bluuebunny/crossref_metadata_2025_split huggingface dataset:
Meningeal Immunity, Learning and Memory, and IL-4: Do T cells Make You Smart? (87.35)
Authors: Derecki Noel , Cardani Amber , Quinnies Kayla , Crihfield Anastasia , Lynch Kevin , Kipnis Jonathan 
Published: January 2023

Abstract
T cells were recently shown to support learning and memory, although the underlying mechanisms for this phenomenon are yet unknown. Here, we show that severe combined immune deficient (SCID) mice, shown to be impaired in assays of learning and memory, display a pronounced pro-inflammatory (M1) skew of meningeal myeloid cells. Acute lymphopenia induced by FTY720, a sphingosine-phosphate receptor agonist, or blockade of T cell migration into the meninges using anti-VLA4 antibodies, results in similar cognitive impairment and M1-skewed meningeal myeloid phenotype in wild type mice. The role of IL-4 is critical: we show accumulation of IL-4+ CD4+ T cells in the meninges of wild type mice after MWM training. Moreover, IL-4 knockout mice exhibit profound cognitive impairment and M1 skew. Transfer of IL-4 knockout bone marrow to wild type mice results in meningeal myeloid inflammation and impaired learning, while transfer of wild type T lymphocytes to IL-4 knockout mice is sufficient to rescue both behavioral phenotype and M1 skew. Our results point to a critical role for meningeal T cells, and for IL-4 in particular, in the regulation of meningeal myeloid cells and concomitant cognitive function. These results might lead to development of new immune-based therapies for cognitive conditions associated with immune decline, such as HIV- and age-associated dementia.
Computational modeling of Hsp90 inhibition illustrates a potential therapy to inhibit immune-mediated inflammation in systemic lupus erythematosus. (87.8)
Authors: Shimp Samuel , Reilly Christopher , Rylander Marissa 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is a chronic inflammatory autoimmune disorder that can affect nearly every organ in the body. Mesangial cells from lupus mice are hyper-responsive to immune stimulation and overproduce inflammatory mediators including IL-6 and nitric oxide (NO). Heat Shock Protein 90 (Hsp90) is known to chaperone numerous signaling client proteins including phosphatidylinositol 3-kinase (PI3K) and Akt, which belong to the immune activated PI3K, Akt, and mTOR (PI3K/Akt/mTOR) inflammatory pathway. Geldanamycin (Geld) inhibits Hsp90 function by binding to the carboxyl (C-) terminal domain resulting in deactivation, destabilization and degradation of Hsp90 inflammatory signaling clients, such as Akt, for decreased inflammation. Our long-term goal is to elucidate and mathematically model the cellular mechanisms of the Hsp90 modulated PI3K/Akt/mTOR pathway in SLE. Lupus mouse mesangial cells were cultured and treated with various concentrations of Geld followed by immune stimulation (LPS/IFN-γ) for 12, 18, and 24 hours. Cell cytokine analysis shows a sharp transition from stimulated levels of NO and IL-6 to significantly reduced NO and IL-6 concentrations at the Geld treatment range of 0.01 µM to 0.1 µM. Based on the measured data an empirical model for predicting the extracellular cytokine concentration following treatment with Geld was formulated. The empirical model suggests that N-terminus inhibition of Hsp90 may be a potential therapy for SLE.
IDENTIFICATION OF CIS-REGULATORY ELEMENTS TARGETING AID-MEDIATED SEQUENCE DIVERSIFICATION TO THE CHICKEN IMMUNOGLOBULIN LIGHT CHAIN GENE (88.2)
Authors: Kothapalli Naga Rama , Norton Darrell , Fugmann Sebastian 
Published: January 2023

Abstract
Somatic hypermutation (SHM) and gene conversion (GCV) are two closely related processes that increase the diversity of the primary immunoglobulin (Ig) repertoire. Both processes are initiated by the activation-induced cytidine deaminase (AID) that converts cytosine residues to uracils in a transcription-dependent manner and processed by direct replication and error-prone DNA repair. It is unknown how this mutagenic activity is targeted almost exclusively to Ig loci. To identify cis-acting elements involved in targeting SHM and GCV to the chicken Ig light chain (IgL), we used DT40 cells and systematically deleted non-coding sequences from the IgL locus using standard gene targeting strategies. We identified a novel as of yet uncharacterized regulatory region (named 3′RR) in the chicken IgL gene, containing a classical transcriptional enhancer and also cis-acting DNA elements essential for targeting AID-mediated sequence diversification. We are continuing our systematic deletion strategy to identify the minimal sequence required for targeting and to determine the trans-acting factors that bind to it. To identify similar targeting elements in the murine Igκ locus, we use a cross-complementation approach replacing the 3′RR in DT40 cells with murine Igκ sequences. Previous mouse transgenic studies indicated that both iEκ and 3'Eκ were necessary for the SHM of Igκ transgenes, and we are currently testing the ability of these enhancers to restore SHM/GCV in the chicken locus.
Th1 bias and gender differences in the ex vivo functional capacity of human NKT cells. (89.17)
Authors: Snyder-Cappione Jennifer , Tincati Camilla , Eccles-James Ijeoma , Cappione Amedeo , Koth Laura , Nixon Douglas 
Published: January 2023

