bluuebunny/crossref_metadata_2025_split huggingface dataset:
FOXG1 is required in thymic epithelial cells for normal development and expression of interleukin 7 (HEM4P.245)
Authors: Jarvis Barton , Wei Qiaozhi , Condie Brian 
Published: January 2023

Abstract
Thymic epithelial cells (TECs) are required in the vertebrate thymus microenvironment for normal differentiation of T-cells. Cortical (cTEC) and medullary TEC (mTEC) are the two subtypes of TECs and each component performs different functions in the production of T-cells. The regulatory pathways that control the specification and differentiation of TEC during development are not completely known. Foxg1 has previously been shown to be expressed in TEC before the onset of differentiation. Analysis in mutants showed that specification and differentiation is initially normal but that differentiation of TEC becomes abnormal at E13.5-E14.5. Further analysis of Foxg1 mutants has shown that the expression of the essential cytokine IL-7 is altered. Postnatal examination of Foxg1 conditional mutants has shown that many of the fetal phenotypes persist into adulthood which suggests a potential role for Foxg1 in TEC maintenance. Our results suggest that Foxg1 is an upstream regulator of IL-7 expression in addition to its role in the development and survival of TEC.
Successful β-selection depends on TCF-1 (HEM4P.228)
Authors: Harly Christelle , Yang Qi , Bhandoola Avinash 
Published: January 2023

Abstract
The transcription factor TCF-1 is expressed very early during thymic development and is important for early T cell specification. TCF-1 expression further increases after b-selection and is maintained at high levels in differentiated T cells, but its function at these later stages has not been examined. We used mice with conditional alleles of TCF-1 (TCF-1f/f) to delete TCF-1 at two time points of T cell development. Whereas deletion of TCF-1 in double-positive thymocytes (CD4-cre TCF-1f/f) had little impact on later T cell populations, the effect of the deletion at double-negative 3 (DN3) stage (CD2-cre TCF-1f/f) was dramatic. Thymic cellularity of CD2-cre TCF-1f/f mice was decreased more than 10-fold compared to littermates control, and competitive BM chimeras clearly revealed a sharp defect at the b-selection checkpoint in these mice. Analysis of TCF-1-deleted DN3 thymocytes did not reveal any defects in T cell gene expression and pre-TCR expression and signaling. However, post b-selection DN3 and DN4 thymocytes failed to display the increase in cell size and proliferation characteristic of successful b-selection. Because these two phenomena have been previously attributed to c-Myc, we examined its expression and found that c-Myc protein is reduced in TCF-1-deleted post b-selection DN3. We are currently examining the mechanism by which TCF-1 controls c-Myc. Our work to date implicates TCF-1 downstream of the pre-TCR during the development of αβ T cells.
Aging of thymic epithelial progenitor pool is determined by the p63-FoxN1 regulatory axis (HEM4P.233)
Authors: Burnley Preston , Ruan Linhui , Coder Brandon , Su Dong-Ming 
Published: January 2023

Abstract
The postnatal thymic epithelial progenitor (TEP) pool is proposed to be regulated by the p63 and FoxN1 genes through proliferation and differentiation, respectively. However, the combined role of these two genes in the aging TEP is still a mystery. Evidence from murine models has elucidated contrasting roles of the p63 isoforms during the aging process. We found that TAp63+, but not ΔNp63+, thymic epithelial cells (TECs) were increased with age, accompanied with increased senescence associated β-gal clusters and p21+ TECs. Senescent clusters also developed after intrathymic infusion of exogenous TAp63 cDNA into young wild-type mice. Using our conditional FoxN1 gene knockout mouse model to disrupt TEP differentiation accelerated this senescent phenotype to early middle age. However, upon infusion of exogenous FoxN1 cDNA into aged wild-type mice resulted in only an increase in ΔNp63+ TECs, but no change in TAp63+ TECs in the partially rejuvenated aged thymus. Interestingly, using a novel FoxN1 transgenic mouse model to enhance TEP differentiation, ΔNp63+ TECs were decreased in young thymus. Additionally, the TAp63+ population contained a high percentage of phosphorylated-p53 and apoptotic TECs, but showed no changes in BrdU-labeled proliferation. As a result, FoxN1 controlled TEC differentiation as a bottleneck to determine TEP pool via affecting TAp63 and ΔNp63 levels. Thus, TEC homeostasis during aging has been determined through the p63-FoxN1 regulatory axis.
PLZF null mice have reduced type 2 pulmonary inflammation due to defective ILC2 effector function (HYP6P.265)
Authors: Verhoef Philip , Constantinides Michael , McDonald Benjamin , Sperling Anne , Bendelac Albert 
Published: January 2023

