bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evidence of T cell lymphopenia in the colonic mucosa in idiopathic CD4 lymphocytopenia patients (HEM4P.250)
Authors: Kovacs Stephen , Sheikh Virginia , Rupert Adam , Thompson William , Estes Jacob , Sandler Netanya , Roby Gregg , Freeman Alexandra , Sereti Irini 
Published: January 2023

Abstract
Idiopathic CD4 lymphocytopenia (ICL) is a syndrome of unknown etiology characterized by low peripheral CD4 T cell counts and susceptibility to opportunistic infections. One hypothesis for its etiology is trapping of T cells in effector sites. 11 ICL patients and 16 healthy controls (HC) underwent colonic biopsies for CD3, CD4, and CD8 T cell enumeration by flow cytometry (FC) and immunohistochemistry (IHC) in lamia propria (LP). Mucosal IL17+ CD4 T cells were assessed after PMA/ionomycin stimulation. Plasma levels of LPS (for microbial translocation), I-FABP (for gut epithelial integrity), sCD14 (for monocyte activation), D-dimer (for coagulation) and CRP, IL-6 and TNFalpha (for inflammation) were tested in 33 ICL patients and 44 HC. ICL patients had fewer CD3 (2.84-fold, P=0.0080), CD4 (3.04-fold, P=0.013) and CD8 (2.28-fold, P=0.043) T cells in the colonic mucosa compared to HC by FC and fewer CD3 (2.31-fold, P=0.036) and CD4 (3.81-fold, P=0.0003) T cells in LP by IHC. The proportion of IL-17+ CD4 T cells was higher in ICL compared to HC (6.04 vs 3.35%, P=0.013). Levels of sCD14 levels were slightly higher in ICL than in HC (1.42 vs 1.21 µg/ml, P=0.003). Levels of LPS, I-FABP, IL-6, TNFalpha, D-Dimer, and CRP levels were not significantly different between ICL and HC (all P&gt;0.05). These data suggest that colonic mucosal lymphopenia accompanies peripheral T cell lymphopenia in ICL, albeit with preserved gut epithelial integrity and a lack of systemic inflammation.
An obesity-associated cytokine profile for immune cell differentiation and activation (HEM4P.253)
Authors: Nikolajczyk Barbara , Zhu Min , Xiao Qiang , Kuchibhatla Ramya , Apovian Caroline 
Published: January 2023

Abstract
Immune cells are dominant sources of the obesity-associated inflammation that predisposes type 2 diabetes (T2D), but triggers of immune cell hyper-responsiveness in obesity/T2D are poorly understood. T cells are key sources of cytokines that promote immune cell differentiation/activation, although T cells generally require contact with other immune cells to produce obesity-associated amounts of cytokines. To test the possibility that obesity associates with cytokine changes that impact immune cell differentiation/activation, we stimulated T cells from lean, obese non-diabetic (ND), or obese pre-diabetic (PD; HbA1c 5.6-6.5) subjects in the context of PBMCs and quantitated cytokines with the Milliplex Human High Sensitivity T Cell Panel. Samples from ND and PD subjects produced more T cell/Th1-supportive IL-2, IL-7 and IL-12p70, myeloid-supportive GM-CSF, and B cell-supportive IL-21 compared to samples from leans. In contrast, anti-inflammatory IL-10 was highest in samples from leans. Based on the anti-inflammatory actions of the T2D drug metformin, we also measured cytokine production by samples from PD subjects that were taking metformin. Metformin partially restored IL-10 production in samples from PD subjects, but had no impact on immune cell-activating cytokines. We conclude that obesity associates with inflammation due to over-expression of cytokines that support immune cell differentiation/activation, and that anti-inflammatory action of in vivo metformin is limited.
Towards understanding the expansion of plasmablasts in dengue virus infections (HEM4P.254)
Authors: Dobbs Andrea , Nascimento Eduardo , Garcia-Bates Tatiana , Smith Amanda , Marques Ernesto , Barratt-Boyes Simon 
Published: January 2023

