bluuebunny/crossref_metadata_2025_split huggingface dataset:
Local autoantibodies in Chronic Rhinosinusitis (CRS) with Nasal Polyps. (48.8)
Authors: Tan Bruce , Li Quan , Suh Lydia , Kato Atsushi , Conley David , Chandra Rakesh , Peters Anju , Norton James , Harris Kathleen , Grammer Lelie , Kern Robert , Mohan Chandra , Schleimer Robert 
Published: January 2023

Abstract
Abstract Introduction: Chronic rhinosinusitis with nasal polyps (CRSwNP) is an inflammatory condition of the nasal passage and paranasal sinuses. It is characterized by a Th2 biased inflammatory infiltrate not currently attributed to autoimmunity. Recent studies from our laboratory demonstrate elevated levels of BAFF, B-lymphocytes, and IgA. The role of these B-cells and the specificities of the immunoglobulins produced are still unknown. Methods: Protein extracts from nasal polyps of 11 patients were compared to nasal tissue obtained from 10 normal controls using autoantibody microarrays. A positive microarray result for anti-double stranded DNA (anti-dsDNA) was confirmed using 15 nasal polyp samples and 10 normal control samples on commercially available ELISAs for Total IgG, and anti-dsDNA IgG. Results: Total IgG levels were higher in nasal polyps relative to normal controls (35μg/mg Total Protein versus 17μg/mg Total Protein, p=0.02). Additionally, Anti-dsDNA IgG levels were higher in nasal polyps relative to normal controls on both the antibody microarray (188nfi/mg Total Protein versus 61nfi/mg Total Protein p&lt;0.01) and ELISA (371U/mg Total Protein versus 66U/mg Total Protein p=0.03). Conclusions: Anti-dsDNA is present at higher levels in nasal polyp tissue than normal controls- suggesting that a local autoimmune reaction is present in nasal polyp tissue. Further studies are needed to test the role of this autoimmune response in the pathogenesis of CRS.
CCR4 improves phenotypic identification of T-regulatory cells; Validation and implementation of clinical test (49.26)
Authors: Gorski Kevin , Sitko John , Thomas John , Gavin Marc , Cottrell Susan , Cox Chris , Vincent Mike , Ferbas John 
Published: January 2023

Abstract
T-regulatory cells (Treg) are critically responsible for controlling unwanted immunological activity. Therefore, potential perturbation of Tregs can be important to consider when studying therapeutic intervention of inflammatory disorders with novel agents. Moreover, reliable assays are needed to allow detection of changes resulting from treatment. In the clinical setting intracellular staining for antigens such as Foxp3 can be unwieldy. Therefore, we chose to identify Treg using cell surface markers. Recent reports indicate that using a CD25hi and CD127dim phenotype on CD4+ T-cells allows identification of an enriched fraction of Treg using surface staining. On average, however, only 85 % of these cells are reported to be Foxp3+ Treg. We evaluated use of CCR4 together with CD25 and CD127 to refine detection of Treg. Foxp3 co-staining was performed as well to characterize the proportion of bonafide Treg. More than 90 % of cells in the CD25hi, CD127dim, CCR4+ CD4+ T-cell gate were found to co-express Foxp3. We then validated a 5-color method incorporating CCR4 as a marker for T-reg enumeration. 26 healthy volunteers and 13 asthmatic subjects were evaluated. Pre-analytical variables and assay performance characteristics will be discussed. We conclude that using CCR4 imparts additional specificity for phenotypic identification of Treg. Implementation is planned for clinical studies where therapeutic intervention has the potential to impact Treg.
Role of Smad and non-Smad signals in the development of Th17 and regulatory T cells (49.3)
Authors: Zheng Song Guo , Lu Ling , Wang Julie , Zhang Feng , Chai Yang , Brand David , Wang Xuehao , Horwitz David , Shi Wei 
Published: January 2023

