bluuebunny/crossref_metadata_2025_split huggingface dataset:
Soluble B7H3 induction in pulmonary fibrosis is associated with disease severity
Authors: Liu Tianju , Rinke Andrew E , Flaherty Kevin , Phan Sem Hin 
Published: January 2023

Abstract
Idiopathic pulmonary fibrosis (IPF) is a progressive disease with limited therapeutic options. Both innate and adaptive immune mechanisms contribute to fibrogenesis. B7 family member B7H3, identified as important regulator of the immune system, has been implicated in cancer, graft-versus host disease, and allergy-related disease. Increased soluble B7H3 (sB7H3) was associated with enhanced lung fibrosis in mice receiving BLM-primed BM. Elevated sB7H3 in bronchoalveolar lavage fluid was associated with acute exacerbation IPF. To investigate whether induced sB7H3 could be used as a biomaker for IPF diagnosis and/or prognosis, plasma sB7H3 from control subjects and IPF patients was measured by ELISA. Its correlation with peripheral blood immune cells, lung function, and drug treatments were evaluated in this study. The results showed that the sB7H3 level was significantly elevated in IPF patients. This induction was associated with increased circulating B7H3+ cells, and significantly correlated with increased granulocytic myeloid-derived suppressive cells (G-MDSCs). This suggests a potential regulatory role of the induced B7H3 in the immune response in IPF. The plasma sB7H3 was negatively correlated with lung function measured as diffusing capacity of the lungs for carbon monoxide (DLCO), indicating the potential utility of plasma sB7H3 as an indicator of disease severity. The slightly elevated sB7H3 in the pirfenidone and/or nintedanib treated IPF patients were not significantly different relative to controls, which was in contrast to the significant elevation noted for the untreated patients. These findings suggest that sB7H3 may be a useful biomarker for clinical assessment of prognosis and response to drug treatment.
Treatment of Experimental Asthma by Lung Restricted Inhibition of Jak1
Authors: Rardin Hart 
Published: January 2023

Abstract
Preclinical and clinical evidence indicates that a subset of asthma is driven by type 2 cytokines such as IL-4 and IL-13 that are dependent on Jak1 for signal transduction. Because many of the asthma-related effects of these cytokines are manifested in the lung, we reasoned that combined targeting of these cytokines by means of an inhaled Jak1 inhibitor with lung restricted exposure may confer therapeutic benefit. In order to test this, we synthesized iJak-381, a novel, inhalable small molecule specifically designed for local Jak1 inhibition in the lung and tested dry powder inhaled delivery of iJak-381 in preclinical PK and efficacy studies. iJak-381 suppressed OVA induced lung inflammation and improved allergen induced airway hyper-responsiveness in mice. In a model driven by human allergens, iJak-381 had a more potent suppressive effect than a systemic corticosteroid on neutrophilic inflammation. While iJak-381 strongly inhibited lung pathology, it did not affect systemic Jak1 activity. Our data suggests that local inhibition of Jak1 in the lung might be effective for treating asthma in the clinic.
Uncovering the allergen-specific modulation in non-allergic laboratory mouse workers – understanding health versus disease
Authors: Yu Esther Dawen , da Silva Antunes Ricardo Filipe , Frazier April , Schulten Véronique , Pham John , Peters Bjoern , Sette Alessandro 
Published: January 2023

Abstract
Mouse allergies are prevalent in inner city households and laboratory workers. Mouse allergens trigger allergic symptoms from atopic dermatitis/conjunctivitis to rhinitis and asthma, thus their research is of great clinical value. The aim of this study was to characterize the mouse-specific T cell response changes in non-allergic laboratory workers before and after vacation. Peripheral blood mononuclear cells (PBMC) were collected pre-break, pre re-exposure (after break), and after re-exposure to mouse, and were stimulated with peptides identified from high molecular weight fractions of mouse allergen extracts (HiMO). Non-allergics had sizeable reacitivity and majority had decreased T cell responses upon re-exposure. In addition, the decreased magnitudes were positively correlated with time of re-exposure but not length of break. Interestingly, the suppressive activity was dominant and specific for mouse responses and did not affect bystander responses. CD4 T cell responses was down-regulated temporally for a period of time (40–160 days) and eventually went back to baseline. Ex vivo immunophenotyping revealed a trend towards down-regulation of B cells and classical Tregs. Mouse-specific CD4 effector cells were reduced while total CD4 effector cells were not. The results indicate that non-allergic status is an active process which involves allergen-specific modulation of responses upon re-exposure to allergens. Uncovering the molecular programs associated with this modulation by transcriptomic study will be the next step to better understand health and allergic diseases.
The role of NK cells during bacterial and viral/bacterial co-infection
Authors: Aguilera Elizabeth Renata , Clark Sarah E. , Lenz Laurel L. 
Published: January 2023

