bluuebunny/crossref_metadata_2025_split huggingface dataset:
Rise in invariant Natural Killer T-17 cells during Bordetella pertussis infection in mice
Authors: Segal-Abramson Tzvia , Santos Daniel 
Published: January 2023

Abstract
Bordetella pertussis is a Gram-negative bacterium known for causing the respiratory disease whooping cough. Whooping cough has recently seen resurgence, especially among children, with the disease’s resolution being delayed. Here, we observed the contribution of invariant Natural Killer T (iNKT) cells in the response to B. pertussis infections. Invariant NKT cells are so called for their invariant chain (Vα14Jα18 in mice) within the T cell receptor, which allows these cells to respond to lipid antigens presented by CD1d molecules. iNKT cells are known to express a variety of transcription factors and release different cytokines, as well as interact with numerous cells of the immune system. Here we hypothesize that IL-17 producing iNKT (iNKT17) cells rise in number during B. pertussis infections to aid in the resolution via an IL-17 driven inflammation. However, due to a pertussis toxin-dependent inhibition, there is a delay in the resolution of the infection. The population of iNKT17 cells, as well as dendritic cells and their expression of CD1d, was assessed via comparison of uninfected to B. parapertussis- and B. pertussis -infected C57BL/6 mice, using a multicolor flow cytometry analysis. The mice infected with pertussis displayed a significant rise in RORγt-positive iNKT cells when compared to uninfected mice, while having reduced IL-17 secretion. These findings indicate a role for iNKT17 cells in the pathogenesis and potential resolution of B. pertussis infection, with the potential for future analysis into iNKT cell knockout studies to determine further understanding of the cell’s function during the infection.
Novel role of intestinal Goblet cells and Goblet Cell Associated Antigen Passages in induction of tolerance to dietary antigens and promoting intestinal homeostasis
Authors: Kulkarni Devesha Hrishikesh , Gustafsson Jenny K. , McDonald Keely G. , Knoop Kathryn A. , Davis Jazmyne , Miller Mark J. , Newberry Rodney D. 
Published: January 2023

Abstract
Mucosal tolerance towards luminal antigens is a process fundamental to intestinal homeostasis, the breakdown of which results in onset of inflammatory bowel disease. How luminal antigens cross the intestinal epithelium to interact with the immune system in a manner capable of induction and maintenance of oral tolerance is unknown. We have identified a new pathway of luminal antigen delivery to dendritic cells (DC) in the lamina propria (LP), which was mediated by goblet cells (GC) associated antigen passages (GAPs). The role of GCs and GAPs in promoting mucosal tolerance is unexplored. We studied tolerogenic responses to dietary antigen, ovalbumin (Ova) in mice where GCs were deleted, or when GAPs are inhibited but GCs remain and compared to corresponding control mice. In the absence of GCs or functional GAPs, LP-DCs could not acquire luminal antigens in a manner capable of inducing antigen specific T cell responses in the draining mesenteric lymph nodes (MLN). Moreover, deletion of GCs/GAPs resulted in loss of induction of regulatory T cells in the MLN and abrogated tolerance to dietary antigens. In addition, loss of GCs/GAPs resulted in rapid reduction of pre-existing induced Treg population in the SI-LP. Notably, the in absence of GCs/GAPs resulted in rapid expansion of IL-17 producing T cells in the small intestine, indicating that GCs and GAPs play a crucial role in balancing the Treg/Th-17 axis in the intestine. Hence, suggesting that GCs and GAPs play a crucial role in the induction and maintenance of oral tolerance. These findings identify a previously unappreciated role of GCs in maintaining the Treg/Th17 balance and that processes resulting in inadequate formation of GAPs may underlie the loss of tolerance to luminal antigens.
Engineering of Targeted Mucoadhesive Chitosan Based Salmonella Nanovaccine for Oral Delivery in Poultry
Authors: Renu Sankar , Markazi Ashley D. , Dhakal Santosh , Lakshmanappa Yashavanth Shaan , Shanmugasundaram Revathi , Selvaraj Ramesh K. , Renukaradhya Gourapura J. 
Published: January 2023