Abstract
Rationale. Pulmonary ILC2s have been recently identified as a source of IL-5 and IL-13 during allergic inflammation and helminth infection. Fate-mapping experiments in our lab revealed that the transcription factor PLZF is upregulated during ILC2 development. We hypothesized that the PLZF null mouse would have an impaired innate type 2 response. Methods. Type 2 pulmonary inflammation was induced by intranasal treatment of PLZF null mice and wild-type littermates with either the protease papain, the cytokine IL-25, or saline for 3 days, followed by bronchoalveolar lavage, cellular isolation, and cytokine assays. PLZF null-wild type mixed bone marrow chimeras were generated to determine if the defects in the type 2 response were cell-intrinsic. Nippostrongylus brasilensis infection was used as a physiologic type 2 inflammatory stimulus. Results. While ILC2s were present in PLZF null mice, pulmonary eosinophilia and type 2 cytokine levels were both significantly reduced in response to papain, IL-25, or Nippostrongylus infection. Mixed chimera experiments indicated that these defects were intrinsic to ILC2s: fewer PLZF-null-derived pulmonary ILC2s were reconstituted and they were less able to produce type 2 cytokines, while eosinophils from wild-type or PLZF null mice responded comparably. Conclusion. Mice lacking PLZF have a defective type 2 immune response in response to various stimuli, due to impaired ILC2 effector function.
Doxycycline exerts multiple anti-allergic effects in vitro and in vivo though modulating PI3K/Akt pathway (HYP6P.269)
Authors: Zheng Song Guo , Su Wenru , Chen Sangxia , Liang Dan 
Published: January 2023

Abstract
Tetracyclines, belonging to an old antibiotic drug family that possesses a striking variety of non-antibiotic properties, have been successfully applied in a wide range of diseases, including asthma. However, their roles in allergic diseases and underlying anti-allergic mechanisms remain elusive.To explore the roles of doxycycline in allergic pathogenesis, we have focused on its role on B cells, mast cells (MCs) and histamine function using and experimental allergic conjunctivitis (EAC) and passive systemic anaphylaxis (PSA) animal models. Our results demonstrated that treatment with doxycycline significantly reduced IgE release of B cells and IgE-mediated activation of MCs. Furthermore, doxycycline treatment significantly inhibited histamine-stimulated vascular hyperpermeability. Mechanistically, doxycycline-mediated inhibition of B cells, mast cells and histamine may be through modulating PI3K/Akt pathway. Using mouse models of EAC, PSA and histamine-induced systemic anaphylaxis like reaction (HISA), we demonstrated that treatment with doxycycline significantly attenuated allergic symptoms. Doxycycline-mediated anti-allergic effects on EAC, PSA and HISA were abrogated when an Akt activator, SC79 was administrated. Our study has suggested that doxycycline inhibits B cell, mast cell and histamine function and attenuates experimental allergic conjunctivitis and systemic anaphylaxis by modulating PI3K/Akt pathway.
Responsiveness to anaphylatoxin C5a delineates hematopoietic lineage of pulmonary dendritic cells (HYP6P.257)
Authors: Nakano Hideki , Moran Timothy , Bortner Carl , Cook Donald 
Published: January 2023

Abstract
The complement subunit, C5a (anaphylatoxin), can either promote or suppress allergic responses through its effects on pulmonary dendritic cells (DCs). However, DCs are heterogeneous, and responsiveness of specific DC subsets to anaphylatoxins has not been well characterized. Two distinct precursors can give rise to DCs, including fms-like tyrosine kinase receptor 3 (FLT3)-dependent preDCs and FLT3-independent monocytes. The integrins CD11b and CD103 are widely used to identify DC subsets in the lung, but CD11bhi DCs are composed of both preDC-derived conventional DCs (cDCs) and monocyte-derived DCs (moDCs), including Ly-6Chi inflammatory DCs and Ly-6CloCD64hi (also CD14hi) resident moDCs. Microarray analysis of FLT3-dependent CD11bhi cDCs and FLT3-independent CD11bhi moDCs revealed that C5aR/CD88 is highly expressed by moDCs. Flow cytometric analysis confirmed that CD88 is displayed on Ly-6Chi inflammatory DCs and CD64hi resident moDCs, but not on CD103+ and CD64lo cDCs. CD88hi lung DCs were FLT3-independent, and adoptively transferred monocytes differentiated exclusively to CD88hi DCs. Calcium-influx analysis of purified lung DCs upon C5a stimulation confirmed that only Ly-6Chi inflammatory DCs and CD64hi moDCs, but not CD103+ DCs and CD64lo cDCs respond to C5a. Our findings demonstrate that CD88 can reliably distinguish moDC from cDCs, and that only moDCs are directly stimulated by anaphylatoxin C5a.
Changes of cell surface markers CD3, CD4, CD44, CD62L on mouse spleen lymphocytes after culture in vitro (HYP7P.303)
Authors: Li Yanchun , Hua Shucheng , Gao Hang , Li Wenxue , Zhu Yaping 
Published: January 2023

