bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of Natural Killer Cells in Sonic Hedgehog driven Medulloblastoma
Authors: Schiavone Francesco , Carlino Claudia , Ascenzi Sonia , Po Agnese , Sanseviero Emilio , Ferretti Elisabetta , Gismondi Angela , Santoni Angela 
Published: January 2023

Abstract
Medulloblastoma (MB) is the most common paediatric neoplasm of the CNS. Standard therapy is mainly curative; however, the severe side effects make immunotherapy an attractive therapeutic option. Natural Killer (NK) cells are innate lymphoid cells that can be used for adoptive cell therapy. Herein, we studied the role of NK cells in MB growth control, focusing on Sonic Hedgehog (SHH) subgroup, using the MB cell line DAOY.
The contribution of SHH pathway on NK cell recruitment to MB was analysed performing an orthotopic xenograft of DAOY in nude mice treated with a pharmacological inhibitor of the SHH pathway. FACS analysis revealed higher percentage of NK cells in cerebella of treated mice compared to control group.
Based on this in vivo evidence, we characterized the mechanism underlying NK cell recruitment. Pharmacological inhibition of the SHH pathway in DAOY raised the expression and secretion of chemokines involved in NK cell migration. Chemotaxis assay confirmed that these secreted molecules were functional.
Successively, we assessed the ability of NK cells to recognize SHH-MB. By FACS analysis we determined the tumor line expression of ligands for the NKG2D and DNAM1 activating receptor. In particular, ULBP3 and ULBP2 were expressed at high levels.
Moreover, we found that NK cells co-culture with DAOY induced degranulation that was reduced in the presence of anti-NKG2D and anti-DNAM1 blocking antibodies, suggesting an involvement of these receptors in triggering degranulation.
Besides, DAOY increased the production of IFNγ in NK cells pre-activated of with suboptimal doses of IL12 and IL15.
Our data provide evidence for a role of NK cells in the control of SHH-MB growth.
Cell surface PCNA is associated with expression of cancer stem cell transcription factors and inhibit NK cell effector function.
Authors: Malaer Joseph D. , Mathew Porunelloor Abraham 
Published: January 2023

Abstract
Natural killer (NK) cell function is regulated through a delicate balance of signals received from activating and inhibitory receptors interacting with ligands on the surface of target cells, playing a vital role in the innate immune response against cancer and infection. Cancer cells may evade NK-mediated killing by expressing ligands for NK cell inhibitory receptors. NKp44, a member of the Natural Cytotoxicity Receptor (NCR) family, functions as an activating or inhibitory receptor. Proliferating cell nuclear antigen (PCNA) associates with Human Leukocyte Antigen (HLA) I and forms the inhibitory ligand for NKp44, resulting in the inhibition of NK function. In this report, we have identified diffuse B cell lymphoma (DB) cells with cell surface PCNA have higher expression of cancer stem cell transcription factors as determined by cell sorting and qRT-PCR. We further demonstrate that the expression of cell surface PCNA is restricted to tumor cells exhibiting a cancer stem cell phenotype. Blocking the NKp44-PCNA interaction enhanced the killing of DB cells displaying cancer stem cell characteristics by NK cells. Characterization of stem cell transcription factors and cell surface PCNA may provide novel immunotherapeutic targets for cancer stem cells.
Ex-Vivo Expanded Human Vγ9Vδ2 T Cells Efficiently Suppress Epithelial Ovarian Cancer Development
Authors: Liu Chao Lien Liu , Miao Carol H. , Yeh Chia Yu 
Published: January 2023

Abstract
The γδ-T cells attract attentions because of their potent cytotoxicity towards tumors. Most γδ-T cells facilitate the major histocompatibility complex (MHC)-independent activation by the interaction between their receptor Natural Killer Group 2 Member D (NKG2D) and MICA/MICB molecules expressed on the tumor cells, leading to tumor death. However, despite of the potent antitumor effects, their therapeutic efficiency and molecular mechanisms remain largely elusive. Ovarian cancer is one of the most lethal and challenging female malignancies worldwide because of the delayed diagnosis and the resistance to traditional chemotherapy, prompting us to investigate the immunotherapeutic effects of γδ-T cells as a new treatment. We successfully expanded γδ-T cells up to ~ 78% from peripheral blood mononuclear cells (PBMCs) with mostly Vγ9Vδ2-T cell subtype. We showed that the expanded γδ-T cells exerted significant cytotoxicity activities towards OVCAR3 and HTB75 cells in-vitro. Furthermore, in the tumor xenografts of NSG mice, γδ-T cells not only suppressed tumor growth but also completely eradicated the preexisting tumors with initial size ~5mm. Thus, we conclude that γδ-T cells possess dramatic cytotoxicity activities towards epithelial ovarian cancers both in-vitro and in-vivo, which strongly support the potential clinical immunotherapeutic application of γδ-T cells to treat this dreadful female malignancy.
Level of expression of vimentin mRNA in 9L gliosarcoma cells compared to microglial cells, astrocytes, and oligodendrocytes isolated from Fischer 344 rats
Authors: Hillman Tyler C , Redd Joan M 
Published: January 2023

