bluuebunny/crossref_metadata_2025_split huggingface dataset:
Gas6/TAM signaling differentially modulates chronic fungal asthma with the expansion of myeloid regulatory cell subsets in mice (57.7)
Authors: Shibata Takehiko , Ismailoglu Ugur , Coelho Ana , Lukacs Nicholas , Kunkel Steven , Moreira Ana , Hogaboam Cory 
Published: January 2023

Abstract
Gas6, a Tyro3, Axl, Mertk (TAM) receptor ligand, is detected in a variety of diseases and has various roles. Herein, we show that Gas6 differentially modulates experimental fungal asthmatic response via the expansion and modulation of myeloid-derived regulatory cells (MDRCs). Aspergillus fumigatus-sensitized mice were challenged with live Aspergillus conidia and received approximately 2 μg (low) or 7 μg (high) of recombinant Gas6 via intranasal installation from days 14 to 28 after conidia challenge. In the low dose Gas6 group, significant airway hyperresponsiveness (AHR), airway remodeling, and whole lung IL-13 were observed compared with the control group. Although high dose Gas6 treatment significantly suppressed AHR and the whole lung levels of inflammatory cytokines compared with control, this treatment exacerbated airway remodeling. MDRCs have both disease enhancing and suppressing cells in asthma. Indeed, low dose Gas6 treatment increased the accumulation of CD11b+F4/80+Ly6C-Ly6G+ MDRC with pro-inflammatory properties into asthmatic lung whereas high dose Gas6 promoted the accumulation of immunosuppressive CD11b+F4/80+Ly6C+Ly6G- MDRC during chronic asthma. Anti-Axl Ab, but not anti-Mer Ab, treatment significantly suppressed not only AHR but also airway remodeling in asthmatic mice compared with IgG control asthmatic groups. Together, these results demonstrate that Gas6-TAM receptor interactions modulate fungal asthma, in part through effects on MDRCs.
Evaluation of three next generation sequencing platforms for immune repertoire sequencing (58.10)
Authors: Wang Chunlin , Levy Shawn , Yang Qunying , Byrne-Steele Miranda , Han Jian 
Published: January 2023

Abstract
The advent of next generation sequencing (NGS) techniques, combined with a semi-quantitative multiplex PCR method, enables us to comprehensively profile the lymphocyte receptor diversity (immune repertoire) of the entire collection of B or T cells from a particular sample. Different NGS platforms vary in read length, throughput, sequencing error rate and error pattern, hands-on time, turnaround time and price. Three popular NGS techniques (Illumina HiSeq, 454 FLX and Ion Torrent) were employed to sequence amplicons from T cell receptor clones generated with arm-PCR in order to evaluate the usage of those techniques on immune repertoire sequencing. The error pattern and rate of the three techniques in the context of T cell receptor sequences were profiled. Based on the differences of the three techniques, we discuss the usage of each NGS platform in the context of immune repertoire sequencing.
IL-27 sustains T-bet expression and promotes the development of terminally differentiated CD4 T cells during tuberculosis (59.5)
Authors: Torrado Egidio , Fountain Jeffrey , Pearl John , Cooper Andrea 
Published: January 2023

Abstract
IL-27 is a member of the IL-12 family of cytokines that was originally described to induce the expression of the transcription factors STAT1 and T-bet, thus promoting Th1 differentiation. Despite this apparent Th1 inducing profile, mice deficient in IL-27R signaling are more resistant to Mtb infection, even though CD4 T cells in the lungs of these mice produce lower amounts of IFN-γ on a per cell basis. Here we show that, in the absence of IL-27 signaling, CD4 T cells express lower levels of the transcription factor T-bet during the chronic phase of infection which corresponds with a reduced accumulation of terminally differentiated CD4 T cells in the lungs of these mice. Mechanistic analysis of the impact of IL-27 signaling in Ag-specific T cells suggest improved competitive fitness of CD4 T cells deficient in IL-27 signaling. Accordingly, the transcription profile of CD4 T cells from the lungs of Mtb-infected wild-type mice suggests that these cells are under oxidative stress, unlike CD4 T cells from the lungs of IL-27R-/- mice. These data suggest that IL-27 has an important impact in the maintenance of effector CD4 T cell responses during tuberculosis.
Cytotoxic effect of IL-17A on neural stem cells as a mechanism underlying Th17-cell-induced proinflammatory demyelination (59.7)
Authors: Zhang Guag-Xian , Lee Eric , Li Ke , Zhu Lin , Kumar Priyanka , Xu Hui , Ciric Bogoljub , Rostami Abdolmohamad 
Published: January 2023

