bluuebunny/crossref_metadata_2025_split huggingface dataset:
Regulation of IL-4 expression by aspirin via a MAPK pathway (139.15)
Authors: Kim Byung Soo , Uhm Tae Gi , Kang Jin Hyun , Park Choon-Sik , Chung Il Yup 
Published: January 2023

Abstract
IL-4, a pleiotropic cytokine produced in Th2 cells, plays a central role in the pathological development of asthma. Aspirin, a widely used nonsteroidal anti-inflammatory drug (NSAID), is reported to have inhibitory effect on IL-4 expression in activated T cells and paradoxically to induce airway hyperreactivity in a subset of asthmatic patients, causing so-called aspirin-intolerant asthma (AIA) by completely unknown mechanism. Here we show the effects of aspirin on IL-4 expression in Jurkat T cells and K562 cells that are shown to be able to express IL-4 mRNA. qPCR analysis revealed that aspirin induced IL-4 mRNA expression in both cell lines. In agreement, aspirin also transactivated IL-4 promoter. Analysis of IL-4 promoter deletion constructs showed that the asprin-mediated IL-4 induction was not attributable to any discrete motif in the IL-4 promoter: all the deletion constructs exhibited the increased reporter activities in response to aspirin. Aspirin was able to activate p38 and ERK1/2, and specific inhibitors for these kinases abolished aspirin-mediated IL-4 induction and IL-4 promoter activity. Aspirin-induced IL-4 expression might implicate airway hyperreactivity found in AIA.
Adenosine receptor A2a inhibits complement-mediated activation of human mast cells by activating Gαs-proteins (139.5)
Authors: Kulka Marianna , Sheen Cecilia , Tancowny Brian P , Schleimer Robert P 
Published: January 2023

Abstract
The complement anaphylatoxin 3a (C3a) and adenosine receptors (AR) are implicated in the inflammatory process in allergic rhinitis although a direct interaction between these pathways has not been demonstrated. To investigate the interaction between these pathways, primary human cultured mast cells (HuMC) and human mast cell line (LAD2) were stimulated with C3a with or without AR agonists and antagonists. The non-selective AR agonist, 5'-N-ethylcarboxamidoadenosine (NECA; 30 uM), inhibited C3a-mediated HuMC and LAD cell degranulation (43 + 4 and 40 + 5 % inhibition respectively compared to control). NECA also blocked C3a-activated production of MCP-1 (57 + 6 %) and C3a-activated chemotaxis (46.7 + 10 %). A selective A2aR agonist, CGS 21680, inhibited C3a-mediated degranulation but the A3R agonist, IB-MECA, had no effect suggesting that inhibition of degranulation was mediated by A2aR. An A2aR antagonist, SCH 58261, blocked the inhibitory effect of NECA but an A2bR and A3R antagonist had no effect. Real-time PCR analysis showed that LAD2 and HuMC expressed mRNA for A2aR, A2bR and A3R but not A1R. Measurements of intracellular cAMP showed that NECA elevated [cAMP]i levels by at least 30% even in C3a-activated cells. Pertussis toxin blocked C3a-activated degranulation. The adenylyl cyclase inhibitor, SQ 22536, had no effect on C3a-activated degranulation but blocked the effect of NECA demonstrating that C3a and NECA mediate their effects through Gαi and Gαs proteins respectively. These results show that adenosine inhibits complement activation of human mast cells through a Gαs-protein pathway.
Funded by a grant from the Ernest S. Bazely Trust and intramural NRC funds
Approaches Against Dust Mite- induced Asthma Through Adaptive T Regulatory Cells (140.12)
Authors: Jin Jin , Hu Yanxin , Liu Qiang , Wang Bin 
Published: January 2023