Abstract
Gliosarcomas are an aggressive brain cancer that can become metastatic. Previously, we used a monoclonal antibody, designated H5, to isolate a tumor-associated antigen (TAA) expressed by 9L, a gliosarcoma line isolated from Fischer 344 (F344) rats. This TAA was tentatively identified as vimentin via mass spectrometry. In this project, we compared the level of vimentin mRNA in 9L cells to that of three types of glial cells: microglial cells, astrocytes, and oligodendrocytes isolated from F344 rats via qPCR. The level of vimentin mRNA in 9L cells was significantly upregulated as compared to the expression levels in all three types of glial cells. Similar research on carcinomas has identified vimentin as being involved in metastatic tumors and is now used as a genetic marker of this specific type of cancer. This research suggests that vimentin mRNA in 9L undergoes similar upregulation to that of carcinomas.
Tumorous niche drives NK exhaustion by disordering metabolic program
Authors: Zheng Xiaohu , Sun Rui , Tian Zhigang , Wei Haiming 
Published: January 2023

Abstract
Natural killer (NK) cells are important effectors of the immune system that can lyse malignancy-transformed cells. In the human liver, NK cells can account up to 50% of the total lymphocyte population. How NK cells lose immune surveillance occurs in hepatocellular carcinoma progression.
In this study, 75 cases from hepatocellular carcinoma (HCC) patients were analyzed by RNA sequence, pathology, flow cytometry and cell culture in vitro. We found a significant enrichment of infiltrating T cells within the tumor tissues of HCC patients but not NK cells, as compared to the adjacent peritumoral tissues. It is interesting that the infiltrating NK cells enrich CD56brightCD49a+ NK subset, like liver-resident NK. Nearly 50% of the entire healthy liver NK cell population is composed of functionally relevant CD56bright CD49a-NK cells, and almost all of the subset was lose. The infiltrating CD56brightCD49a+ NK cells present exhausted phenotype and downregulated expression of efficacy molecules of CD11b, CD226 and IFN-γ. We also found a few of CD56dimCD16+ NK cells were infiltrating, which were similar to their peripheral circulating counterparts in transcription and phenotype, but the lower effect function and viability of infiltrating NK cells compared with normal liver or peripheral circulating counterparts.
Dynamic reprogramming of metabolism is essential for immune cell stable liveness and effector function. Gene Ontology profiling and metabolism analysis revealed the infiltrating NK cells bioenergetic insufficiencies and enrichment of metabolic oxygen specis leading to its subset variations and immune surveillance insensitivity.
Mycoplasma pneumoniae CARDS toxin is a unique virulence factor targeting the Th1/Th2 response for persistent infection
Authors: Segovia Jesus A , Chang Te-Hung , Cagle Marianna P , Winter Vicki T , Coalson Jacqueline J , Kannan Thirumalai R , Baseman Joel B 
Published: January 2023

Abstract
Mycoplasma pneumoniae is a leading cause of bacterial community-acquired pneumonia and is implicated in the initiation and exacerbation of asthma. Over 2 million cases of M. pneumoniae infection occur in the United States each year. However, with no surveillance system in place for reporting infections, the true scope of the problem may be vastly underestimated. One of the hallmark immunological effects of M. pneumoniae infection is an imbalanced Th1/Th2 response. A recently identified M. pneumoniae virulence factor is the Community-Acquired Respiratory Distress Syndrome (CARDS) toxin, an ADP-ribosylating and vacuolating protein produced abundantly during infection that is capable of provoking a Th2 cytokine phenotype in mice. We hypothesize that CARDS toxin is utilized by M. pneumoniae in vivo to dampen the Th1 response and influence the Th2 response in order to establish a strong, persistent infection.
The clinical isolate strain M. pneumoniae S1 abundantly produces CARDS toxin during infection and causes severe pathology. We utilized S1 as a template to generate a CARDS-deficient mutant we designated M7. M7 and S1 were used in vitro and in vivo to identify Th1- and Th2-specific cytokine changes during active infection. We observed significant differences in pathology between M7- and S1-infected mice, indicating that CARDS toxin directly impacted M. pneumoniae disease and persistence. Our findings provide insights to understand the elusive pathological mechanisms behind M. pneumoniae infection and implicate CARDS toxin as a potentially important target for therapeutic intervention during M. pneumoniae infection.
Macrophage galactose-type lectin 1 (MGL-1) regulates neutrophil mobilization during pulmonary Gram negative pneumonia
Authors: Jondle Christopher Ned , Mishra Bibhuti B. , Sharma Jyotika 
Published: January 2023

