peptides, amyloidogenic peptide including serum amyloid A, and inflammatory response-associated peptides such as LL-37. However, the role of FPR-2-expressing cells in mucosal immune inductive area, Peyer’s patch (PP), is still unraveled yet. In this study, we identified the expression of FPR-2 in both M cells located in follicle-associated epithelium of PP and follicular dendritic cells (FDCs) localized in germinal center. In addition, the level of Cxcl13 and TGF-β gene transcripts in FDCs, which are closely associated with recruitment and differentiation of IgA-secreting B cells was substantially increased by treatment of cathelicidin LL-37, one of FPR-2 ligands. Moreover, enhanced immune induction was verified in vivo mouse system through monitoring the antigen-specific mucosal immunity after oral administration of LL-37-conjugated antigen. Treatment of LL-37 has also shown the immune modulatory effects such as recruitment of CD11c+ cells and IL-17-secreting cells together with activation of FPR-2-expressing cells. Collectively, this study suggests that FPR-2-expressing cells are associated with regulation of mucosal immune induction and LL-37 could be used as a modulator for the cells.
S-nitrosothiols (SNO) imported through LAT1 (L-type amino acid transporter type 1) ameliorates phagocytosis in alveolar macrophages and neutrophils (MUC5P.868)
Authors: Brahmajothi Mulugu , Mason Stanely , Auten Richard 
Published: January 2023

Abstract
Phagocytosis is a receptor mediated process where cytoskeletal rearrangement occurs to internalize an organism or particles. Macrophages and neutrophils have this potential as they form the first line of defense in any immune response. During lung inflammation, neutrophils and monocytes are freshly recruited; infiltrating neutrophils outnumber other cells but are short lived. As a result, more focus is on macrophages. Our study was the first to describe LAT1 expression in macrophages. We wanted to see the expression of LAT1 and its effects on SNO uptake in neutrophils. We administered LPS (50µg/kg) intratracheally to induce inflammation in juvenile rats. After 24hrs, BALF was collected by intratracheal aspiration, cells were isolated and enumerated. LAT1 expression was observed in neutrophils, monocytes, red blood cells and macrophages. When these cells were exposed to 100 µm of CysNO, neutrophils and macrophages exhibited remarkable phagosomes, rapidly engulfed particles and other senescing neutrophils. These properties were not observed when L-Leu or JPH 203 (competitor and inhibitor) were used. We observed neutrophils cannibalizing senescing neutrophils and neutrophil clearance by macrophages. This was a preliminary observation and further studies are underway. We propose that s-nitrosylation is involved in post translational modifications of some key signaling and adaptor proteins that may be of therapeutic significance in immune response.
Fully immunomagnetic isolation of untouched purified lymphocyte populations directly from whole blood without the need for red blood cell sedimentation, hypotonic lysis or density gradient centrifugation in just 25 minutes. (TECH2P.871)
Authors: Kokaji Andy , MacDonald G. , Sun C. , Kellerman Drew , Le Fevre Tim , Leung Nathan , Ng Victoria , Tabatabaei-Zavareh Nooshin , McQueen Karina , Fairhurst Maureen , Thomas Terry , Eaves Allen 
Published: January 2023

Abstract
Human whole blood is made up of 93-96% red blood cells (RBCs), 4-7% platelets and 0.1-0.2% leukocytes consisting mainly of granulocytes, lymphocytes and monocytes. Due to the low frequency of lymphocyte subsets within whole blood, their isolation typically requires a pre-processing step such as Lymphoprep™ or hypotonic lysis to deplete RBCs prior to cell isolation. We have therefore developed a new, fully immunomagnetic method for the negative selection of untouched cells directly from whole blood, without the need for any pre-processing steps. The EasySep™ Direct procedure involves labeling RBCs, platelets and unwanted leukocytes present in human whole blood with antibody complexes and magnetic particles. The magnetically labelled unwanted cells are separated from the untouched desired cells using an EasySep™ magnet by simply pouring or pipetting the desired cells into a new tube. In just 25 minutes, purities of 92-99% total lymphocytes, 93-98% T cells, 87-97% CD4+ T cells, 75-91% CD8+ T cells, 91-99% B cells, 87-98% naïve B cells and 65-84% NK cells can be achieved with little to no residual RBC contamination. The isolation of cells directly from whole blood will enable rapid access to highly purified cells immediately ready for downstream functional assays with minimal sample handling.
Nontargeted urinary metabolite profiling of a chronic mouse model of multiple sclerosis (THER3P.884)
Authors: Giri Shailendra , Laila Poisson , Suhail Hamid , Singh Jaspreet , Deshpande Mandar , Datta Indrani , Denic Aleksandar , Rodriguez Moses , Rattan Ramandeep , Mangalam Ashutosh 
Published: January 2023