Abstract
Asthma is a long-lasting lung disease that causes breathing problems. 50 to 80 % of asthma cases may be caused by dust mites and their droppings. This severe reaction is initiated by the allergen-specific T cell responsiveness. Effective antigen-specific treatments or preventions are still lacking. Using a murine asthma model induced by dust mite extract, we first report that co-immunization of DNA and protein vaccines encoding the dust mite specific antigen (Der p1) can prevent allergic responses, including the antigen specific IgE level and infiltration of lymphocytes and mast cells to the site of the allergen challenge. The prevention of Der p1induced asthma is directly related to the induction of a population of Der p1 specific iTreg cells exhibiting a CD4+/CD25-/FoxP3+ phenotype. This subtype of iTreg cells express IL-10 and TGF-beta, but not CTLA-4 and CCR7 which are well-known markers of natural nTreg cells. Adoptive transfer of the iTreg cells is also able to suppress the Der p1-induced asthma, suggesting the inhibition potency of the iTreg cells directly on the asthma causing T cells. The study demonstrates that utility of the co-immunization strategy may present a prophylactive approach clinically and offer us a novel strategy to prevent other allergic and asthma diseases in human and animals.
INTRANASAL CpG THERAPY ATTENUATED EXPERIMENTAL FUNGAL ASTHMA IN A TLR9-DEPENDENT AND INDEPENDENT MANNER (140.4)
Authors: Ramaprakash Hemanth , Hogaboam Cory M. 
Published: January 2023

Abstract
CpG-ODN administration abolishes airway inflammation and remodeling in acute models of allergic airway disease. Herein, we investigated the therapeutic effect of CpG-ODN in a chronic fungal model of asthma. TLR9+/+ and TLR9-/- mice were sensitized to soluble A. fumigatus antigens and challenged with live A. fumigatus conidia. Mice were treated with intraperitoneal (IP) CpG, intranasal (IN) CpG, or left untreated from days 14 to 28-post conidia challenge. All features of allergic airway disease were attenuated in TLR9+/+ mice treated with IN CpG, including airway hyperresponsiveness (AHR), mucus production, and peribronchial fibrosis. TLR9-/- mice treated with IN CpG exhibited attenuated airway remodeling, but not AHR. Whole-lung IL-12 levels were significantly elevated in both TLR9+/+ and TLR9-/- mice receiving IN CpG but not in either group receiving IP CpG. Whole-lung IL-10 levels were significantly elevated in IN CpG-treated TLR9+/+ mice but not in TLR9-/- mice receiving IN CpG. Increased whole-lung transcript and protein levels of the scavenger receptors SR-A and MARCO were observed in TLR9-/- mice compared with TLR9+/+ mice, possibly accounting for the CpG responsiveness in the knockout group. Together, these data show that IN CpG has a therapeutic effect during established fungal asthma, which is TLR9-dependent and independent.
This work was financially supported, in part, by the NIH (grant HL069865 to CMH).
Allergic sensitization in US bakery workers (140.9)
Authors: Biagini Raymond  E. , Page Elena H. , Dowell Chad R. , MacKenzie Barbara  A. , Snawder John E. , Sammons Debbie L. 
Published: January 2023

Abstract
As part of a health hazard evaluation (HHE), the National Institute for Occupational Safety and Health (NIOSH) performed a study at a commercial bakery designed to evaluate allergic sensitization to bakery associated allergens (BAA; alpha-amylase, [AA]; wheat, [W], or flour dust, [FD]). Sera was obtained from 96 employees, dichotomized as having low exposure (N=33) or high (or past) exposure (N=63) to BAA. Workers were evaluated for atopy (AlaTOP®, total IgE), and serum-specific IgE (AA, FD, and W; IMMULITE® 2000). There was no significant difference in the prevalence of atopy between groups by AlaTOP® (47% of high exposure vs. 41% of low exposure, p=0.6) or total IgE (3.9 kU/L [high exposure] compared to 3.7 kU/L [low exposure], p=0.5). At an IMMULITE® specific IgE cutoff of &gt; 0.35 kU/L, there was significantly greater prevalence of positive specific IgE to W. When an IMMULITE® positive cutoff of &gt;0.10 was used, significantly greater prevalence (p&lt;0.05) of specific IgE to FD, AA, and W was observed in the high vs. low exposure groups. Atopics had significantly higher rates of sensitization to W and FD at both the 0.1 and 0.35 kU/L levels, and to AA at the 0.1 level. Exposure to BAA puts workers at a higher risk for sensitization to BAA (specific IgE). As has been shown for other occupational allergens (Annals of Allergy, Asthma, & Immunology, 97:196-202, 2006), using the IMMULITE® at a cutoff of 0.10 kU/L vs. 0.35 kU/L appears to increase the sensitivity of the assay.
Natural Killer cell interactions with dendritic cells and tumor cells are mediated by different NK cell receptors (141.10)
Authors: Wai Lu-En , Martinez Olivia M , Krams Sheri M 
Published: January 2023

