bluuebunny/crossref_metadata_2025_split huggingface dataset:
Predicting Natural Killer Cell Behavior with Mathematical Models
Authors: Millan Alberto Jovani , Sindi Suzanne , Manilay Jennifer 
Published: January 2023

Abstract
Natural killer (NK) cells are specialized lymphocytes with an innate ability to eliminate virally infected and cancerous cells, but the mechanisms that control NK cell development and cytotoxicity are incompletely understood. We hypothesize mathematical models can be used to predict how to direct NK cell development toward mature NK cells. We observed Sostdc1-knockout (KO) mice to display a partial NK cell developmental block. Building from these studies, a best fit model was simulated in which rates of transition and differentiation and cell death was determined by ordinary differential equations of deterministic compartmental models. This approach indicated that NK cell proliferation rates are not necessary to predict WT and KO population outcomes. We further hypothesize that clusters of highly cytotoxic NK cells can be identified computationally, using a combination of Ly49 receptor expression, in vitro and in vivo cytotoxicity assays using β2m−/− targets, and viSNE analysis of flow cytometry data. Sostdc1-KO NK cells also display defective cytotoxicity. We identified that KO splenic CD27+CD11b+ transitional NK cells express lower frequencies of inhibitory Ly49G2, but higher frequencies of activating Ly49H+ and D+ cells. Our viSNE results identified 3 NK cell clusters displaying a “superactive” phenotype that were virtually absent in KO mice; consistent with our observations of their hyporesponsiveness. Taken together, these data support a role for Sostdc1 in the regulation of NK cells and could provide insights into novel biological parameters to expand active NK cell numbers with high killing efficiency for immunotherapies.
Heterogeneity in differentiation capacity among ILC precursors in human peripheral blood and tissues
Authors: Kokkinou Efthymia , Gutierrez-Perez Irene , Pandey Ram Vinay , Rao Anna , Mazzurana Luca , Almer Sven , Höög Charlotte , Lindforss Ulrik , Bryceson Yenan , Mjösberg Jenny 
Published: January 2023

Abstract

Background
The origin of human Innate Lymphoid Cells (ILCs) remains less well characterised compared to mice. Single-cell RNA-sequencing recently revealed a population of ILC precursors (ILCP) expressing CD62L and CD45RA, which shows diversity in frequencies among blood, tonsil and intestine.

Objective
We set out to determine the differentiation capacity and epigenetic landscape of blood, tonsil and intestinal ILCP.

Methods
Blood, tonsil and intestinal ILCP were sort-purified on the basis of CD62L and CD45RA (CD62L+/−CD45RA+/−) and cultured on OP9-DL1 stromal cells under specific cytokine conditions. Additionally, genome-wide chromatin accessibility was measured using ATAC-sequencing.

Results
We show that all ILCP subsets in blood and tonsil were able to differentiate to IFN-γ-producing ILC1/NK cells. Whereas all blood ILCP subsets readily differentiated to IL-13-producing ILC2, no or very few ILC2 were generated from tonsil ILCP. In contrast, tonsil ILCP subsets showed higher ability to generate IL-22-producing ILC3, compared to blood ILCP. These data indicate that ILCP in blood are poised towards ILC1/ILC2 while ILCP in tonsil favours ILC1/ILC3 differentiation. To further dissect their development, we are currently analysing global epigenetic information of tonsil ILCP, aiming to delineate their chromatin landscape. Finally, we investigated the ILCP in intestinal samples and detected accumulation of CD62L−CD45RA+ ILCP in the inflamed mucosa of patients with inflammatory bowel disease (IBD) as compared to the non-inflamed tissue. We are currently analysing the potency of those cells.

Conclusion
Heterogeneity in ILC differentiation in blood and tissues could further reveal novel targets for IBD treatment.
M. tuberculosis antigen-specific T-cell function in breast milk of HIV-infected mothers
Authors: Warr Alex Jay , Shah Javeed , LaCourse Sylvia , Kinuthia Jon , Maleche-Obimbo Elizabeth , Cranmer Lisa , Nguyen Felicia , Matemo Daniel , John-Stewart Grace , Hawn Thomas 
Published: January 2023

Abstract

Objective
Breast milk (BM) is a mucosal compartment containing T cells, however, little is known regarding BM T-cell functional capacity or role in infant immunity. We hypothesized that BM T cells have M. tuberculosis (Mtb)-specific Th1 responses that are compartment-specific and differ from peripheral blood (PB) responses.