Abstract
IL-17A, a Th17 cell-derived proinflammatory molecule, has been found to play an important role in the pathogenesis of autoimmune diseases, including multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE). The IL-17A receptor (IL-17RA), through which IL-17A signals, is expressed in many immune-related cells, including central nervous system resident cells such as microglia and astrocytes. However, whether IL-17A exerts a direct effect on neural stem cells (NSCs) and oligodendrocytes, thus inducing inflammatory demyelination in the central nervous system is not known. Here we report that IL-17RA is constitutively expressed on NSCs, oligodendrocytes and their precursors. IL-17A significantly reduced NSC proliferation, which was mainly due to cytotoxicity, but not to induction of apoptosis. IL-17A did not influence the differentiation of NSCs into neural cells. The effects of IL-17A are similar to those of TNF-α, another proinflammatory cytokine that plays a pathogenic role in EAE; however, a synergistic/additive effect of these two molecules has not been observed. We conclude that IL-17RA is expressed in NSCs, through which IL-17A blocks proliferation of these cells, resulting in reduced numbers of myelinating cells such as oligodendrocytes and their precursors. Thus, in addition to its proinflammatory role in the immune system, IL-17A also plays a direct role in inducing demyelination and blocking neuronal repair in EAE/MS.
The lupus-susceptibility Sle2c1 sublocus regulates T cell expansion through several mechanisms (60.1)
Authors: Xu Zhiwei , Morel Laurence 
Published: January 2023

Abstract
The lupus-susceptibility (278-37)rec1 sublocus is derived from the NZB-derived Sle2 locus found in the NZM2410 lupus-prone mouse. Our previous results showed that the candidate gene p18 in the (278-37)rec1 sublocus demonstrated significantly reduced mRNA expression and the (278-37)rec1 sublocus enlarged spleen, more notably peripheral lymph nodes in Fas-deficient mice. Our current research discovered firstly that the mouse with both p18 gene deletion and lpr mutation displayed expanded peripheral lymph organs as the same as the B6.(278-37)rec1.lpr mouse, meaning an association of p18 deficiency and lymphocyte hyperproliferation. Secondly, the B6.(278-37)rec1.lpr mouse showed significantly more Ki67+ T cells in lymph nodes than B6.lpr mouse, but T cells from both B6.(278-37)rec1.lpr and B6.lpr mice have the same proliferation response in vitro with stimulation by anti-CD3 and anti-CD28. Furthermore, both Treg cell percentage and the ration of Treg/T effective cell in B6.(278-37)rec1.lpr mouse showed markedly decreased in spleen and lymph nodes compared with B6.lpr mouse, but both mouse stains have the same Treg function in inhibiting T cell proliferation. Finally, the T cell apoptosis of B6.(278-37)rec1.lpr mouse seems unimpaired in activation-induced cell death assay. In summary, the T cell hyperproliferation controlled by the (278-37)rec1 sublocus involves several mechanisms, but their interrelation needs further to be elucidated in the future.
Rapid replacement by apoptosis in the draining lymph nodes sustains dynamics of skin-migratory dendritic cells in the steady state. (61.3)
Authors: Tomura Michio , Kabashima Kenji , Kanagawa Osami 
Published: January 2023

Abstract
Although, understanding of skin-DC homeostasis is important issue, its analysis has been delayed, because of the difficulty of labeling skin-DCs. Here we used newly established photoconvertible fluorescent protein, KikGR-KI mice to label DCs in skin and lymph nodes (LN) by exposure with violet light. This system allows us to directory measure replenishment kinetics and migration of skin-migratory DCs in the steady state. CD103-dermal DC (DDC) in draining (d) LN increased rapidly with peak at d 2 after labeling DCs in skin. On the other hand, skin-derived CD103+DDC gradually increased with peak at d 3. LC increased at similar rate to that of CD103+DDC and reached plateau at d 4. In photoconverted skin at d 5, 40% of DDC, 60% of CD103+DDC and none of LC were non-labeled, indicating presence of labeled-DC in the dLN was closely correlated with decrease of corresponding labeled-DCs in skin. Half replenishment time of DDC, CD103+DDC and LC were 0.6, 1.5 and 2 d measured by following decrease of labeled DC after labeling DCs in LN. Only a few emigrated DCs were detected in other LNs, and skin-migratory DCs had high caspase activity cells. Altogether, DC pool in skin is relatively larger than that of LN, and LN is always filled with newly influx DCs. Skin migratory DCs replaced at rapid turnover rate via apoptosis in LN without emigration, which would limit duration of T cell response and enable to transport carried peripheral antigens from skin-migratory DC to LN-resident DCs.
Omalizumab inhibits human mast cell responses by sequestering pre-bound IgE (64.6)
Authors: Kulka Marianna , Catalli Adriana 
Published: January 2023