Abstract
Human NKT cells are a unique T cell subset capable of producing Th1, Th2, or Th17 cytokines. It is hypothesized that the composite cytokine profile of the NKT cell compartment may determine disease pathogenesis. In order to define the functional signature of individual human NKT cells ex vivo with greater resolution, we FACS-Aria sorted NKT cells from fresh PBMC of 12 healthy donors and assessed their functional profiles, on both a population and single cell level, using Luminex and Elispot platforms, respectively. The purified NKT cell populations produced seven of the 14 analytes measured by Elispot, including IL2, IL4, IFNγ, TNFα, MIP1α, MIP1β, and perforin. Surprisingly, IL5, IL6, IL10, and IL17 were not detected from any subjects, and only one subject’s NKT cells produced IL13. Comparison of Elispot numbers with the distribution of several circulating NKT subsets revealed a significant positive association between the percentage of CD4/CD8 Double Negative cells and the number of perforin-secreting cells. NKT functional profiles of age-matched subjects revealed significantly higher frequencies of Th1-producing cells in male donors. These results suggest that 1) the panoply of human NKT Th2 and Th17 responses may not be as pronounced as previously believed, and 2) NKT cells may contribute to the sex bias found in autoimmune and other diseases.
The role of fibroblasts in neonatal intestinal inflammation (89.46)
Authors: Zawahir Shamila , Okogbule-Wonodi Adora , Li Guanghui , Blanchard Thomas 
Published: January 2023

Abstract
Background: Fetal intestinal fibroblast cell lines (FIF) produce more IL-8 in response to LPS compared to non-neonatal intestinal fibroblast lines (NNIF). Necrotizing Enterocolitis (NEC) is the most common gastrointestinal emergency in neonates and occurs more frequently in premature infants than term infants. We propose that fetal fibroblasts are inherently more responsive to stress and contribute to increased inflammation. Method: Primary human FIF and NNIF cell lines were exposed to E. coli LPS for 24 hours and the amount of IL-8 in the supernatants was determined by quantitative ELISA. A membrane blotting cytokine detection assay was also used to establish the cytokine profiles in the supernatants. Supernatants were used to stimulate neutrophils and activation was measured by increased CD11b expression by flow cytometry. Results: LPS stimulation induced a cytokine profile in FIF distinct from NNIF that included MCP-1, IL-6 and GRO-alpha. The supernatants from LPS stimulated FIF also activated neutrophils significantly more than supernatants from LPS treated NNIF. Conclusions: Fetal fibroblasts are more sensitive to stimulation by bacteria endotoxin and produce cytokines that promote inflammation, including the activation of neutrophils. Lamina propria fibroblasts in the developing gut may drive inflammation and put premature infants at an increased risk for developing NEC.
Automated analysis of flow cytometry data for the identification of cellular markers of allergy therapeutic responses (91.2)
Authors: Qian Yu , Kong Megan , Casale Thomas , Campbell John , Sadat Eva , Thomson Elizabeth , Dunn Patrick , Xiang Diane , Dahlke Carl , Scheuermann Richard 
Published: January 2023