Methods
HIV-infected mothers and their infants were enrolled in a randomized clinical trial of isoniazid to prevent Mtb infection in Kisumu, Kenya. Maternal BM and PB were collected at 6–10 weeks postpartum. Cells were stimulated with Mtb whole cell lysate and controls. We measured frequencies of T cells expressing CD4 and CD8 and intracellular cytokines IFN-γ, IL-2, and TNF-α.

Results
Among 12 mothers with &gt;1000 CD3+ cells evaluable, 33% had Mtb-specific IFNγ responses, 66% had Mtb-specific IL2 responses, 66% had Mtb-specific TNF responses, and 25% had combined IFNγ, IL2, and TNF responses. The most common Mtb-specific CD4 cytokine profile was IFNγ-IL2+TNF+ (66%) and the most common CD8 profile was IFNγ+IL2-TNF+ (58%). When compared to PB profiles (N=25), BM had higher polyfunctional CD4 cells expressing IFNγ-IL2+TNF+ (median 0.11 BM vs 0.0 PB, p=.005) and lower frequencies of IFNγ+IL2-TNF− CD4 cells (median 0.0 BM vs 0.03 PB, p=.002). CD8 cells in BM had higher frequencies of IFNγ+IL2-TNF+ cells compared to PB (median 0.05 BM vs 0.0 PB, p=.0002).

Conclusions
Our data demonstrate the presence of Mtb antigen-specific Th1 cells in BM with cytokine profiles distinct from PB responses. Defining BM Mtb-specific immune responses may inform novel vaccine strategies. Studies are ongoing to examine correlations between maternal BM and PB Mtb-specific T-cell responses and infant responses to BCG vaccination.
Differential phenotypes and localization of CD8 T cell responses to acute and chronic viral infection
Authors: Hardman Blair K , Osborne Lisa 
Published: January 2023

Abstract
Norovirus represents the major viral source of gastroenteritis worldwide and the leading cause of foodborne illness in North America. Most infections are acute, symptomatic, and rapidly cleared but some cases persist asymptomatically or induce post-infectious irritable bowel syndrome. Despite the global economic burden of these infections no vaccine to prevent disease exists. Using immune correlates of acute vs chronic infection in mouse models following infection with related strains of murine norovirus, MNV-CW3 and MNV-CR6 respectively, this research interrogates the mechanisms underlying strain-specific differential antiviral CD8 T cell responses. The phenotype and quantity of MNV specific CD8 T cells are analyzed by flow cytometry. Since MNV-CW3 and -CR6 replication is higher in the small and large intestine, respectively, immunofluorescent microscopy is used to determine whether CD8 T cells are preferentially recruited to either tissue. Combining these complementary techniques provides novel insight into mechanisms governing intestinal antiviral T cell responses. Preliminary data shows activated MNV-specific CD8 T cells first accumulate in the MLN, suggesting this is the site of immune activation following oral infection with either virus. However, the magnitude and kinetics of T cell activation are reduced in CR6-infected mice compared to CW3-infected mice. This data suggests that these highly related viral strains induce activation of distinct populations of, or pathways in, APC populations with long-lasting effects on the outcome of infection. These results may have broad impacts on our understanding of how non-latent, chronic viral infections persist within a host.
Mucosal immune mediated protection of the CNS following olfactory viral infection
Authors: Moseman E. Ashley 
Published: January 2023

Abstract
Barrier immunity, particularly at mucosal surfaces, provides a first line of defense against invading pathogens. Unlike classical mucosal barrier surfaces in the gut and respiratory tract, the olfactory epithelium within the nose is a specialized barrier structure containing layers of olfactory sensory neurons (OSNs) dedicated to our sense of smell. Within nasal airways, OSNs are vulnerable to infection but also directly connected to the brain, thus anatomically, the olfactory epithelium functions as a mucosal barrier to the central nervous system (CNS). Upper respiratory viral infections can damage the olfactory neuroepithelium and are associated with loss of olfaction. Neurotropic viruses, such as vesicular stomatitis virus (VSV), readily infect OSNs, yet the olfactory epithelial response to infection is unknown. VSV infection drove a massive influx of hematopoietic cells including antigen presenting cells and antigen specific CD4+ and CD8+ T cells that established residence within the olfactory epithelium after viral clearance. Upon rechallenge, neutralizing antibody completely protected the neuronal tissues of the olfactory mucosa and the CNS, however this protection appeared to be secretory IgA independent. Resident memory T cells within the olfactory mucosa rapidly produced effector cytokines and prevented heterotypic viral challenge from accessing the CNS. Olfactory VSV infection also resulted in widespread viral and immune mediated tissue damage that contributed to permanent changes in airway turbinate structure. Collectively, these studies offer insight into how the olfactory barrier remodels its neural-immune architecture to provide enhanced protection following a viral challenge.
Host immunity modulates efficacy of fecal microbiota transplantation treatment of Clostridium difficile infection
Authors: Abt Michael C , Pamer Eric G. , Littmann Eric R. , Zarin Isma , Carter Rebecca A. 
Published: January 2023

