bluuebunny/crossref_metadata_2025_split huggingface dataset:
Antigen Processing of pneumococcal-CRM197 glycoconjugate vaccine: The polysaccharide component co-localizes with the carrier protein and MHC II on the APC surface (36.8)
Authors: Lai Zengzu , Schreiber John 
Published: January 2023

Abstract
Pneumococcal (Pn) polysaccharides (PS) are T-independent (TI), induce no immunological memory and low Ab titers in infants. Conjugation of the Pn PS to the protein carrier CRM197 induces PS-specfic Ab in infants and memory similar to T dependent (Td) antigens. Glycoconjugates induce a Td PS immune response via antigen processing and presentation of carrier with MHC II, but the fate of the PS attached to the carrier is unclear. To determine the location of the PS of PnPS-CRM197 in the APC, we separately labeled PS and protein and tracked their location in APC. The PS of 19F-CRM197 was labeled by Alexa Fluor ® 594 hydrazide (labels PS red). The CRM197 was separately labeled in a reaction that did not label PS. Labeled 19F PS, 19F-CRM197 and CRM197 were incubated with 1x106 APC for 3–12 hours. APC were fixed and incubated with anti- HLA-DR Ab labeled by Alexa Fluor® 488 (green), followed by confocal microscopy. Labeled CRM197 was taken up and presented on the APC surface co-localized with MHC II (yellow; 42% positive cells). Labeled 19F PS was not internalized or presented on the surface. PS labeled 19F-CRM197 was internalized and co-localized on the surface with MHC II (23% positive). Brefeldin A and chloroquine blocked both CRM197 and 19F-CRM197 from co-localizing on the surface. These data suggest that PS of the 19F-CRM197 glycoconjugate enters the endosome, travels with CRM197 peptides to the APC surface and co-localizes with MHC II.
IgE production is enhanced by novel signaling intermediates activated by stimulation of a G-protein coupled receptor on a B cell (37.12)
Authors: McAlees Jaclyn W. , Sanders Virginia M. 
Published: January 2023

Abstract
The level of a Th2/IL-4-dependent antibody response is regulated by extrinsic homeostatic signals that stimulate a G-protein coupled receptor, the beta2-adrenergic receptor (β2AR), which is expressed on a B cell. We find that stimulation of the β2AR on a CD40L- and IL-4-activated B cell increases the rate of IgE mRNA transcription and the amount of IgE produced per cell, without affecting class switch recombination. Our data also show that β2AR stimulation increases IgE production in a p38 MAPK−, CD23−, and CD21/CD19-dependent manner above that induced by CD40L/IL-4 alone (J. Immunol., 177:2926). However, the mechanism(s) by which β2AR stimulation on a B cell leads to an increase in p38 MAPK activity remains unknown. Our findings suggest that the β2AR-induced increase in p38 MAPK activity occurs in a PKA-dependent, CREB-independent manner that may involve hematopoietic protein tyrosine phosphatase (HePTP) regulation as determined using gene-deficient and -transgenic mice, enzyme activation assays, co-immunoprecipitation, western blot analysis, and pharmacological inhibitors. We also show that p38 MAPK activation is necessary for a release of CD23 from the cell surface (sCD23), which may be mediated by an increase in ADAM8/10 activity. Thus, the β2AR-induced increase in IgE may depend on a p38 MAPK-induced increase in the level of soluble CD23.
Supported by NIH RO1 AI37326.
IL-12 treatment alleviates allergic lung inflammation in T-bet−/− mice (37.19)
Authors: Durrant Douglas Matthew , Metzger Dennis W. 
Published: January 2023