Abstract
Mechanistic studies of biological specimens obtained during the conduct of clinical trials are frequently used to identify markers of therapeutic responses and to understand the underlying physiological processes associated with therapeutic efficacy. The Immunology Database and Analysis Portal (ImmPort; www.immport.org) is a public database resource funded by NIAID to support the management and analysis of clinical and mechanistic data generated by their funded investigators. We demonstrate the use of the ImmPort clinical data management platform and novel data analysis methods in the analysis of the allergen immunotherapy experiments conducted by the Casale group of Immune Tolerance Network. In the Casale study, immune response assays were conducted on samples taken before, during and after a 12-week course of allergen-specific immunotherapy to investigate whether anti-IgE antibody (omalizumab) can enhance the safety and efficacy of rush immunotherapy. Flow cytometry data was re-analyzed with FLOCK, a novel automated flow cytometry analysis pipeline in ImmPort, to determine if the level of any blood cell population correlated with treatment conditions. The analysis has identified a unique cell population expressing CD14, CXCR3 and high levels of the Fc-epsilon receptor (CD23), which is significantly increased in patients undergoing rush immunotherapy. Such data-driven analysis using ImmPort can generate hypotheses for further experimental design and verification.
Immune response profiles after caterpillar exposure (91.3)
Authors: Smith-Norowitz Tamar , Norowitz Kevin , Silverberg Jonathan , Kohlhoff Stephan , Kalra Kaushal , Chice Seto , Nowakowski Maja , Durkin Helen , Bluth Martin 
Published: January 2023

Abstract
Rationale: The role of the immune response to caterpillar exposure is not well described. Methods: Distributions of blood T and B lymphocytes, monocytes (CD1d+), levels of serum Igs(IgM, IgG, IgA, IgE), and cytokines (IFN-γ, IL-4, TNF-α) were studied in an allergic asthmatic serum IgE+ (181 IU/mL) adult who was exposed to the caterpillar, Halysidota harrisii Walsh, 1864. Results: Numbers of CD4+ T cells, CD45 RA+ T cells, CD25 + cells, CD19+ B cells, and CD1d+ monocytes decreased (20-33%) one week post reaction, while CD8+CD60+ T cells and CD23+ cells decreased 48 hrs (33, 74%) post reaction. In contrast, numbers of CD16/56+ NK precursor cells increased (60%) 12 hr, then decreased (65%) 48 hr post reaction; other lymphocyte subsets were unaffected. Serum IgM, IgG and IgA were normal; however, serum IgE levels were elevated 2 hrs post reaction (15%), and returned to pre-exposure levels by 24-48 hrs. High levels of IFN-γ (187-319 pg/mL) and TNF-α (549-749 pg/mL) were detected in serum 24-36 hrs and 3.5-24 hrs post reaction, respectively. In contrast, levels of IL-4 were undetected (&lt;1.0 pg/mL) in serum at all time points. Conclusions. Caterpillar exposure may result in a variety of clinical reactions including increased serum IgE levels, blood CD16/56+ NK precursor cells, and cytokines. Understanding immunomodulation in caterpillar exposure will provide better insight and mechanisms of caterpillar contact and tailor more effective therapeutic intervention.
Roles of TSLP in diverting airway tolerance to Th2 sensitization (91.6)
Authors: Zhou Baohua , Lei Liying , Zhang Yanlu 
Published: January 2023

Abstract
Induction of Foxp3+ regulatory T cells (iTreg) is an important mechanism to generate mucosal tolerance against harmless antigens. On the other hand, mucosal tolerance breakdown is an important step towards allergen sensitization that ultimately leads to allergy and asthma. Recent studies demonstrated that thymic stromal lymphopoietin (TSLP) is essential for the development of allergic inflammation. The precise mechanism by which TSLP induces allergic response remains inadequately understood. Here we report that intranasal treatment of Ovalbumin (OVA) + TSLP was sufficient to induce Th2 sensitization against OVA while inhalation of OVA induced airway tolerance. In fact, elevated airway TSLP was able to suppress the development of antigen-specific iTregs in a dose dependent manner. We further demonstrated that TSLP significantly inhibited the differentiation of Tregs in vitro. This inhibition was TSLP specific as naïve CD4 T cells isolated from TSLP receptor deficient mice showed normal iTreg differentiation in the presence of TSLP. Although TSLP could directly promote Th2 differentiation in vitro, IL-4 or Stat6 deficiency only partially rescued the inhibition of iTreg induction by TSLP. Taken together, our findings suggest that TSLP, in addition to promote Th2 differentiation, can directly suppress antigen-specific iTreg induction in response to inhaled antigens, and thus induces Th2 sensitization and allergic inflammation.
Detection of cardiac myosin-specific CD4 T cells by using MHC II / IAk tetramers in A/J mice (93.12)
Authors: Massilamany Chandirasegaran , Gangaplara Arunakumar , Chapman Nora , Rose Noel , Reddy Jay 
Published: January 2023

