bluuebunny/crossref_metadata_2025_split huggingface dataset:
NKT cells are not required for development of allergic airway disease in allergen-immunized mice (79.1)
Authors: Perkins Charles , Orekov Tatyana , Yanase Noriko , Mattner Jochen , Finkelman Fred D. 
Published: January 2023

Abstract
The role of NKT cells in the pathogenesis of asthma and animal models of asthma is controversial. Although the initial mouse model study failed to identify an NKT cell requirement for induction of allergic airway inflammation or airway hyperresponsiveness (AHR), 2 subsequent mouse studies found both phenomena to be highly NKT cell-dependent. Some studies of human asthma patients also suggest strong involvement of NKT cells in airway inflammation, while others dispute this finding. To re-visit this issue at its origin, we evaluated the ability of ovalbumin (OVA) to induce allergic airway disease in BALB/c wild-type and NKT cell-deficient (CD1-deficient) mice, using a protocol in which mice were initially primed i.p. with OVA/alum, then inoculated intratracheally (i.t.) with OVA, as well as a protocol in which mice were inoculated i.t. with dust mite allergen without priming. Results from both studies demonstrated that wild-type and NKT cell-deficient mice develop similar airway eosinophilia, IL-4 and IL-13 responses, goblet cell hyperplasia and AHR (determined by both invasive and non-invasive methods). Flow cytometric analysis of lung and BAL lymphocytes from saline-, OVA-, and dust mite Ag-inoculated mice failed to demonstrate any specific staining for invariant NKT cells. We conclude that NKT cells are not required for allergen-immunized mice to develop full-blown allergic airway disease.
Expression of IL-19 in Airway Epithelial Cells During Asthmatic Response (79.11)
Authors: Hung Li-Yin , Nguyen Tiffany T , Wu Reen 
Published: January 2023

Abstract
IL-19 is a proinflammatory cytokine that belongs to the IL-10 cytokine family. In addition to its known connection with psoriasis, previous work in our lab, along with others, has also shown its association with asthma. Using cultured primary mouse airway cells, we demonstrated that Th2 cytokines such as IL-4 and IL-13 can induce the expression of IL-19 mRNA. Despite its ability to stimulate IL-19 production in human cells, IL-17A appeared not being involved in this process in the mouse model. With an OVA-induced mouse asthma model, we showed that IL-19 expression is increased in the asthmatic lung tissues at the early stage. The induction is specific to IL-19 rather than other IL-20 family members such as IL-20, IL-22, or IL-24. We further identified that airway epithelial cell, rather than the alveolar macrophage, is the source of IL-19 in the asthma model. These results demonstrate that IL-19 is produced by airway epithelial cell at the early stage of asthmatic response, and may further amplify the Th2 response through acting on T cells at the later stage of asthma.
Annexin-1 deficient mice exhibit enhanced antibody production in a mouse model of asthma (79.18)
Authors: Lim Lina HK , Biswas Subhra K , Mustafa Fatimah B , Kajiji Tasmeen S , Tan Yuwen , Ng Florence SP 
Published: January 2023

Abstract
Allergic asthma is a complex disorder characterized by airway inflammation and reversible airway obstruction and airway hyperresponsiveness. Glucocorticoids are currently the most effective anti-inflammatory therapy for asthma. Annexin-1 is an endogenous anti-inflammatory protein responsible for many anti-inflammatory effects of glucocorticoids. Here we show in a mouse model of asthma, that annexin-1 deficient mice do not exhibit enhanced inflammation, such as eosinophil infiltration, but exhibited higher antibody production. In particular, allergen-specific and total IgE levels, as well as allergen-specific IgG1, IgG2a and IgG2b serum levels were significantly higher in annexin-1 deficient mice. Our results in annexin-1 deficient mice demonstrate that annexin-1 may play a regulatory role in the intrinsic development of allergy.
Thymic Stromal Lymphopoietin (TSLP) Modulates the Balance between Th1 and Th2 Responses in the Skin (79.4)
Authors: Larson Ryan P , Ziegler Steven F 
Published: January 2023

