bluuebunny/crossref_metadata_2025_split huggingface dataset:
Prolonged CD154 expression in pediatric lupus CD4 T cells correlates with increased CD154 transcription, increased NFAT activity, and glomerulonephritis (135.4)
Authors: Cron Randy , Genin Anna , Brunner Michael , Scalzi Lisabeth , Mishra Nilamadhab , Beukelman Timothy , Mehta Jay 
Published: January 2023

Abstract
To explore a mechanism explaining dysregulated CD154 expression in lupus, cell surface CD154 expression was examined, pre- and post-activation, on peripheral blood CD4 T cells from 29 children with lupus and matched controls by flow cytometry. CD154 mRNA levels and transcription rates were studied by real-time RT-PCR and nuclear run-on assays, respectively. NFAT transcriptional activity and NFAT mRNA levels in lupus CD4 T cells were explored by reporter gene analysis and real-time RT-PCR, respectively. We found CD154 surface protein levels were increased 1.44-fold on lupus CD4 T cells compared to controls at one day post-activation ex vivo and this correlated with the presence of nephritis. Increased CD154 protein also correlated with increased CD154 mRNA levels and rates of CD154 transcription, particularly at later time-points post-T cell activation. Reporter gene analyses revealed a trend for increased NFAT, but decreased AP-1 and similar NFkB, activity in lupus CD4 T cell compared to controls. NFAT1 and, in particular, NFAT2 mRNA levels were notably increased in lupus CD4 T cells compared to controls. In summary, following activation, cell surface CD154 is increased on pediatric lupus CD4 T cells compared to controls, and this correlates with the presence of nephritis, increased CD154 transcription rates, and NFAT activity. These results suggest that NFAT/calcineurin inhibitors, such as tacrolimus and cyclosporine, will be beneficial treating lupus nephritis.
Expression of chitinase-like proteins in inflamed tissues of Sjögren’s Syndrome (SS) patients (135.6)
Authors: Greenwell-Wild Teresa , Moutsopoulos N. , Gliozzi M. , Kapsogeorgou E. , Rangel Z. , Munson P. , Moutsopoulos H. , Wahl S. 
Published: January 2023

Abstract
SS represents a chronic autoimmune disease of salivary and lacrimal glands that may be accompanied by multi-organ systemic manifestations. Among the locally involved innate immune defense populations are macrophages, which have been linked to IL-12 and IL-23 that moderate T lymphocyte lineage commitment. To further an understanding of the immunopathologic sequelae associated with SS and to define therapeutic targets, we performed microarray analysis of target tissues from SS patients with severe histopathologic lesions compared to those with sicca symptomatology but negative minor salivary gland biopsy. We identified the expression of multiple genes, including members of the mammalian chitinase family, not previously associated with exocrinopathies, among the most highly expressed SS genes as compared to diseased, but non-SS tissues. Both chitinase-3-like-1 (CHI3L1/YKL-40) and chitinase 1 (CHIT1), highly conserved members of the mammalian chitinase-like glycoprotein family, one with and one lacking enzymatic activity, were evident at the transcriptome levels, as well as detected as glycoproteins within inflamed salivary glands. Monocyte to macrophage differentiation is accompanied by an increase in chitinase expression, which can be augmented by cytokine exposure. Since the elevated expression of these molecules corresponded with more advanced disease in SS patients, these observations suggested a potential immunopathologic involvement.
Positioning of the HA Globular Head within the TLR5 Agonist, Flagellin, Modulates the Immunogenicity and Reactogenicity of a Novel-H1 Pandemic Influenza Vaccine (136.13)
Authors: Umlauf Scott , Gathuru John , Bell Rodney , Hou Fu , Reiserova Lucia , Song Langzhou , Weaver Bruce , Liu Ge , Tussey Lynda 
Published: January 2023