Abstract
Myocarditis is one of the predominant causes of heart failure in young adolescents, and Coxsackievirus B strains (CVB1-6) are common suspects. CVB3 isolates from humans induce myocarditis in A/J mice, but the infectious virus cannot be isolated 18-21 days postinoculation. However viral nucleic acids showing deletions in the 5’ non-translated regions persist. The extent to which continuing low level replication of CVB causes inflammation is not known and an autoimmune response is suspected in late disease progression, and cardiac myosin heavy chain-alpha (MYHC-α) has been identified as a dominant autoantigen. To delineate the contribution of autoimmune response to postinfectious myocarditis, it is critical to demonstrate the generation of pathogenic autoreactive T cells as they expand to a significant proportion to cause disease. To this end, we have created major histocompatibility complex class II / IAk tetramers for MYHC-α, 334-352. By using IAk / RNase 43-56 tetramers as controls, we have optimized the conditions to detect MYHC-α 334-352-specific CD4 T cells in lymph node cell cultures derived from A/J mice immunized with MYHC-α 334-352. Preliminary studies suggest that the reagents can also be used to enumerate the frequency of myosin-specific T cells ex vivo in immunized mice. These reagents will now permit us to determine the generation and functions of myosin-reactive T cells as an indicator of autoimmune response to cardiac myosin in mice infected with CVB3.
Activation of C3 promotes experimental autoimmune thyroiditis in C5 deficient female A/J mice. (93.15)
Authors: Njoku Dolores , Talor Monica , Sharma Rajni , Mellerson Jenelle , Burek Carol , Rose Noel , Njoku Dolores 
Published: January 2023

Abstract
Clinical studies suggest that activated C3, an early product of complement activation which can be triggered by infectious or environmental agents, is associated with more severe autoimmune thyroiditis, but few studies have illuminated mechanisms responsible for this observation. Animal models show that the severity of autoimmune thyroiditis is associated with late complement components, such as C5, and genetic susceptibility at the I-Ak sublocus. To elucidate additional complement-mediated effects on autoimmune thyroiditis we induced thyroiditis in I-Ak positive CBA/J and C5-deficient A/J mice with and without cobra venom factor which would activate the alternative complement pathway. A/J mice as a group developed less severe thyroiditis than CBA/J mice (p &lt; 0.001). Surprisingly, female A/J mice developed less thyroiditis than males (p &lt; 0.01) and had lower serum C3 levels (p &lt; 0.05) during thyroiditis induction while thyroiditis and C3 in male A/J was not different from CBA/J mice. Cobra venom factor increased serum C3 levels, TNFα and thyroiditis but not intra-thyroid C3 deposition in A/J females suggesting that systemic complement activation increased thyroiditis in these mice. Our studies suggest that C3 activation promotes experimental thyroiditis in less susceptible A/J females, and may reveal one mechanism by which infections or other environmental agents promote autoimmune thyroiditis in patients.
A BALB/c locus on chromosome 12 confers resistance to lupus in FcγR2-/- mice. (93.23)
Authors: Tarasenko Tatiana , Bolland Silvia 
Published: January 2023

Abstract
FcγR2B-deficient (R2-/-) mice develop autoantibodies and glomerulonephritis with a pathology resembling human lupus when on the C57Bl/6 background. The same mutation on the BALB/c background does not lead to spontaneous disease, suggesting differences in lupus susceptibility between the BALB/c and B6 strains. An F2 genetic analysis from a B6/BALB cross identified region from the B6 chromosome 12 with positive linkage for autoantibodies. We have generated a congenic strain (B6.R2-/-sbb2) that contains the suppressor allele from the BALB/c centromeric region of chromosome 12 in an otherwise B6.R2-/- background. We have observed that the presence of sbb2 renders B6.R2-/- mice resistant to lupus disease by suppressing all antibody autoreactivity and spontaneous immune cell activation. Mixed bone marrow reconstitution experiments indicated that the sbb2 resistant allele is most likely T cell intrinsic. The sbb2 congenic interval contains several candidate genes that are currently being investigated. We generated transgenic lines containing BAC DNA insertions spanning the sbb2 interval. The new BAC transgenic strains were tested in the induced model of lupus by pristane administration. Newly generated BAC transgenic strains were also bred to B6.R2-/- to test for suppression of lupus in spontaneous disease. The goal of our experiments is to identify a new lupus susceptibility gene within the sbb2 interval that is able to suppress lupus disease by altering T cell function.
Activation of innate immunity influences salivary gland function and development of Sjögren’s Syndrome (93.33)
Authors: Deshmukh Umesh , Nandula Seshagiri-Rao , Scindia Yogesh , Bagavant Harini 
Published: January 2023

