bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mechanisms underlying cytokine receptor suppression of type 1 diabetes (BA11P.127)
Authors: Ukah Tobechukwu , Chen Weirong , Miller Mindy , Barik Subhasis , Ellis Jason , Zaghouani Habib 
Published: January 2023

Abstract
Type 1 or pediatric diabetes (T1D) manifests when the insulin-producing pancreatic beta-cells are destroyed as a consequence of an inflammatory process initiated by lymphocytes of the immune system. IL-4 and IL-13 cytokines which utilize a shared heteroreceptor (HR) comprising IL-4Rα and IL-13Rα1 are viewed as anti-inflammatory. The mechanism by which these cytokines drive suppression of T1D remains undefined. In this study, we used non-obese diabetic (NOD) mice, which develop the disease spontaneously to determine how the cytokines and the shared HR drive modulatory functions. The findings indicate that NOD mice in which the shared HR is made non-functional by ablation of IL-13Rα1 gene resist the disease relative to wild type mice with intact heteroreceptor. In addition, their lymphocytes could not transfer the disease into NOD.Scid mice effectively. This was due to an increase in the frequency of membrane-bound TGF-β T cells expressing intermediate levels of FoxP3 marker (mTGFβ+FoxP3int). Also, there was relocation to the pancreas of anti-inflammatory macrophage (MΦ) expressing the mannose receptor, a marker of M2 phenotype. Thus, the HR modulates both arms of adaptive and innate immune suppressors including Tregs and MΦ. These insights on the role of HR in modulation of T1D will likely inform on the development of therapeutic approaches against the disease.
HIV-1 Infection increases sphingosine-1-phosphate receptor 1 expression in the human thymus (CAM4P.162)
Authors: Resop Rachel , Craft Joshua , Uittenbogaart Christel , Vatakis Dimitrios , Blom Bianca 
Published: January 2023

Abstract
Sphingosine-1-phosphate receptor 1 (S1P-R1) is essential in mice for homing of lymphoid cells as well as egress of mature thymocytes from the thymus to the periphery. We have recently demonstrated that in the human thymus, response of S1P-R1 to its ligand, Sphingosine-1-phosphate (S1P), is required for egress of mature human thymocytes to the periphery. Herein we demonstrate that immune dysregulation accompanying Human Immunodeficiency Virus (HIV) infection alters the expression of S1P-R1 on human thymocytes, increasing expression of the receptor at both the mRNA and protein levels. Moreover, the function of the receptor is maintained, as assayed by flow cytometry based detection of phosphorylated Akt (pAkt). Finally, we found that several cytokines including Tumor Necrosis Factor Alpha and Interferon Alpha are perturbed during infection of the thymus, and exogenous treatment of human thymocytes with these cytokines increases or attenuates S1P-R1 expression. Therefore we have identified a contributing mechanism to alteration of S1P-R1 expression during HIV infection of the human thymus, which may have important implications for therapeutics for HIV infected individuals.
Role of dendritic cell-derived amphiregulin in pulmonary fibrosis (CCR5P.203)
Authors: Liu Tianju , Ding Lin , Wu Zhe , Gonzalez De Los Santos Francina , Phan Sem 
Published: January 2023

Abstract
Recruitment of dendritic cells (DCs) has been implicated in lung fibrosis. Accumulation of BM-derived DC (BMDC) was shown to play an important role in bleomycin (BLM)-induced fibrotic lungs. A microarray analysis of lung DCs derived from control and BLM-treated mice revealed amphiregulin (AREG), an epidermal growth factor receptor ligand was highly upregulated in cells from fibrotic lung. Moreover BLM-induced fibrosis and AREG induction were attenuated when BMDCs were depleted. Thus AREG induced primarily in BMDCs appear to play a significant role in this model of fibrosis. To further confirm the in vivo importance of AREG fibrosis was induced in wild type or AREG knockout (KO) mice by injecting BLM endotracheally, and the fibrotic response was analyzed biochemically and histologically. The results showed that BLM induced significant induction of HYP in WT mice was significantly decreased in AREG KO mice. The fibrotic markers collagen I and α-smooth muscle actin expressions were also significantly reduced compared to the WT mice, along with impaired proinflammatory cytokine/chemokine expression. These fibrotic responses were also reflected by tissue histopathology. Finally, unlike BMDCs isolated from wild type mice, those from AREG KO mice lacked the ability to induce myofibroblast differentiation when co-cultured with lung fibroblasts. Thus BMDC-derived AREG played an important role in pulmonary fibrosis probably by promoting fibroblast proliferation and survival.
Developing soluble mimetics of the chemokine receptor CCR5 for the study of HIV Env/receptor interactions (CCR6P.216)
Authors: Hajduczki Agnes , Bundoc Virgilio , Berger Edward 
Published: January 2023

