bluuebunny/crossref_metadata_2025_split huggingface dataset:
Comparison of intrinsic allergenicity potentials of gliadins from durum and soft white wheats in an adjuvant-free mouse model
Authors: Jorgensen Rick Dean , Gao Haoran , Sundar Saanjh , Chandra Shivam , Raghunath Rajsri , Loy Jaden , Ng Perry , Gangur Venugopal 
Published: August 2023

Abstract
Wheat is one of the major allergenic foods capable of eliciting life-threatening anaphylaxis. Discovering potential hypo/hyper-allergenic wheat classes is imperative to advance our knowledge and manage wheat allergy. Durum wheat (genome AABB) and soft white wheat (genome AABBDD) are two commonly consumed wheats. However, whether there are differences in the intrinsic allergenicity potential between them is unknown. Here, we tested the hypothesis that gliadins extracted from durum and soft white wheats will differ in their intrinsic allergenicity in an adjuvant-free mouse model that uses skin sensitization followed by intraperitoneal (IP) allergen challenge. Balb/c female mice were produced and maintained on a plant protein-free diet during this study. Groups of mice (n=5–10/group) were exposed to gliadins or vehicle via the skin once a week for 9 weeks. IgE responses were analyzed by ELISA to determine sensitization. Systemic anaphylaxis upon IP challenge was quantified by hypothermic shock response (HSR); mucosal mast cell response (MMCR) was quantified by measuring MMCP-1 in the blood. Our results suggest that mice exhibited robust, but comparable levels of specific IgE antibody responses after skin exposures to both wheat gliadins elicited. HSRs induced upon IP challenges with gliadins from the two wheats displayed strikingly similar kinetics. Interestingly, soft white wheat gliadin was significantly more potent in eliciting MMCR compared to the durum wheat gliadin. We demonstrate for the first-time the similarities and differences in intrinsic allergenicity potentials of gliadins obtained from the two wheats.
Supported by grants from USDA/NIFA Hatch Project MICL02486 (Accession Number: 1012322)
A single species bacterial therapy reduces type 2 immune responses in the gut to prevent food allergy
Authors: Hesser Lauren A , Hubbell Jeffrey A , Nagler Cathryn 
Published: August 2023

Abstract
The rising prevalence of food allergy is strongly correlated with dysbiosis of the gut microbiome. Rescuing this dysbiosis by reintroducing specific bacteria is a promising method to prevent or treat food allergy. We have shown that monocolonization with Anaerostipes caccae, a species representative of the healthy infant microbiota, protects against an allergic response to the milk protein b-lactoglobulin (BLG). Recently we developed a synbiotic bacterial therapy (A. caccae plus prebiotic lactulose) which rescues the allergic phenotype of gnotobiotic mice colonized with the fecal bacteria of a dysbiotic, cow’s milk allergic (CMA) infant. Synbiotic treatment minimizes the anaphylactic response to intragastric BLG challenge and reduces circulating BLG-specific IgE and IgG1. To understand how a single bacterium regulates mucosal immunity during the onset of IgE-mediated allergy we first examined the alarmin response. CMA colonized mice express high transcript levels of both IL25 and TLSP in ileal epithelial cells after sensitization with BLG plus cholera toxin. The synbiotic abrogated this alarmin response, potentially via increased mucus production. Synbiotic treatment also reduced the proportion and number of T follicular (Tf) helper cells in the ileal-draining lymph node (LN) and increased Tf-regulatory cells in the colon-draining LN, suggesting a role in T/B cell regulation. Ongoing experiments are examining how the synbiotic affects localization and expansion of IgE producing cells in mucosal tissues including Peyer’s patches and mesenteric LNs. These findings will help to uncover the mechanisms by which specific, protective bacteria prevent food allergy and inform the development of effective live biotherapeutics.
Continued food allergen consumption exacerbates beta-amyloid accumulation in allergen-sensitized AppNL-G-F mice
Authors: Brishti Afrina , Kearney Angela E , Setness Rylan , Combs Colin K , Nagamoto-Combs Kumi 
Published: August 2023