Abstract
NK cells infiltrate solid organ allografts early post-transplant. As NK cells can mediate both maturation and killing of dendritic cells (DC), a better understanding of NK-DC interactions after transplant is warranted. This study investigates the interaction of NK cells with DC and tumor cells, and the specific NK cell activation receptors (NKRs) involved. A rat NK cell line was co-cultured with rat bone marrow-derived DC and rat hepatoma McA-RH7777 (McA). Co-culture of NK cells with DC and McA resulted in significant IFNγ production. This IFNγ production required cell contact suggesting that specific NKRs are involved. To specifically address this possibility, the NKRs, NKp30, NKp46, NKG2D, were knocked down using RNAi.
NK cell activation by DC is mediated by NKp46 as knockdown of this receptor markedly decreased IFNγ production. In contrast, NKp30, not NKp46, was responsible for NK cell activation by tumor. Flow cytometry on DC and McA with NKR fusion proteins indicated the presence of putative ligands. Interestingly, individual NKR knockdown did not markedly diminish NK cell killing suggesting NK cells use multiple receptors and/or other mechanisms to kill cells. In conclusion, both DC and tumor activate NK cells but NKp46 and NKp30 mediate these interactions respectively. These data suggest a means to manipulate specific NK cell interactions after transplantation.
Funding: NIH AI44095, and Agency for Science, Technology and Research (Singapore).
Differences in Bcl-2 Expression by T Cell Subsets Alter Their Balance After In Vivo Irradiation to Favor Regulatory NKT Cells and CD4+ CD25+ T Cells in Wild Type But Not p53-/- Mice (141.19)
Authors: Yao Zhenyu , Jones Jennifer Clare , Liu Yinping , Strober Samuel 
Published: January 2023

Abstract
A more nuanced analysis of the changes in the balance of immune cell subsets is needed to understand the impact of radiation on immune function. We show the balance of T cell subsets changes after increasing single doses of total body irradiation(TBI) or after fractionated irradiation of the lymphoid tissues(TLI) of mice. These changes are due to differences in radioresistance and Bcl-2 expression of the NKT, CD4+CD25+ Treg and conventional T cell subsets to favor CD4+Bcl-2hi NKT and CD4+CD25+Bcl-2hi Treg cells. Reduction of the Bcl-2lo mature T cell subsets was at least 100 fold greater than that of the Bcl-2hi subsets. CD4+ NKT cells upregulated Bcl-2 and developed a Th2 bias after irradiation, while conventional T cells failed to do so. GVHD was more severe after bone marrow transplantation in NKT cell deficient than in wild type hosts and the residual NKT cells were 10 fold more abundant in the wild type spleen after the TLI versus TBI. Although the changes in the balance of T cell subsets and increase in Bcl-2hi cells occurred in irradiated wild type mice, these changes did not occur in p53-/- mice. In conclusion, differences in the level of Bcl-2 and associated differences in resistance of T cell subsets to radiation induced p53 dependent apoptosis results in a marked alteration in the balance of T cell subsets favoring NKT cells and CD4+CD25+ T cells that promote transplantation tolerance.
CCL22 protection of islet allografts in diabetes (141.32)
Authors: Montane Joel , Bischoff Loraine , Soukhatcheva Galina , Tan Rusung , Verchere C Bruce 
Published: January 2023

