bluuebunny/crossref_metadata_2025_split huggingface dataset:
Macrophage depletion decreases inflammation and tumorigenesis in the AOM/DSS mouse model of colon cancer
Authors: Bader Jackie E , Velazquez Kandy T , Enos Reilly T , Cranford Taryn L , Davis J Mark , Murphy E Angela 
Published: January 2023

Abstract
Macrophages have recently emerged as major players in the connection between inflammation and cancer; they represent up to 50% of the tumor mass and produce a wide array of inflammatory mediators with pro-tumoral functions. We used a macrophage depletion technique to examine the role of macrophages in colon cancer. To induce colon cancer, male C57BL/6 mice (n=20) received a single injection of AOM followed by three cycles of DSS supplemented water at concentrations of 2%, 1%, 1% at weeks 1, 4, and 7, respectively. Prior to the final DSS administration (week 7) and twice weekly thereafter for a duration of 5 weeks, mice (n=10/group) were administered 200μl of clodronate (CL2MDP) or PBS encapsulated liposomes via an intra-peritoneal route to deplete macrophages. Body weight, body composition, and symptom severity score were evaluated throughout the macrophage depletion period. Mice were sacrificed at 12 weeks following the initiation of colon cancer and colon tissue was analyzed for tumor burden, macrophage markers, and inflammatory mediators. There were no differences in body weight, body composition, or symptom severity score between the groups. Macrophage depletion decreased tumor number by ~25% and specifically medium tumor number by 50% (p&lt;0.05). This was consistent with a decrease in expression of EMR1 (overall macrophage marker) as well as expression of markers associated with M2 macrophages (IL-13, IL-10, TGFβ & CCL17) (p&lt;0.05) but not M1 markers (NOS2, IL-12a & IL-23). In addition, expression of the inflammatory mediators IL-6 and TNF-α were decreased following macrophage depletion. We demonstrate that macrophages are an important mediator of tumor growth and inflammation in colon cancer.
Ly6G+ neutrophils and polymorphonuclear-myeloid derived suppressor cells promote the survival of tumor cells
Authors: Herlihy Sarah E , Ramachandran Indu , Condamine Thomas , Lin Cindy , Garfall Alfred , Vogl Dan , Gabrilovich Dmitry , Nefedova Yulia 
Published: January 2023

Abstract
Myeloid derived suppressor cells (MDSC) are a heterogeneous group of immature myeloid cells which suppress the immune system. Collectively a MDSC population is comprised of monocyte-like MDSC and polymorphonuclear MDSC (PMN-MDSC). The number of MDSC is increased with the presence of tumors. Our lab has previously shown that PMN-MDSC represent the majority of MDSC in tumor-bearing mice. We found that depletion of myeloid cells in vivo results in increased survival in a mouse model of multiple myeloma (MM). Direct co-culture of neutrophils (CD11b+Gr1+ cells from tumor-free mice) or PMN-MDSC (CD11b+Gr1+ cells from tumor-bearing mice) and MM cells increases the survival of the MM cells from chemotherapeutic-induced death. The protective effect is not due to cell-cell contact as neutrophil and PMN-MDSC supernatants also protect MM cells from cell death. This suggests that the factor protecting tumor cells is soluble. Results in the mouse were confirmed using human MM cells. Similar results have been replicated on breast cancer cells with neutrophils and PMN-MDSC. Several cytokines and growth factors, such as IL-6 and fibroblast growth factor, are implicated in chemoresistance. The role of these neutrophil and PMN-MDSC soluble factors and the identification of novel contributors to chemoresistance are under investigation.
Oxidized lipid bodies block antigen cross-presentation in cancer by preventing trafficking of peptide-MHC class I complexes
Authors: Veglia Filippo , Tuyrin Vladimir A , Mohammadyani Dariush , Murphy Maureen , Gabrilovich Dmitry I 
Published: January 2023

Abstract

Background
Antigen cross presentation is a unique feature of dendritic cells (DCs) and plays an important role in the development of specific CD8+ T cell responses. It is established that the defect in cross-presentation by dendritic cells (DCs) in cancer results in ineffective antitumor immune responses and favors tumor escape. However, the mechanisms behind this phenomenon are unclear.