Abstract
During HIV entry, the surface-exposed gp120 subunit of the envelope glycoprotein (Env) undergoes a series of stepwise conformational changes in response to interactions with the primary receptor CD4, and co-receptors, CCR5 or CXCR4. Obtaining structural information on the various intermediates during viral entry is a key focus of antiviral and vaccine research and could open the doors for more effective treatment and prevention. This project aims to develop recombinant soluble co-receptor mimetics featuring critical determinants of the chemokine receptor, CCR5, using two approaches: fusing the CCR5 moieties to soluble CD4 by flexible polypeptide linkers and using a globular, stable scaffold protein to orient the co-receptor determinants in an optimal conformation to recreate the binding interaction with gp120. We have successfully overexpressed and purified the recombinant proteins from mammalian cells. Characterization of the gp120-binding properties of the variants is underway using a vaccinia-based cell fusion assay where soluble CD4 has been shown to induce membrane fusion between Env-expressing effector cells and target cells bearing CCR5, but no CD4. The CCR5 moieties of the sCD4-fused constructs compete with cell-surface CCR5 for binding to gp120, leading to impaired activation of membrane fusion compared with sCD4. The soluble variants of CCR5 will be used in experiments to elucidate the conformational changes in gp120 that immediately precede membrane fusion.
The role of interleukin-12 in breast cancer (CCR6P.220)
Authors: Nunez Angel , Arroyo Nelly , Godoy Lenin , Dutil Julie 
Published: January 2023

Abstract
Interleukin-12 (IL-12) is a dimeric cytokine which has demonstrated anti-cancer effects in vitro and in vivo. However, the role of this cytokine in breast cancer (BC) has not been studied thoroughly. In this pilot study, the levels of the IL-12 p70 and IL-12 p40 secreted from PBMCs of de novo-BC patients, BC patients in remission, and non-BC controls were measured and compared. Peripheral blood mononuclear cells (PBMCs) from participants (n=8) were stimulated with endotoxin at specific time points to induce IL-12 secretion. We expect to obtain differences on the levels of IL-12 p40 and IL-12 p70 among the study groups. This pilot study will give an initial comparative standpoint to see if there are differences in the levels of IL-12 among the groups. Further studies can be conducted to analyze the mRNA expression and protein levels of other downstream IL-12 signaling pathway components.
Loss of ΔNp63 impairs thymic epithelial progenitor survival and differentiation (HEM7P.234)
Authors: Bain Virginia , Reeh Kaitlin , Carter Carla , Flores Elsa , Richie Ellen 
Published: January 2023

Abstract
The thymic stroma consists primarily of thymic epithelial cells (TECs) that originate from endodermal progenitors in the 3rd pharyngeal pouch (pp). A common epithelial progenitor in fetal thymus differentiates into networks of cortical (cTEC) and medullary (mTEC) cells, which provide distinct maturation signals that support T cell generation. The transcription factor p63 is expressed in cTECs and mTECs and global deletion results in a severely hypoplastic thymus. Alternative promoters generate two p63 isoforms, TAp63 containing a transactivation (TA) domain and ΔNp63, lacking a TA domain. The roles of TAp63 and ΔNp63 in thymus organogenesis have not been determined. We find that TAp63 is undetectable in fetal or steady state adult thymus and deletion of TAp63 in fetal TECs does not affect thymus size or TEC development. In contrast, ΔNp63, is highly expressed by E9.5 and persists in cTECs and mTECs in the postnatal thymus. Using Foxn1Cre to delete ΔNp63fl/fl in TEC progenitors of the 3rd pp results in a severely hypoplastic thymic phenotype by E14.5. Patterning of 3rd pp endoderm into thymus- and parathyroid-fated domains is not disrupted. Plet1+ progenitor TECs are found at a normal frequency at E12.5; however, Plet1- TECs lacking ΔNp63 undergo apoptosis at a higher frequency than wildtype. Furthermore, surviving ΔNp63 null TECs fail to differentiate beyond an early progenitor state. We conclude that ΔNp63 is necessary for survival and differentiation of developing TECs.
MicroRNA-205 regulates thymopoiesis under steady state and stress conditions (HEM7P.222)
Authors: Hoover Ashley , Khan Shaheen , Cleaver Ondine , de la Morena Maite , van Oers Nicolai 
Published: January 2023