Abstract
Allergic asthma is a chronic airway disease characterized by lung inflammation and reversible airway obstruction. Little is known about the underlying mechanisms in the development of asthma, but it is well appreciated that dysregulated CD4+ T helper (Th) cells initiate and perpetuate the disease. To elucidate the potential role of IL-12, a Th1-inducing cytokine, in the treatment of asthma, wild-type and T-bet−/− mice were used in a murine model of ovalbumin (OVA)-induced allergic lung inflammation. T-bet is a Th1-specific T box transcription factor that controls the expression of interferon (IFN)-γ. Thus, T-bet−/− mice have little endogenous IFN-γ production and subsequently exhibit spontaneous lung inflammation. IL-12 treatment reduced pulmonary inflammation in WT mice as expected. Surprisingly, T-bet−/− mice treated with OVA and IL-12 also demonstrated reduced bronchoconstriction compared with T-bet−/− mice treated with OVA only. IL-12 treated T-bet−/− mice also demonstrated reduced airway hyperresponsiveness after methacholine challenge. Intracellular cytokine staining demonstrated the presence of IFN-γ+ CD8+ T cells in the lungs of T-bet deficient mice treated with IL-12. Also, we observed a marked decrease in the levels of the pro-inflammatory cytokine IL-17 in IL-12 treated T-bet−/− mice compared to wild-type, suggesting a role for Th17 cells in this murine model of allergic lung inflammation. Our hypothesis is that the pulmonary immune response is pre-disposed towards a Th17 type in the absence of T-bet regulation and that IL-12 treatment suppresses Th17 cells through a T-bet-independent, yet an IFN-γdependent mechanism. The results of this study will broaden the understanding of pulmonary inflammatory reactions.
Supported by NIH grant RO1 AI41715
The Immunomodulating Effect of AS101 on Interleukin-10 Production and the Involvement of MAPK Signaling Pathway in Atopic Dermatitis (37.3)
Authors: Sredni-Kenigsbuch Dvora , Shohat Michael , Shohat Batia , Ben-Amitai Dan , Chan Chi-Chao , David Michael 
Published: January 2023

Abstract
Atopic dermatitis (AD) is a chronic inflammatory skin disease. Recently it was reported that interleukin 10 (IL-10) is also overexpressed in skin lesions of atopic dermatitis (AD) patients and this ectopic expression is believed to be an important factor in the pathogenesis of the disease. Thus the regulation of IL-10 production is a potential solution for immunotherapeutic intervention in AD. In this study, we determined whether the tellurium immunomodulating compound, ammonium tri-chloro(dioxoethylene-O,O′-) tellurate (AS101) may inhibit IL-10, and thereby affect the profile of cytokines produced by T cells of AD patients. Further, we investigated the possible correlation between changes in IL-10 levels and the expression of mitogen-activated protein (MAP) kinases. We show here that the IL-10 level was higher in AD patients compared to healthy donors, while their IL-2 and IFNγ levels were reduced. The addition of the AS101 inhibited the production of interleukin-10, while increasing the production of IL-2 and IFNγ. These changes correlate with the inhibition of p38 MAP kinase. The immunomodulatory effect of AS101, together with its excellent clinical safety profile in humans, suggests that AS101 has potential as a therapeutic agent for AD.
Characterization of allergen driven type-2 cytokine response in an adjuvant-free mouse model of tree nut allergy: Critical role of IL-4 (37.9)
Authors: Gangur Venu , Parvataneni Sitaram , Birmingham Neal , Fyolek John , Kelly Caleb , Hassan Hanem 
Published: January 2023