Abstract
In search of metabolites signature as biomarkers during EAE disease, we profiled urine from B6 EAE using global untargeted metabolomics. Evaluation of metabolomic profiling of urine from EAE and healthy B6 group by using a combination of high-throughput liquid-and-gas chromatography with mass spectrometry, we found that 132 out of 322 (41%) metabolites were differentially altered (P&lt;0.05) indicating robust alteration in the urine metabolomics profile during disease. Among the perturbed metabolites, 11 were up regulated in EAE urine whereas 121 were down regulated. We conducted pathway analysis of the biochemical pathways of the KEGG and considered both concerted changes in metabolite intensity within the pathway (GlobalTest) and alterations of high impact, and found that a number of pathways were significantly altered including glyoxylate and dicarboxylate, phenylanine metabolism, porphyrin & chlorophyll metabolism, primary bile acid biosynthesis, cysteine & methione metabolism, taurine and hypotaurine metabolism, glycine, serine & threonine metabolism, and beta-alanine metabolism. Alteration in these pathways during EAE disease suggesting that perturbation of certain central metabolites could have impact on multiple metabolic pathways. While some of these metabolite changes could easily be developed as biomarkers, the key to translating metabolomics into therapeutics would require figuring out the central altered metabolic pathway(s), once studied in detail.
GM-CSF-expanded CD1dhiCD5+ B cells suppress experimental autoimmune myasthenia gravis (THER3P.886)
Authors: Sheng Jian Rong , Quan Songhua , Soliven Betty 
Published: January 2023

Abstract
IL-10-competent subset within CD1dhiCD5+ B cells, also known as B10 cells, has been shown to regulate autoimmune diseases. Whether B10 cells can prevent or suppress the development of experimental autoimmune myasthenia gravis (EAMG) has not been studied. In this study, we investigated whether low dose granulocyte macrophage-colony stimulating factor (GM-CSF) can expand B10 cells in vivo, and whether adoptive transfer of CD1dhiCD5+ B cells would prevent or suppress EAMG. We found that treatment of EAMG mice with low-dose GM-CSF increased the proportion of CD1dhiCD5+ B cells and B10 cells. In vitro co-culture studies revealed that CD1dhiCD5+ B cells altered T cell cytokine profile but did not directly inhibit T cell proliferation. On the other hand, CD1dhiCD5+ B cells inhibited B cell proliferation and its autoantibody production in an IL-10-dependent manner. Adoptive transfer of CD1dhiCD5+ B cells to mice could prevent disease and suppress ongoing EAMG. This was associated with downregulation of mature dendritic cell markers and expansion of regulatory T cells resulting in the suppression of acetylcholine receptor (AChR)-specific T cell and B cell responses. Thus, our data have provided significant insights into the mechanisms underlying the tolerogenic effects of B10 cells in EAMG. These observations suggest that in vivo or in vitro expansion of CD1dhiCD5+ B cells or B10 cells may represent an effective strategy in the treatment of human myasthenia gravis.
Profile of circulatory metabolites in chronic mouse model of multiple sclerosis using untargeted global metabolomics (THER3P.883)
Authors: Giri Shailendra , Laila Poisson , Singh Jaspreet , Suhail Hamid , Deshpande Mandar , Datta Indrani , Rodriguez Moses , Rattan Ramandeep , Mangalam Ashutosh 
Published: January 2023