Abstract
Macrophage galactose-type lectin 1 (MGL-1) is a type 2 transmembrane C-type lectin receptor. We recently reported that MGL1-mediated responses are required to mitigate disease severity in Klebsiella pneumoniae (KPn) induced murine pneumosepsis. Here we investigated the role of MGL-1 in neutrophil development and homeostasis during KPn infection. We found that MGL-1−/− mice exhibit severe neutrophilia in their lungs upon KPn infection. Adaptive transfer experiments showed that MGL-1 deficiency on neutrophils significantly increased their capacity to infiltrate the lungs. Analysis of neutrophil numbers and their progenitors in various compartments showed that increased mobilization of mature neutrophils from bone marrow storage pools, but not the progenitor proliferation, accounted for the increased systemic neutrophil counts. This was correlated with elevated levels of neutrophil mobilizing factors G-CSF and MCP-2 in MGL-1−/− mice. Collectively, our findings demonstrate that MGL-1 has a profound role in neutrophil homeostasis during acute inflammatory condition.
The role of ecSOD in phagosomal containment of Listeria monocytogenes by neutrophils
Authors: Okunnu Busola , Witter Alexandra R , Swanta Naomi K , Berg Rance E 
Published: January 2023

Abstract
Extracellular superoxide dismutase (ecSOD) is an enzyme associated with the extracellular matrix that plays a protective role during reactive oxygen species mediated inflammatory responses. Previously, we reported that neutrophils are essential for protection during infection with Listeria monocytogenes (LM), and that high ecSOD activity is detrimental to host resistance during infection, while lack of ecSOD activity is beneficial. Furthermore, using depletion studies, it was determined that neutrophils from ecSOD KO mice are more protective than neutrophils from ecSOD expressing mice during LM infection. To understand how ecSOD modulates neutrophil function during LM infection, bacterial uptake and phagosomal escape were measured. A higher percentage of neutrophils from the spleen and liver of ecSOD KO mice associated with and allowed for phagosomal escape of LM. However, using MFI as an indicator of the relative number of bacteria per neutrophil, we observed that the ratio of cell associated to cytosolic bacteria was higher in ecSOD KO neutrophils in comparison to neutrophils with ecSOD activity. These data suggest that the absence of ecSOD activity enhances neutrophil association with LM and simultaneously limits LM escape from the phagosome. Future studies utilizing neutrophils obtained from other organs as well as from infected mice should help conclusively determine how the lack of ecSOD activity enhances the protective capabilities of neutrophils.
Nociceptor neurons regulate lung infections and bacterial host defense
Authors: Chiu Isaac M , Baral Pankaj 
Published: January 2023

Abstract
The sensory nervous system densely innervates the respiratory tract, mediating breathing, cough and bronchoconstriction. However, the role of the nervous system in lung host defenses has not been defined. Nociceptor neurons sense noxious/harmful stimuli and protect organisms from danger. Here, we find that nociceptor neurons play a critical role in regulating innate immunity against bacterial lung infections and lethal pneumonia. Staphylococcus aureus is a major cause of human disease and pneumonia in hospital acquired infections. We find that S. aureus directly induces nociceptor neuron ionic influx and activation. Nociceptor neurons released the neuropeptide CGRP upon bacterial activation, which suppressed neutrophil recruitment and bacterial clearance. Targeting nociceptor neurons by genetic and chemical strategies led to significantly enhanced innate immunity and survival in a mouse model of S. aureus lethal pneumonia. Therefore, the nervous system plays an important role in lung infections, and targeting sensory neurons could lead to better treatments for bacterial pneumonia.
Eosinophils actively participate in the host response to Mtb infection
Authors: Tocheny Claire , Senbagavalli Prakash , Hu Zhidong , Kauffman Keith , Sakai Shunsuke , Yan Bo , Song Yanzhen , Wong Ka-Wing , Barber Daniel , Klion Amy , Mayer-Barber Katrin 
Published: January 2023