Abstract
The recent appearance of a novel H1N1 (nH1N1) virus has highlighted the importance of rapid response to emerging pandemic threats. VaxInnate has developed an influenza vaccine production system that permits rapid response on a global scale. The vaccine comprises the protective globular head subunit of HA fused covalently to the TLR 5 ligand flagellin. Using animal models that have successfully predicted the tolerated dose range in humans we have identified an optimal position for the HA globular head. Three constructs evaluated the globular head fused to the C terminus of flagellin (C-term), in place of domain 3 (R3) and in both positions (R3.2x). The three formats were tested in mouse immunogenicity and rabbit reactogenicity models. HAI assays were performed using CA/07/2009 virus. Rabbits were monitored for body temperature, food consumption and CRP. In mice, HAI titers &gt;1:40 were achieved with 1 μg doses of all vaccines, but R3 and R3.2x vaccines yielded higher titers. In rabbits, the C-term vaccine elicited elevated temperature and CRP levels and reduced food consumption at doses well below R3 and R3.2x. Replacement of domain 3 of flagellin with the HA globular head, either R3 or R3.2x, proves safe and immunogenic in mice and rabbits. Our clinical experience with other flagellin-linked vaccines has correlated well with the rabbit model. The results predict that the R3 or R3.2x nH1 vaccines would be both safe and efficacious in humans.
Alternative TLR signaling restores Francisella tularensis LPS-mediated protection against Francisella tularenis LVS challenge in TLR2-/- mice (136.17)
Authors: Cole Leah , Vogel Stefanie 
Published: January 2023

Abstract
The Category A select agent Francisella tularensis (Ft) is the causative agent of tularemia, a potentially fatal infectious disease. Intraperitoneal infection of mice with ~10 Ft Live Vaccine Strain (Ft LVS) organisms causes a lethal infection that resembles human tularemia. Immunization with 100 ng Ft LVS lipopolysaccharide (Ft-LPS) provokes an antibody response that protects wild-type (WT) C57BL/6J mice against an otherwise lethal Ft LVS challenge, but offers limited protection to TLR2-/- mice (WT, 85% survival vs. TLR2-/-, &lt;25% survival; p &lt; 0.0001, N = 4 separate experiments). Since macrophage recognition of Ft LVS is overwhelmingly TLR2-dependent (Cole, L. E., et al. 2007. Infect. Immun. 75:4127-37), we hypothesized that treatment of TLR2-/- mice with an alternative MyD88-dependent, TLR agonist would compensate for the failure of Ft LVS to signal in TLR2-/- mice and thereby lead to increased Ft-LPS-mediated protection. Concurrent administration of the TLR4 agonists, E. coli LPS (5μg) or monophosphoryl lipid A (MPL; 100μg), a synthetic non-toxic derivative of E. coli lipid A, with either the Ft-LPS immunization or the Ft LVS challenge, increased the survival of Ft-LPS-pretreated TLR2-/- to 100% (N=4 separate experiments). This suggests that effective Ft-LPS-mediated protection against Ft LVS challenge requires both Ft-LPS recognition and MyD88-dependent, TLR signaling. Supported by NIH AI157168.
In vitro examination of the role of TLR5 in enhancement of antigen presentation. (136.19)
Authors: Atif Shaikh , Letran Shirdi , McSorley Stephen 
Published: January 2023

Abstract
Data from our laboratory suggest that TLR5 can enhance the presentation of flagellin eptiopes to CD4 T cells in vivo. In this present study we have examined this process in vitro using antigen-presenting cells harvested from wild-type and TLR5-deficient mice and examining early activation of flagellin-specific SM1 CD4 T cells. Our in vivo data suggest that TLR5 expression in the spleen is absolutely required for activation of SM1 T cells. Recapitulation of this response in vitro will allow us to identify the TLR5+ antigen presenting cells in this anatomical site. As previous studies have failed to identify any TLR5+ population in the spleen, these experiments have the potential to enhance our understanding of flagellin antigen presentation and splenic dendritic cell subsets.
TLR reporter cell lines for screening TLR agonists and antagonists (136.20)
Authors: Rosenberg Jonathan , Lee Hyun-Ku , Singh Gita , Andrew Kody , Singh Sujay 
Published: January 2023