Abstract
Multiple sclerosis is a chronic inflammatory and demyelinating disease of the CNS. Although, MS is well characterized in terms of the role played by immune cells, cytokines and CNS pathology, nothing is known about the metabolic alterations that occur during the disease process in circulation. Here, we profiled the plasma metabolites of B6 with EAE, a known model of chronic MS using untargeted global metabolomics. Using a combination of high-throughput liquid-and-gas chromatography with mass spectrometry, a total of 326 metabolites were identified, with significant changes observed in 105 metabolites (13 up- and 92 down-regulated), that mapped to lipid and amino acid pathways, followed by the peptide, xenobiotic, carbohydrate, nucleotide and energy pathways (p&lt;0.05). To understand the functional role of these alterations, the KEGG metabolic library was analyzed using Metaboanalyst. The top nine most significant pathways in terms of GlobeTest p-value and impact are 1) fatty acid biosynthesis, 2) lysine degradation, 3) histidine metabolism, 4) beta-alanine metabolism, 5) pentose phosphate pathway, 6) arachidonic acid metabolism, 7) linoleic acid, 8) D-arginine and ornithine. Overall, these metabolic changes could be exploited as biomarkers for EAE/MS disease and to design new treatment paradigms where metabolic interventions could be combined with present and experimental therapeutics to achieve better treatment.
Ex vivo-induction of tumor-specific effector and memory T cells for adoptive T cell therapies using antigen-loaded DCs (TUM4P.910)
Authors: Hansen Morten , Wieckowski Eva , Kalinski Pawel 
Published: January 2023

Abstract
Adoptive transfer of ex vivo expanded tumor infiltrating lymphocytes (TILs), or blood-derived T cells genetically-modified to express specific T cell receptors have been used in adoptive cell therapies (ACT) of melanoma and hematologic malignancies. However, wider application of ACT has been limited by poor T cell infiltration of many cancers, terminal differentiation and limited proliferative and functional capacity of TILs from many lesions, or lack of defined tumor-rejection antigens. Here we show that these limitations can be circumvented by in vitro sensitization (IVS) of blood lymphocytes, using antigen-loaded dendritic cells (DCs) to generate CTLs specific against multiple tumor-specific epitopes. Type-1-polarized DCs (aDC1s) consistently induce high frequencies of functional tumor-specific CTLs from PBMCs of healthy donors and cancer patients. Such IVS-induced CTLs can be further expanded up to 1000-fold, in a single cycle of activation with anti-CD3, irradiated feeder cells and low-dose IL-2. Unexpectedly, presence of aDC1 supernatants during the rapid expansion phase improved not only the functional activity of the expanded CTLs, but also the frequencies of tumor-specific cells, their CD62L and IL-7R expression, and ability to survive and perform killer function in the absence of exogenous IL-2. The current data show a new strategy of generating high numbers of functional CTLs for ACT, with several important advantages compared to high-dose-IL-2 expanded TILs.
Regulation of cell death and antitumor immunity by a natural triterpenoid in malignant lymphoma (TUM4P.930)
Authors: Haque Azizul , Radwan Faisal , God Jason , Hossain Azim , Elvington Michelle , Tomlinson Stephen 
Published: January 2023

Abstract
B-cell lymphoma is a leading cause of hematological malignancies, accounting for almost 10% of new cancer-related deaths in the United States. In this study, we report that Ganoderic acid A (GA-A), a natural triterpenoid from the medicinal mushroom Ganoderma lucidum, induces anti-proliferative activity and apoptotic cell death in human B-cell lymphoma lines and primary B-cell tumors. GA-A treatment induced caspase-dependent apoptotic cell death characterized by a dose-dependent increase in active caspases 9 and 3, up-regulation of pro-apoptotic Bim and Bax proteins, and a subsequent loss of mitochondrial membrane potential with release of cytochrome c. In addition to GA-A’s anti-growth activity, we show that lower doses of GA-A enhance HLA class II-mediated antigen presentation and CD4+ T cell recognition of lymphoma in vitro. The therapeutic relevance of GA-A treatment was tested in vivo using the EL4 syngeneic mouse model of metastatic lymphoma. GA-A-treatment significantly prolonged survival of EL4 challenged mice and decreased tumor metastasis to the liver, an outcome accompanied by a marked reduction of myeloid-derived suppressor cells (MDSC) in the host. Thus, GA-A not only selectively induces apoptosis in tumor cells, but also enhances cell-mediated immune responses by reducing MDSC and elevating Ag presentation and T cell recognition. The demonstrated therapeutic benefit indicates that GA-A is a candidate for future drug design for the treatment of lymphoma.
Monocyte-derived dendritic cells are critical for the induction of anti-tumor immune responses (TUM4P.932)
Authors: Ronchese Franca , Kuhn Sabine , Yang Jianping , Gilfillan Connie 
Published: January 2023

