bluuebunny/crossref_metadata_2025_split huggingface dataset:
Age specific differences in murine alveolar endothelial cells to pathological conditions
Authors: Liu Lunhua , Akkoyunlu Mustafa 
Published: August 2023

Abstract
Neonatal pulmonary viral infections can have diverse outcomes ranging from bronchiolitis and pneumonia in RSV infection to more subdued response in SARS-CoV-2 infection. In this study, we investigated the responses of murine alveolar endothelial cells (MLEC) to neonate mice serum in presence of TLR agonists-treated macrophages, which can mimic SARS-Cov2 induced innate cell activation. We found that when poly I:C and ssRNA40 activated macrophages were cocultured with neonatal mice serum-treated MLECs, the expression of TNFα and IL-6 were significantly decreased. Moreover, macrophage co-culture leads to decreased ACE2 and VEGFR2 in adult MLECs, but macrophage co-culture increased ACE2 expression in neonatal MLECs under Poly I:C but not ssRNA40 stimulation. Addition of neonatal mice serum to MLECs increases the reactive oxygen species but downregulates multiple inflammatory signal pathways. Concurrently, the expression VE-cadherin and Shp2 was downregulated in neonatal mice serum exposed MLECs, which leads to compromised MLEC barrier integrity. Morphologically, neonatal mice serum, but not adult mice serum, triggered the lipid droplets formation and upregulated SR-BI and CD36 receptor expression in MLECs. Taken together, neonatal and adult MLECs differentially regulate cytokine-secretion from activated macrophages, and neonatal serum breaks the tight cell junction between MLECs by down regulating Shp2 and VE-Cadherin expression and induces the lipid droplets formation in MLECs through receptors SR-BI and CD36. The differences in neonatal and adult MLEC responses to pathologic conditions may reflect the age specific pulmonary manifestations to viral infections.
The role of tumor necrosis factor receptor 2 (TNFR2) in EAE
Authors: De Oca Arena Maria Montes , Bartsch Brenden , Schwegman Lance , Abdul-Baki Nawal , Forsthuber Thomas 
Published: August 2023

Abstract
Multiple Sclerosis (MS) is a chronic autoimmune disease of the central nervous system which affects over 2 million people worldwide and is the most common neurological disability in young adults. MS is a heterogenous disease characterized by recurrent, remitting and/or progressive neurological deficits, and clinical manifestations include nerve damage, visual loss, motor and balance impairment, gait disturbance, and others. While several environmental and genetic risk factors have been identified, such as the DR2b (DRB1*1501) human MHC class II allele, the molecular mechanisms underlying the development and severity of MS remain poorly understood. Tumor necrosis factor is a cytokine that exerts its effects through two different surface receptors TNFR1 and TNFR2, and it has been identified as a key player in MS pathogenesis. TNFR2 is thought to have a protective role in experimental autoimmune encephalomyelitis (EAE), a mouse model for MS, by altering the function of astrocytes and regulatory T cells (Tregs). Previous data from our lab demonstrated that in the absence of TNFR2, transgenic mice expressing the DR2b allele and lacking mouse MHC class II (DR2bDR2) develop a more severe and progressive form of EAE, exhibiting astrogliosis. We have previously demonstrated that multiple Treg-associated genes were upregulated in CD4+ T cells from DR2bDR2 mice compared to those from control mice. However, the exact mechanism by which TNFR2 regulates Treg function in EAE is unknown. Here, we show that deletion of TNFR2 impacts the ability of Tregs to expand in vitro as well as their localization within the brains of mice undergoing EAE. Our results provide key insights into the mechanism by which TNFR2 limits and protects against CNS inflammation.
Investigating a novel role of DRAK2 in T cell cytoskeletal regulation and synapse formation
Authors: Wilander Benjamin , Harris Tarsha L , Mandarano Alexandra H , Guy Cliff , Crawford Jeremy , McGargill Maureen A 
Published: August 2023

