bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of H1 receptors in the regulation of IL-4 production (37.16)
Authors: Khan Manzoor M. , Kharmate Geetanjali , Liu Zhongfeng 
Published: January 2023

Abstract
Histamine alters the Th1/Th2 cytokine balance towards the Th2 cytokine profile and consequently play a role in allergic disease and asthma. This study was designed to determine if the effects of histamine on Signal Transducer and Activator of Transcription(STAT)-6 were direct or mediated via release of IL-4 production.The splenocytes from IL-4 knockout C57Bl/6 mice were pretreated with antibody to IL-13 for 30 minutes followed by treatment with histamine. The cells were then stimulated with PMA + ininomycin for 6 hours under standard cell culture conditions. For the control experiments splenocytes from the wild type C57BL/6 mice were used. The cells were lyzed and the total basal and phosphorylated STAT6 were analyzed using the Western Blot Analysis. Furthermore, splenocytes from C57Bl/6 mice were pretreated with antibodies to IL-4 and IL-13 followed by treatment with histamine and PMA + ionomycin and Western blot analysis were performed to determine the phosphorylated Stat6 levels.In the control group histamine caused hyper-phosphorylation of Y641 tyrosine residues of STAT6. These effects were not observed when splenocytes from the IL-4 knockout mice were used or the wildtype splenocytes were pretreated with antibodies to IL-4 and IL-13. These observations suggest that histamine indirectly affected the heper-phosphorylation of STAT6 via its effects on the secretion of IL-4 and H1 receptors played a role in this process, since H1 receptor antagonists blocked these effects. The clinical significance of STAT6 hyper-phosphorylation in the exacerbation of asthma symptoms has not yet been determined.
On Intrinsic Allergenicity. Using Lipocalins To Probe The Mechanisms Of Allergy. (37.20)
Authors: Logdberg Lennart Erik , Akerstrom Bo , Hair Gregory A , Allhorn Maria , Vikulina Tatyana , Kirshenbaum Arnold S , Sundstrom J Bruce 
Published: January 2023

Abstract
What makes an antigen an allergen? It seems obvious that IgE-epitopes of allergens are necessary but not sufficient to cause initial sensitization and further to postulate that allergens contain separate molecular motifs that allow interaction with functionally important non-IgE molecules of the key cellular effectors. This way they actively influence development of the hypersensitive phenotype through “intrinsic allergenicity” of the allergen molecules.. We propose that comparative studies of lipocalin allergens, a collection of distinct, but structurally and functionally homologous molecules from different species, may shed light on the property of intrinsic allergenicity. Our overall hypothesis is that xenogeneic lipocalins, via interaction with components of the innate human immune system, such as mast cells, dendritic cells, lymphocytes, or progenitors or secreted mediators of these cells, can promote the afferent and/or the efferent phase of the allergic response, stimulating development of IgE-, rather than IgG-responses to their major immunogenic epitopes. Using human cultured mast cells, we have shown that these cells express the lipocalin receptor megalin, that megalin is massively induced by cultivating the mast cells with monomeric IgE, and. report on further investigations into its role in development of IgE-dependent lipocalin allergy.
Effect of IL-4 and IL-13 on the chemotaxis of mouse eosinophils (38.2)
Authors: Heller Nicola , Gwinn William M. , Lee James J. , Constant Stephanie L. , Keegan Achsah D. 
Published: January 2023

Abstract
Eosinophils are a major pathological component of the inflammatory cell infiltrate found in lungs as a result of allergic disease. They migrate into lungs in response to a variety of chemokines. The development and effector phases of the allergic response are dependent on the actions of interleukin (IL)-4 and IL-13 on a wide variety of cell types. However, how these two cytokines directly affect eosinophil signaling, function and chemotaxis is largely unknown. Using mouse eosinophils purified from IL-5 transgenic mice and a transwell migration assay, eotaxin-1 (30nM) induced eosinophil migration 6–11x over background. IL-4 alone was able to induce eosinophil migration 2x over background. More eosinophils migrated in response to eotaxin when IL-14 or IL-13 was also placed in the bottom chamber. IL-4 appeared to be more potent than IL-13. This effect was dependent upon cytokine concentration. Interestingly, when eosinophils were pretreated with IL-4 for 30 minutes prior to chemotaxis, migration to eotaxin-1 was enhanced 4-fold. However, pretreatment with IL-13 did not enhance chemotaxis to eotaxin-1. These data suggest that IL-4 and IL-13 have direct and rapid effects on the chemotactic function of eosinophils with some differences. We are investigating the mechanisms of these effects, as well as other signaling and functional responses of eosinophils to these cytokines. (AI38985 and HLT32007698)
Insights into mast cell signaling from rigid trivalent DNA-based ligands (38.5)
Authors: Sil Dwaipayan , Lee Jong B. , Luo Dan , Holowka David , Baird Barbara 
Published: January 2023