Abstract
Alzheimer’s disease (AD) is the most common neurodegenerative disease, with β-amyloid (Aβ) plaque deposition being one of the hallmark pathologies. However, the etiology of AD remains elusive. While chronic inflammation from recurrent infections or injury seems to contribute to AD development, it is unclear whether atopic diseases, such as allergies, are associated with AD. We previously reported that continuous consumption of a whey protein (WP)-containing diet led to lasting neuroinflammation in C57BL/6J mice that were sensitized but tolerant to a bovine milk allergen, β-lactoglobulin (BLG; Bos d 5). Thus, we hypothesized that the persisting neuroinflammation due to repeated allergen consumption would exacerbate AD pathology over time in genetically predisposed, allergen-tolerant individuals. To address this hypothesis, we sensitized 1-month-old male and female transgenic AppNL-G-F knock-in mice to BLG and subsequently fed them either a whey-free control or a WP diet for 6 months. Despite their lack of overt allergic reactions, BLG-sensitized mice showed elevated plasma levels of BLG-specific IgE and IgG1. Leukocyte infiltration was observed in the hippocampus of sensitized mice, and WP-fed sham mice to a lesser extent. In contrast, mast cell accumulation was apparent in the dura of sensitized mice regardless of the diet. Importantly, Aβ plaque load and Aβ peptide levels were greater in the hippocampus of BLG-sensitized mice on the WP diet compared to the respective sham group. These results indicated that continuous allergen consumption exacerbated AD-like pathology in BLG-sensitized AppNL-G-F mice, suggesting that chronic food allergen exposure may promote the progression of AD in susceptible individuals.
Supported by the National Institute of Health/National Institute on Aging (NIH/NIA) under grant number R21AG070412
Role of CCR2 in Staphylococcus Enterotoxin B-induced Acute Respiratory Distress Syndrome
Authors: Wilson Kiesha Maria , Carter Taylor , Aladhami Ahmed , Nagarkatti Prakash S , Nagarkatti Mitzi 
Published: August 2023

Abstract
Acute Respiratory Distress Syndrome (ARDS) is a fatal condition in which damage to the lungs results in fluid buildup and eventually hypoxia. Elucidating the immunological mechanism of this disorder is vital for reducing fatality. Staphylococcus enterotoxin B (SEB)-induced ARDS mimics the cytokine storm presented in patients with severe COVID-19. In previous studies, we found that SEB induced the infiltration of multiple CCR2+ macrophage subgroups and neutrophils in the lungs. Here, we investigated the role of CCR2 in the regulation of lung inflammation in SEB-induced ARDS, by utilizing CCR2-deficient mice. Mice were administered 50μg i.n. of SEB and evaluated for clinical distress and lung function over 72 hours. We performed flow cytometry and scRNA-seq on cells isolated from the whole lung of CCR2+/+ and CCR2−/−mice. We found a decrease in Ly6c+monocytes, macrophages, and CCR2+ macrophages in CCR2−/−mice, using flow and scRNA-seq analysis. Interestingly, alveolar macrophages were increased in the CCR2−/−mice. The CCR2−/−mice illustrated significant differences in their lung immune cell landscape. Mice exhibited increases in multiple T cell populations not observed in the wild-type mice. CCR2−/−mice expressed novel naïve T cells and a population of T cells which expressed genes associated with chromosome condensation (Hist1h2ap, Hist1h2ae, and Hist1h4d). Overall, top genes expressed in CCR2−/−mice such as Tcf7, Trbc2, and Stmn1, have been associated with T cell differentiation and tolerance induction. These results suggest that CCR2 deficiency prevents the migration of inflammatory monocytes to the lung but in return increase the presence of multiple unique T cell populations which alters the innate response in ARDS.
NIH P01AT003961, P20GM103641, R01AI129788, R01ES030144, R01AI123947, and K99GM147910
Discovery of a molecular clock that controls CD8+ T cell function and exhaustion
Authors: Mann Thomas Holladay , Ma Shixin , Furgiuele Jesse , Globig Anna-Maria , Chung Hokyung K , McDonald Bryan , Kaech Susan 
Published: August 2023

Abstract
During cancer and chronic viral infections, the persistence of antigen progressively causes CD8+ T cells to differentiate into a dysfunctional PD1+ “exhausted” state, with reduced production of inflammatory cytokines relative to effector cells that form during acute infections. Antigen and costimulation signals activate kinase cascades to induce distinct T cell transcription programs, but how T cells distinguish acute and chronic signals to program the exhausted or effector transcriptional states remains poorly understood. We found that members of the protein kinase C (PKC) family function together as a “molecular clock,” sensing acute or chronic agonism to drive distinct transcriptional programs. Continuous stimulation of PKC induces many features of T cell exhaustion, including a loss of production of the cytokines IFNγ and TNF, upregulation of inhibitory receptors and TOX, and altered expression of the proteins in the AP-1 family. Mechanistically, CD8+ T cells express several different PKC proteins, and chronic agonism of PKC leads to degradation of multiple family members and selective maintenance of only one PKC protein, PKC-η. This “PKC switch” alters downstream signaling to support the transcriptional reprogramming of T cells into a terminally exhausted state. In summary, continuous signaling through PKCs causes changes in the output from these kinases initially at the protein level, driving transcriptional changes downstream of PKC targets in the AP-1 transcription factor family and thus allowing further widespread transcriptional and functional changes that characterize T cell exhaustion.
Supported by a Damon Runyon Cancer Research Fellowship (DRG-2358-19) and an NIH training grant, T32-CA009370.
M. tuberculosis induced alterations in Siglecs’ expression on neutrophils predicts susceptibility to infection
Authors: Sankar Poornima , Saqib Mohd , Nafiz Tanvir Noor , Mishra Bibhuti B 
Published: August 2023