Abstract
DRAK2 is a serine/threonine kinase expressed highly in T cells and a potential therapeutic target for autoimmune disease. Drak2−/− mice are resistant to autoimmune disease in models of type 1 diabetes and multiple sclerosis. This resistance is partly attributed to diminished accumulation of autoreactive T cells in target organs of Drak2−/− mice. Interestingly, Drak2−/− T cells can effectively eliminate tumors and pathogens similar to wildtype T cells, which makes DRAK2 an ideal target to inhibit autoreactive, but not pathogen or tumor-specific T cells. Additionally, Drak2−/− T cells are hypersensitive to suboptimal stimulation, indicating that DRAK2 impacts T cell function. These changes in Drak2−/− T cell function may be due to alterations in T cell activation, survival, or migration. However, the distinct role of DRAK2 in T cells remains unclear. To address this, we conducted RNA-seq analysis on wildtype and Drak2−/− T cells at rest and following activation. We identified significant changes in expression of genes within pathways associated with synapse formation, and cytoskeletal regulation. Thus, we examined whether DRAK2 impacts the T cell cytoskeleton or synapse formation. Our data show that Drak2−/− T cells have significantly reduced polymerized actin, but not monomeric actin compared to wildtype T cells. Additionally, we observed that synapse area was significantly increased while the concentration of TCR in the synapse was decreased in the absence of Drak2. Drak2−/− T cells also had more malformed synapses and formed fewer conjugates with antigen presenting cells than wildtype T cells. Ongoing experiments will address how DRAK2 mechanistically regulates T cell actin polymerization and synapse formation.
Supported by ALSAC and St. Jude Graduate School of Biomedical Sciences.
Gads is differentially required for the thymic development of conventional and innate-like T cells
Authors: Yablonski Deborah , Shalah Rose , Hallumi Enas , Klopstock Naama 
Published: August 2023

Abstract
Thymocyte development is tightly regulated by TCR signaling. Gads bridges a TCR-inducible interaction between SLP-76 and LAT, two adaptors that are essential for T cell development. In contrast, Gads-deficient mice have moderately impaired thymic development and accumulate non-naive peripheral T cells of unclear origin.
To illuminate the regulatory roles of Gads, we developed a model for tamoxifen-induced ablation of Gads (GadsiKO), accompanied by the expression of tdTomato. Using this model, we followed the fate of GadsiKO (Tom+) and Gads+ (Tom−) thymocytes within the same mouse. Compared to Gads+ (Tom−) DN thymocytes, GadsiKO (Tom+) DN thymocytes were less likely to undergo conventional β-selection; yet their ability to develop into γδ or NKT cells was not impaired.
A key feature of thymocyte development is the TCR-induced upregulation of CD5, which occurs during β-selection and further increases upon positive selection. Nevertheless, most GadsiKO DP thymocytes did not express CD5, nor did they increase calcium upon TCR ligation. These observations suggest that a TCR-independent, innate-like thymic developmental pathway may be unmasked in the absence of Gads.
GadsiKO SP thymocytes were heterogeneous: some were CD5−TCRlo, while others were CD5+TCRhi. Among the CD5+ GadsiKO CD4 SP thymocytes, a large proportion exhibited innate-like characteristics such CD44hiCD62Llo and/or NK1.1+. In contrast, peripheral GadsiKO T cells maintained a naive CD44loCD69Lhi phenotype for up to five weeks. Our results suggest that Gads is dispensable for peripheral quiescence, but regulates thymic development, by promoting conventional αβ T cell development, while suppressing innate-like developmental fates.
Supported by a grant from the Israel Science Foundation, and by a short-term grant from the Rappaport Institute for Research in the Medical Sciences
Trisomy 8-associated Autoinflammatory Disease (TRIAD) is Characterized by Dysregulated Myeloid Cells
Authors: Manthiram Kalpana , Xu Qin , Wu Zhijie , Bowes Mary , Gao Shouguo , Oguz Cihan , Elkahloun Abdel , Kalsi Shelley , Dulau-Florea Alina , Romeo Tina , Shi Lihong , Cassini Thomas , Deuitch Natalie , Kirby Martha , Anderson Stacie , Bruns Deborah , Ombrello Amanda , Barron Karyl , Remmers Elaine , Donovan Frank , Peterson William , Cochran Jill , Laxer Ronald , Greenberg Jessica , Zein Wadih , Brooks Brian , Perazzio Sandro , Kone-Paut Isabelle , Al-Mutiari Nora , Thors Valtyr Stefánsson , Pálmadóttir Johanna Guðrún , Aeschlimann Florence , Quartier Pierre , Murphy Karen , Chandrasekarappa Settara , Torgerson Troy , Schwartzberg Pamela L , Kastner Daniel 
Published: August 2023