Abstract
Salmonellosis in poultry is a serious economic burden. The major concern is the public health hazard by consumption of salmonella contaminated poultry meat and egg. Currently used salmonella vaccines are ineffective in combating the Salmonellosis of chicken thus warranting the need of a potent vaccine, especially an oral vaccine that can elicit robust local intestinal immunity. Biodegradable and biocompatible natural polymers are FDA approved vehicles for vaccine delivery. We prepared a candidate subunit chitosan nanoparticle vaccine containing the immunogenic outer membrane proteins (OMPs) and flagellar (F) protein of salmonella, and surface decorated with F-protein (OMPs-F-CS NPs). Physicochemical and biocompatibility properties of OMPs-F-CS NPs were studied in detail including its stability in stomach pH conditions. Salmonella targets the microfold (M) cells in the ileum Peyer’s patches (PPs) of chicken. We designed the oral OMPs-F-CS NPs vaccine to target ileal PPs to induce specific local intestinal immunity, and demonstrated that by ex vivo and in vivo studies. Layer chickens vaccinated orally with OMPs-F-CS NPs had significantly higher OMPs- specific intestinal mucosal antibody and OMPs-specific lymphocytes proliferation responses. Further, OMPs-F-CS NPs vaccination upregulated the expression of toll-like receptor (TLR) -2, TLR-4, IFN-g, IL-4 and GATA-3 mRNA levels in cecal tonsils of birds. In conclusion, we engineered chitosan based oral salmonella nanovaccine targeting intestinal PPs immune cells of birds, and demonstrated its ability to induce antigen specific mucosal antibody and T cell responses. Thus, our candidate oral salmonella vaccine has the potential to mitigate Salmonellosis in poultry.
Self-tolerance to an ileal antigen requires both IL-17A and IFNγ in mice predisposed to spontaneous ileocolitis
Authors: Jeschke Jonathan C. , Mayne Christopher G. , DeCiantis Christopher L. , Kumar Suresh N. , Drobyski William R. , Salzman Nita H. , Williams Calvin B. 
Published: January 2023

Abstract
Homeostasis in the ileum involves ongoing immune responses. To study how homeostatic processes of the ileum impact CD4+T cell responses, we used TCR transgenic tools to breed “Bigenic” mice that spontaneously produced CD4+T cells reactive to a model antigen expressed in the ileum. Bigenic mice were grossly similar to control littermates at weaning. By DOL 50, the Bigenic cohort showed reduced weight, increased circulating CD4+TEM frequencies and TH17-associated ileal hyperplasia. An extended natural history study to DOL 150 revealed that only half of Bigenic mice exhibit chronic wasting disease marked by mucosal infiltration by TH17 and TH1 cells expressing non-transgenic clonotypes, colitis and loss of ileal crypt hyperplasia. By contrast, adult Bigenic mice with normal growth maintained the TH17-associated ileal crypt hyperplasia phenotype and additionally accumulated ileal-reactive Treg cells. Adoptive transfer of nTreg cells prevented both ileal crypt hyperplasia and ileal-reactive Treg cell accumulation. Both IL-17A and IFNγ were required for ileal-reactive Treg accumulation and regulation of ileal-reactive CD4+Tconv cells. Ifng−/− Bigenic mice also exhibited an IL-23R-dependent, acutely progressive wasting disease that was &gt;95% penetrant by DOL 50. Thus, our studies identify an IL-17A and IFNγ-dependent homeostatic inflammatory process that mobilizes ileal-reactive Treg cells and is disrupted by IL-23.
Single-cell RNA-seq identifies the neuropeptide NMU as a novel regulator of ILC2 function
Authors: Wallrapp Antonia , Riesenfeld Samantha J. , Burkett Patrick R. , Abdulnour Raja-Elie E. , Nyman Jackson , Dionne Danielle , Hofree Matan , Cuoco Michael S. , Rodman Christopher , Farouq Daneyal , Haas Brian J. , Tickle Timothy L. , Trombetta John J. , Baral Pankaj , Klose Christoph S. N. , Mahlakõiv Tanel , Artis David , Rozenblatt-Rosen Orit , Chiu Isaac M. , Levy Bruce D. , Kowalczyk Monika S. , Regev Aviv , Kuchroo Vijay K. 
Published: January 2023