Abstract
The Presence of neutrophils in tuberculosis (TB) lung lesions is associated with bacterial growth. Despite being potent microbicidal cells, why neutrophils in TB lesions fail to control bacterial replication is poorly understood. By infecting mice with a hypervirulent replication reporter Mycobacterium tuberculosis (Mtb) W-Beijing Strain HN878, we found elevated neutrophils containing bacteria, associated with host susceptibility to infection. Phenotypic characterization of neutrophils from infected lungs revealed a downregulation of canonical neutrophil marker Siglec E (SigE) with concomitant upregulation of Siglec H (SigH), a marker generally used to define plasmacytoid dendritic cells. SigH expression on lung neutrophils increased with disease progression and predicts susceptibility to TB. Importantly, these non-conventional Ly6G+SigH+ neutrophils harbored more replicating bacteria compared to Ly6G+SigH-SigE+ lung neutrophils. In-vitro infection of naïve bone marrow neutrophils showed enhanced SigH expression with increased infectivity, and this was dependent on glycolysis and fatty acid metabolism. Treatment of immunocompetent mice with neutralizing antibodies for SigH reduced live cell bacterial burden, neutrophil infiltration, restricted bacterial replication and increased neutrophil expression of SigE. Further, SigH neutralized mice also showed elevated T cell numbers, indicating protective immunity. Collectively, our data suggests a model where Mtb induces SigH expression on neutrophils to survive within them, as a potential immune evasion strategy. Our future studies will determine the functional significance of this alteration in Siglecs on neutrophils and its role in TB pathogenesis.
R56 AI148239-01A1 and R21AI16359 (sub-award) from NIH/NIAID toDr. Bibhuti B Mishra
Allergen inhalation transforms the lungs into a permissive environment for local IgE responses
Authors: Nelson Alexander J , Wu Yee Ling 
Published: August 2023

Abstract
Immunoglobulin E (IgE) is the primary cause of allergic diseases and must be tightly regulated. Production of IgE occurs when activated B cells undergo class switch recombination (CSR) to IgE. These key events are rare, thus questions on which B cell subsets undergo IgE CSR, where IgE CSR predominantly occurs, and which molecular and cellular signals promote IgE CSR remain unresolved.
We modeled allergic asthma by intranasal administration of ragweed pollen and ovalbumin (OVA) in mice, and found that allergen inhalation induced the formation of lymphoid aggregates and production of OVA-specific IgE in the lungs. Using fluorescent mice reporting IgE switching, we revealed that allergen-sensitized lungs were a major site of IgE CSR with IgG1+ memory B cells (MBCs) being the dominant IgE-switching cells. Single-cell transcriptomics and flow cytometry analyses revealed a tight correlation between CCR6 expression and lung IgG1+ MBCs, potentially explaining their recruitment/retention in the respiratory mucosa. Furthermore, lung MBCs underwent IgE CSR more frequently than MBCs in the draining lymph nodes or spleen. Since IgE CSR requires interleukin 4 (IL-4), we used IL-4 reporter mice and found that IL-4-producing TH2 cells were enriched in the lungs compared with lymphoid organs. Co-culture experiments using purified lung MBCs from IgE reporter mice with lung or splenic effector CD4+ T cells revealed that IgE CSR was elevated in the presence of lung T cells, suggesting lung-infiltrating TH2 cells drive elevated IgE CSR in the lungs.
This study clarifies that the respiratory mucosa is a key site of IgE CSR – in allergic asthma the lung environment supports IgE CSR with IgG1+ MBCs and CD4+ T cells collaborating to promote local IgE responses.
Supported by grants from NIH (F31 HL156459, T32 AI007508, R01 HL165120, R21 AI159456)
Modulation of mucosal immunity and tissue resident memory formation by NS1-deficient influenza A virus
Authors: Tam Rachel C Y , Liao Yingyin , Mok Bobo Wing-yee , Chen Honglin 
Published: August 2023