Abstract
CD4+CD25+ T regulatory (Treg) cells prevent spontaneous development of autoimmune diabetes and may delay islet allograft rejection. Tumour cells attract Treg cells to foster immune privilege by producing the chemokine CCL22, which promotes trafficking of Treg cells to the tumour. Therefore, manipulating Treg migration with CCL22 may provide a means of preventing islet transplant rejection in autoimmune diabetes. To drive islet-specific expression of CCL22 we created an adenovirus (Ad-CCL22) in which a mouse CCL22 cDNA is expressed downstream of the CMV promoter. Isolated islets transduced with Ad-CCL22 (10 MOI) produced and secreted high levels of CCL22 in vitro whereas CCL22 production by non-transduced islets was undetectable. To determine whether CCL22 expression prevents islet allograft rejection, donor Balb/c islets were isolated and transduced overnight with Ad-CCL22 (n = 6) or Ad-lacZ (n = 5). Islets (300 per recipient) were then transplanted into the left renal subcapsular space of age-matched female C57/BL6 diabetic recipients. By 5 weeks post-transplant, only 1 of 7 CCL22-expressing islet grafts had been rejected whereas 4 of 5 Ad-LacZ-transduced grafts were rejected. Induced expression of CCL22 by transplanted islets thus delays islet allograft rejection. Manipulation of Tregs cells by CCL-22 in transplanted islets may be a novel therapeutic strategy for diabetes.
Induced Tolerance to Liver Allografts Involves Apoptosis and the Generation of Regulatory T Cells (141.40)
Authors: Fujiki Masato , Esquivel Carlos O , Martinez Olivia M. , Strober Samuel , Krams Sheri M. 
Published: January 2023

Abstract
Post-transplant total lymphoid irradiation (TLI) is a non-myeloablative regimen that has been studied in rodent models for induction of tolerance to allografts. We show in a rat model of orthotopic liver transplantation that 88% of Lewis recipients of DA livers (n=16) survive more than 100 days (and accept donor skin grafts) after receiving post-transplant TLI alone whereas the control group (liver transplantation without host conditioning) rejected their liver allografts within 14 days.
To establish the mechanism of this induced tolerance, lymphocytes were isolated from the livers of groups (n=3-5) of Lewis recipients of Lewis or DA liver grafts, on day 7 post-transplant, and analyzed for apoptosis. Apoptosis of T cells in allografts was increased by TLI treatment (Allo+TLI vs. Allo, CD4+ 41.5 % vs 13.4%, p=0.002, CD8+, 22.5% vs. 9.7% p=0.034). Syngeneic grafts from recipients treated with TLI had minimal apoptosis similar to the levels detected in normal livers.
Further, intragraft CD4+CD25+FoxP3+ cells were increased in the TLI group in the first week post-transplant and remained elevated in the graft and in the spleen. Importantly, adoptive transfer of splenocytes from day &gt;100 liver allograft recipients that received TLI prolonged the survival of donor, but not third-party, heart grafts in naïve recipients. Depletion of CD4+CD25+ cells from transferred splenocytes abrogated this prolongation. We conclude that post-transplant TLI significantly increases apoptosis of infiltrating T cells in the allograft during the first week post-transplant. We suggest that apoptosis of host lymphocytes helps to promote an environment whereby CD4+CD25+FoxP3+ T regulatory cells can develop and accumulate in the graft and periphery, and maintain graft non-responsiveness.
Supported in part by NIH AI044095, AI037683
Pancreatic islets engineered to display on their surface a novel form of FasL protein survive indefinitely by inducing tolerance through phagocytes/CD4+CD25+FoxP3+ Treg cell axis (141.47)
Authors: Zhao Hong , Yolcu Esma S. , Bandura-Morgan Laura , Shirwan Haval 
Published: January 2023