Abstract
Object To Study the changes of cell surface markers CD3, CD4, CD44 and CD62L on mouse spleen lymphocytes after culture. Method Spleen cells were separated from Balb/c mice and asthmatic mice and lymphocytes were isolated by using lymphocyte separation liquid. We cultured lymphocytes with RPMI 1640 medium(10% FBS) in cell incubator for 3 days. Then the four cell surface markers were detected by using flow cytometry. Results In normal mice, 19.09% of lymphocytes stained positively for CD3 and CD4, among these cells 98.61% was CD44+ T cell and 68.71% was CD62L+ T cell before the culture. After culture with ConA for 3 days, CD3+CD4+ and CD3+CD4+CD44+ T cell population was down regulated to 8.96% and 71.82%, respectively. While CD62L almost disappeared, accounting for 11.27% only. The similar results were obtained even cells cultured without ConA present. For asthmatic mice, CD3+CD4+ T cell accounted for 20.33% in total lymphocytes, among which 97.72% was CD44+ and 75.74% was CD62L+ before the culture. After culture, CD3+CD4+T cell accounted for 10.4%, among which CD44+ only 55.5% and CD62L+ only 2.63%. The similar results were obtained when cells were cultured with OVA. Conclusion In both normal and asthmatic mice, CD3,CD4,CD44 and CD62L on spleen lymphocytes were down regulated after culture in vitro regardless with or without stimulating factors, especially CD62L, almost disappeared. This should be given full consideration when these markers are detected after cell culture.
Effect of extrusion processing on in vivo allergenicity of hazelnut in an adjuvant-free mouse model (HYP7P.313)
Authors: Ortiz Tina , Para Radhakrishna , Gonipeta Babu , Ravi Ramasamy , Reitmeyer Mike , He Yingli , Srkalovic Ines , Ng Perry , Gangur Venugopal 
Published: January 2023

Abstract
Life-threatening nut allergy is a growing public health problem in many countries including the USA. Causes underlying this alarming trend are incompletely understood and methods are urgently needed to prevent such immune reactions. Here, we tested the hypothesis that extrusion processing of hazelnuts will reduce in vivo allergenicity using an adjuvant-free mouse model of hazelnut allergy established in our lab. Groups of mice received transdermal exposure (TDE) to extrusion processed hazelnut protein (EHNP) versus raw hazelnut protein (RHNP) extract preparations. Mice were evaluated for systemic IgE antibody responses (sIgER), hypothermia shock response (HSR), and mucosal mast cell responses by measuring plasma mouse mast cell protease-1 (mMCP-1) levels. Results showed that EHNP elicited a reduced sIgER compared to RHNP after 4 TDE but similar responses after 5 TDE. However, EHNP induced less HSR compared to RHNP upon oral challenge. Both protein preparations induced similar HSR upon i.p., challenge. Finally, we measured plasma mMCP-1 levels after oral and i.p., injection challenges. We found that EHNP elicited reduced mMCP-1 response compared to RHNP upon both oral as well as i.p., challenges. This is the first study demonstrating the utility of a mouse model to evaluate the effect of food processing on in vivo food allergenicity in general and hazelnut in particular.
Role of α-melanocyte stimulating hormone in the induction of regulatory T cells in ocular allergy (HYP7P.318)
Authors: Galicia-Carreón Jorge , Rodríguez-Lemus Karen Alejandra , Ayala Balboa Julio , Pérez-Tapia Sonia Mayra , Jimenez-Martinez Maria 
Published: January 2023