Abstract
Antigen-mediated crosslinking of IgE bound to its receptor, FcεRI, stimulates degranulation and other responses in mast cells and basophils to initiate allergic responses. To investigate possible roles for structural constraints of crosslinked receptors on signaling, we utilized double-stranded DNA spacers to synthesize ligands with tunable lengths and valencies. We produced and characterized a series of symmetric trivalent ligands with rigid spacing between 2,4-dinitrophenyl (DNP) haptenic groups in range of 4.5–13.5 nm. These ligands all bind to anti-DNP IgE on RBL mast cells with similar avidity, and they all crosslink IgE receptor complexes effectively. They initiate length-dependent tyrosine phosphorylation of FcεRI β and γ subunits and the adaptor protein LAT that are ~5–10-fold less for the longest ligand compared to the shortest. Receptor-mediated Ca+2 responses exhibit different kinetics and magnitudes as a function of ligand length, and degranulation responses parallel differences in these Ca+2 responses and in FcεRI tyrosine phosphorylation. In contrast, tyrosine phosphorylation of phospholipase Cγ1 and Ca2+ release from stores are relatively length independent, indicating differences in signaling pathways important for Ca+2 influx and release from stores. The length dependence of FcεRI phosphorylation that we observe is consistent with a transphosphorylation model of receptor activation leading to degranulation, whereas length-independent activation of PLCγ1 reveals unanticipated divergence from conventional signaling pathways for this process.
Regional increases of CD103+ Tregs corresponds with resolution of murine allergic airway disease (39.10)
Authors: McNamara Jeffrey Thomas , Carson William F , Guernsey Linda A , Secor Eric R , Singh Anurag , Schramm Craig M , Thrall Roger S 
Published: January 2023

Abstract
Mice sensitized to ovalbumin (OVA) develop allergic airway disease (AAD) with short-term OVA aerosol challenge (7 days); however, airway inflammation resolves and local inhalational tolerance (LIT) results with long-term continuous aerosol challenge (42 days). We have previously seen that regulatory T cell (Treg) populations that are CD4+CD25+Foxp3+ increase locally in the lung at AAD and expand further at LIT. A recent role for CD4+CD25+Foxp3+ CD103+ Tregs (CD103+Tregs) has been defined in mucosal models of inflammation; this subset of Treg cells has been demonstrated to have greater functionality in suppressing inflammation. The current study was designed to determine if the numbers of CD103+Treg cells in HLN correlate with the development of LIT. HLN and inguinal lymph nodes (ILN) were harvested from groups (n=6) of naive, sensitized, AAD and LIT mice. Flow cytometric analysis on the cells isolated from HLN and ILN demonstrated a significant (p&lt;.001) increase in the numbers and percentages of CD103+ Treg in HLN at LIT (7x103+ 4 HLN; 2x103+0.5 ILN ) compared to AAD (2x103+ 1 HLN; 1x103+0.5 ILN ). There was no significant increase of CD103+Treg in HLN between naïve, sensitized and AAD mice. The increase of CD103+ Tregs was seen regionally in HLN but not in ILN. In summary, the data suggests a novel in vivo role for CD103+Tregs in the subsequent resolution of a Th2 driven inflammatory response.
Funded by NIH/AIRO1 HL-43573
A CD1d-dependent antagonist inhibits the activation of iNKT cells and prevents development of allergen-induced airway hyperreactivity (39.9)
Authors: Akbari Omid , Yang Wen , Meyer Everett , Umetsu Dale T , Wilson S. Brian 
Published: January 2023