Abstract
Trisomy 8 mosaicism (T8M) has been associated with a Behçet’s-like inflammatory disease, but immunologic features and treatment responses are not well-characterized. Here, we characterize 20 individuals with constitutional T8M and inflammatory disease. Most participants had congenital dysmorphologies and developmental delay. Two developed hematologic malignancies, two had bleeding diathesis due to platelet dense granule deficiency, and most had macrocytosis. The majority had recurrent fever and severe oral ulcerations, while nearly half had genital ulcers or rash. Colchicine, apremilast, and IL-1 and TNFa inhibitors were effective therapies. With ddPCR on sorted cell populations, we found that cells from the myeloid lineage (monocytes, neutrophils, megakaryocytes, erythroid progenitors) had a significantly higher percentage of trisomy 8 mosaicism compared to those from the lymphoid lineage (T and B cells) in both the peripheral blood and bone marrow, suggesting that trisomy 8 is tolerated to a greater degree by myeloid cells. Furthermore, we found that participants with T8M had more classical monocytes and had upregulation of genes associated with activated neutrophils and monocytes in their whole blood compared to healthy controls. With single cell RNAseq, we identifed which cells were trisomy 8 and disomy based on chromosome 8 gene expression and found that trisomy 8 monocytes had distinct transcriptional signatures and alteration of innate immune genes. Our findings suggest that patients with T8M are prone to a distinct autoinflammatory disease which we propose calling trisomy 8 associated autoinflammatory disease (TRIAD) and have complications due to dysregulation of cells arising from the myeloid lineage.
This work was supported by the Intramural Research Program of NIAID, NHGRI, and NHLBI, NIH.
Deciphering the functional role of commensal-induced T-DC clustering along sensory nerves in the lung
Authors: Ichise Hiroshi , Hor Jyh Liang , Arroyo-Mejías Armando J , Belkaid Yasmine , Germain Ronald N 
Published: August 2023

Abstract
The airway mucosa is a primary site of the immune response to inhaled antigens. However, how immune homeostasis is maintained in this organ during continuous exposure to airborne particles and microbial antigens is not known. Previous studies have shown that the spatial organization of immune components in relationship to each other and to non-hematopoietic components in tissues is critical to their proper function, as is communication with commensal microbes. To interrogate such relationships in the lung, especially the possible connections between immune cells and the densely innervated pulmonary airway, 3D information is required. However, current methods for 3D imaging of optically cleared tissue do not provide the marker depth afforded by high-content 2D imaging.
To overcome this constraint, we combined a fast hydrophilic tissue clearing technique, Ce3D, with the recently developed IBEX technique, and created Ce3D-IBEX. To date, up to 20 markers have been visualized in a single sample. Using Ce3D-IBEX, we identified a previously uncharacterized clustering of T cells and dendritic cells in proximity to the pulmonary sensory nerves in naïve mice. Agonistic denervation of the sensory nerves with resiniferatoxin reduced the number of clusters, and the cluster formation was disrupted in germ-free mice as well as OT-I and OT-II TCR-Tg mice. These findings indicate that the clustering requires sensory nerve-derived signals and TCR-mediated recognition of bacterial antigens. Future studies will be aimed at elucidating the functional role of the clusters in homeostasis and disease conditions and determining the molecular basis for neuro-immune communication that regulates cluster formation and activity.
All studies were done with approval from the ICUC of the NIH/NIAID. This work was supported by the Intramural Research Program of NIAID, NIH.
Respiratory syncytial virus promotes allergic predisposition via gastrointestinal microbiome alteration
Authors: Ethridge Alexander Dale , Martinez Llilian Arzola , Rasky Andrew J , Huffnagle Gary B , Lukacs Nicholas W 
Published: August 2023

Abstract
Respiratory syncytial virus (RSV) is a leading cause of childhood hospitalizations and increases the relative risk for the development of wheezing and asthma following acute bronchiolitis. Previously, we observed increased allergic predisposition following early life RSV infection in response to cockroach allergen (CRA) with a concurrent alteration in the gastrointestinal microbiome in mouse models of RSV infection and allergy. Importantly, differences in the gastrointestinal microbiome have been reported in children at a high risk for asthma. However, the effect of viral infection induced gastrointestinal microbiome alteration on allergic predisposition has not been studied. Our initial studies showed that early life RSV infection led to altered microbiome development that corresponded with disease development. We hypothesized that RSV-induced alteration of the microbiome can predispose infected animals to enhanced allergic inflammation. To address this, neonatal germ-free mice were colonized with normal microbiome from specific pathogen free mice with subsequent RSV infection followed by CRA modeling at 4 weeks post-infection. Colonization of germ-free neonatal mice protected the development of severe allergic asthma later in life, while an early life RSV infection of colonized mice reduced the protective effect of the microbiome by increasing airway hyperreactivity and T helper 2 induced responses. Taken together, our data indicate that early life RSV infection alters the neonatal microbiome leading to enhanced allergic predisposition.
Supported by grants from NIH (R35 HL150682, R01 AI138348)
Ablation of regulatory T cells lead to compromised quality of humoral immunity to SA14-14-2 live attenuated Japanese encephalitis vaccine
Authors: Nikam Bhushan Sanjay , Kalia Anurag , Pandit Suvechchha , Gupta Nimesh 
Published: August 2023