Abstract
Objective: Pancreatic islets engineered to display on their surface a chimeric form of FasL (SA-FasL) protein survive indefinitely in allogeneic recipients. In this study, we tested the role of phagocytes and CD4+CD25+FoxP3+ Treg cells in the induced tolerance.
Methods: To study the function of Treg cells, OT-I transgenic and rag-/- B6 mice were adoptively transferred with 1 x 106 CD4+ Teff cells from naïve B6 together with 2-5x103 Tregs from na ïve, rejected, or tolerant animals one day prior to BALB/c islet Tx. The function of phagocytes in the induced tolerance was investigated using liposome-encapsulated clodronate for depletion.
Results: OT-I mice did not reject BALB/c mice islet grafts (n= 5; &gt;100 days) due to the restricted TCR repertoire for ovalbumin. Adoptive transfer with sorted 1x106 CD4+ Teff cells from naïve B6 mice promptly rejected allogeneic islets (n=5; MST=29.0±9.2 days). Cotransfer of 2-5x103 Tregs from long-term (n=5; &gt;100 days), but not rejecting (n=5; MST=34.4±3.5 days), animals prevented graft rejection. Treatment with clodronate resulted in depletion of phagocytes and acute rejection of SA-FasL-engineered islets (n=5; MST=22.7±10.4 days). In marked contrast, 5/6 mice treated with control liposome accepted their grafts (MST&gt;100 days).
Conclusions: Tolerance to SA-FasL-engineered pancreatic islets is maintained by Treg cells that require phagocytes for induction.
NIH (R21 DK61333, R01 AI47864, R21 HL080108), ADA (1-05-JF-56), and AHA Fellowship 0725348B.
Temporal variability of cellular alloreactivity in kidney transplant candidates (141.8)
Authors: Kramer Kristen L , Siegel Hanni , Lalli Peter , Baldwin William , Fairchild Robert , Poggio Emilio 
Published: January 2023

Abstract
Assessment of preexistent cellular alloimmunity (CA) in transplant candidates aids in understanding individual immunological risk. We tested the extent of CA over time and characterized factors associated with it. Blood samples were obtained every 3 mo from 21 hemodialysis (HD) pts and 5 healthy controls over 1 yr. CA was tested by IFN- ELISPOT using a panel of 6 HLA-typed B cell lines as stimulators (panel of reactive T cells, PRT). For each time point, PRT was calculated and reactivity to &gt;75% of the panel was considered highly reactive. In HD pts, the PRT % increased over baseline with time but was stable in healthy subjects. African-American (AA) pts had significantly higher PRT values at any time point than non-AA, and pts on HD&gt;5 yrs had a trend to higher PRT values. PRT was not different based on gender or age. By logistic regression analysis, race was found to invariably be independently associated with higher PRT values at any time point (OR:1.50, p&lt;0.01), while there was a trend to higher PRT values for pts with longer HD. These data expand upon our previous reports that AA race is a significant risk factor for cellular alloreactivity, potentially accounting for the worse posttransplant outcomes in this population. Serial measurements of preexisting cellular anti-HLA immunity in transplant candidates by PRT is warranted as results can vary in an individual candidate over time.
WIP is critical for T cell responsiveness to IL-2 (33.1)
Authors: Le Bras Severine , Massaad Michel , Suresh Koduru , Kumar Lalit , Oyoshi Michiko , Hartwig John , Geha Raif S 
Published: January 2023

Abstract
The WASP-interacting protein WIP stabilizes WASP, the product of the gene mutated in Wiskott-Aldrich syndrome. WIP-deficient T cells have low WASP levels, limiting the usefulness of WIP KO mice in defining the role of WIP in T cell function. To define this role, we compared WIP/WASP double knockout (DKO) mice to WASP KO mice on DO11.10 background. T cell development was normal in both strains, but peripheral T cell numbers were significantly decreased in DKO mice. WASP KO T cells proliferated and secreted IL-2 normally in response to OVA peptide (OVAp). In contrast, T cells from DKO mice proliferated poorly in response to OVAp in vitro and cutaneous hapten hypersensitivity was deficient in these mice. DKO T cells up-regulated CD25 expression and secreted normal amounts of IL-2 after antigen stimulation, but had defective response to IL-2, evidenced by failure to further up-regulate CD25 expression, phosphorylate STAT5, and induce expression of STAT5 dependent genes. DKO, but not WASP KO, T cells had a disrupted subcortical actin cytoskeleton and impaired actin polymerization following TCR ligation. These results indicate that WIP is essential for IL-2 signaling and responsiveness in T cells, possibly because of its critical role in TCR-triggered actin cytoskeletal reorganization.
IL-2 differentially regulates perforin and FasL mediated cytotoxicity in CD4 T cells (33.10)
Authors: Brown Deborah M. , Harris Leah , Moore Tyler , Swain Susan L. , Condon Shirley A. 
Published: January 2023