Abstract
Sjögren’s syndrome (SS) is a chronic autoimmune disorder characterized by progressive lymphocytic infiltration within the exocrine glands. Loss of salivary gland (SG) function is a major feature of the disease. This study was undertaken to investigate the role of innate immunity activation in pathogenesis of SS. Mice were repeatedly injected with TLR3 agonist poly(I:C), and monitored for SG function and sialadenitis. Poly(I:C) treatment rapidly induced SG hypofunction. However, SG function was restored, once poly(I:C) treatment was terminated. This phenomenon was independent of the genetic background of the mice, and did not require T and B cells. Mice deficient in IFNAR1 and thereby lacking type I IFN signaling, did not loose SG function to the same extent as the wild type mice. In the autoimmune-prone NZB/W F1 mice, poly(I:C) treatment induced severe and accelerated sialadenitis. The SG in these mice showed presence of T-B cell aggregates and germinal center formation. Some of the T cells in these infiltrates were CXCR5+ICOS+ Tfh cells. Significant up-regulation in the expression of IL-21 and Il-21R genes was seen in the SG of poly(I:C) treated mice. This study demonstrates that activation of innate immunity in a genetically susceptible individual modulates the development of SS. This study also shows that type I IFN can adversely affect SG function and that IL-21 pathway and Tfh cells might be important in the pathogenesis of SS.
Mechanisms behind the induction of Ttc7 transcription during B lymphocyte development(93.35)
Authors: Tait Heidi 
Published: January 2023

Abstract
Changes in Ttc7 (tetratricopeptide repeat domain 7) transcription have been associated with changes in B lymphocyte signaling through changes in both the lymphopoietic environment and homeostasis of B lymphocyte development in mice with the Ttc7fsn/fsn(flaky skin) mutation. We have demonstrated that young Ttc7fsn/fsn mice exhibit decreased naive B lymphocyte populations in the bone marrow and spleen as compared to their wild type littermates, and that this disparity is exaggerated with age. The Ttc7fsn/fsn mutation also leads to excessive production of harmful B1b lymphocytes causing autoimmune disease closely related to SLE. This is significant as both autoimmune and aging human populations share similar B lymphocyte profiles. The evaluation of signaling events upstream of Ttc7 transcription will shed light on stage-specific B cell developmental signaling mechanisms in the bone marrow and spleen that cause immunological dysfunction. We have begun to classify B lymphocyte stages at which Ttc7 transcription rates fluctuate in pursuit of these signaling mechanisms. We have found Ttc7 to be transcribed in the bone marrow at early B lymphocyte stages and have evidence that its transcription is repressed at later stages in the spleen. To further support the hypothesis that induction of Ttc7 transcription during B cell development occurs in early B lymphocytes, we have also found that a 1kb segment of the promoter induces a higher rate of transcription in the pre-B 70Z/3 cell line.
Pre-exposure to uninfected sand fly bites protects rhesus monkeys against cutaneous leishmaniasis (94.14)
Authors: Oliveira Fabiano , Rowton Edgar , Gomes Regis , Lawyer Phillip , Teixeira Clarissa , Aslan Hamide , Valenzuela Jesus , Kamhawi Shaden 
Published: January 2023

Abstract
Leishmaniasis is a vector-borne disease that distresses millions in the poorest regions of the globe. There are no anti-Leishmania vaccines approved for human use. Mice pre-exposed to uninfected sand fly bites (USFB) are protected against cutaneous leishmaniasis (CL). To test if pre-exposure to USFB protects rhesus monkeys against CL, we first developed a model where monkeys challenged with 50 L. major-infected Phlebotomus duboscqi sand flies developed self-healing cutaneous lesions mimicking human CL. In a following experiment, we exposed animals 3 times, every 21 days, to 20 uninfected P. duboscqi. We found that 70% of the monkeys reacted to USFB(DTH reaction and antibodies). Twenty-one days after the last exposure to USFB naïve, pre-exposed reactive (PE-R) and pre-exposed non-reactive monkeys were challenged with 50 L. major-infected P. duboscqi. We observed a reduction in disease burden and healing time in the PE-R group. This protection correlated with a two log reduction in parasite load and the PE-R animals showed a reduced maximum lesion diameter indicative of diminished disease. Skin biopsies two weeks post-infection showed a higher expression of IFN-γ in the PE-R group compared to naïve monkeys. Altogether, these data show that pre-exposure to uninfected P. duboscqi bites can protect monkeys against L. major infection. Identification of the salivary proteins responsible for the observed protection may lead to the discovery of vaccine candidates against human CL.