Abstract
Mycobacterium tuberculosis (Mtb) is the leading cause of mortality worldwide due to a single infectious agent. Mtb resides in pulmonary macrophages, neutrophils and phagocytes, which are also key effector cells that limit bacterial growth. The role of other granulocytic phagocytes during Mtb infection is largely unknown. Eosinophils have comparable phagocytic function to neutrophils and an overlapping repertoire of granular contents capable of limiting bacterial growth. Here we characterized eosinophil responses to Mtb in vitro and in vivo. Eosinophils from healthy human donors responded to Mtb in vitro by CD69 up-regulation and CD62L down-regulation. Mtb induced the release of the eosinophil-specific granule proteins EDN and ECP. Mtb exposure resulted in secretion of IL-1a, TNFa, MIP-1a, MIP-1b, and IL-8. These results suggest that Mtb directly affects eosinophil degranulation, and that eosinophils are able to contribute to Mtb-driven inflammation. To investigate whether eosinophils actively participate in the cellular immune response to Mtb in vivo we followed the eosinophilic response after pulmonary Mtb infection in non-human primates. Eosinophils were significantly increased in the BAL fluid of Mtb infected rhesus macaques, providing evidence that eosinophils are being actively recruited to the lung in response to Mtb in vivo. Indeed, when we analyzed Mtb infected lung tissue from patients who had undergone lung resection, eosinophils were enriched in fibrotic TB consolidations, cavity wall and necrotic caseum.
We are currently investigating the biological relevance of the eosinophilic response during Mtb infection in the mouse model.
This work was supported by the intramural research programs of NIAID, NIH.
CD4+ T cells induce homeostatic expression of the Treg-attracting chemokine CCL22 in dendritic cells
Authors: Piseddu Ignazio , Kühnemuth Benjamin , Knott Maximilian , Vetter Viola , Layritz Patrick , Endres Stefan , Anz David 
Published: January 2023

Abstract
Regulatory T cells (Tregs) are highly potent modulators of immune responses by various mechanisms including direct interaction with dendritic cells. Thereby, Tregs play a critical role not only in prevention of autoimmunity but also in tumor immune escape. By binding to its only known receptor CCR4 the chemokine CCL22 is a potent chemoattractant for Tregs, influencing their localization and activity. However, the regulation of CCL22 expression is still poorly understood.
Here we demonstrate that under homeostatic conditions cells of the dendritic cell lineage are the main producers of CCL22. In vitro, coincubation of dendritic cells with CD4+ CD25− T cells strongly induced CCL22 expression in a paracrine manner. Using neutralizing antibodies we identified the cytokine family IL-3, IL-5 and GM-CSF, as well as IL-4 to be the main inducers of CCL22 in dendritic cells. In vivo, transfer of CD4+ T cells into Rag−/−-mice lacking mature T and B cells was able to increase CCL22 levels in several lymphoid organs.
In summary, we identified the aforementioned T cell-derived factors as the main inducers of the Treg-associated chemokine CCL22, in vitro and in vivo. Therefore, our data suggests that paracrine signalling of T cells might indirectly enhance Treg activity by induction of CCL22 expression in dendritic cells. In conclusion, the data presented here potentially shows a self-limiting mechanism of adaptive immune responses.
Vasoactive intestinal peptide induced epithelial to mesenchymal transition activities in PC3 prostate cancer cells
Authors: Laney Caleb N , Bains Manpreet , Dorsam Glenn 
Published: January 2023

Abstract
Epithelial to mesenchymal transition (EMT) is a fundamental cellular process preceding movement. Malignant transformation of epithelial cells also undergo EMT leading to metastasis and a worse prognosis for human cancer patients. Understanding molecular pathways that are linked to EMT may provide additional drug targets to treat epithelial cancers like prostate cancer. Vasoactive intestinal peptide signaling through one of its naturally occurring G-protein coupled receptors, termed VPAC1, induces non-tumor and tumorigenic epithelial cell lines (RWPE-1, LnCaP and PC3 cells) to undergo EMT and downregulate E-cadherin, a hallmark for EMT. Curcumin, an anti-inflammatory molecule, blocked these VIP EMT effects suggesting a role for NF-kB signaling. Our group has uncovered EMT-associated molecular mRNA targets induced by VIP treatment of primary, resting CD4 T cells, including the EMT master regulator transcription factor, SNAIL1. We hypothesize that EMT transition effects by VIP signaling may also involve the upregulation of SNAIL1 protein. To this end, we plan on treating PC3 cells with varying levels of VIP over time (0–24 hr) and measure by immunoblots the extent to which Snail1 protein, and up to 10 additional EMT-associated proteins, are differentially altered. Successful completion of this research is expected to uncover important VIP protein targets that is expected to better explain its epithelial tumorigenic potential.
The allergic T cell response against mouse proteins
Authors: Schulten Véronique MJ , Westernberg Luise , Peters Bjoern , Sette Alessandro 
Published: January 2023