Abstract
We have developed novel TLR/NF-κB SEAPorter™ HEK 293 cell lines each expressing an intact Toll-like Receptor (TLR) as well as a secreted alkaline phosphatase (SEAP) reporter whose expression is under control of an NF-κB response element. These cell lines are suitable for TLR-dependent functional assays and are an ideal fit for high-throughput screening of TLR agonists or antagonists. TLR expression of each cell line has been validated by Flow Cytometry. Using a 96-well plate format, functional activity has been evaluated by criteria including cell number titration, ligand dose response, cell stimulation time and DMSO tolerance. Validation results yield an average signal-to-background (S/B) ratio of 16.9±0.7 as well as Z' value of 0.75±0.05 for ligand dose-response tests. Assay validation is based on the 96-well plate format screening assay, but are also suitable for 384- or 1536-well plate format screening assays. For TLR agonist screening, each compound in a library can be directly applied to cells and induced SEAP in culture media can be measured without additional steps. TLR antagonist screening can be performed as agonist screening except the addition of TLR ligands is done before or after treatment with library compounds. Consequently, the TLR-specific cell-based high-throughput feature of our novel TLR/NF-κB SEAPorter™ HEK 293 cell lines will provide a fast, flexible and convenient means for lead compound screening in the pharmaceutical drug development fields.
Evidence that gC1qR and DC-SIGN associate on the surface of immature dendritic cells (136.24)
Authors: Hosszu Kinga , Vinayagasundaram Uma , Vinayagasundaram Rama , Santiago-Schwarz Frances , Peerschke Ellinor , Ghebrehiwet Berhane 
Published: January 2023

Abstract
Complement protein C1q is involved in many immunoregulatory functions, such as modulating T-cell proliferation and regulating dendritic cell (DC) differentiation and maturation. C1q has at least two multi-functional cell surface receptors, cC1qR and gC1qR. While these receptors ostensibly lack a direct conduit into intracellular elements, they are known to form docking/signaling complexes with transmembrane partners. DC-SIGN (dendritic cell-specific-ICAM-grabbing-non integrin) is a C-type lectin expressed on the surface of DCs and involved in T-cell activation and endocytosis. Because SIGN-R1, a C-type lectin related to DC-SIGN has been shown to bind directly to C1q, we investigated whether DC-SIGN also binds to C1q and if DC-SIGN may act as a transmembrane partner for gC1qR and/or cC1qR. Using flow cytometry, we found that DC-SIGN, gC1qR and cC1qR are all present on the surface of human immature DCs (iDC) grown in culture. While antigen-capture ELISA experiments using iDC lysates showed that C1q, gC1qR and cC1qR directly or indirectly associate with DC-SIGN, immunofluorescence microscopy revealed that C1q and gC1qR, but not cC1qR co-localize with DC-SIGN on the surface of iDCs. The data suggest that DC-SIGN and gC1qR may directly associate on the surface of iDCs and regulate DC differentiation in a stage specific manner. Since both gC1qR and DC-SIGN have been shown to bind HIV, it is possible that the two receptors may play a role in the pathogenesis of HIV-AIDS.
TLR5 functions as a scavenger receptor to enhance flagellin-specific CD4 T cell Responses (136.29)
Authors: Letran Shirdi , Lee Seung-Joo , Atif Shaikh , McSorley Stephen 
Published: January 2023