Abstract
Food allergy is a growing problem of serious proportions with potential for fatality. Specific role of type-2 cytokines in food allergy is not completely understood. Our laboratory has previously developed an adjuvant-free mouse model to study food allergy using hazelnut as a model tree nut. Using this model, the present study was conducted to elucidate the role of selected type-2 cytokines in this model. BALB/c mice were transdermally sensitized with hazelnut protein and systemic IgE antibody responses were confirmed. Subsequently, using short term ex vivo spleen cell culture, allergen driven type-2 cytokine (IL-4, IL-5 and IL-13) responses were studied using ELISA. Here, robust allergen driven, dose dependent, recall IL-4, IL-5 and IL-13 responses were observed. Further, all three type-2 cytokine responses could be largely blocked using anti-CD4 antibodies. Furthermore, BALB/c Stat6 knockout (KO) mice lacked an IL-4 but not IL-5 or IL-13 recall response to hazelnut. In addition, BALB/c Il4 KO mice did not elicit an IgE response to transdermal hazelnut exposure and were resistant to systemic anaphylaxis following oral hazelnut challenge. These data together suggest that:
transdermal exposure to hazelnut results in activation of IL-4, IL-5 and IL-13 by CD4+spleen cells; andonly IL-4, which is STAT-6 dependent, is required for food allergy phenotypes studied in this model.
Funding: Michigan State University, Michigan Agricultural Experiment Station.
Endotoxin Tolerance Abrogates Eotaxin-2 Production and Eosinophil Recruitment While Increasing Airways Hyperresponsiveness In Murine Asthma (39.1)
Authors: Natarajan Sudha , Kim Jiyoun , Remick Daniel G. 
Published: January 2023

Abstract
Studies suggest that exposure to high levels of endotoxin (E) at sensitization results in reduced inflammation upon allergen challenge. We hypothesized that persistent E exposure results in endotoxin tolerance (E-T), modulates allergen sensitization and results in reduced inflammation. E-T was induced by exposing female BABL/c mice to 4 consecutive days of 1μg E intratracheally. Non-E-T mice received PBS. 24h later, mice were sensitized with 2μg cockroach allergen, rechallenged 14 and 21 days later and sacrificed 24h post final challenge. Pulmonary Eotaxin-2 levels in E-T mice (92pg/mL) were significantly reduced compared to non-E-T mice (1965pg/mL) and pulmonary eosinophils in E-T mice (13,800) were significantly reduced compared to non-E-T mice (92,800). A significant reduction in pulmonary levels of TNFα (E-T vs. non-E-T 44pg/mL vs.65pg/mL), IL-17 (82 vs. 217) and KC (145 vs. 299) was also seen. Interestingly, airways hyperresponsiveness (AHR) was increased in E-T mice (215% above baseline) compared to non-E-T mice (83%). Reduced inflammation along with increased AHR suggests that these pathways are modulated by divergent mechanisms. Our data indicates that E-T significantly blunts the inflammatory response to allergen challenge, thereby providing protection against severe asthmatic inflammation. NIH Grant ES13528.
Complex role of STAT6 in allergic airway inflammation (39.11)
Authors: Chapoval Svetlana P , Kelly-Welch Ann E , Smith Elizabeth , Keegan Achsah D 
Published: January 2023

Abstract
STAT6 plays a critical role in Th2 cell differentiation and in allergic lung inflammation. Using a chimeric mouse model, we observed alternative lung pathology in STAT6 KO mice even when WT bone marrow or Th2 cells were provided. Thus, we hypothesized that STAT6 contributes to inflammation in a complex manner. To detail STAT6 function, WT and STAT6 KO mice were subjected to OVA priming and challenges. Broncho-alveolar lavage (BAL) cell composition, lung histology, and FACS analysis of digested lungs were assessed 48h after the last challenge. As expected, eosinophils composed a majority of BAL cells in WT mice and less than 2% in STAT6 KO mice. The OVA-induced inflammation in STAT6 KO lungs was composed mainly of macrophages with small fractions of neutrophils and lymphocytes. The OVA-treated WT lungs showed strong although divergent expression of F4/80, Mac-2, and CD11b molecules; this was significantly reduced in STAT6 KO mice. However, the numbers of dendritic cells, B cells, CD4+ and CD8+ T cells in the lungs of OVA-treated mice were unaffected by STAT6 deficiency. Interestingly, STAT6 KO mice showed enhanced basal airway reactivity to methacholine and numbers of Mac-2+ cells as compared to WT mice, suggesting that STAT6 deficiency altered lung homeostasis. Taken together, our studies demonstrate STAT6-dependent and –independent features of asthma phenotype which may impact treatments targeting STAT6. (AI38985)
ICOS/ICOSL interaction is required for CD4+ NKT cells function and for the induction of airway hyperreactivity (39.4)
Authors: Akbari Omid , Meyer Everett H , Freeman Gordon J , Sharpe Arlene H , DeKruyff Rosemarie H , Umetsu Dale T 
Published: January 2023