Abstract
While TGF-β is essential for the development of peripherally induced Foxp3+ regulatory T (iTreg) and Th17 cells, the intracellular signaling mechanism by which TGF-β regulates development of both cell subsets is less well understood. Using Smad2 lymphocyte conditional knock out and Smad3 knock out mice, we report that neither Smad2 nor Smad3 deficiency abrogates TGF-β-dependent iTreg induction by a deacetylase inhibitor Trichostatin A (TsA) in vivo although loss of Smad2 or Smad3 partially reduces iTreg induction in vitro. Similarly, Smad2 and Smad3 have a redundant role in development of Th17 in vitro and in experimental autoimmune encephalomyelitis (EAE). In addition, ERK and/or JNK pathways were shown to be involved in regulating iTregs, while the p38 pathway predominately modulates Th17 and EAE induction. Therefore, selective targeting of these intracellular TGF-β signaling pathways during iTreg and Th17 cell development may lead to the development of therapies in treating autoimmune and other chronic inflammatory diseases.
Hepatic stellate cells regulate T cell responses in cooperation with Kupffer cells. (49.9)
Authors: Ichikawa Shintaro , Mucida Daniel , Kronenberg Mitchell , Cheroutre Hilde 
Published: January 2023

Abstract
Induced regulatory T cells (iTreg) are important regulators of mucosal effecter T cell responses. TGF-β is essential for iTreg differentiation, however, in the presence of interleukin-6, it promotes pro-inflammatory T helper 17 (Th17) cells. We have shown that vitamin A metabolite; retinoic acid (RA), acts as a key controller of TGF-β-dependent immune responses, inhibiting Th17 differentiation and promoting Foxp3+ Treg induction. Hepatic stellate cells (HSC) are liver resident cells that store the majority of vitamin A and RA in our body. Because the liver is a tolerogenic organ, in spite of its direct contact with intestinal antigens, we explored if HSCs are involved in the tolerogenic nature of the liver. Although HSC can prime naïve T cells we found they were less effective than the neighboring Kupffer cells (KC), which are major antigen presenting cells in the liver. Because KCs reside in the proximity of the HSCs within the liver sinusoide, we rendered it possible that HSCs could regulate the differentiation of KC-primed T cells. We show here that HSCs indeed promote Foxp3 induction by KC-primed OT-II T cells and interestingly, they inhibited Th17 differentiation, similar to the RA-mediated reciprocal regulation of TGF-β-dependent T cell polarization. These data suggest that liver resident KCs and HSCs synergize to cooperatively regulate liver T cell responses.
Reorganizing the signaling molecules in CD4+ T cells by TCR-MHC class II interactions in the thymic medulla prevents T cell hyperactivation (50.1)
Authors: Stephen Tom , Wilson Bridget , Laufer Terri 
Published: January 2023

Abstract
Immature thymocytes that are positively selected based upon their response to self-peptide-MHC complexes develop into mature T cells that are not overtly reactive to those same complexes. Developmental tuning is the active process through which TCR-dependent activation of single-positive thymocytes is attenuated in response to the self peptide-MHC molecules that will be encountered in the periphery. Mature “tuned” thymocytes lose responsiveness to low affinity ligands. We previously reported that tuning of CD4 T cells requires interactions with MHC II expressing thymic medullary epithelium and thymic DCs. Tuning is mediated by MHC II interaction with the TCR but not with the CD4-coreceptor, even though mature CD4 T cells depend on CD4 co-receptor associated Lck for TCR signaling. The increased threshold for TCR stimulation in tuned CD4SP thymocytes is associated with increased expression of the SHP-1 phosphatase and inhibition of ERK activation by weak ligands. Immature CD4 T cells contain a “free pool” of Lck that is not co-receptor associated. This pool of Lck localizes with TCR and may lower the activation threshold for weak ligands. Tuning limits this TCR associated pool of Lck. Thus, TCR signaling in mature T cells depends on CD4 associated Lck for activation. Taken together, our results suggest that reorganization of the TCR signalasome during thymic medullary residency defines the activation threshold of CD4 T cells and prevents hyperactivity to weak self antigens.
TcR-stimulation upregulates MS4a4B expression through induction of AP-1 transcription factor during T cell activation (50.14)
Authors: Yan Yaping , Zhang Guang-Xian , Rostami Abdolmohamad , Xu Hui 
Published: January 2023