Abstract
Type 2 innate lymphoid cells (ILC2s) are important for both mucosal homeostasis as well as for initiation and progression of allergy and asthma. Alarmin cytokines produced by damaged or stressed epithelial cells activate ILC2s, however, the role of other pathways regulating the homeostatic versus inflammatory functions of ILC2s remain poorly defined. We generated transcriptional profiles of thousands of lung-resident ILCs isolated from mice challenged with either PBS or the alarmin cytokines IL-25 or IL-33 using massively parallel droplet-based single-cell RNA sequencing (scRNA-seq). We found significant transcriptional heterogeneity within activated ILCs and identified differentially expressed genes as potential novel regulators of ILC function. One of the genes, neuromedin U receptor 1 (NMUR1), is a receptor for the neuropeptide neuromedin U (NMU) and is specifically expressed by ILC2s and not by other immune cells. Nmur1 is particularly expressed by ILC2s at steady-state and after IL-25 treatment. In vivo NMU synergized with IL-25 to induce more severe airway inflammation than was seen with either IL-25 or NMU alone and this effect was dependent on ILCs. Mechanistically, IL-25 + NMU expanded highly proliferative and inflammatory ILCs with distinct transcriptional profiles and increased expression of IL-5 and IL-13. We observed reduced ILC2 frequency and function in NMU- or NMUR1-deficient mice after allergen challenge and found that ILCs from these mice had a lower score for an inflammatory ILC2 gene signature. Our data therefore establish a novel neuro-immune pathway that amplifies ILC2-driven airway inflammation.
Bioengineered organoid models of human germinal centers
Authors: Quizon Nicolas , Kwak Kihyuck , Shah Shivem , Singh Ankur , Pierce Susan 
Published: January 2023

Abstract
B cells undergo affinity maturation to T cell-dependent antigens within specialized microenvironments in secondary lymphoid organs termed germinal centers (GCs) resulting in the selective proliferation and differentiation of high-affinity B cells. Selection is driven by the ability of antigen-specific GC B cells to extract antigen presented on follicular dendritic cells and subsequently recruit stimuli from T follicular helper cells. Successful GC B cells ultimately differentiate into plasma cells that secrete large amounts of antigen-specific antibody, or memory B cells that contribute to long-term immunity. However, the factors that influence selection and differentiation of GC B cells in humans are not yet well understood. One challenge arises from the rapid loss of viability of human primary GC B cells ex vivo under conventional cell culture conditions. To overcome this, we bioengineered organoids using a customizable matrix along with modified feeder cell lines. When encapsulated in these organoids, primary human GC B cells exhibit enhanced viability similar to that of B cells in GCs in vivo. The low auto-fluorescent properties of these organoids allow for imaging of cellular interactions and migration. Furthermore, the customizable chemistry of the organoid matrix allows us to screen for factors that influence the differentiation and fate of human GC B cells. Improving our understanding of these processes may provide important insights in areas such as therapeutics and vaccine development.
The potential application of ctDNA technology as a cancer diagnostic tool within the Socialist Republic of Vietnam
Authors: Sika-Paotonu Dianne , Guilford Parry 
Published: January 2023

Abstract
Small fragments of tumor DNA can be found in the blood circulation and can be monitored and analysed as part of a novel cancer detection method known as circulating tumor DNA (ctDNA). These tumor DNA fragments can be isolated and quantified to produce detailed information related to cancer progression and treatment responses in patients. The possibility of utilizing a simple blood sample to allow detection and monitoring of cancer growth and behavior is highly desirable given that gaining access to and obtaining biopsy samples from solid cancers in people is not always possible or as straightforward as needed. It is anticipated this simple method of detecting and monitoring cancer will enhance the precision of cancer care for patients, and will eventuate into a particularly useful tool within resource constrained environments. The purpose of this work was to identify the diagnostic methods, treatment and pharmacological options currently available for cancer patients within the Socialist Republic of Vietnam and to explore the potential utility and applicability of ctDNA as a mechanism to support the management, treatment and diagnosis of cancer patients. Favorable interest was expressed by medical, health, community and lay persons regarding the development of ctDNA as a potential diagnostic tool within the health setting in the Socialist Republic of Vietnam and remains an attractive tool with potential to support cancer patients in Vietnam.
The application of ctDNA technology for early Cancer diagnostics in Samoa
Authors: Sika-Paotonu Dianne , Viali Satupa’itea , Naseri Leausa Toleafoa Take K. , So’o Fui Le’apai Tu’ua Ilaoa Asofou , Lameko Toleafoa Viali , Punivalu Monalisa , Burich Shelley , Maiava Fuatai , Ah-Ching Salausa John , Guilford Parry 
Published: January 2023