Abstract
The thymus comprises an interconnected meshwork of epithelial cells (TECs) and thymocytes. This organ is exceedingly sensitive to stress, undergoing rapid involution in response to infection, trauma, radiation exposure, and/or glucocorticoid treatments. The nature of the stress predicates whether the epithelial cells, developing thymocytes, and/or both are affected. MicroRNAs are small noncoding RNAs that regulate cellular responses to stress and during regeneration. We discovered that miR-205 is a stress responsive, epithelial-specific miR that modulates thymopoiesis. Mice lacking miR-205 selectively in TECs exhibited a significant decline in thymopoiesis beginning at 8 weeks of age. When stressed with a dsRNA mimic (polyI:C), these mice exhibited a much more severe thymic atrophy compared to littermate controls. Quantification of the TEC subsets revealed a defect in the proliferative recovery of the cortical TECs lacking miR-205. This contributed to a significant reduction in the number of single positive CD4 and CD8 T cells, indicating that the capacity of the TECs to support selection processes at the CD4+CD8+ stage of thymopoiesis is compromised. Current experiments to determine how miR-205 regulates TEC function and proliferation are ongoing. The findings reveal an important role for miR-205 in thymopoiesis, which may be of therapeutic importance.
Wnt signaling during thymic involution and reconstitution. (HEM7P.226)
Authors: Singh Varan , Schwartz Carla , Jamil Nawshin , Hoque Mohammed , Osada Masako , Pezzano Mark 
Published: January 2023

Abstract
Despite its critical role in T-cell development, the thymus undergoes profound age-related involution. Inhibition of Wnt signaling in thymic epithelium (TECs) leads to rapid involution. Lack of a Wnt reporter that was functional in TECs hindered characterization of Wnt signaling during TEC ontogeny. Using the TCF/LEF-H2BGFP Wnt reporter mouse, TECs undergoing active Wnt signaling were characterized during different stages of thymic aging and recovery. In the postnatal thymus, H2BGFP expression was restricted to TECs in the medulla and CMJ including 60% of the Aire-expressing mTECs, suggesting a role for Wnt signaling in the development of this critical TEC subset. Comparison of the TECs undergoing Wnt signaling in old versus young thymus revealed an increase in the frequency of TECs undergoing Wnt signaling even though overall TEC numbers decreased significantly with age. The decrease in both total TECs and TECs undergoing Wnt signaling was most apparent in the UEA1hi mTEC subset and suggests that the age related decline in AIRE expression is correlated with the decline in mTECs undergoing Wnt signaling. SCID/TCF/LEF1-H2BGFP mice were used to observe Wnt signaling during thymic reconstitution induced by either bone marrow transplant (BMT) (full thymic development) or injection of mature T cells (mTEC development). During thymic reconstitution induced by both BMT and mature T-cells, Wnt signaling was primarily associated with Aire expressing mTECs, critical for self-tolerance.
a conversed gene regulatory pathway underwrites the development of innate lymphoid cells and adaptive T cells (HEM8P.239)
Authors: Yang Qi , Xue Hai-Hui Howard , Bhandoola Avinash 
Published: January 2023

Abstract
Innate lymphoid cells (ILC) are a specialized family of lymphocytes that functionally mirror subsets of effector T cells but lack adaptive antigen receptors. ILC express many T-lineage characteristic genes, and are closer to T cells than to other leukocytes at the transcriptome level. Here we show that the transcription factor TCF-1 drives a shared T/ILC lineage gene pathway that underwrites the development of both ILC and T lineages. TCF-1-/- bone marrow progenitors failed to give rise to all known adult ILC and T lineage subsets as well as their precursors. Ectopic expression of TCF-1 elicited the expression of many key genes shared by ILC and T lineages. Like in early T cell development, Notch signaling promoted pulmonary ILC2 development in part through TCF-1, and TCF-1 acted through both GATA-3 dependent and independent mechanisms. Our data suggested that a shared gene regulatory circuit underwrites the development of innate lymphoid cells and T cells.
Atg7 plays a critical role in the development of iNKT cells with altered PLZF expression. (HEM8P.242)
Authors: Jeong Dongjin , Jin Young-Hee , Chung Doo Hyun 
Published: January 2023