Abstract
Japanese encephalitis (JE) virus is the leading cause of encephalitis in Asia and the western Pacific. Most people infected with JE do not develop symptoms or have only mild symptoms. However, few infected people develop encephalitis and about 1 in 4 cases are fatal. Currently there is no treatment for JE however the vaccines are available. One of the most widely used vaccine against JE is live attenuated SA14-14-2, which is recommended for children and adults. However, less is known about the mechanism by which the vaccine induces protective immunity. We previously found that vaccine-induced CD4+ T cells play crucial role both in protection and in the development of protective humoral immunity. Here, we attempted to understand the role of CD4+ T cell subsets, mainly regulatory T cells (Tregs), in the mechanism of protection conferred by the vaccine. To address this, we used the Treg depletion mouse model and the TCRb−/− mouse as challenge model. We observed that depletion of Tregs lead to enhancement in germinal centre (GC) reaction to vaccine. We found a significant increase in GC-Tfh cells and GC-B cells in Treg ablation condition. We also found an increase in the magnitude of antibodies in response to vaccine particularly the IgG1 isotype. However, as marked by FRNT and the challenge studies, the neutralizing potential of the antibodies was compromised in Treg depleted condition. Altogether, the data suggest that the exclusion of Tregs may enhance the cellular and humoral response, however, that will negatively impact the protective quality of humoral immunity. Overall understanding the immunological mechanism of protective immunity conferred by this vaccine will be essential key for rational design of vaccine against JE or other flaviviruses.
NII
Correlates of immunity in SARS-CoV-2 post-vaccine infections in the Prospective Assessment of SARS-CoV-2 Seroconversion (PASS) Study
Authors: Goguet Emilie , Weiss Carol , Olsen Cara , Powers John , Coggins Si’Ana , Tribble David , Davies Julian , Meyer William , Ansari Sara , Conner Tonia , Illinik Luca , Lusvarghi Sabrina , Edwards Margaret Sanchez , Jackson-Thompson Belinda , Hollis-Perry Monique , Pollett Simon , Wang Gregory , Alcorta Yolanda , Wong Mimi , Saunders David , Mohammed Roshila , Ortega Orlando , Parmelee Edward , Lindrose Alyssa , Haines-Hull Hannah , Moser Matthew , Samuels Emily , Tso Marana , Graydon Elizabeth , Malloy Allison M W , Schully Kevin , Burgess Timothy , Broder Christopher , Laing Eric , Mitre Edward 
Published: August 2023

Abstract

Background
We sought to determine pre-infection correlates of immunity against SARS-CoV-2 post-vaccine infections.

Methods
Serum and saliva samples from 176 BNT162b2-vaccinated adults in the Prospective Assessment of SARS-CoV-2 Seroconversion study were collected between October and December 2021 and assessed for serum and saliva levels of WT SARS-CoV-2 Spike (S)-specific IgG and IgA antibodies using a microsphere-based multiplex immunoassay. Serum samples were also assessed for neutralization activity against D614G, Delta (617.2), and Omicron BA.1 and BA.1.1 variants using a lentiviral pseudovirus neutralization assay. After the fall visit, participants reported all positive PCR and/or antigen tests for SARS-CoV-2. Duration, severity, and type of symptoms, as well as risk exposures and adherence to precautionary measures, were assessed by questionnaires during the spring 2022 visit.

Results
Thirty-two participants (18.2%) had infections between December 7, 2021 and April 1, 2022. Pre-infection anti-S IgG antibody levels and neutralizing titers against BA.1 and BA.1.1 were higher in individuals that did not develop infection (p values 0.0098, 0.0313, and 0.021, respectively). No individuals with an anti-S level greater than 15,000 binding antibody units (BAU/ml) developed a post-vaccine infection.