Abstract
Influenza virus is a large burden to the human population and a leading cause of death worldwide. Much of this high morbidity and mortality is due to co-infections with pulmonary bacterial pathogens, such as Streptococcus pneumoniae. Prior studies indicate that a primary influenza infection exacerbates secondary infection of S. pneumoniae in murine models. In part, this is due to increased bacterial burdens in the lung driven by pro- or anti-inflammatory cytokines such as IFNγ and IL-10. However, the cellular source(s) of these cytokines and their roles during infection is unclear. Previous work in our lab has shown that during systemic Listeria monocytogenes infection, NK cells are a critical source of early IFNγ with delayed IL-10 production and that IL-10 specifically facilitates pathogenesis. Here, we report that NK cells also increase susceptibility to pulmonary S. pneumoniae infection, as demonstrated by reduced bacterial burdens in organs following NK cell depletion. Using an in vitro BMDC/NK cell co-culture system, we further show that S. pneumoniae infection can directly elicit NK cell IL-10 production. Therefore, we hypothesize that NK cells play an important role during S. pneumoniae infection. To test whether NK cells also influence influenza/S. pneumoniae co-infection, we established a mouse model where S. pneumoniae bacterial burdens are increased in organs of co-infected mice. We are currently investigating how NK cells affect susceptibility to secondary S. pneumoniae infection and specifically the role of NK cell IL-10. Ultimately, this work has the potential to further define the mechanism(s) by which NK cells contribute to the control or pathogenesis of S. pneumoniae and influenza virus/S. pneumoniae co-infection.
Transcriptional reprogramming drives neutrophil phenotype and function in chronic lung disease
Authors: Margaroli Camilla , Giraldo Diego Moncada , Arafat-Gulick Dalia , Dobosh Brian , Giacalone Vincent , Kissick Haydn , Forrest Osric A , Gu Chunhui , Gibson Gregory , Tirouvanziam Rabindra M 
Published: January 2023

Abstract

Rationale
Neutrophils are often considered the first line of defense against bacterial infections. In cystic fibrosis (CF), massive neutrophil presence is concomitant with phenotypical changes and prolonged bacterial infections. Here, we sought to determine whether this paradox was dependent on transcriptional reprogramming of airway neutrophils.

Methods
Blood neutrophils were transmigrated in an in vitro model recapitulating the in vivo phenotype of CF airway neutrophils. Blood and airway neutrophils from the in vitro model and from CF patients were analyzed by flow cytometry, microarrays and RNA sequencing, as well as used for bacterial killing assays.

Results
Airway neutrophils in vivo and in vitro showed increased total RNA content and profound shifts in their transcriptional profile for survival, anabolic, and antimicrobial genes. Furthermore, transcriptional blockade by α-amanitin in airway neutrophils in vitro and ex vivo fully restored their bacterial killing capacity.

Conclusions
These findings show that some canonical functions in neutrophils, including bacterial clearance, depend on the active repression of RNA transcription, which can be modulated by the local microenvironment at the site of inflammation, challenging the paradigm holding these cells as terminally differentiated.
Neuropeptide coordination of mucosal immunity by regulating ILC3 activity
Authors: Belz Gabrielle T 
Published: January 2023