Abstract
TSLP is an IL-7 related cytokine produced by epithelial cells in response to tissue damage, inflammation, or TLR ligation. Overexpression of TSLP in mouse skin or lung leads to the development of atopic dermatitis and asthma, respectively; however, these models fail to elucidate which cell types are acted on by TSLP. TSLP has also been hypothesized to inhibit aberrant Th1 responses by inhibiting the production of IL-12 family members by DCs. TSLP receptor deficiency significantly inhibits the development of Th2-type contact hypersensitivity (CHS) response induced by the irritant dibutyl phthalate in combination with the hapten fluorescein isothiocyanate (FITC). Specifically, FITC-sensitized TSLPR-/- mice do not experience swelling or eosinophilia at the site of challenge; nor do they experience increased serum IgE, all hallmarks of Th2-type CHS observed in WT mice. The reduced CHS response is due in part to the significant reduction in frequency and number of skin-derived antigen-bearing FITC+CD11c+ DC in skin-draining lymph nodes 24 hours post FITC sensitization of TSLPR-/- mice. Furthermore, skin-derived TSLPR-/- FITC+CD11c+ DCs are reduced in their ability to drive proliferation of naïve antigen specific CD4+ T cells compared to WT counterparts. In addition, we have found that there is no requirement for CD4+ T cells to respond to TSLP for the development of FITC-induced CHS. Interestingly, TSLPR-/- mice experience an enhanced Th1-type CHS response to the Th1-inducing contact sensitizing agent 2,4-dinitrofluorobenzene, compared to WT. These results support a role for TSLP acting on DC in vivo resulting in the development of Th2 allergic response and potentially playing a regulatory role by inhibiting Th1 CHS responses.
Coordinate posttranscriptional regulation of asthma genes by the RNA binding protein HuR (79.6)
Authors: Gubin Matthew M , Hollingsworth John D , Calaluce R , Magee Joseph D , Schwartz David A , Atasoy Ulus 
Published: January 2023

Abstract
Traditional microarray analysis has uncovered a variety of genes involved in diseases such as asthma. However, it may potentially miss important genes due to the discordance between steady state mRNA levels and protein products. Posttranscriptional events conferred by RNA binding proteins (RBPs) contribute to this phenomenon. We have developed novel techniques known as RIP-on-Chip (microchip) which allow for the immunoprecipitation and identification of in vivo mRNA targets bound to RBPs. We study the RBP HuR, which regulates many early response genes including cytokines and chemokines. Using RIP-on-Chip, HuR has been demonstrated to associate and regulate many asthma relevant genes including recently identified genes IL-4 and IL-13.
We have extended this assay to solid organ tissues. Employing the standard ovalbumin model of allergic airway inflammation, coupled to genome-wide arrays (47,000 genes) we have identified in vivo HuR targets in lungs of naïve, alum only or ova challenged mice. Remarkably, there are only 12 HuR targets (2 fold or greater) up regulated in ova and alum immunized over alum alone mice. Some are known HuR target genes such as TLR4 and IL-13; others are potentially novel asthma genes. Furthermore, we have created a transgenic mouse which over expresses HuR in CD4+ T cells. In the allergic airway inflammation model these mice have an increase in lung lymphocyte infiltration when compared to wild-type littermates, further implicating coordinate posttranscriptional gene regulation and HuR in allergic airway inflammation.
Selective stimulation of IL-4 receptor on smooth muscle induces airway hyperresponsiveness in mice (79.9)
Authors: Finkelman Fred D. , Perkins Charles , Smulian George , Gildea Lucy , Orekov Tatyana , Potter Crystal , Brombacher Frank , Wills-Karp Marsha 
Published: January 2023

Abstract
Hyperresponsiveness to cholinergic stimulation of increased airway resistance is a defining characteristic of asthma. Both IL-4 and IL-13 activate Stat6 via IL-4Rα to induce airway hyperresponsiveness (AHR) in mouse models of asthma and increase mouse and human smooth muscle contractility in vitro. However, the relatively small size of mouse vs. human airways, differences in airway smooth muscle cell distribution between mouse and human, and observations that selective IL-13 stimulation of Stat6 in airway epithelium is sufficient to induce AHR in mice have raised questions about the importance of direct IL-4/IL-13 effects on smooth muscle in murine models of allergic airway disease and whether these models are appropriate for studying AHR in human asthma. To determine whether direct effects of IL-4/IL-13 on smooth muscle contribute to murine allergic airway disease, we produced mice that express IL-4Rα only on smooth muscle and evaluated the effects of intratracheal IL-4, IL-13 and allergen on their airways. We demonstrate that IL-4 and IL-13 induce AHR in these mice, without stimulating goblet cell hyperplasia or airway eosinophilia. Similar results were observed following airway inoculation of these mice with a potent allergen. Thus, allergen-stimulated IL-4/IL-13 can induce AHR in mice through direct effects on airway smooth muscle.
Aspergillus-induced eosinophil recruitment to murine lung is dependent on cell wall polysaccharides (80.9)
Authors: Van Dyken Steven , Reese Tiffany A. , Huang Xiaozhu , Corry David B. , Locksley Richard M. 
Published: January 2023