Abstract
Recurrent influenza A virus (IAV) epidemics and occasional pandemics as a result of viral genome reassortment and human adaptation pose significant health burdens globally. The non-structural protein 1 (NS1) mediates attenuation of host response and facilitates viral replication in the nucleus. While intranasal vaccination using NS1-deleted live attenuated virus has been shown to induce pulmonary mucosal immunity in animal models, the immunological details have not been fully studied. In vivo mouse infection using wild type (WSN/WT) and NS1-deleted WSN (WSN/Del) showed WSN/Del induced massive type I interferon response in murine lungs, whereas pulmonary inflammation was minimal. Skewed migration of CD103+ dendritic cells (DC) to mediastinal lymph nodes upon WSN/Del infection was observed with enhanced formation and persistence of tissue resident influenza-specific memory CD8 T cells (TRM). After rechallenge, while influenza-specific TRM in both groups expanded comparably, more polyfunctionality was seen in TRM from WSN/Del immunized mice than WSN/WT recovered mice. Single cell RNA-seq of migratory DC in initial immunization showed differential gene programs associated with WSN/Del infection. Ongoing work is undertaken to identify the signals provided by DC that may lead to preferential priming of T cells to tissue resident lineage. Since TRM have been illustrated to confer heterosubtypic protection, knowledge on the specific physiological cues that facilitate TRM formation and retention will be useful to vaccine development.
CD8+ TRM-derived IFNγ is a critical activator of human lung epithelial cells
Authors: Mattingly Cameron , Elliott Jennifer , Thomas Jeronay , Lobby Jenna , Michalets Sarah , Kost Kirsten N , Kohlmeier Jacob E 
Published: August 2023

Abstract
Due to their position in the pulmonary mucosa, CD8+ tissue resident memory T cells (TRM) act as sentinels that rapidly respond to, and mediate protection against, respiratory viruses. In mice, TRM have been shown to mediate protection at barrier sites by producing cytokines, chemokines, and performing cell lysis. However, in humans, less is known about the effector functions of virus-specific lung CD8+ TRM. Using cells from healthy human lung donors, we first identified and quantified the frequency of antigen-specific cells against common respiratory viruses by performing intracellular cytokine staining. We next investigated the polyfunctionality and residency profiles of responding antigen-specific CD8+ T cells and found that that CD69+ CD103+ TRM comprise a larger portion of the antigen-specific CD8+ T cells for viruses largely restricted to the respiratory epithelium when compared to cells specific for viruses with a broader tissue tropism. Next, by performing a series of in vitro peptide stimulation and cytokine neutralization experiments, we investigated how antigen-specific CD8+ T cells impact local innate and epithelial cells. When stimulated with their cognate antigen, lung CD8+TRM-derived IFNγ strongly correlated with epithelial cell activation. Finally, we stimulated lung CD8+ TRMwith cognate antigen +/− anti-IFNγ and performed RNAseq on responding antigen-specific CD8+ TRM (CD25+CD137+) and epithelial cells to determine exactly how TRM-derived IFNγ alters the cellular program of these cells. Results of this study suggest that human lung CD8+ TRM act to rapidly reprogram local immune cells, and these data will ultimately help us understand how CD8+ TRM fit into the overall immune response to respiratory viruses.
This project is supported by R35 HL150803 and the Emory Center of Excellence for Influenza Research and Response (CEIRR) 75N93019R0028.
γδ T cells mediate protective immune responses following an insect virus-based Chikungunya virus chimera vaccine
Authors: Rodriguez Leslie , Adam Awadalkareem , Luo Huanle , Osman Samantha R , Weaver Scott C , Wang Tian 
Published: August 2023