Abstract
Allergic conjunctivitis (AC) is one of the most common eye disorders in ophthalmology. AC is a disorder mediated by Th2 cells, where the eye inflammation is caused by a type I hypersensitivity. It has been suggested that some neuropeptides are increased (CGRP, SP and VIP) in AC contributing to local damage. In other hand, α-MSH plays an immunomodulatory role in ocular tissues; however, its role in ocular allergic disease is unknown, thus it was the aim of this study. Methods: Peripheral blood mononuclear cells (PBMC) were isolated from blood samples of healthy donors (HD) and AC-patients skin reactive to Dermatophagoides pteronyssinus (Der p), then PBMC were cultured with different stimuli: Der p, α-MSH, or Der p plus α-MSH. After 7 days, PBMC were harvested, labeled with mAbs against CD4-PECy5, CD25-PE, FOXP3-FITC and analyzed by flow cytometry. Results. Der p or α-MSH stimulation did not induced significant changes in the frequency of Tregs; interestingly, after α-MSH plus Der p stimulation a significant higher frequency of Tregs cells was observed than Der p alone. Conclusions: Tregs differentiation in patients with AC is dependent of Ag and addition of α-MSH, if α-MSH is involved in local Treg differentiation is not known and needs further investigation.
Alpha-tocopherol inhibits the development of select dendritic cell subsets and lung inflammation during allergic responses in neonates (HYP7P.282)
Authors: Cook-Mills Joan , Abdala-Valencia Hiam , Berdnikovs Sergejs , Soveg Frank 
Published: January 2023

Abstract
Offspring from allergic mothers develop allergic responses to suboptimal allergen challenge. This is mediated by dendritic cell responses in the offspring. Dendritic cell development and responses are dependent on protein kinase Cα (PKCα) and we reported that recombinant PKCα is inhibited by binding of the vitamin E isoform α-tocopherol (αT). However, it is not known whether αT regulates development of dendritic cell subsets or development of allergic responses. Therefore, we determined whether αT supplementation inhibited development of dendritic cell subsets and allergic responses in offspring of allergic female mice. αT supplementation of allergic female mice during pregnancy/lactation resulted in a dose-dependent reduction in allergic lung inflammation in the pups challenged with the antigen OVA. Interestingly, in these pups, there was a reduction in lung CD11b+ but not CD11b- subsets of dendritic cells. There was also a reduction in allergen-induced lung mRNA expression of indoleamine 2,3-dioxygenase, IL-4, IL-33, TSLP, CCL11 and CCL24. In conclusion, supplementation with αT reduced development of CD11b+ dendritic cells and allergic responses in offspring from allergic mothers. These results have implications for supplementation of allergic mothers with αT.
In vivo bisphenol-A exposure in combination with high-fat diet modulates murine innate immune responses (INC6P.345)
Authors: Walser-Kuntz Debby , Cho John , Rab Rashed , Wu Ivy , Iroz Cassandra , Kansakar Shamir 
Published: January 2023

Abstract
The recent obesity epidemic has led to an expansion in our understanding of the link between adipose tissue and inflammation. The prevalent use of bisphenol-A (BPA) in food containers and other products provides an additional environmental challenge to the immune system. BPA disrupts the action of physiologic estrogens by binding to estrogen receptors and has previously been shown to impact immune responses. We explored the combined effects of high-fat diet and in vivo BPA exposure on spleen innate immune mediators. BPA treatment (2.5 or 25.0 ug/kg body weight per day; 4 weeks) reduced the LPS-induced secretion of KC chemokine, a neutrophil chemotaxin, by 25% and 40% respectively in normal diet mice, with a more dramatic BPA-induced decrease in KC secretion observed in high-fat fed mice. High-fat diet increased TNF-α mRNA expression overall compared to normal diet, and high dose-BPA reduced LPS-stimulated TNF-α mRNA expression in both normal (19%) and high-fat fed (34%) mice. High-fat fed mice also had a greater increase in LPS-induced COX-2 mRNA expression than normal diet mice, with BPA modulating expression in both groups in a dose dependent manner. High-fat diet and low dose BPA exposure increased non-fasting blood glucose levels by 30% on average when compared to control normal diet mice. These data demonstrate that peripubertal BPA exposure dampens aspects of innate immune function and that BPA combined with high-fat diet may disrupt glucose metabolism.
Differential regulation of CD8 T cell responses by surface and endosomal TLRs (INC6P.347)
Authors: Mandraju Rajakumar , Murray Sean , Forman James , Pasare Chandrashekhar 
Published: January 2023