Abstract
The DC subsets that are required for the induction of effective anti-tumor immune responses and the success of immunotherapy are unknown. We used mouse models to compare different immune-activating agents for their impact on tumor growth, tumor-specific T cells, cytokine production and DC activation. Treatments with poly I:C or a combination of monososdium-urate crystals (MSU) + Mycobacterium smegmatis delayed growth of B16 melanoma and 4T1 mammary carcinoma tumors, whereas LPS, M. smegmatis or MSU alone were ineffective. Effective treatments required both CD8+ and NK1.1+ cells, and increased CD8+ T cell priming, tumor-infiltration by effector cells and effector cytokine production. In addition, only the successful treatments induced the appearance of a population of monocyte-derived DC in tumor-draining lymph nodes. Recent experiments revealed that immunotherapy was required for the differentiation of blood monocytes into DC in the lymph node, and that these monocyte-derived DC were both necessary and sufficient for successful immunotherapy of B16 melanoma tumors. Further experiments are required to clarify the precise function of this DC population. We conclude that the activation of existing DC subsets is not sufficient for the induction of anti-tumor immunity, whereas newly recruited inflammatory DC are critical mediators of the successful activation of anti-tumour immune responses.
Semaphorin7A is up-regulated in breast cancer and promotes macrophage production of angiogenic molecules (TUM4P.933)
Authors: Garcia Areas Ramon , Libreros Stephania , Amat Samantha , Iragavarapu Vijaya 
Published: January 2023

Abstract
Semaphorins are a large family of molecules involved in the axonal guidance and development of the nervous system. In addition, they also play important roles in immunomodulation. Semaphorin 7A (SEMA7A) has been reported to be a modulator of monocytes and macrophages via α1β1integrins. SEMA7A promotes chemotaxis of monocytes and induces them to produce proinflammatory mediators. In this study we explored the role of SEMA7A in breast cancer. We first show that SEMA7A is highly expressed by DA-3, EO771 and 4T1 mammary tumor cells in comparison to normal mammary cells (EpH4). Furthermore, peritoneal elicited macrophages from mammary tumor-bearing mice express higher levels of SEMA7A compared to those from normal mice. We also show that murine macrophages treated with recombinant murine SEMA7A significantly increased their expression of angiogenic chemokine CXCL2 via activation of the MAPK pathway. Peritoneal elicited macrophages from mice bearing SEMA7A-silenced tumors produce significantly lower levels of CXCL2, CXCL1 and MMP-9, compared to macrophages from control tumor bearers. Gene silencing of Sema7A in macrophages from tumor-bearers decreased in their angiogenic potential. We postulate that SEMA7A in mammary carcinomas may skew monocytes into a pro-tumorigenic phenotype to support tumor growth.
Natural cytotoxic antibodies against tumor N-glycolilated ganglioside decrease with age and are absent in NSCLC patients (TUM4P.906)
Authors: Hernandez Ana , Zhurbenko Nely , Martinez Darel , Rondon Teresa , Ramirez Tania 
Published: January 2023