Abstract
Mast cells are key effectors of allergic inflammation. IgE-mediated mast cell activation induces degranulation and inflammatory mediator release. Omalizumab (Xolair; Genentech Inc.) is a recombinant humanized monoclonal anti-IgE antibody that prevents IgE binding to its receptor, FcϵRI. Objective: We investigated the effects of omalizumab on IgE pre-sensitized human LAD2 mast cells. Methods: LAD2 degranulation was determined by β-hexosaminidase assay. Chemokine expression and prostaglandin synthesis was measured by quantitative PCR analysis and ELISA, respectively. IgE binding and FcϵRI expression was determined by flow cytometry. Results: Omalizumab pretreatment inhibited IgE binding to LAD2 cells and entirely prevented IgE-dependent upregulation of FcϵRI expression. In addition, omalizumab removed FcϵRI-prebound IgE as early as 24 hrs after treatment. After 5 days, bound IgE was reduced by 92%. Furthermore, omalizumab concomitantly reversed IgE-dependent FcϵRI upregulation by 49% 48 hrs post treatment and by 93% 5 days post treatment. Consequently, omalizumab attenuated ongoing IgE-mediated responses, reducing degranulation by 34%, chemokine expression up to 79% and prostaglandin synthesis by 34% after 7 days of omalizumab treatment. Conclusions: Omalizumab is able to remove pre-bound IgE from sensitized mast cells thereby reducing ongoing response to FcϵRI-dependent signals. This data suggests that omalizumab is an effective inhibitor of sensitized human mast cells.
Glucocorticoids repress the type-I-IFN system and accelerate osteoporosis during Pneumocystis lung infection (70.8)
Authors: Searles Stephen , Gauss Katherine , Mozer Michelle , Meissner Nicole 
Published: January 2023

Abstract
Pneumocystis (PC) infection can range from asymptomatic colonization to fatal pneumonia in severely immunocompromised individuals. Type-I-IFNs are critical regulators of immune responses to PC. In mice lacking type-I-IFN signaling (IFNAR-/-), PC clearance is delayed and transient bone marrow (BM) depression and accelerated osteoclastogenesis are observed. Lack of type-I-IFN-signaling in lymphocyte-deficient mice (IFrag-/-) augments systemic symptoms causing progressive BM failure and significant loss of bone mass due to accelerated osteoclastogenesis. These data highlight the importance of the type-I-IFN system in regulating local and systemic responses to PC infection. Glucocorticoids (GCs) are used to treat chronic inflammatory lung diseases and are immunosuppressive in part by inhibiting type-I-IFN signaling. Interestingly, osteoporosis is a systemic manifestation of chronic inflammatory lung disease and a side-effect of GC treatment. Here we show that GCs inhibit the PC-induced systemic type-I-IFN response in a mouse model. Mice infected with PC and treated with GC have significantly reduce trabecular bone mass compared to uninfected, GC treated mice, mimicking the IFrag-/- phenotype. BM numbers were also reduced and the osteoclastogenic potential increased in PC infected, GC treated mice. These data suggest that PC could act as a co-morbidity factor during GC treatment, ultimately augmenting side effects such as osteoporosis.
Pulmonary endotoxin tolerance increases costimulatory molecule expression on dendritic cells and antigen migration to the lung draining lymph nodes in a mouse model (71.2)
Authors: Natarajan Sudha , Beal Dominic , Kim Jiyoun , Remick Daniel 
Published: January 2023

Abstract
It has been shown that lung-specific endotoxin tolerance (ET) attenuates allergic responses in an environmentally relevant model of cockroach-allergen (CRA) induced asthma. The mechanisms of this response have not yet been elucidated. We hypothesized that ET would reduce antigen migration to lung draining lymph nodes, thereby suppressing development of the adaptive immune response to allergen. ET was induced in outbred (CD-1) mice by direct intratracheal installation of 100ng (low dose) or 1μg (high dose) endotoxin for 4 consecutive days. Controls received PBS. The following day, mice received 2μg of FITC-conjugated CRA and were sacrificed 24h later. Lungs, mediastinal and cervical lymph nodes were collected for FACS analysis. Results showed a dose-dependent increase in the number of antigen-carrying CD11c+FITC+ dendritic cells in the lymph nodes (PBS vs. 100ng LPS vs. 1ug LPS; 1.4*106 cells vs. 2.3 vs. 5.3). An increase was seen in the number of dendritic cells expressing costimulatory molecules CD80 (2.4*104 cells vs. 5.5 vs. 13.0), CD86 (5.1*104 cells vs. 12.7 vs. 30.1) and MHC class II (10.5*104 cells vs. 14.0 vs. 45.7). An increase in the MFI for CD86 was observed for both ET groups compared to control. MFI for MHC class II expression was also elevated in the high-dose ET group compared to controls. These data suggest that dendritic cells in ET mice have an increased capacity for upregulation of costimulatory molecules and antigen transport to the draining lymph nodes.
Aerosolized Influenza Infection During Late Gestation Pregnancy Results in Severe Clinical Outcomes in the Ferret Model (76.7)
Authors: Gage Emily , Reed Doug , Trichel Anita , Dunsmore Tammy , Beigi Richard , Stefano Cole Kelly 
Published: January 2023