Abstract
Innate immune activation via Toll-like Receptors (TLRs) causes dendritic cell maturation and the production of inflammatory mediators. Our data also demonstrate that TLR5 functions as a scavenger receptor for flagellin, enhancing the activation of flagellin-specific CD4 T cells. Flagellin-specific CD4 T cells were found to expand poorly in TLR5-deficient mice immunized with flagellin and this deficiency persisted even when additional TLR agonists were provided. In marked contrast, TLR5-deficient mice generated a robust flagellin-specific T cell response to processed flagellin peptide. The deficiency in flagellin-specific T cell activation was also detected in TLR5-deficient mice that were orally infected with a flagellated pathogen, demonstrating that TLR5 is responsible for directing a dominant CD4 response to flagellin in the context of infectious disease. Together, these data demonstrate that in addition to its function as an initiator of innate inflammation, TLR5 functions as a scavenger receptor allowing optimal presentation of flagellin epitopes to CD4 T cells.
Novel signaling interactions between proteinase-activated receptor 2 and Toll-like receptors in vitro and in vivo (136.36)
Authors: Nhu Quan , Rallabhandi Prasad , Shirey Kari Ann , Teijaro John , Farber Donna , Netzel-Arnett Sarah , Antalis Toni , Fasano Alessio , Vogel Stefanie 
Published: January 2023

Abstract
Toll-like receptors (TLRs) and proteinase-activated receptors (PARs) function as innate immune biosensors in mucosal epithelial cells (ECs). We previously reported the functional and physical interactions between TLR4 and PAR2, and more recently, between PAR2 and the TLR adapter, TRIF, and established a novel mechanism of “receptor cooperativity” between PAR2 and TLR4. We hypothesized that intracellular signaling pathways utilized by TLRs and PAR2 would converge either cooperatively or non-cooperatively when co-engaged. We demonstrate herein the cooperation between PAR2 and TLR2, TLR3, or TLR4 for activation of NF-κB-dependent signaling in mucosal EC lines. In contrast, activation of PAR2 negatively regulated TLR3-dependent antiviral pathway, blunting the expression of TLR3/IRF-3-driven genes, as well as activation of IRF-3 and STAT1. Similarly, TLR4-TRIF-activated, IRF-3-driven genes were also attenuated by PAR2 co-activation. Consistent with these in vitro observations, PAR2−/− and TLR4−/− mice, which were refractory to footpad edema induced by PAR2 agonist peptide, were protected from mouse-adapted H1N1 influenza A virus-induced lethality when compared to wild-type mice. These data support and extend our recently described, novel model of PAR2-TLR4 “receptor cooperativity” and highlight the complexity of signaling integration between heterologous innate immune biosensors.
Role of PI3K and autophagy in virus-stimulated IFN-α production by human plasmacytoid dendritic cells (136.46)
Authors: Deng Jing , Singh Sukhwinder , Fitzgerald-Bocarsly Patricia 
Published: January 2023

Abstract
Human plasmacytoid dendritic cells (pDC) produce large amount of IFN-α in response to viral stimuli through TLR7 and TLR9. Phosphatidylinositol-3 kinase (PI3K) has been shown to be activated by TLR stimulation in many cell types and Class I and class III PI3K are essential in the regulation of autophagy. We aimed to study the role of autophagy in both the processing of virus and induction of IFN-α in human pDC, which have not been described. We investigated the effects of Class I and class III PI3K inhibitors wortmannin, 3-methyladenine (3MA) and LY294002 on IFN-α production by HSV, Flu, SeV, CpG A and HIV-MN/AT2-stimulated human pDC. pDC were pretreated with the inhibitors then stimulated with virus for 6hr for detection of intracellular IFN-α by intracellular flow cytometry or overnight for stimulation and release of IFN-α, which was detected by ELISA. Inhibition of PI3K with wortmannin, 3MA and LY294002 inhibited IFN-α production by pDC. The PI3K inhibitors did not affect endocytosis or macropinocytosis. In addition, HSV dramatically induced autophagosome double-membrane structures in pDC as visualized by TEM and LC3B aggregation as seen in imaging flow cytometry. All of the inhibitors inhibited HSV-induced LC3B aggregation in pDC. These results support key roles for PI3K and autophagy in mediating IFN-α secretion and virus detection by human pDC. Supported by AI26806.
TIM-4 mediated phagocytosis of apoptotic cells induces regulatory cytokine secretion by mouse peritoneal macrophages (136.47)
Authors: Bu Xia , Zhu Baogong , Umetsu Dale , DeKruyff Rosemarie , Freeman Gordon 
Published: January 2023