Abstract
MS4a4B is a novel member of the MS4A family expressed in T-cells. We have shown previously that expression of MS4a4B in T cells is upregulated upon T cell activation. However, the mechanism that regulates MS4a4B expression is unknown. In this study, we explored the mechanism underlying TcR-stimulation-induced expression of MS4a4B by promoter analysis. We cloned 2495 bp of 5’-flanking region upstream of MS4a4B start coden and inserted the DNA fragment into pGL4.20 reporter plasmid. To analyze promoter activity, we transiently transfected EL4 thymoma cells and the T32 cell line with reporter plasmids. Expression of reporter gene was determined by dual-luciferase assay. Potential activator- and repressor-binding sites were analyzed by serial length of 5’-deletion. We have identified at least two potential activator binding regions and two repressor binding regions. The activator binding sites have been localized to two fragments, which are a 442-base pair region (region A) positioned from -1176 to -735, and a 119-base pair region (region B) positioned -188 to -70 respectively. MatInspector analysis showed that region A contains consensus binding motif of Jun/Fos (AP-1) family of transcription factors. EMSA showed that nuclear proteins extracted from CD3/CD28-activated T cells strongly bind to the AP-1 binding element, suggesting that TcR-stimulation may enhance MS4a4B expression through transactivation of AP-1 in T cells.
A critical role of GRAIL in T cell activation and tolerance (50.18)
Authors: Nurieva Roza , Zheng Shuling , Jin Wei , Sun Shao-Cong , Lozano Guillermina , Dong Chen 
Published: January 2023

Abstract
T lymphocyte activation is tightly regulated to ensure effective elimination of invading pathogens as well as maintaining tolerance against self-tissues. Gene related to anergy in lymphocytes (GRAIL) is a type I transmembrane protein localized to endosomal compartment with homology to RING finger proteins whose expression was previously associated with induction of T cell tolerance in vitro. In order to understand the physiological function of GRAIL in immune regulation, we have generated and analyzed a Grail-deficient mouse model. We found that naïve T cells lacking GRAIL exhibited greatly enhanced proliferation and cytokine production after TcR activation and did not depend on CD28 and ICOS for their effector cytokine expression. We also determined that lack of GRAIL abrogated suppressive function of regulatory T cells. Both naïve and regulatory T cells from Grail-deficient mice were less efficient in down-regulation of their TcR/CD3 expression. In vivo, Grail-deficient mice were resistant to immune tolerance induction and when compared to the wild-type mice, exhibited greater susceptibility to autoimmune diseases. Our results thus indicate GRAIL as an essential regulator of T cell tolerance by regulating naïve T cell tolerance and regulatory T cell function.
Sustained A1 and Mcl-1 expression from costimulatory signals promotes T cell survival (50.20)
Authors: Song Jim 
Published: January 2023

Abstract
Costimulation is essential for mediating efficient T cell responses. In recent years, advances have been made in studies of costimulation as potential therapies in the treatment of diseases. However, the intracellular signals by which costimulation affects the function of T cells still needs to be elucidated. A1 and Mcl-1, two bcl-2 family proteins are up-regulated in effector T cells by ligation with an agonist antibody to the costimulatory molecule OX40 (CD134), which is essential for long-term T cell survival. To understand whether they are molecular targets of OX40, we examined A1 and Mcl-1 expression in primary T cells from wild-type and OX40-deficient mice, and found A1 and Mcl-1 were up-regulated after early T cell activation, but not be maintained in T cells from OX40-deficient mice. In addition, we used retrovirus-mediated transduction to introduce A1 or Mcl-1 into antigen (Ag)-responding T cells from OX40-deficent mice. We found that exogenous expression of A1 or Mcl-1 reversed the survival defect of OX40-deficient T cells both in vitro and in vivo. These results indicate that A1 and Mcl-1 are molecular targets of OX40, which regulate T cell survival. The data provide new insights into the mechanisms by which costimualtion regulates T cell survival.
Partial rescue of Itk-/- iNKT cell development by Txk/Rlk reveals a unique role for Itk in the survival of iNKT cells. (50.40)
Authors: Qi Qian , Bai Yuting , Schwartzberg Pamela , August Avery 
Published: January 2023