Abstract
Invariant Natural Killer T (iNKT) cell is a distinct subset of T cells that plays a regulatory role in various immune diseases. Although many studies have reported that various factors regulate development of iNKT cells, the role of autophagy in the development is still unknown. Here, we report that autophagy-related protein 7(Atg7) is required for development of iNKT cells in the thymus. iNKT cells of the thymus and liver expressed Atg7 and their development is completely attenuated in the thymus of CD4-specific Atg7 knock-out (CD4-Cre/Atg7f/f) mice compared with WT mice. The levels of PLZF expression were altered in thymocytes from CD4-Cre/Atg7f/f mice as compared with WT mice. CD4-Cre/Atg7f/f mice showed a few iNKT cells in peripheral organs, which led to functional deficient in iNKT cell-specific activation in vitro and in vivo. Accordingly, autoantibody-induced arthritis was attenuated in CD4-Cre/Atg7f/f mice. Therefore, our findings suggest that Atg7 regulates iNKT cell development with altered PLZF expression.
Pregnancy does not attenuate the antibody or plasmablast response to inactivated influenza vaccine (HUM6P.247)
Authors: Bayless Nicholas , Kay Alexander , Fukuayama Julia , Aziz Natali , Dekker Cornelia , Mackey Sally , Swan Gary , Davis Mark , Holmes Susan , Blish Catherine 
Published: January 2023

Abstract
Background. Inactivated influenza vaccine (IIV) is recommended during pregnancy to prevent influenza infection and its complications in pregnant women and their infants. However, the extent to which pregnancy modifies the antibody response to vaccination remains unclear, and prior studies have focused primarily on hemagglutinin inhibition (HI) titers. A more comprehensive understanding of how pregnancy modifies the humoral immune response to influenza vaccination will aid in maximizing vaccine efficacy. Methods. Healthy pregnant women and control women were studied prior to, 7 days after and 28 days after vaccination with IIV. HI titers, microneutralization (MN) titers and the frequency of circulating plasmablasts were evaluated in pregnant vs. control women. Results. Pregnant women and control women mount similarly robust serologic immune responses to IIV, with no significant differences for any influenza strain in post-vaccination geometric mean HI or MN titers. HI and MN titers correlate, though MN titers demonstrate more robust changes pre- vs. post-vaccination. The induction of circulating plasmablasts is increased in pregnant women vs. controls (median fold-change 2.60 vs. 1.49 [Interquartile Range 0.94-7.53 vs. 0.63-2.67]; p = 0.03). Conclusion. Pregnant women do not have impaired humoral immune responses to IIV, and may have increased circulating plasmablast production compared to control women.
Early exposure to phosphorylcholine-bearing microbes dampens the development of house dust mite allergy during adult life (HYP7P.263)
Authors: Patel Preeyam 
Published: January 2023

Abstract
A suggested mechanism to explain the dramatic rise in the incidence of allergic disease among individuals living in developed countries has been an imbalance in the TH1/TH2 subsets; however, a majority of neonatal pathogens evoke potent B cell responses. These B cell clones, with shared specificity for allergens, can be protective against the development of allergic disease. Streptococcus pneumoniae and house dust mite (HDM) bear similar phosphorylcholine epitopes. Therefore, we hypothesized that B cell clones generated as a result of early pneumococcal exposure would dampen the development of HDM allergy during adult life. To test our hypothesis, we immunized neonatal mice with PC-deficient or PC-bearing pneumococcus and then challenged these mice with HDM as adults. Mice receiving the PC-bearing pneumococcus had a large frequency of PC-specific B cells that resided in their spleen and continually secreted antibodies. Upon pulmonary exposure to PC-bearing HDM, these B cells migrated to the lung and secreted antibody locally into the pulmonary space. As a result, mice immunized with PC-bearing pneumococcus, presented with dampened production of serum IgE and TH2-associated cytokines, development of airway hyperreactivity, and infiltration of allergic mediators into the lung as compared to mice exposed to PC-devoid pneumococcus as neonates. We further demonstrated that neonatal exposure to bacteria was not sufficient enough to disrupt the balance in the TH1/TH2 subsets.
Increased survival and exocytosis of toxic primary granules by neutrophils in an in vitro model of Cystic Fibrosis airway disease (INC6P.310)
Authors: Forrest Osric , Ingersoll Sarah , Preininger Marcela , Laval Julie , Brown Milton , Tirouvanziam Rabindra 
Published: January 2023