Abstract
TIM-4 is a member of the T cell immunoglobubulin mucin (TIM) family and is expressed on macrophage and dendritic cell populations. TIM-4 specifically binds phosphatidylserine (PtdSer) on the surface of apoptotic cells and mediates phagocytosis of apoptotic cells by macrophages. NIH3T3 cells transfected with TIM-4 can phagocytose apoptotic thymocytes and this phagocytosis can be blocked by PtdSer liposomes, indicating that TIM-4 is sufficient to mediate phagocytosis of apoptotic cells in a PtdSer-dependent manner. We examined cytokine secretion by mouse peritoneal macrophage after phagocytosis of apoptotic thymocytes and found that production of a set of cytokines including TGF-β, IL-10, and others was increased dramatically. We performed qRT-PCR assays to detect the mRNA level of these cytokines upon peritoneal macrophage phagocytosis of apoptotic thymocytes. We found that the increased cytokine mRNA levels peaked from 30 minutes to 3 hours and protein production peaked at 24h. The anti-mouse TIM-4 antibody, 21H12, can block TIM-4 recognition of PtdSer and reduce phagocytosis of apoptotic cells by peritoneal macrophages. The 21H12 antibody effectively suppressed the cytokine mRNAs induced by TIM-4-mediated phagocytosis of apoptotic cells. These findings indicate that TIM-4 plays an important role in phagocytosis of apoptotic cells and secretion of regulatory cytokines by mouse peritoneal macrophages and may play an important role in immune regulation through these cytokines.
Immunomodulatory effects of Lactobacillus plantarum and Lactobacillus acidophilus strains on a rat intestinal epithelial cell line (137.21)
Authors: Duersteler Megan , Novak Kimberly , Davis Ellen , Smith Alexandra , Rehberger Thomas 
Published: January 2023

Abstract
Lactobacillus plantarum and Lactobacillus acidophilus have the potential to be immunomodulatory probiotics. An experiment was conducted to determine the effects of three L. plantarum and three L. acidophilus strains on the expression of five inflammatory cytokine genes (TNF-α, MIP-2, IL-1β, IL-6, and IL-18) by the IEC-6 rat intestinal epithelial cell line. The Lactobacillus strains had unique genotypic profiles based on RAPD PCR. IEC-6 cells were exposed to 107 cfu viable bacterial cells for one hour with and without lipopolysaccharide (LPS) stimulation. Expression of inflammatory cytokines was analyzed by quantitative PCR. L. plantarum 7-12 and L. acidophilus La-11 had an inflammatory effect on unstimulated cells as indicated by up-regulation of IL-18 by 7-12, and up-regulation of MIP-2 and IL-6 by La-11. L. plantarum 5-39 and L. acidophilus NCFM had an anti-inflammatory effect as indicated by down-regulation of IL-1β. L. plantarum CI15 and L. acidophilus La1 did not affect cytokine expression. LPS increased the expression of TNF-α and MIP-2 in IEC-6 cells &gt; 60 fold. All strains except L. plantarum 7-12 down-regulated expression of TNF-α in cells stimulated with LPS. L. plantarum CI15 and 5-39, as well as L. acidophilus NCFM and La1 down-regulated MIP-2 expression in LPS-stimulated cells. These results indicate that L. plantarum and L. acidophilus have immunomodulatory properties, and the effects on cytokine expression are strain specific.
A role for regulatory T cells and TGF-β in intestinal IgA to rotavirus (137.22)
Authors: Miller Amber , Blutt Sarah , Conner Margaret 
Published: January 2023