Abstract
Asthma is a major public health problem that has increased markedly in prevalence in the past two decades. In several mouse models, natural killer T cells (NKT) have recently been found to be required for the development of airway hyperreactivity (AHR), a cardinal feature of asthma. Moreover, in patients with persistent severe asthma, a significant number of NKT cells are present in the lungs. While T cell receptor signaling is required to activate NKT cells, the costimulatory requirements for NKT cell activation and function are not clear. In this study we found that ICOS was highly expressed on both naïve and activated NKT cells and that expression of ICOS was much higher on the CD4+ NKT than CD4− NKT cells. The function of NKT cells from ICOS−/− mice was impaired, as adoptive transfer of these cells into NKT cell deficient Ja18−/− mice failed to restore AHR, whereas transfer of wildtype NKT cells fully restored AHR in the Ja18−/− recipients. The failure of ICOS−/− NKT to induce AHR may be due to a great extent to the absence of cytokine production by ICOS−/− NKT cells. Additionally, ICOS−/− mice challenged with α-GalCer did not develop AHR, but adoptive transfer of WT NKT cells into ICOS−/− mice restored AHR. Finally, ICOS may be required for CD4+ NKT cell survival in the periphery, as the number of CD4+ NKT cells was significantly lower in spleen and liver of ICOS−/− mice. These results indicate that costimulation by ICOS is required for the function and homeostasis of CD4+ NKT cells, which play a major role in development of AHR.
Stem cell factor and recruitment of bone marrow derived fibroblasts in chronic asthma airway remodeling. (39.5)
Authors: Dolgachev Vladislav A , Ullenbruch Matthew R , Lukacs Nicholas W , Phan Sem H 
Published: January 2023

Abstract
Stem cell factor (SCF) is recently shown to be important in airway remodeling in asthma. Moreover bone marrow progenitors contribute to the lung fibroblast population in response to injury. To assess whether SCF is involved in the recruitment and differentiation of these bone marrow derived progenitor cells, GFP bone marrow chimera mice were created. These mice were then sensitized and chronically challenged with cockroach antigen (CRA) to induce the chronic airway disease model. As previously shown these mice exhibited significant airway remodeling with increased numbers of fibroblasts or fibroblast-like cells. Fluorescence microscopy revealed the presence of significant numbers of GFP expressing fibroblasts in these airways, indicative of their bone marrow origin. This was confirmed by flow cytometric analysis, which showed the presence of GFP and collagen I double positive cells. Interestingly the GFP positive cells did not express alpha-smooth muscle actin, thus suggesting that myofibroblasts were not derived from the bone marrow. Furthermore &gt;20% of these cells expressed c-kit, which represented &gt;60% of all bone marrow-derived c-kit+ cells. Most of the latter cells (&gt;90%) were also positive for telomerase reverse transcriptase. Treatment with neutralizing antibodies to SCF significantly reduced airway remodeling and suppressed recruitment of these bone marrow-derived cells to the lung. Thus SCF may play a significant role in airway remodeling by promoting the recruitment of bone marrow-derived fibroblast precursors into the lung.
Smooth muscle responsiveness to IL-13 is sufficient for IL-13 induction of airway hyperresponsiveness (AHR) (39.6)
Authors: Finkelman Fred D. , Perkins Charles , Orekov Tatyana , Potter Crystal , Smulian George , Gildea Lucy , Wills-Karp Marsha , Brombacher Frank 
Published: January 2023