Abstract
The prevalence of asthma continues to increase and its optimal treatment remains a significant therapeutic challenge. Recently, CD1d-restricted iNKT cells were found to play a critical role in the induction of airway hyperreactivity (AHR) in rodents and are associated with severe peristent asthma in humans. To test whether iNKT cell-targeted therapy could be used to treat allergen-induced airway hyperreactivity disease, mice sensitized with ovalbumin were treated with a CD1d-binding lipid antagonist, DPPE-PEG. A single dose of DPPE-PEG prevented the development of AHR and pulmonary infiltrates after ovalbumin challenge, but not the development of ovalbumin-specific Th2 responses. Because iNKT cells play a critical role in the development of AHR, the inhibition of iNKT activation by DPPE-PEG suggests a novel therapy for iNKT cell-mediated diseases such as asthma.
Intestinal macrophages in human Crohn disease produce excess IL-23 in response to the enteric bacteria (40.7)
Authors: Kamada Nobuhiko , Hisamatsu Tadakazu , Chinen Hiroshi , Kobayashi Taku , Okamoto Susumu , Hibi Toshifumi 
Published: January 2023

Abstract
We have previously reported that murine intestinal macrophages (Mφ) show anti-inflammatory phenotypes, which produced IL-10 but not IL-12/23 and negatively regulated the excess immune response to enteric bacteria. In contrast, intestinal Mφ from IL-10 deficient mice, which spontaneously develop Th1 dominant chronic colitis, produced abundant levels of IL-12/23 in response to bacteria and these abnormal responses of intestinal Mφ contribute to Th1 polarization. In the present study, we examined the pathogenic role of intestinal Mφ in human Crohn disease (CD). We identified CD14+CD33+ Mφ subset in human intestine. These subsets produce larger amount of pro-inflammatory cytokines than CD14−CD33+ intestinal Mφ. The number of CD14+CD33+ intestinal Mφ were increased in CD patients compared to normal controls (NL) and ulcerative colitis (UC) patients. Moreover, these CD14+CD33+ intestinal Mφ from CD patients produced larger amount of IL-12/23p40 and IL-23 in response to enteric bacteria compared to NL and UC. Thus, intestinal Mφ may contribute the pathophysiologies of human CD via excess production of IL-12/23p40 and IL-23 in response to enteric bacteria.
Resident CNS cells functionally express NOD2 (40.8)
Authors: Marriott Ian , Sterka David 
Published: January 2023

Abstract
There is a growing appreciation that resident glial cells such as astrocytes and microglia can initiate and/or augment inflammation following trauma or infection. These cells are uniquely positioned to detect and respond to invading pathogens, and the mechanisms by which resident CNS cells perceive microbial challenges are now becoming apparent. We have previously shown that astrocytes and microglia express NOD2, a member of the nucleotide-binding oligomerization domain (NOD) family of proteins that serves as an intracellular receptor for bacterial peptidoglycan motifs and that can act in a cooperative manner with Toll-like receptors (TLR). In the present study, we have performed experiments to confirm the functional significance of NOD2 expression in resident CNS cells. We demonstrate that exposure of glia to the clinically relevant CNS pathogens, B. burgdorferi and N. meningitidis, elicits interaction between NOD2 and its downstream effector molecule Rip2 kinase as assessed by co-immunoprecipitation. In addition, we have shown that siRNA for NOD2 can diminish the robust constitutive expression of NOD2 in both microglia and astrocytes as assessed by Western blot analysis. Finally, we have demonstrated that the exacerbation of TLR ligand-induced inflammatory cytokine production by microglia in the presence of the NOD2 ligand, MDP, is attenuated by siRNA for NOD2. Taken together, these data indicate that NOD2 is functionally expressed by resident glia and may therefore represent an important component in the initiation and/or regulation of bacterially-induced inflammation in the CNS.
Substance P augments bacterially induced inflammatory cytokine production by resident CNS cells (40.9)
Authors: Chauhan Vinita Singh , Sterka David , Marriott Ian 
Published: January 2023