Abstract
Group 3 innate lymphoid cells (ILC3)-mediated production of the cytokine IL-22 is critical for the maintenance of immune homeostasis in the gastrointestinal tract. How this occurs is not well defined. We found that the function of ILC3s was not constant across the day, but instead oscillated between active phases and resting phases. Coordinate responsiveness of ILC3s in the intestine depended on the food-induced expression of the neuropeptide VIP. Intestinal ILC3s expressed high levels of the G protein–coupled receptor VIPR2 and activation by VIP markedly enhanced the production of IL-22 and the barrier function of the epithelium. Conversely, deficiency in signalling through VIPR2 led to impaired production of IL-22 by ILC3s and increased susceptibility to inflammation-induced gut injury. Thus, intrinsic cellular rhythms acted in synergy with the cyclic patterns of food intake to drive the production of IL-22 and synchronize protection of the intestinal epithelium through a VIP–VIPR2 pathway in ILC3s.
High dimensional cellular profiling of the myeloid compartment in the geriatric mouse model
Authors: Krishnarajah Sinduya , Ingelfinger Florian , Friebel Ekaterina , Cansever Dilay , Becher Burkhard 
Published: January 2023

Abstract
Aging, and age-related physiological changes, have been heavily implicated in declining immune functions. Many of these changes to the immune system include changes to the myeloid subsets, which have been poorly studied so far. Myeloid cells play crucial roles in acute infection and are involved in antigen presentation to cells of the adaptive immune system. Consequently, deciphering these age-related mechanisms holds great potential for targeting age-related changes in immunity. Here, we developed a model to map age related phenotypic changes in the myeloid compartment in all immunologically relevant murine organs. Using mass cytometry analysis (CyTOF), we assessed over 35 cell surface parameters on myeloid cells using a geriatric healthy mouse model. Results indicates age-related changes affecting the frequency and cell surface density of lineage markers on myeloid cells. Concretely, we see significant changes to cellular frequencies and marker expression within various resident myeloid populations, and exemplar organs will be presented here. Such age-related patterns may contribute to the impaired immune decline observed in aging. We are currently continually expanding this study and will validate the implications of these findings using spectral flow cytometry analysis.
Inverse relationship between LPS concentration and monocyte and DC inflammatory response
Authors: Perros Alexis J , Flower Robert L , Dean Melinda M 
Published: January 2023

Abstract
Dendritic cells (DC) and monocytes, which link the innate and adaptive immune response, can be stimulated with lipopolysaccharide (LPS) via toll-like receptor 4 recognition. However, understanding of human cell-specific responses to different doses of stimuli (LPS) is limited. We investigated the monocyte and DC specific, as well as the overall inflammatory response following exposure to varying doses of LPS. Fresh peripheral whole blood (n=8) was used in an in-vitro whole blood culture model to assess DC and monocyte responses in co-culture with varying doses of LPS (0.25 μg/mL, 0.5 μg/mL, 0.75 μg/mL, 1 μg/mL). DC and monocyte cytokine responses were measured via flow cytometry. Supernatants collected from the in-vitro model were used to assess the overall inflammatory response using a cytometric bead array. Exposure to all doses of LPS tested increased monocyte and DC specific cytokine production, and the overall leucocyte response. DC and monocyte cytokine production decreased in a dose-dependent manner when exposed to higher LPS doses. Our data suggests LPS concentration impacts on cell-subset specific responses more than the overall inflammatory response. Assays that assess the overall inflammatory response may lack the sensitivity required to elucidate underlying pathophysiology of infection and inflammation.
Cross-reactive dengue immune complexes modulate innate antiviral responses to Zika virus infection in human placental macrophages
Authors: Zimmerman Matthew G , Eddins Devon , Muhammad Ali , Register Emery , Kauffman Robert , Cordes Sarah , Jean Valerie , Haddad Lisa , Wrammert Jens , Suthar Mehul S 
Published: January 2023

Abstract
Zika virus (ZIKV) is a mosquito-borne flavivirus that has recently emerged in the Americas and is a pathogen of significant public health concern. Unique amongst flaviviruses, ZIKV can be vertically transmitted from an infected mother to the developing fetus in utero, in some cases resulting in adverse pregnancy outcomes, including spontaneous abortion and fetal brain abnormalities. We recently found that Hofbauer cells (HCs), a fetal-derived placental macrophage, are primary targets for ZIKV infection. Further, cross-reactive dengue antibodies facilitate ZIKV transcytosis across the placental barrier to seed infection within HCs. However, little is known about the innate immune mechanisms of HCs to limit virus replication and spread within the placenta. Here, we report that HCs express the RIG-I-like receptors, RIG-I, MDA5, and LGP2, and the cytosolic DNA sensor, cGAS, and can induce a potent innate immune response characterized by production of monocyte and macrophage chemoattractants, MCP-1, MIP-1α, and MIP-1β. In contrast, HCs infected in the presence or absence of ZIKV immune complexes induced little to no type I interferon (IFN), pro-inflammatory cytokines, or chemokines. Notably, HCs infected with ZIKV immune complexes displayed reduced antiviral gene induction as compared to HCs infected in the absence of immune complexes despite similar kinetics of virus replication. We are currently performing RNA sequencing analyses to determine how binding of viral immune complexes can alter the antiviral gene signature within HCs to enhance productive ZIKV infection. These studies will further our understanding of innate immunologic mechanisms and modulation during vertical transmission of flaviviruses in the placenta.
Analysis of PBMC Activation Using Parallel Bead and Cellular Assays
Authors: Clor Julie L. , Tyagarajan Kamala 
Published: January 2023