Abstract
IL-13 is the primary inducer of AHR in mouse models of asthma and increases contractility of isolated muscle cells in vitro and the contractility of intestinal smooth muscle in worm-infected mice. Consequently, we evaluated whether smooth muscle responsiveness to IL-13 is sufficient for development of AHR in mice inoculated intratracheally with IL-13. The smooth muscle-specific promoter SMP8 was used to generate mice that express IL-4Rα only on smooth muscle cells (SMonly mice, produced by crossing SMP8-IL-4Rα transgenic mice to IL-4Rα-deficient mice). SMonly mice failed to develop goblet cell hyperplasia or BAL eosinophilia in response to intratracheal IL-13 but developed AHR to inhaled methacholine to the same extent as wild-type mice when tested by barometric plethysmography with a Buxco apparatus and significant, albeit reduced AHR to injected acetylcholine when anesthetized, intubated and evaluated by the airway time-pressure index (APTI) technique. Thus, smooth muscle expression of IL-4Rα is sufficient for IL-13 induction of AHR and this is not dependent on goblet cell hyperplasia or pulmonary eosinophilia. Studies designed to determine whether smooth muscle expression of IL-4Rα is necessary for IL-13 induction of AHR are in progress.
Supported by the US Dept of Veterans Affairs and NIH.
Indoleamine 2,3-Dioxygenase is not required for tolerance induction in the airways but is important for allergic airway disease (39.8)
Authors: Xu Hui , Oriss Timothy B , Melgert Barbro N , Henry Adam C , Chen Li , Mellor Andrew L , Munn David H , Irvin Charles G , Ray Prabir , Ray Anuradha 
Published: January 2023

Abstract
Indoleamine 2,3 dioxgenase (IDO) has emerged as an important negative regulator of immune responses. However, the role of IDO in Th2 cell-mediated diseases is not completely clear. We investigated the role of IDO in the induction of inflammation or tolerance in the airways in response to ovalbumin (OVA). The results of these experiments show that IDO−/− mice are susceptible to tolerance induction. However, IDO−/− mice displayed significantly lower airway hyperresponsiveness (AHR), lower serum OVA-specific IgE and lower Th2 type cytokines (IL-5 and IL-13) in comparison to wt mice. In an OVA-induced chronic airway inflammation model, the IDO−/− mice also showed lower AHR and Th2 type cytokines in the lung and less infiltration of inflammatory cells in the airways. To dissect the mechanism underlying IDO-mediated promotion of allergic airway disease, we stimulated lung DC by allergens in vitro. The expression of important co-stimulatory molecules and MHC-II was lower in DCs from IDO−/− mice in comparison to those in DCs from wt control mice suggesting that the reduced airway Th2 type immune response in IDO−/− mice may be due, at least in part, to less effective co-stimulation between DCs and T cells. In summary, our data suggest that IDO is not required for the induction of airway tolerance, but may play a role in promoting Th2-induced allergic airway disease.
Supported by NIH grants HL 77430 (to A.R) and HL60207 (to P.R.)
Role of adaptive immunity in Helicobacter-associated gastric adenocarcinoma (40.1)
Authors: Schmitz Julia Michelle , Lorenz Robin G. 
Published: January 2023