Abstract
Development of allergic inflammation in response to fungal organisms involves innate immunity, but initial molecular recognition events remain unclear. We sought to identify elements of Aspergillus essential to initiating allergic inflammation using IL-4-GFP reporter (4get) mice. Intranasal administration of an Aspergillus preparation resulted in robust recruitment of innate immune cells to the lungs of 4get mice, independent of protease activity, implicating cell wall components. The major cell wall polysaccharides of Aspergillus include chitin, along with linear and branched β-glucan polymers, both of which can initiate innate immune responses in mammals. Enzymatic degradation of either chitin or β-glucans led to significant reductions in the magnitude of eosinophil recruitment to the lungs of 4get mice after intranasal Aspergillus administration, an effect that was enhanced additively by degradation of both polysaccharides. The specific contribution of chitin was examined using 4get-SPAM mice, which express elevated levels of acidic mammalian chitinase in the lungs and thus possess an enhanced ability to degrade chitin in vivo. After Aspergillus challenge, lung eosinophils numbers in 4get-SPAM mice were significantly diminished compared to 4get-WT littermates, in both acute and extended dosing regimens, while eosinophils remained at WT levels when challenged similarly with ovalbumin. These data reveal a previously unidentified link between specific polysaccharide recognition and eosinophil recruitment during Aspergillus-induced allergic lung inflammation.
Ad5 delivery of IL-15 enhances porcine natural killer cell activity (81.11)
Authors: Patch Jared R. , Toka Felix N. , Zarlenga Dante , Golde William T. 
Published: January 2023

Abstract
Foot-and-mouth disease virus (FMDV) is a highly infectious picornavirus that causes disease in cloven-hoofed animals, and remains an international economic threat to the livestock industry. Although a protective killed-virus vaccine exists, its utility is limited by the window of susceptibility present between vaccination and the induction of adaptive immunity. Natural killer (NK) cells play an important role in innate immunity by eliminating virus-infected cells. We have previously shown that interleukin-15 (IL-15) plays a critical role in the development, maintenance, and activation of NK cells in swine, and represents a potential method for improving porcine NK cell killing activity. We have constructed a replication-defective human Ad5 vector expressing the gene for porcine IL-15 (Ad5-pIL-15). Culture supernatants of cells infected with Ad5-pIL-15 were capable of stimulating porcine NK cells in vitro as indicated by elevated target cell killing. This suggests that Ad5-pIL-15 has the potential to stimulate porcine NK cell killing activity in vivo. Further experiments will focus on Ad5-pIL-15 and whether it can inhibit FMDV pathogenesis in pigs.
Onset of Antibody Repertoire Diversification in the Rabbit Appendix (81.17)
Authors: Lanning Dennis K. , Knight Katherine L. 
Published: January 2023

Abstract
Unlike mice and humans, rabbits generate their primary antibody repertoire by somatically diversifying V(D)J genes in gut-associated lymphoid tissues (GALT). Rabbit V(D)J genes are diversified by somatic hyper-point mutation and somatic gene conversion, both of which require activation-induced cytidine deaminase (AID). To determine when repertoire diversification begins, we examined AID expression in neonatal rabbit appendix by in situ hybridization. We first detected AID mRNA expression at one week of age, in B cells in the basolateral region of appendix follicles. We asked whether AID+ B cells were undergoing V(D)J gene diversification by using laser capture microdissection (LCM) to isolate AID+ and AID- B cells from 1-week-old appendix follicles. We RT-PCR-amplified and cloned VDJ genes from the two B cell populations, and searched for evidence of somatic diversification by comparing their nucleotide sequences to those of the appropriate germline genes. We found somatic gene conversion events in the VDJ genes from AID+ B cells, but not in those from AID- B cells. These results demonstrate that diversification of the rabbit primary antibody repertoire begins around 1 week of age, in B cells residing at the basolateral region of appendix follicles. This work was supported by NIH Grant 1 RO1 AI49458-01A1.
The role of Kruppel-like factor 2 (KLF2) in thymic emigration and trafficking of non-conventional T cell lineages (82.16)
Authors: Odumade Oludare A , Weinreich Michael A. , Takada Kensuke , McCaughtry Tom , Carlson Corey M. , Lingrel Jerry , Elewaut Dirk , Jameson Stephen C. , Hogquist Kristin A. 
Published: January 2023