Abstract
Chikungunya virus (CHIKV) is a re-emerging alphavirus that causes acute chikungunya fever (CHIKF) which often is accompanied by severe and persistent arthralgia. There are currently no licensed human vaccines against CHIKV. Eilat virus (EILV), a mosquito-specific alphavirus, replicates well in mosquito cells, but not in vertebrate cells. EILV/CHIKV is a chimeric CHIKV vaccine candidate that contains the nonstructural proteins of EILV and the structural proteins of CHIKV. This chimera vaccine candidate has been shown to protect against wild-type (WT) CHIKV challenge in mice and non-human primates (NHPs), yet the immune mechanism of the vaccine-induced host protection remains unknown. γδ T cells are known to react to WT CHIKV infection by controlling CHIKV inflammation, cell infiltration, and tissue damage. We found that γδ T cells are expanded following vaccination with EILV/CHIKV. TCRδ−/− mice, which are deficient of γδ T cells, exhibited a reduced CD8+ T cell effector function response at different timepoints compared to WT mice. This reduced CD8+ T cell response was also observed 7 days post WT CHIKV infection in the vaccinated mice. A significant reduction of CHIKV-specific IgM titers was also observed 7 days post WT CHIKV challenge in the vaccinated mice. Lastly, we noted that the vaccinated TCRδ−/− mice had higher levels of viremia than vaccinate WT mice following challenge with WT CHIKV. Overall, our results suggest that γδ T cells contribute to the host protection by promoting vaccine induced CD8+ T cell and antibody responses against CHIKV infection.
B cells shape naive CD8 T cell programming and fate decisions
Authors: Klarquist Jared , Guerrero Miguel Moreno , Manes Cameron , Cimons Jennifer , Getahun Andrew , Kedl Ross 
Published: August 2023

Abstract
Renewed interest in the relationship between B cells and CD8 T cells has revealed direct interactions between these cells, despite long-prevailing views that they function independently from one another. In fact, an IL-21-producing “helper” CD8 T cell subset has been described; yet, whether B cells have the reciprocal capacity to support CD8 T cells is less well-studied. Here, we find B cells influence the phenotype of naive, effector, and memory CD8 T cells. In MD4 mice, which have a fixed BCR repertoire, naive CD8 T cells exhibit an activated/effector phenotype, including relatively high pAkt, p70-s6K, and 4e-bp1, and low Foxo1 and Eomes. Primary CD8 T cell responses to subunit vaccination in MD4 or B cell-deficient mice are numerically reduced and effector-skewed. Importantly, the magnitude and function of memory CD8 T cells 3 months after vaccination are impaired in MD4 mice, leading to increased susceptibility to bacterial challenge. Adoptive transfer of CD8 T cells from WT into MD4 mice results in a similar effector-skewing upon vaccination, demonstrating that these phenotypic changes result from T cell-extrinsic factors. Furthermore, CAR T cell-mediated B cell depletion in 6-week-old WT mice also recapitulated the reductions in primary CD8 T cell numbers and their effector-skewing after vaccination. In agreement with prior literature, peak T cell responses after Listeria m. infection are of equivalent magnitude in WT and B cell-deficient mice, yet here we also observe reductions in their CD127+ memory phenotype. Together, these data indicate that B cells perform an important, broad role in the establishment of the naive CD8 T cell pool and subsequent regulation of CD8 T cell programming following subunit vaccination or infection.
Canine osteosarcoma as a solid tumor model for evaluation of dual valent B7H3-CXCR2 CAR T cell therapy
Authors: Cao Jennifer Wang , Lake Jessica , Chow Lyndah , Ammons Dylan , Perez Luisanny , Impastato Renata , Kurihara Jade , Verneris Michael , Dow Steven 
Published: August 2023

Abstract
To develop more effective solid tumor CAR T cell therapies, we utilized the spontaneous canine metastatic osteosarcoma (OS) animal model. Our studies evaluated the relative activity of single B7H3 and dual-valent B7H3-CXCR2 CAR constructs. Studies were also designed to determine if modifying the tumor microenvironment (TME) with oral immunotherapy improved CAR activity, based on recent studies of losartan and propranolol in dogs with OS and glioma. We evaluated B7H3 expression on canine OS cell lines using flow cytometry and OS biopsies using IHC. High levels of B7-H3 were observed on OS cell lines and OS biopsies, whereas normal canine tissues were negative or low. To assess the utility of the dual CAR we determined that canine OS cell lines secreted high IL8 spontaneously. To assess the functionality of canine B7-H3 CAR T cells, we generated canine CAR T cells from whole blood from dogs with cancer. We next determined that canine dual CAR T cells exerted specific killing and cytokine activity against B7H3+ canine OS cell lines. To further establish functionality, we evaluated the two CAR constructs in a mouse xenograft model of canine OS. We found that the dual CAR construct was significantly more active than the single CAR construct in slowing tumor growth. We found that losartan and propranolol blocked migration of mouse inflammatory monocytes to xenografted canine OS tumors. Future steps will be to evaluate the effectiveness of dual CAR T cells with TME modification in dogs with metastatic OS, to investigate the translational relevance in human patients. Results from these studies in dogs will likely give more clinically relevant results for evaluating new CAR T cell therapies and bridging results between mouse models and human clinical trials.
The V Foundation, The Shipley Family Foundation, and NIH T32 training grant