Results
In this study we found that tumor-derived factors impaired cross-presentation by differentiated conventional DCs without affecting expression of MHC class I, class II, or costimulatory molecules. The ability of DCs to stimulate antigen-specific T cells after loading with short peptide (directly bound to MHC class I) was fully preserved. We found that tumor-derived factors blocked trafficking of peptide-MHC class I (pMHC) complexes to the cell surface. These complexes were found to be entrapped into the lysosomes. This effect was caused by the accumulation of lipid bodies (LB) containing oxidized (ox) triglycerides (TAG) in DCs. Importantly LB containing non-oxidized TAG did not affect cross-presentation. Further experiments have identified the specific molecular mechanism responsible for oxTAG containing LB mediated block of cross-presentation in DCs.

Conclusions
Our results suggest a novel mechanism of regulation of cross-presentation in cancer
Glycolysis promotes MDSCs and cancer stemness through the interaction between Sirt1 and C/EBPb
Authors: LI WEI , Kryczek Ilona , Maj Tomasz , Wu Ke , Peng Dongjun , Zhao Ende , Xia Houjun , Lei Zhang , Tao Kaixiong , Wang Guobin , Zou Weiping 
Published: January 2023

Abstract
Glycolysis is considered a hallmark of cancer. However, its involvement in tumor immune regulation and breast cancer stem cell biology is poorly understood. Here we investigated how glycolysis affected the development of myeloid derived suppressor cells (MDSCs) and in turn controlled breast cancer stemness in breast cancer. G-CSF promotes MDSC development in 4T1 breast cancer model. We found that knock down of LDH-A in 4T1 cells resulted in decreased G-CSF production and reduced MDSC numbers. Accompanied with these findings was the inhibited tumor glycolysis, reduced breast cancer stemness potential, and increased mouse survival. Molecular mechanistic studies demonstrated that glycolysis regulated G-CSF expression, MDSCs and cancer stemness by controlling the interaction between Sirt1 and C/EBPb via NAD+-dependent manner. Moreover LDH-A expression is positively associated with breast cancer patient outcome. Collectively, our study has revealed that glycolytic metabolism is associated MDSCs and linked to breast cancer stemness.
Combined nanoemulsion and RIG-I agonist adjuvants-activation of diverse pathways for a broad spectrum influenza vaccine.
Authors: Wong Pamela T , Goff Peter H , O’Konek Jessica J , Landers Jeffrey J , Janczak Katarzyna W , Leroueil Pascale R , Palese Peter , Baker James R 
Published: January 2023

Abstract
Current influenza vaccines are limited by low immunogenicity, short duration of protection, and narrow cross-strain efficacy. Broad-based cellular immune responses and mucosal immunity are key to an effective vaccine. However, most single adjuvants exhibit only some of these qualities. Here we combine two adjuvants, an oil-in-water nanoemulsion (NE), and a synthetic RNA from Sendai virus (IVT DI), each individually shown to elicit protective immunity to influenza via distinct mechanisms. Intranasal (IN) immunization with NE and influenza hemagglutinin induces mucosal and systemic antibodies, activates NF-κB via TLR signaling, and induces a balanced TH1/TH2/TH17 cellular response. NE is also an antigen carrier, facilitating cellular uptake and processing. IVT DI activates the RIG-I receptor inducing type-I interferon and TH1 immunity. We sought to determine if combining the complementary properties of NE and IVT DI improves the strength of the immune response and affords new activities not seen with the single adjuvants. Combined adjuvant was evaluated with inactivated H1N1 virus (PR8) as antigen. While IN coadmin of NE and IVT DI did not improve humoral responses in mice, cellular recall responses were much enhanced relative to either adjuvant alone. Separate sequential admin of NE and IVT DI (2 wks apart) resulted in very high humoral and mucosal antibody responses and increased cytokine production. Interestingly, the cytokine pattern for sequential vs. coadmin of NE and IVT DI was distinct. Notably, both methods elicited a TH17 response not seen with IVT DI alone. Combining complementary adjuvants may be a promising approach for generating a broader, more optimal immune response for influenza when used with diverse HA antigens.
Ex vivo evaluation of HIV envelope specific human naive B cells
Authors: DeBuysscher Blair L , Steach Holly R , Stamatatos Leonidas , Taylor Justin J. 
Published: January 2023