Abstract
An increased prevalence of dengue has resulted in its emergence as an important public health threat. Pre-immunity to any of the 4 dengue virus (DENV) serotypes is the main risk factor for developing severe disease during heterologous DENV infections. We have recently published that during secondary DENV infections there is a major expansion of DENV-specific antibody secreting plasmablasts (CD19+CD27+CD38+) that correlates with the development of severe disease. The mechanisms driving this response remain unknown and may be important in understanding the pathogenesis of DENV infections. We are using in vitro approaches to define the possible mechanisms responsible for driving expansion of plasmablasts during heterologous DENV exposures. PBMCs from a cohort of healthy DENV seropositive and seronegative donors were collected for this study. We used flow cytometry to assess the differentiation of B-cells into plasmablasts 7 days after stimulating PBMCs with each DENV serotype. Preliminary in vitro studies suggest that B-cells from DENV immune donors are more apt to differentiate into plasmablasts when stimulated with DENV with possible variances between the different serotypes. Our results with isolated B-cells also suggest that the presence of other cell populations, potentially monocytes, is required for the expansion of plasmablasts. Ongoing experiments will reveal further cellular requirements for the in vitro expansion of plasmablasts after exposure to heterologous DENV.
Vitamin D augments house dust mite-mediated Th2 responses in human pulmonary dendritic cells (HYP6P.267)
Authors: Hrusch Cara , Sullivan Meghan , Tjota Melissa , Williams Jesse , Blaine Kelly , Sperling Anne 
Published: January 2023

Abstract
Low serum levels of vitamin D (VD3) have been implicated in multiple immune disorders including atopic asthma. Other studies have shown that VD3 promotes monocyte-derived dendritic cell (MDDC) differentiation into tolerogenic DCs, which in turn induce regulatory T cells. However, we have found that VD3 actually augments allergen-induced Th2 cytokines. Specifically, cells isolated from human lung draining lymph nodes produced more IL-13 when treated with a combination of VD3 and the common allergen house dust mite (HDM) than either treatment alone. Similarly, when peripheral blood-derived MDDCs were co-treated with VD3 and HDM, these cells were able to induce less IFNγ, but more IL-4, from allogeneic CD4 T cells in a mixed lymphocyte reaction. In both MDDCs and pulmonary DCs, co-treatment with VitD and HDM led to a phenotype distinct from either treatment alone, as the DCs expressed both the stimulatory molecules HLA-DR and CD86 as well as the VD3-induced molecule thrombomodulin (CD141). Additionally, while the inhibitory receptors ILT3 and ILT4 are induced in MDDCs by VD3, we found that HDM was able to block this induction. Interestingly, the ILT4 ligand HLA-G was increased in this population. Future studies will explore the role of DC-expressed HLA-G and its receptors on DC function. Together, these data indicate that while VD3 may induce markers of tolerance, common allergens abrogate this response in DCs to augment the induction of a Th2 response.
Exploring the role of miRNA 210 and regulatory T cells during TDI sensitization (HYP6P.270)
Authors: Long Carrie , Marshall Nikki , Siegel Paul , Meade Barbera , Lukomska Ewa , Anderson Katie , Beezhold Donald , Anderson Stacey 
Published: January 2023

Abstract
TDI (Toluene 2, 4-diisocyanate) is one of the leading causes of chemical-induced occupational asthma. MicroRNAs are small, noncoding RNAs that exhibit profound functional significance through the inhibition of gene expression of mRNA targets. Recently, the robust regulatory potential of miRNAs has become apparent, including their functional role in the regulation of allergic disease. Our laboratory has previously showed that miRNA 210 expression is significantly augmented in the draining lymph nodes of mice dermally dosed with TDI at various time points post exposure, however, the specific role of miRNA 210 in chemical sensitization is unknown. In this work, to elucidate the functional role of miRNA 210 during TDI sensitization, BALB/c mice were dermally exposed to TDI (4% v/v) or vehicle and endpoints were evaluated via RT-PCR, magnetic cell separation, and flow cytometry. Draining lymph node CD4+ cells showed significantly increased miRNA 210 expression following a single exposure to TDI. Confirmed (foxp3) and predicted (runx1t1, runx3, smad4, and stat6) miRNA 210 transcription factor target expression was evaluated and augmentations in foxp3 cellular protein expression and decreases in runx1 and foxp3 mRNA were observed. This data indicates that miRNA 210 has a functional role in TDI sensitization which may be related to regulatory T cell differentiation and function.
Aspirin induces IL-4 mRNA expression via activation of MAPKs pathway (HYP6P.272)
Authors: Kong Su-Kang , Chung Il Yup 
Published: January 2023