Abstract
Regulatory T cells (Tregs) play an important role in suppression of effector T cell function and inflammation. Recently Tregs have been shown to play a role in mucosal IgA production through production of TGF-β1, suggesting they also can induce B cell responses. During rotavirus (RV) infection, there is little effector T cell activation but a profound intestinal IgA response. Using flow cytometric based approaches, we observed a significant increase in total number of FoxP3+CD25+CD4+ T cells in both MLNs (5 fold) and PPs (2 fold) 2-4 days post RV infection compared to uninfected mice. We also detected a significant increase in the total number of TGF-β1+ cells and the total number of TGF-β1+CD4+ cells in both tissues isolated from infected mice compared to uninfected mice. The increased number of FOXP3+ Tregs and CD4+/TGF-β1 expression in PP and MLN during RV infection concurrent with previously observed B cell activation and rotavirus-specific IgA production suggests that Tregs might play an important role in the induction of rotavirus specific intestinal IgA in these immune tissues through production of the critical switch factor, TGF-β1.
ANALYSIS OF THE RESPONSE OF HUMAN NEONATAL DENDRITICS CELLS AND MACROPHAGES AGAINST ROTAVIRUS (137.26)
Authors: Rosales Denisse , Gutierrez-Xicotencatl Lourdes , Santana-Calderon Angelica , Ramirez-Pliego Oscar , Cortez-Gomez Rudyard , Esquivel-Guadarrama Fernando 
Published: January 2023

Abstract
Rotavirus (RV) is the most common cause of infectious diarrhea. Dendritic cells (DCs) and macrophages play a crucial role in the immune response against viral infections; therefore it is important to dissect the interaction of RV with these cells. Here we analyzed the interaction of RV with neonatal DCs and macrophages in an in vitro model. Cord blood-derived mononuclear cells were culture in vitro for 6 days in the presence of either hGM-CSF/IL-4 or hM-CSF to generated enriched DCs and macrophages, respectively. These cells were cultured for 24 hrs with rotavirus and the levels of the activation markers CD40, CD86 and MHC class-II, and the intracellular cytokines IL-6, IL-12 and IL-10 were analyzed by flow cytometry. TGF-β was evaluated by ELISA. Finally, the induction of allogenic T-celll proliferation was analyzed. We found that RV could induce the activation of the neonatal DCs and macrophages since all the activation markers analyzed were up regulated, albeit at different levels. As result of this activation, the neonatal DCs and macrophages produced high levels of the regulatory cytokines, but moderate levels of pro-inflammatory cytokines. Finally, the activated DCs and macrophages were able to induce a strong allogenic T-cell proliferation. These results showed that neonatal DCs and macrophages could be activated by RV and that this activation was sufficient to induce the priming of allogenic T-cells. Supported by grant SEP-CONACYT (80149) and DGAPA, UNAM.
The role of TGFβ in the development of Th17 and Treg cells during influenza viral infection in neonatal mice (137.6)
Authors: Hoskins Samantha , Lines Janet , Garvy Beth 
Published: January 2023

Abstract
Influenza virus is responsible for more infant hospitalizations and deaths than any other vaccine preventable disease in America. Our laboratory has established a neonatal murine model to influenza and have reported that T cells in the neonatal lung are less activated than in adults and do not infiltrate the alveolar spaces. It is known that TGFβ is elevated in the lungs of neonatal mice, and since neonatal mice do not respond with a typical Th1 response to influenza virus, the dynamics of T regulatory (Treg) cells and the Th17 response was evaluated. Our current studies show that neonatal mice have higher levels of T regulatory cells (Treg) than adults, and these elevated levels persist during influenza viral infection. However, in the absence of Treg modulation, neonatal T cells were not more activated and did not infiltrate into the alveolar spaces during an influenza viral infection. Larger numbers of neonatal CD4 T cells produced IL-17A than adults during viral clearance. Th17 cells may contribute to the clearance of influenza in the immunosuppressive environment of the developing neonatal lung. Investigation of the importance of TGFβ signaling on T cell activation and migration is currently underway using conditional knockout mice.