Abstract
Recent isolation of HIV-1 broadly neutralizing antibodies (bNAbs) has led to vaccine strategies focusing on inducing bNAb responses. Interestingly, not all infected individuals develop bNAbs. Current vaccine candidates regrettable, only induce antibodies that are protective against narrow subsets of strains. This could be because vaccine design is based largely on assumptions that the desired B cells are present in the host, will respond, and are protective. However, recent results suggest that a limited number of B cells respond to vaccination and may require multiple rounds of somatic hypermutation to gain neutralizing breadth. We aim to study pre-vaccination B cell repertoires in hopes of illuminating HIV-1 specific B cell responses to vaccination. Using an HIV envelope (Env)-tetramer enrichment method, we have begun to assess human Env-specific B cells in the blood of HIV-unexposed individuals. This approach allows for careful analysis of the population of naive B cells able to bind to various Env variants, allowing us to compare the germ-line antibody sequences utilized by these cells to the predicted germline sequences of bNAbs. In the course of this work, we have found naive B cells able to cross-react with structurally distinct Envs derived from different clades. Ongoing work is focused on gauging whether naive Env-cross-reactive B cells express antibodies able to broadly neutralize HIV, or can gain this ability after somatic hypermutation.
Predicting vaccine efficacy against tuberculosis using immunologic and radiologic measures in the non-human primate model
Authors: DiFazio Robert M , Maiello Pauline , Tomko Jaime , Frye Lonnie , Bigbee Carolyn , Chedrick Chelsea , Updike Cassaundra , Lin Philana , Agger Else Marie , Andersen Peter , Flynn JoAnne L 
Published: January 2023

Abstract
The only licensed vaccine (BCG) for tuberculosis (TB) has variable efficacy and does not protect against pulmonary TB. New vaccines are in development and in human clinical trials. However, vaccine trials are long and expensive without useful surrogate markers of protection. A problem thus emerges: what does a protective response to TB look like? This project’s goal is to identify markers of vaccine efficacy that can further be developed as new clinical endpoints. We posit that exploiting differences in vaccine outcome (protected vs. unprotected) will identify novel markers that correlate with outcome, and that serial PET/CT will predict outcome early in infection, as well as identify sites of protection. Cynomolgus macaques vaccinated with BCG and a multistage vaccine (H56 in CAF01) have more sterile thoracic lymph nodes (LN) 6 months post-Mtb challenge, compared to unvaccinated controls, indicating that LN play an important role in H56-conferred protection. BCG/H56 prevents dissemination but not establishment of infection; vaccinated animals also have fewer CFU per granuloma than controls. Total FDG avidity by PET scan in lungs (a surrogate for inflammation and disease) and individual granuloma FDG avidity of vaccinated animals are significantly lower than controls at both early and late time points. Total FDG avidity can also be used to predict vaccine efficacy, as the animals protected by BCG/H56 had less FDG in lungs than unprotected vaccinated and latent controls at early time points post challenge, and will be further developed as a surrogate marker of protection at early time points in pre-clinical trials.
Phenotypic characterization of islet antigen-specific CD4 T cell subsets in mice of varying susceptibilities to Type 1 Diabetes
Authors: Martinov Tijana , Pauken Kristen E , Spanier Justin , Heffernan James R , Osum Kevin C , Fife Brian T 
Published: January 2023

Abstract
Type 1 diabetes (T1D) results from lymphocyte-mediated destruction of insulin-producing pancreatic beta cells. Islet-reactive CD4 T cells are critical mediators of disease. Here, we investigated the phenotype and regulation of insulin-specific and bulk CD4 T cells in diabetes-prone (NOD) and diabetes-resistant (NOR or B6.g7) mice. We utilized dual color tetramer staining with enriment techniques to enumerate and phenotype insulin-specific CD4 T cells in NOD, NOR and B6.g7 mice. To test whether there are inherent differences in the CD4 T cell compartment between the strains, we stimulated sorted CD4 T cells in vitro with αCD3/CD28 and performed transcriptional analysis of &gt;40 targets. Additionally, we analyzed Th1/Th2/Th9/Th17/Th22/Treg cytokine production. Both insulin-specific CD4 T cells and bulk polyclonal CD4 T cells displayed a strong Th1 skewing in NOD and not in NOR or B6.g7 mice, indicating that different disease susceptibilities could be in part due to inherent differences in CD4 T cell activation. Experiments are underway to determine the transcriptional profile of insulin-specific CD4 T cells in these strains using an unbiased approach, and identify the peripheral tolerance mechanisms underlying diabetes resistance.
The role of the orphan nuclear receptor NR4A1 in erythro-myelopoiesis.
Authors: Mumau Melanie , Golec Sophia , Vanderbeck Ashley , Lynch Elizabeth , Punt Jennifer A , Emerson Stephen 
Published: January 2023