Abstract
Invariant natural killer T (iNKT) cells are a distinct population of innate T lymphocytes that play important role in the immune response. Itk and Txk/Rlk are Tec family kinases that are expressed in iNKT cells, with the expression level of Itk about 7-fold higher than that of Txk. It has been reported that the Itk null mice have reduced iNKT cell frequency and numbers, as well as defects in iNKT cell development and cytokine secretion, all of which are exacerbated in the Itk/Txk DKO mice. By contrast, the phenotype of iNKT cells in Txk null mice is similar to that from WT mice. In order to determine whether Itk and Txk plays distinct roles in iNKT cell development and function, we examined the iNKT cells in Tg(Lck-Txk)/Itk-/- mice, which over express Txk in T cells to the similar level as Itk. Over expression of Txk rescues the maturation and cytokine secretion of iNKT cells in Itk null mice. It also rescues the altered expression of transcription factor T-bet, eomesodermin and PLZF. By contrast, over expression of Txk does not rescue the reduced iNKT cell numbers in Itk null mice, which is due to the failure to rescue the increased apoptosis observed in Itk-/- iNKT cells. These data suggest that Txk plays an overlapping role with Itk in iNKT cell development and function, but that Itk also has a unique function in the survival of iNKT cells.
Evaluation Of The Immune Response Against The Newcastle Disease Virus When Co-Administered With Beta Subunit Of The Cholera Toxin (52.17)
Authors: Franco García Julio , Moreno Salcedo Yazmín , González Hernández Carlos , León Kempis Maria 
Published: January 2023

Abstract
METHODS:4 groups of 10 SPF chicken each were included: Positive control, vaccinated subcutaneously with a commercial ND vaccine, negative control -non-vaccinated- and two more groups were vaccinated with ND+CTB, one subcutaneously and the fourth group was vaccinated via intranasal. Vaccination at 3 wks of age and challenge 21 days post-vaccination. Sera were collected before vaccination and in a weekly manner during 6 wks post-vaccination in order to evaluate serological response and UHI titers. Mortality and clinical signs were evaluated during 21 days post-challenge. RESULTS. Chickens vaccinated subcutaneously with ND-CTB reached the highest antibody titers when compared to the other experimental groups followed by chickens vaccinated with the commercial vaccine, then the intranasally vaccinated group and then the negative control. The group immunized subcutaneously with CTB also showed an earlier onset of serological response against ND and the duration of immunity was similar to the positive control. Mortality post challenge could be observed in the negative control as well as in intranasaly immunized birds, where all birds died as opposed as in groups of the positive control and the subcutaneously vaccinated, where all the birds survived. CONCLUSIONS: CTB seemed to contribute towards an earlier onset of humoral response in the ND model
Infectious challenge of Plasmodium berghei γ-spz immunized mice rescues effector CD8+ T cells, thus assuring protracted protection (52.9)
Authors: Krzych Urszula , Dalai Sarat , Ehrler Lindsey , Jobe Ousman , Egner Lisa 
Published: January 2023

Abstract
Protracted protection against experimental Plasmodia sporozoite (spz) challenge has been achieved in human and murine malaria by multiple immunizations with radiation-attenuated Plasmodia sporozoites (γ-spz). In B6 mice, long-lasting protection induced by Plasmodium berghei (Pb) γ-spz is linked to liver CD8+ T effector memory (EM) and central memory (CM) cell subsets. Protracted protection also depends on the interval between the last boost immunization and the infectious spz challenge. An interval of 6 months or longer results in loss of protection. In this study we asked whether a delayed challenge causes a decline of either CD8+ TEM or TCM or both cell subsets. We measured IL-2 and INF-γ producing polyclonal CD8+ TEM (CD44hiCD62Llo KLRGintCD107+CD122lo) and CD8+ TCM cells (CD44hiCD62Lhi KLRG-CD122hi) and observed that the frequencies of cytokine producing KLRGintCD107+ CD8+ TEM cells fell precipitously at 210 days after the last boost with γ-spz and that these mice were not protected when challenged 7 months after the last boost immunization. In contrast, mice challenged 2 weeks after the last boost maintained high frequencies of both CD8+ TEM and CD8+ TCM subsets when tested at 210 days following the last boost immunization with γ-spz. These mice were also protected upon a re-challenge (210 days) and remained protected beyond day 548. We are currently investigating the rescuing mechanism of CD8+ TEM cells by infectious spz challenge.
Abnormal morphology of melanosomes in the autoimmune vitiligo-prone Smyth line chicken does not appear to be due to alteration in lipid composition (83.16)
Authors: Stepicheva Nadezda , Liyanage Rohana , Lay Jackson , Dienglewicz Robert , Erf Gisela 
Published: January 2023