Abstract
Activation of human peripheral blood mononuclear cells (PBMCs) leads to expression of cytokine receptors, cytokine secretion and the expression of specific cell surface molecules resulting in divergent immune responses. The correlation of cell surface activation and cellular health markers from the lymphocyte subpopulations with cytokine levels can vary across donors and can provide valuable information to evaluate secretion levels. Bead-based multiplexed assays using Luminex xMAP® platforms provide a powerful tool for quantifying and studying several secreted cytokine levels in a single assay. Performing cellular assays performed in with flow cytometry provide information in immune cell population percentages, counts, and overall health. In this study, we present the combined cellular and secreted cytokine data on human PBMCs stimulated by the lectin phytohaemagglutinin (PHA) or the endotoxin lipopolysaccharides (LPS). Time course data from multiple donors will be presented. Combining data from these two approaches gives a comprehensive picture of the impact of stimulation on the various PBMC populations and the correlation of the secreted cytokine levels to the number of cells.
Investigating nuclear organization of the inactive X in female lymphocytes
Authors: Sierra Isabel , Nguyen Son , Joyce Eric , Anguera Montserrat C. 
Published: January 2023

Abstract
Autoimmune diseases are devastating, incurable disorders that affect over 20 million people in the US, and many of these disorders have a striking female bias. The genetic basis for this sex-bias arises from the X chromosome. Female mammals, with two X chromosomes, utilize the epigenetic process of X Chromosome Inactivation (XCI), which transcriptionally silences one X. Expression of the long noncoding RNA Xist from the inactive X (Xi) reorganizes the Xi into a compact, bipartite structure, and targets the Xi to the nuclear periphery in somatic cells. The paradigm of XCI maintenance in somatic cells is stable association of Xist RNA and heterochromatic marks on the Xi, which function to maintain the Xi structure. Recently, we found a dynamic mechanism of XCI maintenance in female lymphocytes. In naïve female B cells, Xist RNA is not localized to the Xi, but after stimulation Xist RNA and heterochromatin marks return to the Xi. How this dynamic XCI process impacts Xi structure in lymphocytes is unknown, including which factors regulate Xi nuclear organization in this system. Using a novel allele-specific imaging system, we will present new findings on the compaction and organization of the Xi in lymphocytes. Additionally, we are investigating the regulatory roles of chromatin organization and nuclear structure proteins for lymphocyte-specific XCI maintenance, and how these factors regulate X-linked gene expression. Together, these results underscore the importance of Xi structure for the fidelity of XCI in female lymphocytes.
A germinal center B cell-specific long non-coding RNA regulates the germinal center response
Authors: Wikenheiser Daniel J , Weisel Florian , Chikina Maria , Shlomchik Mark J 
Published: January 2023