Abstract
Clostridium difficile is an opportunistic pathogen that infects the large intestine following perturbation of the intestinal microbiota causing epithelial damage and debilitating, potentially fatal, colitis. Current antibiotic treatment options result in high recurrence rates, highlighting the need to identify alternative approaches to control this disease. Fecal microbiota transplantation (FMT) is clinically proven to efficiently clear recurrent C. difficile infection. Despite remarkable efficacy, implementation of FMT therapy is limited due to inadequate understanding of FMT mechanism of action. We use a murine model of persistent C. difficile infection to demonstrate a critical role for the host’s immune system in supporting efficacy of FMT. Immunodeficient Rag1−/− and littermate Rag1HET mice infected with C. difficile exhibit comparable establishment of persistent infection with no difference in bacterial burden or toxin levels in the cecum. Following FMT, Rag1HET mice resolve C. difficile infection while cohoused Rag1−/− mice fail to clear the infection. Analysis of intestinal microbial communities following FMT demonstrates that the microbiota of Rag1HET mice assimilate toward the composition of the FMT donor, while Rag1−/− mice exhibit impaired engraftment of bacterial species associated with 2° bile acid biosynthesis. Conversion of 1° bile acids to 2° bile acids in the cecum drives resolution of C. difficile. Metabolomic analysis of cecal content from Rag1HET mice following FMT reveals restoration of 2° bile acids while the cecal content from FMT-treated Rag1−/−mice remain comprised of 1° bile acids. These data support a role for host immunity in determining efficacy of microbiota-based therapeutics.
Protective effect of resveratrol on the integrity of alveolar and intestinal epithelial barrier in SEB-induced acute lung injury
Authors: Alghetaa Hasan F , Mohammed Amira , Nagarkatti Mitzi , Nagarkatti Prakash S 
Published: January 2023

Abstract
Acute lung injury (ALI) is a state of inflammation that breaks down the lung endothelial and epithelial cell barriers. In the current study, we investigated the role of resveratrol (RES) in regulating the expression and functions of tight junction proteins (TJP) in epithelial cell responses following exposure to this superantigen. To this end, C3H mice were given resveratrol orally twice prior to intranasal challenge with lethal SEB doses. 16S rRNA results showed that there were microbes transported in the blood in addition to the lung and colonic tissues. For this purpose, we used a reporter E. coli-GFP labeled bacterium to monitor and examine the viability of this bacterium in case of leak from lung into blood and other body compartments during the peak of ALI. The flow cytometry results showed significant reduction in copy numbers of this reporter in the blood in RES-treated mice in comparison to vehicle-treated mice. However, bacteriological examination for viability by culture of the blood showed all were negative for all experimental groups. Mouse transcriptome array showed increase in TJP gene expression in colonic and alveolar epithelial cells of RES-treated mice. Some of these genes were validated with qPCR. We found that there was an increase in the expression of some TJP genes following RES treatment. Commensals and probiotics are also known to decrease intestinal barrier dysfunction caused by inflammatory cytokines. Therefore, mass spectrometry (MS) analysis showed increase in the metabolome in colons of RES-treated group that help in strengthening of intestinal barrier. In conclusion, RES treatment increased directly or indirectly the rigidity of cell boundaries resulting in amelioration of ALI-dependent complications.
Skin-recirculating NK cells express α4β1 integrin, E-selectin ligand, and produce IFNγ
Authors: Riley John T , Hunka Julia , Debes Gudrun F. 
Published: January 2023