Abstract
The neuropeptide, substance P (SP), can augment inflammatory responses at peripheral sites such as the gastrointestinal tract and skin. We have recently demonstrated the expression of high affinity receptors for SP (NK-1R) on microglia and shown that this tachykinin can significantly elevate inflammatory prostanoid production by these cells following bacterial challenge. Here, we have investigated the ability of SP to augment the production of inflammatory cytokines by microglia following challenge with B. burgdorferi or N. meningitidis. Furthermore, we have assessed the in vivo relevancy of these findings by directly administering these pathogens in into the CNS in NK-1R knockout mice or normal animals in the absence or presence of the NK-1R antagonist, L703,606. Our results indicate that substance P significantly augments the production of TNF-a and IL-6 by bacterially challenged microglia in vitro. Importantly, we show that in vivo expression of these inflammatory cytokines is significantly diminished in both NK-1R KO animals and mice given L703,606. Together, these results suggest that SP augments the immune responses of resident glia to disparate and clinically relevant bacterial pathogens, which could serve to exacerbate damaging inflammation that occurs following CNS infection.
Induction of mucosal immunity by hsp gp96 (41.2)
Authors: Strbo Natasa , Pahwa Savita , Kolber Michael , Podack Eckhard 
Published: January 2023

Abstract
Targeting immune responses to the mucosal entry site before pathogen dissemination is critical for protection. We have generated a secreted gp96-fusion protein (gp96-Ig) that mediated strong, antigen specific CD8-CTL expansion in vivo.
The aim of this study was to evaluate the effect of the site of gp96 secretion on mucosal immunity.
Expansion and migration of antigen specific CD8 T cells (adoptively transferred gfp-OT-I responder cells) were measured in response to immunization. Following different routes of administration of gp96-Ig secreting cells (EG7/3T3-OVA) the OT-I response was measured in the peritoneal cavity, spleen, mesenteric lymph nodes and gut mucosa.
OT-I home significantly more efficiently to the gut mucosa if the gp96-Ig was delivered by the intraperitoneal route. Vaginal administration of gp96 also mediated mucosal immunity while subcutaneous and intradermal immunization delivered the weakest response. Secreted gp96-ova complexes recruited OT-I to Peyer’s Patches, intraepithelial compartment and lamina propria within five days. Mucosal OT-I express alpha4beta7, alphaEbeta7/CD103, CCR9, granzyme B, IFNgamma and exhibited effector memory phenotype.
Our results indicate that peritoneally or vaginally cell secreted gp96-Ig induces migration of effector memory cells equipped with cytotoxic molecules into the mucosal compartment by inducing selective expression of gut-homing markers.
Interleukin-17 plays a role in successful resolution of Trypanosoma cruzi infection. (43.17)
Authors: Yoshida Hiroki , Wang Sen , Iwakura Yoichiro , Miyazaki Yoshiyuki 
Published: January 2023

Abstract
Interleukin-17 (IL-17) is a proinflammatory cytokine produced mainly by T lymphocytes. Production of IL-17 is activated by IL-23, an IL-12 cytokine family member, and its over-production causes inflammatory disorders such as rheumatoid arthritis. In contrast, signaling through IL-27, another IL-12-related cytokine, and its receptor (WSX-1) participated in suppression of Th17 development and excessive IL-17 production during Trypanosoma cruzi infection. While IL-17 has been reported to be important for resolution of some bacterial infection, its role in defense against infection of intracellular protozoan parasites remains to be elucidated. Therefore, we investigated the effect of IL-17-deficiency on T. cruzi infection. C57BL/6 (wild-type; WT) and IL-17-deficient (IL-17−/−) mice were intraperitoneally injected with the plasma containing trypomastigotes, and the number of parasites in the blood was microscopically counted. IL-17−/− mice showed prolonged higher parasitemia over WT mice. As acute T. cruzi infection causes liver injury, we measured serum levels of liver enzymes, ALT and AST. The levels of ALT and AST increased along with the parasite expansion, and these were higher in IL-17−/− mice as compared with WT mice. Furthermore, mortality of the infected IL-17−/− mice was higher than that of WT mice. These results demonstrated that IL-17−/− mice were more susceptible to T. cruzi than were WT mice. Therefore, it is suggested that IL-17 plays an important role in resolution of T. cruzi infection. Detail functions of IL-17 in regulation of the protective immunity are currently under investigation.
Functional impairment of CD8 T cells following acute virus infections (43.32)
Authors: Fulton Ross Bane , Varga Steven M 
Published: January 2023