Abstract
In 2009, the emergence of pandemic A influenza virus (pH1N1) demonstrated greater severity in pregnant women as recognized previously in other influenza pandemics. The Centers for Disease Control and Prevention reported a death rate of 20% in pregnant women in the United States who were hospitalized with influenza-like symptoms. To understand the immunological basis for the enhanced influenza-associated morbidity and mortality observed in pregnant women, pregnant and nonpregnant ferrets were challenged with aerosolized pH1N1 strain A/California/04/09. The pregnant ferrets had poorer gestational outcomes and presented a more rapid and severe clinical disease when compared to the nonpregnant ferret group. Viral load and tissue tropism were similar for both groups as determined by quantitative RT-PCR. Influenza virus was detected in the brains of both groups. Gross pathological analysis and H&E stains identified interstitial pneumonia in the lungs, with no significant pathology in any other tissues. Virus was also detected in placental tissue and kits of the pregnant ferrets suggesting transmission across the placental barrier. This model will be useful in discerning the differences in the host response to infection that results in greater morbidity and mortality during pregnancy.
Characterizing the T cell receptor repertoire of beryllium-responsive CD4+ T cells (P5021)
Authors: Bowerman Natalie , Falta Michael , Mack Douglas , Crawford Frances , Kappler John , Fontenot Andrew 
Published: January 2023

Abstract
Multiple public T cell receptor (TCR) repertoires exist in CD4+ T cells derived from the lung of patients with chronic beryllium disease (CBD). Here, we characterize the TCR Vβ5 repertoire which involves expression of a conserved glutamine (Q)-containing motif that is essential for TCR recognition of HLA-DP2/peptide/Be2+. Using a panel of human TCR Vβ antibodies, we show that beryllium-responsive, Vβ5.1+CD4+ T cell expansions exist in all DP2+ CBD patients tested to date. Sequence analyses revealed oligoclonal expansions consisting of Vβ5+ TCR genes that express a conserved motif composed of identical lengths, conserved joining region expression (Jβ2.5/1.4), and homologous amino acid (Ala/Gly) residues surrounding an essential Q residue. The existence of a public Vβ5+ TCR repertoire suggests selection by an identical antigen in lung, which is likely composed of HLA-DP2, peptide, and Be2+. As expected, these Vβ5+ T cells recognize the same antigen, as revealed by positive staining of CBD T cell lines with a soluble tetrameric HLA-DP2/peptide/Be2+ molecule. Interestingly, this MHCII tetramer was used to identify an additional public Vβ18+ T cell repertoire which expresses the conserved CDR3 motif found in the predominant Vβ5+ T cell population. The presence of beryllium-specific, Vβ5.1+CD4+ T cells in the lung of all DP2+ CBD patients suggests that the MHCII tetramers can be used as an early method of detection of CBD development in beryllium-exposed workers.
Measuring T cell activation in T cell-APC conjugates using the ImageStream imaging flow cytometer (P5043)
Authors: Friend Sherree , Kong Raymond , Hall Brian , Basiji David , Haley Pugsley 
Published: January 2023