Abstract
Mouse allergy is rising in prevalence, affecting inner-city households and laboratory workers. Increased exposure to mice in inner-city households is a strong predictor for asthmatic disease. Despite a strong link between mouse exposure and asthmatic disease, the allergic immune response to mouse has been significantly understudied. To date, only one major allergen in mouse, Mus m 1, has been identified and very little is known about the targets of the allergic immune response against mouse.
Using a proteomic/transcriptomic approach, we sought to identify T cell targets in 24 mouse allergic and asthmatic patients. Mouse urine and epithelial extracts were analyzed by 2D-IgE/IgG immunoblots using pooled sera from mouse-sensitized donors. Mass spectrometry of selected protein spots identified 30 novel antibody reactive proteins. Predicted MHC binding peptides from these novel proteins and mouse homologs to mammalian allergens were screened for T cell reactivity in PBMCs from mouse allergic patients. Overlapping peptides from the major mouse allergen Mus m 1 and its major urinary protein isoforms were screened in parallel. T cell epitope mapping experiments showed that responses to Mus m 1 peptides were dominant overall. However, reactivity to mouse peptides homologous to other mammalian allergens, specifically guinea pig, was also detected. In summary, our data demonstrates that the cellular and serological targets of the allergic response overlap, with Mus m 1 being the major target for both T cells and IgE antibodies. However, on a serological level, other proteins are also recognized. To the best of our knowledge, this is one of the first comprehensive studies of T cell epitope targets in mouse allergy.
Transplantation of Human Skin Microbiota in Models of Atopic Dermatitis
Authors: Myles Ian Antheni 
Published: January 2023

Abstract
Atopic dermatitis (AD) is characterized by reduced barrier function, reduced innate immune activation, and susceptibility to Staphylococcus aureus. Host susceptibility factors are suggested by monogenic disorders associated with AD-like phenotypes, and can be medically modulated. S. aureus contributes to AD pathogenesis and can be mitigated by antibiotics. Recent work has revealed that the skin microbiome differs significantly between healthy controls and patients with AD, including decreased Gram-negative bacteria in AD. However, little is known about the potential therapeutic benefit of microbiome modulation. To evaluate if parameters of AD pathogenesis are altered after exposure to different culturable Gram-negative (CGN) bacteria collected from human skin. CGN bacteria were collected from healthy controls and patients with AD. Impacts on cellular and culture-based models of immune, epithelial, and bacterial function were evaluated. Representative strains were evaluated in the MC903 mouse model of AD. We found that CGN bacteria taken from healthy volunteers but not from patients with AD were associated with enhanced barrier function, innate immunity activation, and control of S. aureus. Treatment with CGN from healthy controls improved outcomes in a mouse model of AD. These findings suggest that a live-biotherapeutic approach may hold promise for treatment of patients with AD.
Tyrosine Phosphorylation of NLRP3 by Lyn Suppresses NLRP3 Inflammasome Activation
Authors: Lin Guoxin , Tang Juan , Guo Hui , Xiao Yun , Gupta Neetu , Tang Na , Zhang Jian 
Published: January 2023

Abstract
The NLRP3 inflammasome is a multi-protein complex that triggers the activation of inflammatory caspase-1 and the maturation of IL-1β and IL-18 in response to microbes and danger signals in the cytosol of host cells. However, how the NLRP3 inflammasome is regulated is not well defined. Here, we showed that NLRP3 undergoes tyrosine phosphorylation in macrophages upon stimulation with ATP, a NLRP3 inflammsome stimulus, and the tyrosine phosphorylation of NLRP3 correlates with its ubiquitination. Pretreating the macrophages with a specific Src kinase inhibitor inhibits NLRP3 tyrosine phosphorylation and ubiquitination, which leads to enhanced production of IL-1β. Further we found that Src family PTK Lyn phosphorylates NLRP3 at Tyr918 which facilitates its ubiquitination and proteasome-mediated degradation. Consistent with these data, NLRP3 tyrosine phosphorylation and ubiquitination is abrogated in macrophages lacking Lyn, which correlates with heightened-activation of NLRP3 inflammasome. Therefore, our data demonstrates that the Lyn-mediated tyrosine phosphorylation of NLRP3 is prerequisite for its ubiquitination, thus dampening the NLRP3 inflammasome activity.