Abstract
Studies of hematopoietic stem cells (HSCs) have advanced our understanding of the intracellular signals and microenvironmental interactions that influence HSC fate. Our lab is interested in the role of orphan nuclear receptor NR4A1, an immediate response gene sensitive to external stimuli, in HSC development. NR4A1 regulates the development of specific, mature hematopoietic cell lineages from both the innate and adaptive immune system including patrolling monocytes, a mature myeloid cell subset. More recently, we have shown that NR4A1 expression also identifies a subpopulation of myeloid-biased long-term HSCs in the bone marrow. Given that NR4A1 directs both immature and differentiated cell types, we investigated its role in myeloid cell maturation. Within the bone marrow and spleen progenitor cell compartments, we find that NR4A1 is exclusively expressed by myeloid progenitors and not by more restricted megakaryocyte, erythroid, or common monocyte progenitors. Nr4a1−/−mice exhibit skewed myeloid progenitor cell populations, exhibiting a 2-fold increase in CD105+CD150− erythroid progenitors and cKit+Ter119+CD71+ pro-erythroblasts in the spleen but not in the bone marrow. In vitro cultures of Nr4a1−/−splenic myeloid progenitors also show that NR4A1 restricts the frequency of erythroid cells, yet bone marrow transplant studies show similar erythroid progenitor reconstitution from both WT and Nr4a1−/− donors. Taken together, our data reveal a new role for NR4A1 as both a direct and indirect modulator of erythropoiesis.
RNA binding protein HuR posttranscriptionally regulates antigen-specific T cell proliferation and allergen induced airway inflammation
Authors: Ellis Jason S , Techasintana Patsharaporn , Glascock Jacqueline J , Ridenhour Suzanne , Atasoy Ulus 
Published: January 2023

Abstract
RNA binding protein HuR (ELAVL1) governs transcript stability of IL-4, IL-13 and GATA-3 mRNAs, thereby controlling Th2 differentiation. We hypothesized that HuR plays a role in antigen induced T cell activation and initiation of allergic airway inflammation. We conditionally ablated HuR in T cells prior to activation (distal lck-Cre ROSA HuRfl/fl mouse). Half of mature CD4+ T cells were HuR deficient in these mice. To determine the effects of HuR KO in T cells, mice were immunized with OVA, boosted 10 days later, and antigen specific proliferative responses were measured in vitro. Despite being fully capable of proliferation upon anti-CD3/CD28 stimulation, YFP+ (HuR KO) T cells were unable to proliferate in response to antigen while YFP− control cells had no proliferative defects. We then used the OVA challenge model of airway inflammation to ascertain effects of HuR KO in vivo. HuR KO mice had significantly less total cellular inflammation, as well as significant reductions in BAL IL-13. To determine the mechanism of suppression, we conducted in vitro activation experiments. HuR KO CD4+ T cells were sorted into YFP+ and YFP− pools and activated. YFP+ T cells had significant decreases in IL-4, IL-5, IL-13 but not IFNγ protein. Gata-3, IL-4 and IL-2Ra (CD25) transcription were significantly reduced. KO T cells have reductions in pSTAT5 signaling which is required for robust Th2 differentiation. While only 50% of CD4+ T cells lack HuR, there was total suppression of Th2 differentiation in airway inflammation. These data elucidate a critical role for HuR in regulating Th2 differentiation and allergic airway inflammation.
Aeroallergen-induced IL-33 predisposes to respiratory virus-induced asthma by dampening anti-viral immunity
Authors: Phipps Simon 
Published: January 2023

Abstract
Frequent viral lower respiratory infections in early-life are an independent risk factor for asthma onset. This risk, and the development of persistent asthma, is significantly greater in children who later become sensitized. Here, we sought to elucidate the pathogenic processes that underlie the synergistic interplay between allergen exposures and viral infections. Mice were inoculated with a Pneumovirus (Pneumonia virus of mice; PVM) and exposed to low-dose cockroach extract (CRE) in early- and later-life, and airway inflammation, remodeling and hyperreactivity assessed. Mice were treated with anti-IL-33 or apyrase to neutralize or block the release of IL-33. PVM infection or CRE exposure alone failed to induce disease whereas PVM/CRE co-exposure acted synergistically to induce the features of asthma. CRE exposure during virus infection in early-life induced a biphasic IL-33 response and impaired IFN-a/l production, which in turn increased epithelial viral burden, airway smooth muscle growth, and type-2 inflammation. These features were ameliorated when CRE-induced IL-33 release was blocked or neutralized, while substitution of CRE with exogenous IL-33 recapitulated the phenotype observed in PVM/CRE co-exposed mice. Mechanistically, IL-33 down-regulated viperin and IFN regulatory factor 7 gene expression, and rapidly degraded IRAK1 expression in pDC, leading to TLR7 hypo-responsiveness and impaired IFN-a production. We identify a hitherto unrecognized function of IL-33 as a potent suppressor of innate antiviral immunity, and demonstrate that IL-33 contributes significantly to the synergistic interplay between respiratory virus and allergen exposures in the onset and progression of asthma.
Oncostatin M is produced by neutrophils, and promotes epithelial barrier dysfunction in mucosal airways disease
Authors: Pothoven Kathryn L , Norton James E , Suh Lydia A , Carter Roderick G , Welch Kevin C , Shintani-Smith Stephanie , Conley David B , Grammer Leslie , Peters Anju T , Liu Mark C , Kato Atsushi , Kern Robert C , Tan Bruce K , Schleimer Robert P 
Published: January 2023