Abstract
Long non-coding RNAs are a unique class of molecules involved in an exceptional variety of cellular processes, including transcriptional, translational, and epigenetic regulation. Here, we report the initial characterization of a lncRNA expressed specifically in the germinal center--organized sites of B cell proliferation, somatic hyper-mutation, and cellular differentiation that develop in response to antigenic challenge. The precise signals dictating how and when differentiated cells, such as memory B cells and long-lived plasma cells, exit the GC reaction remain incompletely understood. Thus, additional levels of molecular regulation are likely at work in the coordination of these diverse processes. We identify GCLnc1--a novel, nuclear-localized lncRNA--as a regulator of the GC reaction. As GCLnc1 is located adjacent to the murine Bcl6 locus, this lncRNA has the potential to modulate expression of key transcription factors controlling B cell identity and/or differentiation. Interestingly, forced over-expression of GCLnc1 in B cells led to upregulation of TFs associated with plasma cell identity, such as IRF4 and Blimp-1. When adoptively transferred in vivo, over-expression of GCLnc1 led to transduced B cells predominantly adopting a non-GC phenotype at the typical peak GC response, in addition to enhancing expression of IRF4 and Blimp-1. Genetic deletion of GCLnc1 led to reduced frequency of GC B cells, decreased Bcl6 expression, and dysregulated light zone/dark zone distribution in response to NP-KLH immunization. Collectively, we suggest GCLnc1 plays a key role in the integration of signaling events during the GC reaction, and may modulate the processes that drive GC exit.
Downstream effectors of BCR signaling control transcription of autoimmunity-associated transcription factor Ets1
Authors: Kearly Alyssa , Saelee Prontip , Garrett-Sinha Lee Ann 
Published: January 2023

Abstract
The transcription factor (TF) Ets1 is a critical regulator of B cell quiescence. Its expression must be maintained at a high level in resting B cells to prevent inappropriate differentiation and autoimmune responses, and its downregulation in response to antigen receptor stimulation is vital for proper B cell responses. How Ets1 expression is controlled remains unstudied. Quantification of newly-transcribed Ets1 pre-mRNA shows that Ets1 gene transcription is reduced upon BCR stimulation. Inhibition of new protein synthesis did not prevent repression of Ets1 transcription in response to stimulation, suggesting that the phenomenon depends on modulation of existing TF binding to Ets1 regulatory elements (REs) rather than de novo synthesis of a repressor protein. Epigenetic analyses identified numerous putative REs in the mouse Ets1 locus. A 217 kb BAC transgenic construct containing potential RE sequences drives eGFP reporter expression in a manner that recapitulates that of endogenous Ets1. To identify REs that may be important for regulating Ets1 expression and stimulation-induced repression, we performed ATAC-seq in untreated and BCR-stimulated B cells, and observed potentially meaningful changes to chromatin accessibility. Motifs for highly relevant TFs are enriched in accessible regions in the Ets1 locus, including members of the Smad, ETS, AP-1, and NFκB families. Following up on NFκB as a potential regulator of Ets1 expression, downregulation of Ets1 transcription was found to be dependent upon IKK2 activity but did not require RelA. These analyses suggest a potentially complex regulatory network behind the expression of Ets1 as it safeguards the resting B cell state.
Funded by NIH R01 AI122720 and Lupus Research Alliance.
Robust serum-free expansion of human B cells with an animal component-free cell culture supplement
Authors: Antignano Frann , Babic Sandra , Eaves Allen C , Brown Mark A 
Published: January 2023

Abstract
Human B cells are utilized in workflows for discovering therapeutic antibodies and in the developing field of cellular therapy. The ability to expand the B cell population in blood samples from pathogen-exposed or naive patients offers the opportunity to improve the recovery of antibody sequences, increase the diversity of the B cell repertoire, or achieve de novo immunization in vitro. It can be difficult, however, to obtain robust expansion of B cells in cultures. Addition of serum improves performance, albeit with high lot-to-lot variability, and introduces the risk of contamination by adventitious agents. We have developed an animal component-free (ACF) supplement for culturing human B cells that does not require the use of serum, feeder cells, or specialized culture plates to achieve robust in vitro expansion. The ACF supplement is a 50X concentrate comprising non-animal-derived recombinant proteins and factors that can be added to a suitable base medium of choice to promote B cell activation and expansion. Human pan-B cells isolated from peripheral blood mononuclear cells (leukopaks) by immunomagnetic enrichment could be cultured and expanded in standard 24-well tissue culture plates for over 30 days, starting with seeding densities of 1 × 105 cells/well and passaging every few days from day 7 (± 1 day) onwards. Though pronounced inter-donor cell variability was observed in the rate of proliferation, an average ~55-fold, ~110-fold and ~160-fold expansion of viable B cells was obtained after 7, 11 and 14 days of culture, respectively (n = 21 donors; range 27- to 1090-fold at day 14 ± 1 day). Differentiation to plasma cells was apparent by flow cytometric analysis of CD138 and CD20.