Abstract
NK cells are increasingly recognized as important players of skin immunity in health and during skin diseases, including skin cancers and autoimmunity. Despite their importance, little is known about the functional and migratory properties of skin-associated NK cells. Here, we employed afferent lymph vessel cannulation in sheep to assess NK cells re-entering the lymphatic system from the skin. NK cells (CD3−Nkp46+) consistently comprised 2–5% of total lymphocytes traveling in afferent lymph. The majority of skin-recirculating NK cells expressed perforin and readily produced IFNγ upon stimulation, indicating continuous recirculation of pro-inflammatory NK cells through unperturbed skin. Skin lymph NK cells expressed high levels of α4β1 integrin, similar to other circulating NK cells including those in blood and mesenteric lymph. Furthermore, a subset of lymph-borne NK cells expressed E-selectin ligand (CLA), suggesting a preference to return to skin in a manner similar to skin-homing T cells. Consistent with organ-selective as opposed to ubiquitous homing we observed that skin-recirculating NK cells lacked the gut-homing molecule, α4β7 integrin. In chemotaxis assays, skin-draining NK cells responded strongly to CCL21, a ligand for ‘tissue exit receptor’ CCR7, suggesting that NK cells use CCR7 to egress from skin via lymph. Our results support a model in which skin-specific NK cells with proinflammatory properties continuously survey the skin and are readily available for potential NK memory responses. Additional studies are underway to further assess the function and tissue tropism of skin-draining NK cells.
Aryl hydrocarbon receptor activation does not impact the number of myeloid-derived suppressor cells in the spleens of mice immunized with cholera toxin
Authors: King Aspen E , Schoenherr Elleanour M , Metten Madeline R , DeKrey Gregory K 
Published: January 2023

Abstract
The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that regulates immune responses. Previous studies in this laboratory have found that AhR activation with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in mice suppresses the anti-cholera toxin (CT) IgA response, but fecal antibody levels recover at least two weeks earlier than serum levels after repeated immunization. Studies by others have shown that TCDD exposure increases splenic myeloid-derived suppressor cell (MDSC) numbers in mice, and MDSC have been associated with enhanced IgA responses. In this study, we hypothesized that TCDD exposure would increase MDSC numbers in CT-immunized mice. Female C57Bl/6 mice were administered peanut oil (vehicle) or TCDD (40 μg/Kg) orally on day 0, and each animal was orally immunized with cholera toxin (CT, 10 μg/Kg) on days 1 and 8. On day 14, CT-specific serum IgA levels (ELISA) were significantly suppressed by TCDD exposure to 21.6% of control (P &lt; 0.05), but fecal IgA levels were not significantly different. Total spleen cell numbers were significantly reduced by TCDD exposure to 72.9% of control (P &lt; 0.05), but neither the percent nor number of MDSC (CD11b+ Gr1+, by flow cytometry) per spleen was significantly altered: 2.21% ± 0.48% and 1.91% ± 0.23% (450,000 ± 98,000 and 290,000 ± 34,000) for control and TCDD-treated, respectively (mean ± SEM). These results suggest that AhR activation differentially impacts antigen-specific IgA responses between serum and feces in cholera toxin-immunized mice, but not the number of MDSCs.
Formation of Human T Cell Mucosal Resident Memory Starts in Early Life with the Intestines
Authors: Connors Thomas , Thapa Puspa , Yopes Margot C , Guyer Rebecca S , Li Mei Mei , Snyder Mark E , Farber Donna L 
Published: January 2023

Abstract
Memory T cells are essential for optimal protection from previously encountered pathogens. Immune responses in infants lack the benefit of memory and are intrinsically distinct from those in adults. Early life adaptive immune responses have been characterized by increased generation of terminal effector subsets with decreased production of long lived memory. Here we investigate the origin of memory T cells utilizing flow cytometry and immunohistochemistry to characterize proportions and localization of T cell subsets in mucosal (intestine/lung) tissue from infant organ donors (n=12). At birth lung T cells are predominantly naïve and localized to the interstitium of the airways with memory accumulating over the first year of life. In contrast, significant proportions of memory phenotype T cells can be found in the lamina propria and intraepithelial layer of the intestine shortly after birth along with naïve cells. Effector memory subsets could be detected in robust proportions, highest in the jejunum, from 3 organ donors all less than 8 days of age. Additionally these memory T cells displayed high levels of canonical resident memory markers (CD69 and CD103). We conclude that mucosal tissues are host to naïve cells which cannot be attributable to circulating populations and that memory subsets are generated within these mucosal sites early in life. Importantly T cells in the youngest donors (ages 0–8 days) displayed resident memory phenotypes raising the possibility of prenatal development of adaptive memory responses populating the intestine. These results warrant additional investigation to determine the persistence and functionality of these subsets and emphasize the importance of the intestine to early life adaptive immunity.
Effect of Muscarine and Atropine on the production of myeloperoxidase and proinflammatory cytokines in small intestine
Authors: RESENDIZ-ALBOR ALDO ARTURO , Arciniega-Martinez Ivonne Maciel , Santamaria-Chavez Monica Miguel , Rebollar-Ruiz Xochitl Abril , Campos-Rodriguez Rafael 
Published: January 2023