Abstract
Immune function is dependent on the proper development of T cells in the thymus and emigration of these cells to the periphery. We recently showed the transcription factor KLF2 is required for emigration via regulation of S1P1 (sphingosine 1-phosphate receptor 1) and CD62L in conventional αβ T cells. However, the role of KLF2 and S1P1 in trafficking of non-conventional T cell subsets (γδ T cells, Treg, NKT, and Gut intraepithelial lymphocytes -IEL) has yet to be described. We show here that KLF2 is differentially expressed within the T cells subsets studied using both mRNA expression via quantitative PCR and/or a novel KLF2-GFP reporter mouse model. While KLF2 and S1P1 can be detected in sorted non-conventional thymic T cells and Gut IEL αβ T cells, we report no detectable KLF2 in Gut IEL γδ T cells. Using several KLF2 deficient mouse models, trafficking defects were observed in Treg, NKT, and Gut-IEL (αβ and γδ) populations. Finally, the data suggest that lack of KLF2 alters both the distribution and the phenotype of γδ T cells in secondary lymphoid organs. Overall, our results suggest that KLF2 is a common means by which different T cell lineages regulate trafficking and/or entry.
Funding provided by the National Institutes of Health (AI038903 to SCJ and AI039560 to KAH).
The energy sensitive kinase mTOR regulates T-bet mediated heritable type 1 effector cell fate of CD8 T cells (83.1)
Authors: Rao Rajesh  R , Shrikant Protul A 
Published: January 2023

Abstract
The inflammatory cytokine; IL-12, provides an essential third signal for CD8 effector cell fate determination. The molecular mechanisms by which IL-12 regulates STAT4 and T-bet activity for instructing effector cell fate remains unclear. The energy sensitive kinase; mTOR (mammalian target of rapamycin), a target of both the PI3K-Akt and/or AMPK signaling pathways has recently been shown to affect trafficking of effector CD8 T cells in vivo, however its role in cytokine determined effector cell fate remains untested. Using latex microspheres; H-2Kb/SIINFEKL and mB7.1, as surrogate antigen presenting constructs, we have established that the presence of IL-12 enhances and sustains antigen/B7.1 induced mTOR activity in naïve OT-1 T cells. The augmented mTOR activity requires PI3K as well as STAT4 and inhibition of mTOR; rapamycin or temsirolimus, leads to the loss of IL-12 induced sustained T-bet expression and subsequent loss of type I (IFNg) CD8 effector cell fate. Interestingly, inhibition of mTOR in IL-12 conditioned OT-1 cells produced only marginal decreases in cytolytic activity and increases in CD69, L-selectin and Eomesodermin expression. Hence, we have demonstrated a novel role for mTOR in integrating instructions to regulate effector cell fate in naïve CD8 T cells and characterized its mechanisms of action.
Supported by NIH and RPCI Alliance Foundation
The expansion of parasite-specific CD8+ T cells during infection is regulated by the transcription factor T-bet (83.7)
Authors: Smeltz Ronald , Cobb Dustin , Guo Siqi , Lara Ana , Manque Patricio , Buck Gregory 
Published: January 2023

Abstract
T-bet is a transcription factor that promotes Type I immunity to pathogens through effects involving T cells and dendritic cells. However, many of the functions of T-bet in CD8+ T cells can be compensated for by Eomes. Therefore we investigated the role of T-bet in immunity to T. cruzi, an intracellular pathogen that causes Chagas disease and which requires IFN-γ-secreting CD8+ T cells for resistance. T-bet-deficient mice (t-bet-/-) were highly susceptible to T. cruzi infection. However, susceptibility was not secondary to a lack of T cell activation, as T cells displayed typical markers of activation, including CD44, CD62L, and CD25. In contrast, we observed a profound reduction in the number of antigen-specific CD8+ T cells in t-bet-/- mice as determined by MHC tetramer analysis. Failure to generate sufficient numbers of antigen-specific T cells was not due to impaired APC functions, as we observed no differences in co-stimulatory molecule expression or in antigen-driven proliferation of wild-type CD8+ T cells in infected t-bet-/- mice. However, transfer of naïve T cells from t-bet-/- donors into infected rag-2-/- mice (t-bet+/+) demonstrated a similar quantitative defect, as CD8+ T cells failed to expand in vivo. These data demonstrate that T-bet contributes to resistance to T. cruzi infection partially via regulating the expansion of T. cruzi-specific CD8+ T cells in a T cell-intrinsic manner. They also serve to further illustrate the multi-faceted functions of T-bet in controlling quantitative aspects of T cell immunity, in addition to qualitative components such as Th1/Th17 immunity.
Deltex1: a novel regulator of T cell activation and T cell anergy (84.12)
Authors: Liu Wen-Hsien , Hsiao Huey-Wen , Lai Ming-Zong 
Published: January 2023