Abstract
Fragmented pieces of tumor DNA can be found in the human blood circulation. These tumor DNA fragments can be isolated and quantified to produce detailed information related to cancer progression and treatment responses in patients. This monitoring and analysis comprises a novel cancer detection method known as circulating tumor DNA (ctDNA). Given that gaining access to and obtaining biopsy samples from solid cancers in people is not always possible or as straightforward as needed, the utilization of a simple blood sample to allow detection and monitoring of cancer growth and behavior is highly desirable. This simple detection and monitoring technology is likely enhance the precision of cancer care for patients, and support early cancer detection. The purpose of this work was to explore in detail the potential for ctDNA to be utilized as an early cancer detection tool within the healthcare setting within the Pacific in Samoa. Consultation was sought with senior Government officials, Medical, Nursing, Health and Community research staff concerning the development and implementation of ctDNA as a diagnostic tool within the clinical health care setting throughout Samoa. The application of the ctDNA technology as an early cancer detection tool within the clinical healthcare setting was explored in depth and received with approval, given issues that currently exist concerning resource constraints and late cancer presentations. It is anticipated that the utility of ctDNA as an early cancer detection tool will support improved cancer management within the health care setting in Samoa.
A simple and rapid method for measuring human natural killer cell function in whole blood by anti-NKp30-conjugated nanoparticles
Authors: Kato Kazunori , Moriya Masaaki , Kato Eri 
Published: January 2023

Abstract
Natural killer (NK) cells play a central role as innate immune effector cells to virus infected cells and tumor cells. The standard assay to evaluate NK activity is 51Cr release assay using radio-labeled K562 cells, however, new methods using fluorescence-labeled targets or flow cytometry to examine degranulation markers have been developed recently. In addition to cytotoxicity, NK cells enable to produce various cytokines to regulate the adaptive immune responses in infection, inflammation and cancer. Here we generated nanoparticles conjugated with various antibody for NK receptors and examined to stimulate human whole blood with these nanoparticles instead of K562 target cells. We found that various cytokines such as IFN-γ, IL-6, IL-1β and CCL2 were produced by NK cells in whole blood by anti-NKp30-conjugated nanoparticles in dose-dependent manner more than by anti-NKp46 or anti-NKG2D nanoparticles. Production of IFN-γ was detected within 4 hours and reached maximum level at 24 hours after stimulation. IFN-γ production was relatively high at incubation temperature of 37°C, but that was significantly decreased over 39°C. Importantly, we could evaluate the reproducible cytokine production in same donors at another blood collection date because of the stable stimulus of nanoparticles and less variation data by human manipulation for lymphocyte separation. Additionally, cytokine secretion did not correlate with NK cell-mediated lysis of K562 target cells and NK cell numbers in whole blood. These data indicated that this method using NKp30 nanoparticles might be useful for measurement of individual differences of NK function especially correlated in adaptive immune response and inflammation.
Gene Therapy induced Tregs: A treatment for relapsing-remitting MS in mice
Authors: Keeler Geoffrey D. , Hoffman Brad E. , Cote Isabelle , Gaddie Cristina D. 
Published: January 2023