Abstract
The cholinergic system it’s known as a modulator of the intestinal immune system. The aim of this work was to analyze the impact of muscarine and atropine on the production of myeloperoxidase (MPO) on lamina propia of small intestine as well as other local effects, like stimulation of goblet cells and mucus secretion. Three groups of male BALB/c mice were administred by i.p. route: a) control (SS), b) atropine (4 mg/kg) and c) muscarine (50 ng/kg). At 7 day of treatment, histological samples were obtained for Naphtol AS-D method to MPO activity analysis, immunofluorescence assay to localize MPO secreting cells and Flow Cytometry for intracellular TNF-α, IFN-γ, IL-2 and IL-6 measures. In the analysis of the histological samples it was observed that in the group treated with muscarine the number and size of goblet cells and mucus were augmented in the lumen. The number of neutrophils in the villus was also augmented in the muscarine group in comparison of treated with atropine. On the other hand, the immunofluorescence had an increased number of positive cells and activity in tissue to MPO in the muscarine group. In the Flow Cytometry analysis for lamina propia and Peyer’s patches, IFN-γ and TNF-α were augmented in the muscarine group, while IL-2 and IL-6 decreased. In this study, the MPO can be regulates immune functions such as increasing cytokines in macrophages and endothelial cells, also suppressing DC’s function of antigen presentation and switch to adaptive immunity which can lead to more innate response, and increased production of proinflamatory cytokines by lymphocytes.
The E3 Ligase VHL Controls Lung Inflammation by Regulating Epithelium-ILC2 Cross-talk
Authors: Han Jihye , Wan Qingqing , el Baghdady Sarah , Lee Jeeho , Liu Yun-Cai 
Published: January 2023

Abstract
Group 2 innate lymphoid cells (ILC2) are a specialized subset of lymphoid effector cells that play an important role in allergic inflammation in response to host-derived cytokines and alarmins. Pulmonary epithelium play a critical role in the initiation of type 2 immunity by production of cytokines, such as TSLP, IL-25 and IL-33 after exposure to allergens. However, the epithelium-derived signals and their regulatory mechanisms that directly regulate ILC2 remain unclear. Here we report that conditional deletion of the E3 ubiquitin ligase VHL in lung epithelial secretory club cells caused a selective increase of ILC2 under steady state and during pulmonary inflammation, resulting in exacerbated type 2 immune responses accompanied by increased eosinophil infiltration. VHL deficiency in lung epithelial cells upregulated the number and activity of ILC2, which was rescued by hypoxia-inducible factor 1α(HIF1α) ablation. Our findings indicate that VHL-HIF pathways control allergic inflammation in the lung epithelium by regulating ILC2, but the mechanisms of their regulation need to be identified.
TLR4 signaling regulates an inflammatory response to food allergens
Authors: Campbell Evelyn , Nagler Cathryn 
Published: January 2023

Abstract
Allergies have become increasingly more prevalent over recent years, and this increase has been found to correlate with dysbiosis of the gut microbiota. Bacterial signaling through TLR4 is known to be play a role in allergic sensitization, but the specific host-bacterial interactions and cellular immunological networks have not been elucidated. Preliminary data from our lab has shown that colonization of germ-free mice with feces from a subset of cow’s milk allergic (CMA) infant induces Serum-Amyloid A1 (Saa1) expression in the small intestine epithelium. Saa1 is an acute-phase protein that is part of a transcriptome signature associated with a Th17 response induced by adherent bacteria in the small intestine. Moreover, we show that in mice with a global ablation of TLR4, upregulation of Saa1 in abolished. Because dendritic cells (DCs) are known to be important for mediating intestinal immune responses, we sought to determine the roll of CD11c+ cells in mediating Saa1 upregulation. Antibiotic-treated CD11ccre TLR4fl/fl SPF mice were colonized with CMA feces and intestinal epithelial cells from the ileum were isolated to determine relative expression of Saa1 compared to antibiotic-treated SPF mice. Results from these experiments show that ablation of TLR4 signaling in CD11c+ cells reduces Saa1 upregulation in the ileal epithelium. Future directions of this work include examining immune cell populations in the small intestine draining lymph nodes and lamina propria to determine if this phenotype increases type 3 inflammation to exacerbate an allergic response.