Abstract
CD4 T cell effectors can promote survival against lethal influenza virus via perforin mediated cytolytic mechanisms, however; our understanding of how naïve CD4 cells differentiate into class II restricted killers remains obscure. To address this, TCR Tg CD4 cells were activated in vitro and examined for their ability to lyse target cells. We found that cytokine polarized CD4 T cell effectors displayed cytolytic activity with the hierarchy Th0&gt;Th1&gt;Th2&gt;Th17; however, IL-6, TGF-β, IL-10, IL-12 or TNF-α were not required for inducing cytolytic activity in CD4 effectors. Antigen dose had a marked effect on cytotoxicity: low concentrations of peptide induced more potent cytolytic activity than relatively high concentrations. At low peptide concentration, exogenous IL-2 was necessary to drive granzyme B (GrB) expression and perforin mediated lysis. Using IL-2 deficient T cells, we found that IL-2 signaling was required in the first 24 h of culture, but increasing concentrations of IL-2 did not increase cytolytic activity. Studies are underway to determine whether Stat5 signaling is required for induction of cytolytic activity in CD4 T cells. Thus, low antigen dose and early activation signals via IL-2 direct the CD4 T cell response toward effectors with perforin mediated cytolytic potential.
3D cell-matrix contact prevents IFN-γ induced upregulation of endothelial MHC II expression in high glucose environment (33.17)
Authors: Methe Heiko , Kim Jong-Oh 
Published: January 2023

Abstract
Cell matrix interactions are critical to function. Embedding endothelial cells (EC) within 3D matrices (3D-EC) retains biosecretory function and limits cell immunogenicity. Cardiovascular risk factors induce alterations in matrix composition and EC functionality. We examined how glucose levels dictate IFN-γ induced MHC II expression in EC.
EC cultured on polystyrene plates (TC-EC) and 3D-EC were grown to confluence in 0-50 mM glucose with or without 1000 U/ml IFN-γ. Intracellular signaling and MHC II expression were analyzed via PCR, Western Blot, and flow cytometry.
Without glucose phosphorylation of JAK- and STAT-proteins upon IFN-γ stimulation were attenuated 3.5-fold (p&lt;0.001) in 3D-EC compared to TC-EC. This was mirrored by reduced MHC II expression in 3D-EC (PCR: 0.3±0.1 vs. 0.1±0.1 relative units, p&lt;0.005). Suppressor of cytokine signaling protein (SOCS)-1 and -3 expression were 2.5-fold higher in unstimulated 3D-EC (p&lt;0.002). Matrix-embedding amplified upregulation of SOCS by IFN-γ. Low and high glucose medium significantly upregulated IFN-γ signaling pathways and MHC II expression (25 mM: 0.7±0.2, 50 mM: 0.9±0.2 relative units, p&lt;0.0001 vs. no glucose) in TC-EC but had no significant effect on SOCS and MHC II expression (0.2± 0.1 relative units) in 3D-EC.
Glucose amplifies IFN-γ dependent signaling in TC-EC. In contrast, increased SOCS expression in 3D-EC associates with a higher threshold at which IFN-γ induces MHC II, even in states of hyperglycemia. Matrix-embedding presents EC with the near-physiological state of substrate adherence and confluence. Intact cell-matrix contact shields EC from immune recognition and atherosclerotic risk factors like hyperglycemia.