Abstract
We have previously shown that Oncostatin M (OSM) was elevated in nasal polyp (NP) tissue from chronic rhinosinusitis (CRS) patients, and in BAL fluid following segmental allergen challenge (SAC) in allergic asthmatics (AA). Levels of OSM detected in vivo were sufficient to impair epithelial barrier function in ex vivo cultured airway epithelium. To determine which cell type(s) was producing OSM, NP sections were stained for OSM and hematopoietic cell specific markers. OSM showed co-localization with neutrophil elastase(n=10), but not with markers for eosinophils, macrophages, T cells or B cells (n=3–5). OSM mRNA correlated with mRNA for neutrophil markers CD16 (r=.61, p&lt;.01) and cathepsin G (r=.56, p&lt;.01) in mRNA from CRS patients and controls. OSM also correlated with the percent neutrophils in the BAL of AA following SAC (r=.50, p&lt;.05), but did not correlate with total cell counts or eosinophil counts. Flow cytometric analysis of cells isolated from NP (n=10) showed that 14±6% of CD45+ cells were OSM+, and 85±6% of OSM+ cells were CD16+Siglec 8-, indicating neutrophil lineage. GM-CSF and FSTL-1 have been shown to induce OSM, and both were found to be elevated in NP. Additionally, GM-CSF, alone or in combination with FSTL-1, was sufficient to induce both OSM mRNA in ex vivo cultured blood neutrophils and OSM protein in the culture supernatants. These data suggest that neutrophils are the major producers of OSM in NP, and may potentially mediate epithelial barrier dysfunction in mucosal airways disease.
A heterozygous RAB27A mutation causes delayed cytolytic granule polarization and hemophagocytic lymphohistiocytosis in 2 unrelated teenagers
Authors: Cron Randy Q , Zhang Mingce , Bracaglia Claudia , Prencipe Giusi , Bemrich-Stolz Christina J , Beukelman Timothy , Dimmitt Reed A , Chatham Winn , Zhang Kejian , Li Hao , Walter Mark R , De Benedetti Fabrizio , Grom Alexei A 
Published: January 2023

Abstract
Frequently fatal, primary hemophagocytic lymphohistiocytosis (HLH) occurs in infancy resulting from homozygous mutations in natural killer (NK) and CD8 T cell cytolytic pathway genes. Secondary HLH presents after infancy and may be associated with heterozygous mutations in HLH genes. We report 2 unrelated teenagers with HLH and an identical heterozygous RAB27A mutation (259 G&gt;C). The contribution of this Rab27A missense (A87P) mutation on NK cell cytotoxicity was studied by cloning it into a lentiviral expression vector prior to introduction into the human NK-92 cell line. NK cell degranulation (CD107a expression), target cell conjugation, and K562 target cell lysis was compared between mutant and wild-type (WT) transduced NK-92 cells. Polarization of granzyme B to the immunologic synapse and interaction of mutant Rab27A (A87P) with Munc13-4 were explored by confocal microscopy and proximity ligation assay (PLA), respectively. Over-expression of the RAB27A mutation had no effect on cell conjugate formation between the NK and target cells but decreased NK cell cytolytic activity and degranulation. Moreover, the mutant Rab27A protein was predicted to disrupt binding to WT Munc13-4 by crystal structure modelling, and decreased interaction of Rab27A (A87P) with WT Munc13-4 was shown by PLA in situ and by co-immunoprecipiation in vitro. Finally, Rab27A (A87P) over-expression in NK-92 cells delayed granzyme B polarization toward the immunologic synapse, as noted by confocal microscopy, with a resulting increase in interferon-γ production, a cytokine responsible for HLH. This heterozygous RAB27A mutation blurs the genetic distinction between primary and secondary HLH by contributing to HLH via a partial dominant-negative effect.