Abstract
Following acute respiratory syncytial virus (RSV) infection the virus-specific CD8 T cells from the spleen show a 1:1 correspondence between tetramer staining and in vitro cytokine production after peptide stimulation. In contrast, pulmonary virus-specific CD8 T cells are deficient in IFN-γ and TNF-α production and show impaired ex vivo cytotoxicity compared to CD8 T cells from the spleen. We have studied CD8 T cell functional impairment in viral infections outside the Paramyxoviridae family and show that similar dysregulation of pulmonary CD8 T cells also occurs after respiratory infections by lymphocytic choriomeningitis virus (LCMV) and by vaccinia virus expressing the RSV M2 protein (vvM2). In addition, LCMV and vvM2-specific CD8 T cells in the lungs demonstrate impaired degranulation relative to splenic CD8 T cells as measured by CD107a staining following ex vivo peptide stimulation. Importantly, we show here that pulmonary CD8 T cell IFN-γ and TNF-α production can be recovered by stimulating these cells in vitro using P815 cells pulsed with peptide as a source of exogenous antigen presenting cells (APCs). Our results indicate that the CD8 T cells in the lung are fully functional and that the apparent functional impairment may be caused by the APC milieu within the lung tissue.
A quantitative analysis of the variables affecting the repertoire of T cell specificities recognized following VACV infection (43.56)
Authors: Assarsson Erika , Sidney John , Oseroff Carla , Pasquetto Valerie , Bui Huynh-Hoa , Peters Bjoern , Grey Howard , Sette Alessandro 
Published: January 2023

Abstract
Many components contribute to immunodominance in the response to a complex virus, but their relative importance is unclear. This was addressed using vaccinia virus and HLA-A*0201 as the model system. A comprehensive analysis of 18 viral proteins recognized by CD8+ T cell responses demonstrated that about 1/40 of all possible 9–10 mer peptides were high-affinity A*0201 binders. Peptide immunization and T cell recognition data generated from 91 peptides indicated that about half of the binders were capable of eliciting T cell responses, and that 1/7 immunogenic peptides are generated by natural processing. Based on these results, we estimate that vaccinia virus encodes about 150 dominant and subdominant epitopes restricted in the context of HLA-A*0201. However, of all these potential epitopes, only 15 are immunodominant and actually recognized in vivo during vaccinia virus infection of HLA-A*0201 transgenic mice. Neither peptide binding affinity, complex stability, TCR avidity nor amount of processed epitope appeared to strictly correlate with immunodominance status. Further experiments suggested that vaccinia infection impairs the development of responses directed against subdominant epitopes. This suggested that immunoregulatory factors restrict the repertoire of T cell specificities following vaccinia infection by at least a factor of ten.
Length of Antigenic Stimulation During Expansion Dictates Commitment to CD8 T Cell Memory Subsets (43.6)
Authors: Sarkar Surojit , Kalia Vandana , Haining W. Nicholas , Konieczny Bogumila T. , Subramaniam Shruti , Ahmed Rafi 
Published: January 2023

Abstract
There is considerable interest in understanding early events that drive CD8 T cells towards diverse cell fate decisions: differentiation into central memory (TCM), effector memory (TEM) or terminal effector (TE) subsets. Although, a brief antigenic encounter is sufficient to trigger a cell autonomous program of clonal expansion and differentiation of naïve cells into effector and memory cells, how the ensuing differentiation program is impacted by subsequent antigenic stimulation in the setting of an acute infection is unclear. Here we show that commitment to the TCM, TEM and TE subsets occurs early during the induction and expansion phase of CD8 T cell responses. Repetitive antigen encounters during expansion and the extent of proliferation conditioned by it play a major role in dictating this memory subset commitment, such that increasing antigenic encounter favors TE&gt;&gt;TEM&gt;TCM differentiation. Interestingly, early effectors that preferentially differentiate into TCM, exhibit a novel signature; in addition to elaborating characteristic cytotoxic function, they exhibit strong IL-2 production, a property typically ascribed to naïve and TCM cells. Gene profiling of this novel IL-2+ effector subset revealed that they represent a slightly less differentiated effector subset compared to the non-IL-2 producing counterparts, even though their ex-vivo and in-vivo cytotoxic potential were comparable. These studies underscore the importance of the length of antigenic stimulation, in regulating the commitment to various memory lineages, and will likely provide leads upon which tailored vaccines can be designed.
Supported by EGPAIDS Foundation Scholar Awards to SS & VK, & NIH & Gates Foundation Grants to RA