Abstract
Aspirin-exacerbated respiratory disease is characterized by asthma, nasal polyps, and intolerance to aspirin. IL-4 is mainly produced by allergic cells, including Th2 cells, mast cells, and eosinophils, and plays a central role in pathogenesis of allergic diseases such as asthma. In the present study, we demonstrated that aspirin induced IL-4 expression. Aspirin dose-dependently induced IL-4 mRNA and IL-4 promoter reporter activity in these three cell types. Treatment with pharmacologic inhibitors showed that the aspirin-induced IL-4 expression occurred through the MAPK ERK1/2 pathway, which was consistent with the observation that aspirin induced ERK phosphorylation. Analysis of serially deleted IL-4 promoter reporter showed that aspirin elevated the promoter activities regardless of the deleted constructs. When nuclear extract from these cells treated with aspirin were reacted with the probes harboring P0, P1, P2, P3, P4, P5 elements, and a region between P1 and P2 elements, DNA-protein complexes were remarkably augmented in the probes. Some of the DNA-protein complexes were supershifted by anti-NFAT. Collectively, these data indicate that aspirin induces IL-4 expression via ERK1/2 activation and availability of transcription factors that bind critical cis-acting elements in IL-4 promoter, thereby contributing to pathogenesis of aspirin-intolerant asthma.
Inhibition of LPS-induced acute lung inflammation in mice by adenovirus mediated Foxp3 expression (HYP6P.275)
Authors: Chung Hwan-Suck , Park Soojin , Baek Hyunjung , Bae Hyunsu 
Published: January 2023

Abstract
Forkhead transcription factor 3 (Foxp3) is critical transcription factor in CD4(+)CD25(+) regulatory T cells. There are increasing evidences that the functions of Foxp3 proteins are not restricted to regulatory T cells. The aim of this study is to evaluate the functions of Foxp3 proteins in LPS induced lung inflammation. In this study, LPS was administrated to Balb/c mice to induce a disease that resembles COPD. Two days prior to LPS administration, mice were infected once with the Foxp3-GFP or control GFP expression adenovirus to intra-trachea. The total cells and neutrophil numbers in bronchoalveolar lavage (BAL) fluid were counted and pro-inflammatory cytokine levels in BAL fluid were measured after 3 days later from LPS administration. Lung tissues were stained with hematoxylin and eosin (H&E) for histologic evaluation. Total cells, including eosinophil, macrophage and lymphocytes was significantly reduced in the foxp3 expression adenovirus infected mouse BAL fluid compared with the control adenovirus infected mouse BAL fluid. Foxp3-GFP or GFP was only detected in the lung epithelial cells by confocal microscopy. Using a mouse lung epithelial cell line, LA-4, Foxp3 adenovirus infected cells reduced the expression of chemokines such as CCL3, CXCL2 and CXCL5 by LPS stimulation. These results suggest that Foxp3 expression in lung epithelial cells suppresses inflammation via inhibition of chemokines secretion in lung inflammation.
House dust mite increases IL-33 and thymic stromal lymphopoietin (TSLP) gene expression in response to human rhinovirus 1B in murine bone marrow-derived macrophages. (HYP6P.276)
Authors: Akbarshahi Hamid , Menzel Mandy , Uller Lena 
Published: January 2023

Abstract
Acute viral infections are the most frequent cause of asthma exacerbations, and are therefore responsible for the majority of healthcare costs attributed to asthma. Human rhinovirus is the most common identified one in children and adults. It has been shown that IL-33- driven mechanism initiates TH2 responses to the clinically relevant allergens house dust mite (HDM). We have shown that TSLP is increased in asthma exacerbation. Alveolar macrophages have higher expression of IL-33 and TSLP in asthma. To determine whether HDM can have an effect on IL-33 and TSLP gene expression in response to human rhinovirus (RV), we treated bone marrow-derived macrophages (BMM) with HDM in the presence of M-CSF and infected them with RV1B. RV1B induced expression of IL-33 (2-fold, P-value&lt;0.001) and TSLP (2-fold P-value&lt;0.05) in HDM- treated macrophages after 24 hours compared to HDM non-treated BMM. RV1B infection caused an increase in IL-33 in IL-33 (P-value&lt;0.01) and TSLP ( P-value&lt;0.01) in HDM treated BMM compared to UV-treated RV1B group. There was no increase in IL-33 and TSLP gene expression in response to RV1B in the HDM-treated BMM at eight hours compared to UV-treated RV1B group, while HDM non-treated BMM showed increase in IL-33 (P-value&lt;0.01) and TSLP (P-value=0.059 ) in response to RV1B compared to UV-treated RV1B. These results suggest that HDM increases IL-33 and TSLP gene expression in macrophages in response to RV1B which can play a role in asthma exacerbations.
Chronic helminth infection protects mice from type III hypersensitivity (HYP6P.277)
Authors: Evans Holly , Mitre Edward 
Published: January 2023