Abstract
The majority of gastric cancer is associated with Helicobacter pylori infection, however the mechanism is unknown. Our laboratory has previously demonstrated that T cells are critical to Helicobacter-induced gastric pathology. This involvement could be through T cell cytokine induced alterations in the protective gastric surface mucous layer. C57BL/6 mice, after infection with H. felis, develop severe gastritis which progresses to a gastric adenocarcinoma that is associated with a decrease in the production of the mucin, muc5ac. C57BL/6.Rag-1−/− mice, which lack both T and B cells, do not develop gastritis, adenocarcinoma, or changes in muc5ac expression after infection. An analysis of gastric cytokine expression by RT-PCR has demonstrated that infected C57B6.RAG-1−/− mice produce less inflammatory cytokines, IL-17, TNFα, and IFNγ, while producing more IL-6 and equal levels of IL-10 and IL-1β. This suggests that B and T cells are only responsible for the secretion of inflammatory cytokines during infection, but not innate or regulatory cytokines. These results also indicate that there is an association between the inflammatory cytokines, IL-17, TNFα, and IFNγ, made by the adaptive immune response and alterations in the protective gastric mucous layer. Further studies will be required to investigate the exact role of each cytokine in the progression to gastric cancer.
This work was supported in part by ACS RSG MBC-103892, NIH R01 DK59911, NIH T32 AI07041, and R24 DK064400.
Promoting dendritic cell migration by mouse airway epithelial cells through IL-17A-induced mouse CCL20 expression (40.10)
Authors: Hung Li-Yin , Wu Reen 
Published: January 2023

Abstract
Dendritic cells are the antigen presenting cells that play a central role in regulating immune response. Their migration on airway epithelium in response to antigen challenge is incompletely understood. In this study, we demonstrated that mouse IL-17A is a potent stimulator for primary mouse airway epithelium to secrete chemotactic factors for the migration of bone marrow-derived dendritic cells (BMDC) along its gradient. Neutralizing antibody study with anti-mCCL20 demonstrated the inhibition of the migration. Real-time quantitative RT-PCR study demonstrated time- and dose-dependent effects of IL-17A on mCCL20 expression, but not the induction of hBD-2 equivalent messages, in mouse airway epithelial cells. The lack of induction of hBD-2 equivalent messages by IL-17A on mouse cell system is in contrast to that was seen in human, suggesting a species difference in response to IL-17A. Using inhibitor, we observed the inhibition of induced CCL20 expression by an NF-κB inhibitor, isohelenin, suggesting an NF-κB mediated transcriptional mechanism in the induction. Based on these observations, we suggest that IL-17A may be responsible for in vivo dendritic cell migration on airway epithelium through the induction of mCCL20 expression.
This study is supported by NIH grant HL077315 and HL077902
Intestinal Bacteria Induce T Cell-Independent IgA2 Class Switching by Linking Epithelial Cells with Lamina Propria B Cells through APRIL (40.3)
Authors: He Bing , Xu Weifeng , Santini Paul A. , Polydorides Alexandros D. , Chiu April , Chadburn Amy , Knowles Daniel M. , Rescigno Maria , Cerutti Andrea 
Published: January 2023

Abstract
Bacteria colonize the intestine shortly after birth and thereafter exert several beneficial functions, including induction of protective IgA antibodies. The distal intestine contains IgA2, which is more resistant to bacterial proteases than IgA1. The mechanism by which B cells undergo IgA2 class switching remains unknown. We found that human intestinal epithelial cells (IECs) triggered IgA2 class switching by activating B cells, including IgA1-expressing effector B cells, through a T cell-independent pathway involving a proliferation-inducing ligand (APRIL). IECs released APRIL after sensing whole bacteria or bacterial products, such as lipopolysaccharide or flagellin, through a Toll-like receptor (TLR)-mediated MyD88-dependent pathway. Bacteria-activated IECs further increased APRIL production by cross talking with dendritic cells via thymic stromal lymphopoietin (TSLP), an IL-7-like cytokine. Our findings indicate that bacteria and their products elicit IgA1-to-IgA2 diversification by linking lamina propria effector B cells arriving from mucosal lymphoid follicles with IECs through a TLR-inducible APRIL-dependent signaling program. Our data also suggest that vaccines should activate IECs to effectively induce protective IgA2 responses in the intestinal mucosa.
This work was supported by NIH grant AI057653 (to A.C.) and NIH T32 training grant AI07621.