Abstract
Cystic Fibrosis (CF) lung disease is characterized by the massive recruitment of neutrophils (PMNs) into the bronchiolar lumen. Extracellular neutrophil elastase (NE) that is released from PMN primary granules, is a strong predictor of lung function and survival in CF patients. However, the process by which CF airway PMNs exocytose HNE is not well understood and not reproduced in animal models of CF airway disease. Methods: We used a surface collagen-coated 3D porous polystyrene scaffold (Alvetex) to mimic the airway lamina propria and grew human bronchiolar H441 cells at air-liquid interface on top. In the transepithelial migration assay, blood PMNs were placed on the basal side and allowed to migrate towards to CF airway fluid placed apically, after which PMNs were analyzed by flow cytometry for rate of migration, survival, and degranulation. Results: Using this model we show that (i) CF airway fluid induces rapid transepithelial migration and survival of neutrophils, (ii) live PMNs actively release of primary granules upon exposure to CF airway fluid, and (iii) migration towards CF airway fluid induces PMN reprogramming allowing for the activation of the inflammasome and pinocytosis. Conclusions: These results suggest a primary role for the CF microenviroment in inducing increased survival and phenotypic reprogramming of PMNs and establish our model as a robust platform for deconstructing CF airway PMN dysfunction and developing new therapies for CF inflammation.
Alternatively activated macrophages attenuate influenza-induced acute lung injury in mice heterozygous for the F508del mutation in CFTR. (INC6P.314)
Authors: Woods Parker , Davis Ian 
Published: January 2023

Abstract
Infection with influenza A virus can result in acute lung injury (ALI), characterized by pulmonary edema and hypoxemia. We found that influenza-induced ALI in C57BL/6 (WT) mice was associated with increased bronchoalveolar epithelial cell Cl- secretion via the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel. Influenza-induced ALI was highly attenuated in C57BL/6-congenic mice that are heterozygous for the F508del CFTR mutation (HETs), which exhibit a 50% reduction in CFTR expression and CFTR-mediated Cl- transport. This beneficial phenotype was alveolar macrophage (AM)-dependent and was not due to differences in viral replication between mouse strains. The aim of this study was to evaluate the role of HET AMs in the attenuation of influenza-induced ALI. We hypothesized that ALI attenuation in influenza-infected HET mice results from “alternative” activation (M2 polarization) of HET AMs in response to infection. Compared to WT AMs, Arginase 1 (Arg1) protein expression was higher while inducible nitric oxide synthase (iNOS) protein was lower in HET AMs following infection. Arg1 enzymatic activity was also higher in HET AM lysates. BALF from HET mice contained higher levels of urea and lower levels of nitrate/nitrite (end-products of Arg1 and iNOS, respectively) than WT. These findings indicate that reduced CFTR expression/activity promotes an anti-inflammatory, M2 AM polarization response to influenza infection, which contributes to ALI attenuation.
Human CCL1 antisense oligodeoxynucleotide (ODN) screened as a potent inhibitor of M2b monocytes (INC6P.322)
Authors: Ito Ichiaki , Bhopale Kamlesh , Nishiguchi Tomoki , Loucas Bradford , Suzuki Sumihiro , Kobayashi Makiko , Suzuki Fujio 
Published: January 2023

Abstract
In a murine model of γ-irradiation, bacterial translocation and subsequent sepsis have been effectively controlled by CCL1 antisense ODN, a M2b macrophage reverter. In the present study, we tried to obtain human CCL1 antisense ODN which is inhibitory on human M2b macrophages. Twenty human CCL1 antisense ODNs were synthesized based on the prediction of CCL1 mRNA secondary structure, calculation of GC content, and binding energy of ODNs. To evaluate the activity of these ODNs, an amount of CCL1 in culture fluids of M2b monocytes cultured with the ODNs was determined by ELISA. M2b monocytes were generated in vitro from healthy donor peripheral blood monocytes after stimulation with IL-1β and IgG in combination. In the results, 2 ODNs (HCA-15 ; CAACATCTGGAGAAGGGT : HCA-17 ; GGGCTGTGGTGATGATC), at doses of 20 and 100 µg/ml, suppressed 70-80% or more of CCL1 production by M2b monocytes. Also, decreased expression of IL-10 and CD163 was shown in M2b monocytes cultured with HCA-15 and HCA-17. The other 18 ODNs were not significantly active on the M2b macrophage reversion. These results suggest that HCA-15 and HCA-17 may be beneficial in controlling bacterial translocation and subsequent sepsis in patients with γ-irradiation.