Abstract
In this study we evaluated the effects chronic helminth infection has on type III hypersensitivity, an allergic reaction driven by antigen/antibody immune complex deposition. Mice were sensitized to ovalbumin (OVA) and 4 weeks later infected with Litomosoides sigmodontis for 10 weeks. Chronically infected animals exhibited significantly reduced ear swelling 12-24 hours after intradermal ear challenge with OVA. Although total IgG levels were higher in sensitized + infected mice than sensitized mice, OVA-specific IgG1 and IgG2a were all significantly reduced in mice that were chronically infected. Immune complexes in the ear tissue were detected by fluorescence microscopy and were found to be larger and more focal in infected animals. Histological analysis of infected mice showed reductions in edema, necrosis, and cellular recruitment compared to sensitized animals. Furthermore, a chemokine array performed on ear tissue 6 hours after OVA challenge showed that infected animals had reduced production of neutrophil-specific chemokines MIP-2 and KC. These data demonstrate that a 10 week L. sigmodontis infection protects against type III hypersensitivity responses. Future studies aim to further elucidate the immunological pathways mediating protection against type III hypersensitivity, and to evaluate whether worm-derived therapies can be useful in protection against diseases in which immune complex deposition is involved such as lupus nephritis.
A distinct immune response in steroid-refractory severe asthma and its modeling in mice (HYP7P.288)
Authors: Raundhal Mahesh , Morse Christina , Oriss Timothy , Khare Anupriya , Krishnamoorthy Nandini , Kolls Jay , Wenzel Sally , Ray Anuradha , Ray Prabir 
Published: January 2023

Abstract
Severe asthma (SA) constitutes only 5-10% of all asthmatics but accounts for 50% of the healthcare costs and morbidity associated with the disease. SA patients remain symptomatic despite treatment with high doses of corticosteroids (CS), the mainstay of asthma therapy. The presence of neutrophils in asthma has been correlated with refractoriness to CS, fatal exacerbations and severity. To date, our understanding of the pathological mechanisms central to neutrophil-prominent SA is minimal primarily because of the lack of a suitable animal model, limiting the development and testing of new therapeutics. Studies in our lab have shown that unlike patients with milder disease, SA patients consistently have a mixed Th1/Th2/Th17 immune phenotype in their airways despite high dose CS treatment. With the association of viral and bacterial infections with severity and frequency of asthma exacerbations, we hypothesized that the combination of an allergen with agents that mimic infection would induce a SA phenotype in mice. As in human severe asthmatics, the immune response in this mouse model elicits mixed granulocytic airway inflammation and airway hyperresponsiveness that are poorly responsive to CS. Using RNA sequencing and bioinformatics analysis tools, we have identified molecules that may play a central role in the pathogenesis of SA that can be targeted for developing novel therapies for this disease.
The addition of CpG to alum-adjuvanted vaccines provides a bystander effect that reduces the severity of peanut allergy in mice. (HYP7P.311)
Authors: Johnson-Weaver Brandi , Burks A. , Staats Herman 
Published: January 2023

Abstract
Rationale: The “hygiene hypothesis” predicts that a “clean” environment increases the risk of developing allergic disease while endotoxin exposure is protective. The vaccine adjuvant alum may alter immune responses to enhance the induction or severity of allergic disease. A mouse model of peanut allergy was utilized to determine if endotoxin or alum exposure influence the severity of allergic disease. Methods: C3H/HeN mice were housed in high or low endotoxin environments. Mice received three biweekly ovalbumin (Ova) immunizations adjuvanted with alum or alum combined with MPL or CpG 1826. Peanut sensitization was achieved by gastric gavage of peanut and cholera toxin weekly for 4 weeks beginning one day after immunization with Ova + adjuvants. One week after the last immunization all mice were challenged with peanut and monitored for hypothermia and allergy clinical symptoms. Results: Mice immunized with Ova + Alum + CpG exhibited mild allergic symptoms after peanut challenge (-2.0°C and 1.8 score) that did not statistically differ from challenged unsensitized mice (0.68°C and 0 score). All other immunization groups exhibited severe allergic symptoms after challenge (&lt; -2.5°C and &gt;2.5 score) and were significantly different when compared to challenged unsensitized mice. Conclusion: Endotoxin and alum exposure did not increase the severity of peanut allergy. The addition of CpG 1826 to the Ova + Alum vaccine reduced the severity of peanut-specific allergic disease.
Repeated allergen challenge provokes mechanical sensitivity, hyper-innervation and mast cell accumulation in the vulvar tissue of mice (HYP7P.319)
Authors: Chatterjea Devavani , Swanson Linnea , Ashbaugh Alyssa , Martinov Tijana , Tonc Elena , Benck Charles , Daughters Randy 
Published: January 2023