Abstract
Adaptive immune responses require antigen presentation to antigen-specific effector cells. Upon interaction between the antigen presenting cell (APC), specific antigen and T cell there is a reorganization of the cytoskeleton and recruitment of adhesive and signaling molecules to the site of intercellular contact forming an immunological synapse (IS). Upon formation of a functional IS, signaling molecules join the clusters and contribute to the signaling cascade leading to translocation of nuclear factors from the cytoplasm to the nucleus culminating in gene activation. Recruitment of these molecules is measured by imaging cell conjugates directly. To analyze a statistically significant number of specific conjugates we used the ImageStream imaging flow cytometer with a 60X objective, which objectively and rapidly collects large numbers of images and provides quantitative image analysis tools to evaluate subcellular localization of molecules. In this study we evaluate the specific location of the adhesion and signaling molecules LFA-1 and Lck within the IS complex in T cells when presented with SEB pulsed APC. In addition we measured the translocation of NFkB in the T cell from the cytoplasm to the nucleus showing the activation of the T cells when conjugated to APC.
Cytosolic flagellin-induced lysosomal pathway regulates inflammasome-dependent and independent macrophage responses (P4141)
Authors: Bortoluci Karina , Lage Silvia , Buzzo Carina , Amaral Eduardo , Matteucci Kely , Massis Liliana , Ferreira Luis Carlos , Rodriques Mauricio , D'Império Lima Regina , Amarante Mendes Gustavo 
Published: January 2023

Abstract
NAIP5/NLRC4 inflammasome activation by cytosolic flagellin results in caspase-1-mediated processing and secretion of IL-1β/IL-18 and pyroptosis, an inflammatory cell death pathway. Here, we found that although NLRC4, ASC and caspase-1 are required for IL-1β secretion in response to cytosolic flagellin, cell death nevertheless occurs in the absence of these molecules. Cytosolic flagellin-induced inflammasome-independent cell death was accompanied by IL-1α secretion and was temporally correlated with the restriction of S. Typhimurium infection. Despite displaying some apoptotic features, this peculiar form of cell death did not require caspase activation but was regulated by a lysosomal pathway, in which cathepsin B and cathepsin D play redundant roles. Moreover, cathepsin B contributed to NAIP5/NLRC4-inflammasome-induced pyroptosis, and IL-1α and IL-1β production in response to cytosolic flagellin. Together, our data describe a novel pathway induced by cytosolic flagellin that induces a peculiar form of cell death and regulates inflammasome-mediated effector mechanisms of macrophages.
The immune system and cancer stemness cross-talk can predict therapeutic response and patient outcome (P3030)
Authors: Kryczek Ilona , Tanikawa Tkashi , Szeliga Wojciech , Vatan Linda , Zou Weiping 
Published: January 2023

Abstract
Myeloid-derived suppressor cells (MDSCs) inhibit anti-tumor immunity. However, their involvement in cancer stem cell (CSC) biology, and their molecular, clinical and therapeutic relevance of this involvement have not been studied. Here, we investigated MDSCs in human breast cancer. We show that MDSCs prompt breast cancer stemness through enhance CSC gene expression, sphere formation and cancer metastasis. The effects depend on the cross-talk between interleukin (IL)-6/signal transducer and activator of transcription 3 (STAT3) and nitric oxide (NO)/NOTCH signaling pathways. MDSCs initiate STAT3 phosphorylation through IL-6, and activate Notch and sustain prolonged STAT3 phosphorylation via NO in CSCs. Furthermore, MDSCs positively correlate with CSC numbers and signature genes in breast cancer, and serve as a negative predictor for patient survival and herceptin response. Collectively, our study has revealed novel extrinsic cellular and molecular mechanisms controlling cancer stemness, and suggests that targeting both MDSCs and cancer is essential for engendering potent therapeutic efficacy in cancer patients. Efforts to target this network might be a valuable strategy in anti-cancer therapy.
IL-4-dependent regulation of skin barrier function. (P3042)
Authors: Serezani Ana Paula , Riley Jon , Spandau Dan , Goenca Shreevrat , Kaplan Mark 
Published: January 2023

Abstract
Atopic dermatitis (AD), a common manifestation of allergic disease, can be an early indicator of subsequent atopic phenotypes. Mutations in the Filaggrin gene, a component of the Epidermal Differentiation Complex (EDC) has emerged as a predisposing factor for the development of AD. EDC genes are critical for barrier function in the skin, but the Th2-type cytokines IL-4 and IL-13 inhibit expression of EDC genes in keratinocytes. We hypothesized that pro-allergic cytokines alter skin barrier function and facilitate allergen exposure by altering keratinocyte differentiation. Thus, to define the mechanisms of IL-4-dependent changes in gene expression, the activation of STAT6 was investigated in normal human keratinocytes (HK). We observed that STAT6+ cells were induced by IL-4, resulting in a peak of positive cells after 30 minutes of stimulation. Moreover, IL-4 significantly inhibited EDC genes expression in HK differentiated in vitro. IL-4 also resulted in a significant increase in the expression of the chemokine CCL26 (Eotaxin3) and the matricellular protein periostin, which is involved in persistent allergic inflammation. Thus, IL-4-dependent changes in keratinocyte gene expression are likely a key component in AD pathogenesis.