Abstract
N-glycolylated (NeuGc) gangliosides are not naturally expressed in normal human tissues but are overexpressed in several tumors and have immunosuppressive capacity, contributing to cancer progression. The present work shows the existence of a natural antibody response against NeuGcGM3 in healthy donors. These antibodies specifically recognized NeuGcGM3 and killed tumor cells expressing the antigen by a complement mediated mechanism. Interestingly, even after complement inactivation 17% of the positive sera showed a direct cytotoxic effect over the tumor cells. This cytotoxicity was dependent on the presence of the antigen on the tumor cell and resembles an oncotic necrosis kind of cell death. The levels of anti-NeuGcGM3 antibodies in the sera of healthy donors and the percentage of donors with this natural immunity decreases with age. Furthermore we could not detect this reactivity in the sera of 51 non-small cell lung cancer (NSCLC) patients that matched in sex and age the assessed healthy donors. Interestingly the level of sera IgM, but not IgG, was significantly lower in cancer patients in comparison with healthy donors. Also, the capacity of patients B1 cells to spontaneously secrete IgM and to respond to DAMPs, like CpG, seems to be affected. Defects in the natural surveillance against tumor antigens could increase the risk of elderly donors to develop cancer and affect the capacity of cancer patients to effectively fight against them.
The role of danger signals in tumor elicited inflammation (TUM5P.934)
Authors: Francescone Ralph , Hou Vivi , Dmitrieva Oxana , Posocco David , Grivennikov Sergei 
Published: January 2023

Abstract
Sporadic colorectal cancer (CRC) has traditionally been considered a genetic disease, with inactivation of classic tumor suppressors, such as APC and P53, driving tumorigenesis. Recently, however, it has come to light that inflammation plays a major role in the progression of CRC, and that tumor cells can induce a massive infiltration of immune cells into the tumor. Danger Associated Molecular Patterns (DAMPs) are proteins that are released from dying or stressed cells, which promotes an immune response. While these so called “danger signals” are often up-regulated in cancer, the mechanisms behind their role in tumor progression and inflammation require further exploration. Using a number of mouse models of spontaneous CRC, we are beginning to uncover which inflammatory cues induce danger signals, and which immune cells are the major producers. We have found that S100A8/A9 and HMGB-1 are significantly up-regulated in tumor tissue and mesenteric lymph nodes (MLN) of mice with CRC, compared to adjacent normal colon tissue. Additionally, using FACS to sort individual immune populations within the mouse tissue, intraepithelial monocytes and neutrophils seem to be the major immune cell producers of danger signals. Finally, in a kRas inducible model, to simulate a more aggressive cancer, danger signals are elevated even further in tumor tissue and MLN. Future work will focus on elucidating the precise signaling between immune modulators and danger signals.
A B cell riddle: a new tool to study the link between malaria and Burkitt’s Lymphoma (TUM5P.935)
Authors: Hart Geoffrey , Wei Chungwen , Chida Asiya , Crompton Peter , Sanz Ignacio , Pierce Susan 
Published: January 2023

Abstract
Burkitt’s lymphomas (BLs) are B cell lineage tumors that are the most common childhood cancer in the P. falciparum malaria endemic areas of Africa. The molecular basis of malaria infection causing endemic BLs (eBLs) remains unknown. Recent data has confirmed that remarkably 30% of eBLs are enriched for one particular heavy chain V region, VH4-34. The US frequency in normal individuals is 4%, however it was unknown in endemic regions of Africa. This high frequency of VH4-34 in eBLs initiated the use of the anti-VH4-34 antibody, ‘9G4’, to study VH4-34 at the cellular level in Africa. Here we report that in adults and children living in a malaria endemic area of Mali, VH4-34 expressing B cells are present in frequencies similar to those in individuals in the U.S, however more B cells are found in memory B cell subpopulations. We have also found that VH4.34 circulating antibody levels increase when infected with malaria but do not significantly increase with age unlike other malaria specific antibodies. This suggests the VH4-34 heavy chain is possibly important for malaria, but is decreased from the repertoire after infection is controlled. We are initiating studies to determine: the frequency of VH4-34 expressing B cells latently infected with EBV to see if it enriched, whether VH4-34 Abs are protective in malaria, and elucidating a molecular mechanism for transformation. This data will start an avenue to better understand the complex mechanisms behind this childhood cancer.