Abstract
Deltex1 (DTX1) is a target of Notch signaling, but its physiological function in T cells is unclear. Here we show that Dtx1 is a novel transcription target of NFAT involved in T cell anergy. DTX1 was upregulated during both in vitro and in vivo T cell anergy induction. Transgenic expression of Dtx1 attenuated T cell activation but does not alter T cell development. DTX1 transgenic mice were resistant to EAE induction. In the present study, we generated DTX1-deficient mice to study the function of DTX1 in T cell anergy and T cell development. Dtx1 knockout did not interfere with thymocyte development. Deficiency of DTX1 augmented T cell activation, conferred resistance to anergy induction, enhanced autoantibody generation, and increased inflammation and autoimmunity. DTX1 therefore represents an interactive component downstream of calcium/NFAT that regulates T cell anergy.
Protein phosphatase 2A (PP2A) is involved in the age-related down-regulation of E47 mRNA stability (84.14)
Authors: Blomberg Bonnie B. , Landin Ana Marie , Diaz Alain , Riley Richard L. , Frasca Daniela 
Published: January 2023

Abstract
We have previously shown that the transcription factor E47, which regulates activation-induced cytidine deaminase (AID) and Immunoglubulin class switch in splenic B cells, is down-regulated in aged murine B cells due to increased E47 mRNA decay. At least part of the decreased stability of E47 mRNA seen in aged B cells is mediated by tristetraprolin (TTP), a physiological regulator of mRNA stability. We found that TTP mRNA and protein expression are higher in old than in young B cells. Both TTP protein expression and function in B cells are dependent upon p38 MAPK and there is less phospho-p38 in old activated B cells. Also there is less phospho-TTP in old than in young splenic activated B cells which leads to more binding of TTP to the 3'-UTR of E47 mRNA, therefore decreasing its stability. PP2A is a serine/threonine protein phosphatase that plays an important role in the regulation of a number of major signaling pathways. We found that not only the amount but also the activity of PP2A is increased in old B cells. By specific inhibition of PP2A we have shown less phospho-p38 MAPK and phospho-TTP in old B cells. PP2A dephosphorylation of either p38 MAPK and/or TTP may account for more binding of the hypophosphorylated TTP to the 3'-UTR of the E47 mRNA, inducing its degradation. This mechanism may be at least in part responsible for the age-related decrease in antibody class switch.
This work is supported by NIH AG-23717 (BBB) and by NIH AI-064591 (RLR).
Th2 specific immunity and function of peripheral T cells requires the p56Lck SH3 domain (84.15)
Authors: Straus David B. , Finkelman Fred D , McCoy Margaret E. 
Published: January 2023

Abstract
T-cell activation and effector function is essential for immunity. T-cell antigen receptor (TCR) signals can regulate the outcome of differentiation, but the mechanisms are unclear. We have found that the Src family tyrosine kinase, p56Lck, specifically links TCR signals to activation of the Mitogen Protein Kinase (MAPK) pathway through the action of its SH3 domain and controls T cell activation and Th2 immunity. We have utilized an Lck SH3 mutant knock-in mouse line, LckW97A, to investigate the potential role of this regulatory domain in regulation of T-lymphocyte activation and effector function. Our results demonstrate that the p56Lck SH3 domain is required for normal activation of T-lymphocytes. LckW97A T-cells have reduced IL-2 production, CD69 induction and proliferation following TCR stimulation. Biochemical studies show that activation of the MAPK pathway is selectively altered as P-ERK1/2 induction is significantly reduced but phospho-PLCγ1 induction is unaffected. In vivo experiments demonstrate significantly impaired Th2 immunity with reduced serum levels of IgG1, IgE and IL-4 following immunization with DNP-KLH or infection with the helminth Nippostrongylus brasiliensis. These data indicate that p56Lck SH3 domain regulates activation of T-lymphocytes by controlling MAPK pathway induction and demonstrate a critical role for p56Lck in the regulation of Th2-type immunity.