Abstract
Epidemiological evidence suggests a link between a history of environmental allergies and the risk of developing vulvodynia - a chronic provoked vulvar pain condition affecting up to 20% of women in populates assessed. We have previously shown that acute labiar exposure to a hapten allergen provokes increased mechanical sensitivity in the vulvar tissue of sensitized mice. Here, we show repeated challenge with contact hypersensitivity allergen oxazolone produces sustained mechanical hyperalgesia in the vulvar region of previously sensitized female mice for 10-14 days after final allergen exposure. Acute inflammatory responses including labiar neutrophil influx, T and B cell amplification in the draining iliac lymph nodes, labiar expression of inflammatory cytokine genes (IL-6, IL-1β and Cxcl-2) are all resolved within 5 days after final allergen exposure while increased densities of both peptidergic nerve fibers and mast cells in the labiar tissue are observed. Our findings provide the first evidence of the induction of measurable and sustained pain persisting after the resolution of overt inflammatory events following an allergic response in the labiar/vulvar tissue in mice. We have established the first model of chronic allergy-provoked vulvar pain in mice that can be used to elucidate the underlying etiology of subsets of vulvar pain that present without accompanying inflammation and are associated with histories of allergic skin responses.
Roles of scaffolding protein Gab1 in regulation of myeloid dentritic cell-mediated allergic asthma (HYP7P.281)
Authors: Zhang Yun , Xu Yun , Cen Dong , Xu Jia , Zhang Xue , Ke Yue 
Published: January 2023

Abstract
Scaffolding proteins Gab1 is ubiquitously expressed and belongs to the GAB family (GABs), which integrates multiple signal events. This protein family has recently attracted interest that genetic variants of human GAB1 are considered a risk factor for asthma. Our previous research has revealed a novel function of Gab2 in mediating mucin gene expression and Goblet cell hyperplasia, which is an important process in asthma. However, the function of Gab1 in asthma remains unclear. In asthma exacerbated patients, we found a prominent elevated level of Gab1 in peripheral blood monouclear cells compared to that of convalescence patients. Next, we generated conditional Gab1 deletion mice in myeloid cells (Gab1 KO mice) to determinate physiological the role of Gab1 in asthma. Lung histology and mucus metaplasia indicated that disruption of Gab1 in dentritic cells ) leads to alleviated asthmatic inflammation. Future investigation showed that loss of Gab1 results in impaired DC migration to lymph node, while has no influences on DC maturation, antigen uptake and presentation. Mechanically, Gab1 is essentially involved in CCL19-mediated AKT activation. Together, these finding has characterized a novel of Gab1 in mediating DC migration and that deregulation of Gab1 signal in lungs predisposes asthmatic inflammation.
Live two-photon imaging reveals distinct contribution of neutrophilic and stromal Toll-like receptor 4 in hyper-acute bone marrow response to systemic lipopolysaccharide insult (INC5P.324)
Authors: Huang Alex , Myers Jay 
Published: January 2023

Abstract
Produced in the bone marrow (BM) in large quantities, neutrophils are rapidly mobilized and released into circulation in response to inflammation and infection. How BM neutrophils are released as a result of CXCL12 and CXCL1/CXCL2 gradients have been studied at a long time scale (hours to days) following lipopolysaccharide (LPS) challenge. However, little is known about how neutrophil and BM stroma react during the hyper-acute (minutes to hours) phase. We performed intravital microscopy on mice that express GFP under the Lysozyme M (LysM) promoter. BM chimeras were generated to differentiate TLR4-mediated responses in hematopoietic and radio-resistant stroma. We observed a 2.5-fold increase in the speed and the formation of dynamic cellular clusters in 50% of BM neutrophils 30-60 minutes after LPS injection. The neutrophils achieved a peak average speed of 8.4 um/min, accompanied by their disappearance into circulation as evidenced by a 40% reduction in the GFP fluorescence after 90 minutes. BM chimera experiments revealed that while intra-marrow migration and cellular clustering are dependent on TLR4 signaling in neutrophils, their release into systemic circulation is dependent on stromal TLR4 signaling. Blocking CXCR2 signaling had the same effect as chimeras expressing TLR4-defective stroma, further revealing that BM stroma modulate TLR4-mediated neutrophil mobilization and release via CXCL1/CXCL2 chemokine signaling during the hyper-acute phase of marrow response to LPS.