Abstract
Sensing of pathogen/pathogen derived products through Toll like receptors (TLRs) induces dendritic cell (DC) maturation characterized by the expression of co-stimulatory molecules and secretion of pro-inflammatory cytokines. These two signals play a critical role in activation and differentiation of CD4 T cells. However, since the role of TLRs in the regulation of CD8 T cell responses is not clear we sought to address their role in this process. Our results show that although most TLRs induce the CD8 T cell proliferation in-vitro, there are significant differences in the CD8 T cell priming in-vivo to pathogenic infections. Endosomal TLRs (TLR3 and TLR9) that sense nucleic acids show a remarkable ability to induce CD8 T cell responses. However, surface TLRs (TLR2 and TLR4) that sense bacterial ligands were incapable of inducing CD8 T cell responses. Moreover, surface TLRs dominantly suppress the CD8 T cell activation induced by endosomal TLRs. Cell migration experiments show that surface TLR activation skewed lymphoid (CD11c+, CD8+) to myeloid (CD11c+, CD11b+ CD8-) DC populations in the draining lymph nodes. We also found that DC intrinsic TLR2 and TLR4, acting in a MyD88-dependent manner, altered the ability of DCs to prime CD8 T cells. In summary, our results demonstrate that bacterial co-infection dampens the ability of DCs to induce efficient anti-viral CD8 T cell responses.
Tetramer identification of functional mouse mucosal associated invariant T cells (INC6P.352)
Authors: Sakala Isaac , Kjer-Nielsen Lars , Eickhoff Christopher , Wang Xiaoli , Liu Ligong , Fairlie David , Rossjohn Jamie , Hoft Daniel , Hansen Ted 
Published: January 2023

Abstract
Mucosal associated invariant T (MAIT) cells have a semi-invariant TCR Vα chain, and their development is dependent upon the commensal flora and the expression of MR1. MAIT cells are activated in vitro in an MR1-restricted manner by cells infected with diverse strains of bacteria suggesting a widely shared antigen. Such an antigen was recently defined by the observation that human MR1 binds bacterial riboflavin metabolites (ribityllumazines, RL antigens) that are capable of activating MAIT cells. Recently MR1/RL tetramers were constructed allowing us to compare MR1-dependency and function of mouse MAIT cell subsets in invariant TCR Vα19-Jα33 (Vα19i) transgenic (Tg) mice. Although significant numbers of tetramer+ cells were detected in both MR1+/+ and MR1-/- Vα19i Tg mice, only tetramer+ cells from MR1+/+ and not MR1-/- Vα19i Tg mice had predominant expression of CD69, NK1.1 and Vβ6/8 and displayed robust in vitro responses to IL-12+IL-18 or RL Ag. In addition, tetramer+ MAIT cells expressing CD4, CD8 or neither developing in MR1+/+ Vα19i Tg mice had disparate cytokine profiles in response to RL Ag. Most notably, after mycobacterial infection all MAIT cell subsets decreased in the blood regardless of whether they express NK1.1 and/or Vβ6/8; however, only the NK1.1+Vβ6/8+ subset predominated in the lung airways of infected MR1+/+ Vα19i Tg mice. These findings present evidence for the functional diversity of MAIT cell responses to innate cytokines, RL Ag and bacterial infection.
IFNγ drives human primary skin mast cells to function as antigen presenting cells. (INC6P.341)
Authors: Lotfi-Emran Sahar , Ward Brant , Woodfolk Judith , Manjili Masoud , Schwartz Lawrence 
Published: January 2023

Abstract
We assessed the antigen presenting ability of mature primary human skin mast cells (MCs) in vitro. MCs do not express HLA II at baseline, but do so when stimulated with IFNγ. IFNγ-primed, but not un-primed, MCs facilitated activation of Jurkat T cells by superantigen. Unlike polyclonal primary CD4+ T cells, which harbor a subset of IFNγ producing cells, Jurkat cells do not spontaneously produce IFNγ. Co-culture of un-primed MCs with CD4+ T cells for 2 days upregulated HLA II expression on MCs and subsequent superantigen mediated T cell activation. MC HLA II upregulation is primarily due to the Th1 cell subset and is prevented by IFNγ neutralizing and/or receptor blocking antibody. Superantigen bypasses HLA II-loaded peptide:T cell receptor interactions; therefore, we assessed the ability of MCs to present cytomegalovirus antigen. MCs primed ±cytomegalovirus antigen exposure for 3 days then ±IFNγ for the last 2 of those days were washed thoroughly and co-cultured with HLA-matched CD4+ cells. After one week, CD4+ T cells from cytomegalovirus seropositive, but not seronegative, individuals showed proliferation (CFSE dilution), activation (CD25high and CD69high expression), and appropriate intracellular cytokine production (IFNγ and IL-4, but not IL-17). Thus, MCs can take up, process, and present antigen in an HLA II-restricted manner, causing antigen specific activation and proliferation of CD4+ T cells.