Abstract
Autoimmune disease is caused by a breakdown of tolerance against endogenous protein(s). New therapies are being developed that utilize the natural abilities of regulatory T (Treg) cells to suppress autoimmune effector cells in an antigen-specific manner. Recently, we demonstrated a clinically relevant in vivo gene immunotherapy approach for restoring Ag-specific tolerance using hepatic adeno-associated virus (AAV) gene transfer. This approach was shown to be remarkably successful at preventing and reversing a monophasic, MOG-induced EAE disease. Additionally, we have shown the dynamic ability of this treatment to prevent and/or reverse disease induced with various MOG epitopes, in genetically diverse mouse strains. However, it is unclear if this approach would be effective in the more complicated, PLP induced relapsing-remitting disease model. Thus, we re-engineered our therapeutic vector to contain the full-coding sequence of PLP. This is critically important as the established RR-EAE (SJL) model is characterized by epitope spreading; a phenomenon that may more closely resemble multiple sclerosis. SJL mice received a single peripheral injection of AAV. PLP vector, or control, 2 weeks prior to PLP 139–151 induced EAE. By day 14, control mice developed significant neurological deficits followed by a relapsing-remitting disease course. In contrast, the treated mice failed to exhibit any neurological symptoms of disease throughout the duration of the experiment, ~75 days. The long-term prevention of PLP induced disease, provides the first evidence of an effective Treg based therapy for combating epitope drift in RR-EAE, and potentially a new paradigm for treating the autoimmune disease multiple sclerosis.
B Cell Maturation Antigen impacts the efficacy of B-cell targeting therapies in neuro-inflammation.
Authors: Kumar Gaurav , Ko Rose M. , Axtell Robert C. 
Published: January 2023

Abstract
B cells have emerged as a therapeutic target for multiple sclerosis (MS). Depleting B cells with anti-CD20 antibodies are effective in treating MS. Yet, blockade of BAFF and APRIL, two cytokines important for B cell development and function, paradoxically increased disease activity in MS patients. The stark differences in clinical outcomes with these therapies demonstrate that B cells have both inflammatory and regulatory effects in MS. B cell Maturation Antigen (BCMA), a receptor that binds both BAFF and APRIL, plays a prominent role in B cell function. The role BCMA plays in MS remains enigmatic and therefore we designed this study to explore the impact BCMA deficiency has on the development and progression of experimental autoimmune encephalomyelitis (EAE), a rodent model of MS. We found that EAE induced with MOG35–55 was more severe in BCMA−/− mice compared to BCMA+/+ mice. The increased disease was associated with increased CNS inflammation and a skewing of B-cells toward a mature/inflammatory phenotype. Using bone marrow chimera strategies, we found that BCMA deficiency in B-cells was responsible for the increase in disease severity. Next, we found that anti-CD20 treatment significantly reduced EAE in BCMA−/− mice but had no effects on BCMA+/+ mice. Conversely, we found that TACI-Ig treatment had no effect on BCMA−/− mice but did reduce EAE in BCMA+/+ mice. Our data demonstrate that BAFF and APRIL constrains the development and function of inflammatory B-cells through BCMA. Furthermore, our data provide clues into the paradoxical results of clinical trials with anti-CD20 and TACI-Ig in MS.
Immune modulation by combination type I interferon and checkpoint blockade therapy in murine urothelial carcinoma
Authors: Plote Devin E , Choi Woonyoung , Mokkapati Sharada , McConkey David , Schluns Kimberly S , Dinney Colin 
Published: January 2023

Abstract
Type I interferon (IFN) has potent antitumor effects in bladder cancer and provides an essential alternative treatment option for patients who do not respond to Bacillus Calmette-Guerin (BCG). However, the mechanism of the IFN-stimulated immune response in bladder cancer is not well understood. We hypothesized that type I IFN enhances recruitment and activation of immune cells in murine bladder cancer tumors, and makes them more responsive to anti-PD-1 mAb therapy. To incite an IFN-driven inflammatory response, poly(I:C) was given to murine MB49 bladder cancer tumors peritumorally alone and in combination with anti-PD-1 mAb to determine effects on tumor growth and animal survival. IFN induction in MB49 tumors significantly inhibited tumor growth. This response was reliant on both innate and adaptive immune subsets, shown in the increased influx of intratumoral Ly6G+ neutrophils and the increase in IFNy+CD8 T cells. In addition, depleting specific immune cell subsets indicated that no one type of immune cell was critical for the IFN-mediated anti-tumor response. When used in combination with anti-PD-1 mAb, poly(I:C) prolonged mouse survival and enriched gene pathways involved in metabolism, extracellular matrix formation and organization, and PI3K and AKT signaling. To examine effects of IFN-I in BCG-unresponsive patients, tumor samples were analyzed for RNA expression and immunohistochemical (IHC) staining both before and after treatment with adenoviral interferon-α. Among these samples, 25% showed increased expression of T cell and checkpoint markers, similarly reflected in IHC analysis. Our findings suggest type I IFN grooms the tumor-immune landscape in bladder cancer by activating both innate and adaptive immune cells.