Conclusion
High serum anti-S IgG antibody levels, high neutralization titers against Omicron subvariants, and low home risk scores correlated with protection against post-vaccine infections during the initial Omicron wave. BAU levels greater than 15,000 in the fall of 2021 were associated with complete protection during the initial Omicron wave.

This work was supported in whole, or in part, with federal funds from the Defense Health Program (HU00012020067, HU00012020094) and the Immunization Healthcare Branch (HU00012120104) of the Defense Health Agency, United States Department of Defense, and the National Institute of Allergy and Infectious Disease (HU00011920111), under Inter-Agency Agreement Y1-AI-5072.
Myeloid-KLF2 knockout neonatal mice experience mortality from lipopolysaccharide in an age-dependent and cell-specific manner
Authors: Mukherjee Devashis , Satyavolu Sriram , Thomas Asha , Jain Mukesh , Nayak Lalitha 
Published: August 2023

Abstract

Background
Preterm neonates exhibit increased mortality from Gram negative sepsis as compared to term neonates. The innate immune system is at a critical phase of development during the neonatal period. Mechanisms underlying activation of the preterm neonatal innate immune response are poorly understood. Our prior data has shown that Kruppel-like factor (KLF)-2 is a critical transcription factor to maintain murine myeloid cells in a quiescent state. We hypothesize that myeloid KLF2 plays a critical role in the neonatal immune response in Gram negative sepsis.

Methods
Preterm neonatal (P4) and term neonatal (P12) myeloid KLF2 knockout (MyK2KO) and age matched Cre mice were given 5ug/gm of E. coli O55:B5 lipopolysachharide (LPS) via intraperitoneal (i.p.) injection. For cell depletion studies, antibodies were injected i.p. 24 hours prior to LPS injection, followed by a repeat dose 24h after LPS. Ly6G antibody was used for neutrophil depletion, CD115 for monocyte depletion and equal dose of both for depletion of both cell lines.

Results
P4 MyK2KO mice have significantly increased mortality within 48 h of LPS exposure as compared to P4 Cre mice (81% vs 41%, n=19, p&lt;0.01 χ2 test). At P12, there is 100% survival of both groups of mice. Depletion of either neutrophils, monocytes or both together, decreased the mortality in P4 MyK2KO mice from 81% to 55%.

Conclusion
Preterm neonatal mice lacking myeloid KLF2 have an exaggerated response to LPS as compared to term neonatal MyK2KO or age-matched control mice. In vivo depletion of neutrophils, monocytes or both prior to LPS attenuates this response. Future studies will focus on the mechanism underlying the postnatal age-specific and cell-specific role of KLF2 in the neonatal innate immune response.

Supported by grants from Rainbow Faculty Pilot Grant (Mukherjee PI) and NIH (R01 HL142647 Nayak PI)
Propolis inhibits IgE-mediated mast cell activation by suppressing SYK/MAPK signaling pathway
Authors: Van Nguyen Thi , Jiang Yuna , Yu Zhen Nan , Jin Juan , Han Eui Hyeog , Kim Hyoung Tae , Song Chang Ho , Hur Yong Kap , Chai Ok Hee 
Published: August 2023

Abstract
Mast cells play a critical role in the effector phase of immediate hypersensitivity and allergic reactions. Propolis exerts multiple biological effects and exhibits immunomodulatory and antioxidant properties. However, its role in antidinitrophenyl (DNP) immunoglobulin E (IgE)-activated mast cells has not been investigated. In this study, we examined the mechanism underlying the inhibitory effect of propolis (Unibee powder propolis, Uniquebiotech, Co., Ltd., Iksan) on DNP-IgE-mediated mast cell degranulation, with a specific focus on histamine and cytokines release, using rat peritoneal mast cells. Propolis protected mast cell degranulation and inhibited histamine release triggered by anti-DNP IgE in rat peritoneal mast cells (RPMC) in vitro. Interestingly, propolis also decreased the level of TNF-a, IL-4, and IL-13 released from anti-DNP IgE-activated RPMC. Moreover, the SYK and its downstream-MAPK signaling pathway also were suppressed in the anti-DNP IgE-activated RPMC by the incubation with propolis. Together, these results suggest that propolis extract effectively inhibits IgE-mediated mast cell anaphylaxis-like reactions.
This research was supported by the 2022 Collaborative R&BD Program of The Food Industry Promotional Agency of Korea.