Abstract
The Smyth line (SL) chicken is an important animal model for autoimmune vitiligo. SL vitiligo is multifactorial in nature including genetic susceptibility, immune system components and environmental triggers. Genetic susceptibility to SL vitiligo appears to be manifested in part by an inherent melanocyte defect. SL melanosomes (pigment-carrying organelles) have irregular-shaped membranes with pigmented extensions that may be due to abnormal lipid composition. Lipids can affect protein topology and trafficking (and hence protein degradation and presentation), membrane stability and oxidative stress resistance. To compare melanosomal lipids from SL and control lines of chickens, melanosomes were isolated from growing feathers of 4-week-old SL, Brown line (parental control) and Light Brown Leghorn (vitiligo-resistant) chickens and lipids were extracted. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) combined with thin-layer chromatography (TLC) was used to determine expression differences in different classes of lipids by using TLC-MALDI imaging technique. Total fatty acid composition was examined by gas chromatography-mass spectrometry (GC-MS) of fatty acid methyl esters. Statistical analysis of preliminary data shows no significant differences in total fatty acid and phospholipid composition in melanosomes from SL and control chickens. Hence, lipid composition may not explain the melanosomal abnormalities observed in SL melanocytes.
Global transcriptome analysis of feathers from Smyth line chickens revealed different gene-expression profiles at various stages of autoimmune vitiligo (83.17)
Authors: Shi Fengying , Lee Jeong Yoon , Song Joon Jin , Kong Byung-Whi , Dienglewicz Robert , Erf Gisela 
Published: January 2023

Abstract
The Smyth line (SL) chicken is an animal model for autoimmune vitiligo (SLV). SL chickens develop spontaneous, post-hatch, autoimmune melanocyte loss in growing feathers (GF). With repeatable access to GF and the high incidence (85-95%) of SLV, SLV development can be examined in the same bird. A microarray study was conducted to investigate the global transcriptomic profile in GF from SL and Brown line (BL, parental control) chickens. RNA pools were made from GF of BL chickens, SL chickens that never developed SLV (S0), and SL chickens before (S1), during (S2) and at complete (S3) SLV. Complementary RNA probes were generated from BL and SL (S0~S3) RNA, labeled with Cy3 and Cy5, respectively and hybridized on a chicken 4x44 K Agilent microarray. The slide was scanned and normalized signal intensities were analyzed to identify differentially expressed (DE, &gt;2x) genes between BL and SL samples. There were 504, 518, 1124, and 1151 DE genes containing 262, 226, 658, and 680 unique DE genes in S0, S1, S2 and S3, respectively. S0 and S1 had similar proportions of up-and down-regulated DE genes while up-regulated DE genes predominated in S2 and S3. The highest expressed DE genes were related to immune activity and cell death. Ingenuity Pathway Analysis software was used to determine biological functions and pathways of DE genes. The study provides comprehensive information from global gene expression to functionalities and biological networks of DE genes in the process of SLV.
Pigmented growing feathers from autoimmune vitiligo-prone Smyth line chickens have heightened sensitivity to 4-tertiary butyl phenol (83.19)
Authors: Dong Lei , Dienglewicz Robert , Erf Gisela 
Published: January 2023

Abstract
The Smyth line (SL) chicken model for vitiligo exhibits spontaneous, autoimmune loss of melanocytes in growing feathers. Onset of SL vitiligo (SLV) can be observed between 6 and 20 wks of age and affects 85-95% of SL chickens. Multiple factors contribute to the melanocytes’ cell death, including genetic susceptibility to vitiligo as well as environmental factors that trigger autoimmune loss of SL melanocytes (e.g., live turkey herpesvirus (HVT) administration at hatch). In humans, phenolic derivatives, such as 4-tertiary butyl phenol (4-TBP), have been shown to trigger vitiligo in susceptible individuals. 4-TBP is structurally similar to tyrosine, the initial substrate of tyrosinase in melanin synthesis. In addition to interfering with melanin production, 4-TBP exposure results in oxidative stress. To examine the effect of 4-TBP in SL and control chickens, pigmented growing feathers were collected from 1, 4 and 7 week-old SL, Brown Line (BL, parental control, &lt;1% vitiligo) and Light Brown Leghorn (LBL, vitiligo resistant) chickens that did or did not receive HVT. The newest three mm growth (the melanocyte-containing, active pigmentation site) of the feathers was cultured with 4-TPB (200 μM) and the production of oxidative radicals was monitored in vitro. Independent of HVT treatment, 4-TBP induced more oxidative stress (P &lt; 0.05) in feathers from SL than BL and LBL chickens, suggesting heightened sensitivity of melanocytes to 4-TBP in vitiligo-prone SL chickens.