bluuebunny/crossref_metadata_2025_split huggingface dataset:
Flow-cytometric evaluation of T cell responses elicited by an alphavirus replicon particle vaccine for cytomegalovirus (CMV) in healthy adults (132.14)
Authors: Reap Elizabeth A , Katen Kevin S , Watson Aubrey D , Smith Kaitlin M , Norberg Pamela K , Chulay Jeffrey D , Olmsted Robert A 
Published: January 2023

Abstract
We recently completed a Phase I evaluation of a bivalent, virus-like replicon particle (VRP) vaccine (AVX601) expressing three CMV proteins in CMV seronegative adults. One VRP component expressed the pp65 and IE1 genes as a polyprotein and the second expressed gB. AVX601 was well-tolerated, with no significant safety issues identified. Analysis of the immune responses by IFN-γ ELISPOT and CMV neutralization showed the vaccine to be highly immunogenic. In this study, multiparameter flow cytometry was used to further characterize the AVX601-induced T cell responses. Overall, the CD8 T cell responses to all 3 antigens were of higher magnitude than the CD4 T cell responses. The highest levels of CD8 and CD4 antigen-specific responses detected were 0.97% and 0.17% for pp65, 0.12% and 0.10% for IE1, and 0.18% and 0.08% for gB. Antigen-specific T cells were analyzed for their ability to produce IFNγ, TNFα and/or IL-2 after CMV peptide stimulation. Polyfunctional subsets of both CD4 and CD8 lineages specific for all 3 antigens were detected at all time points tested. The maximum % of CD8 T cells positive for 2 or 3 cytokines at any time point studied was 0.58% for pp65, 0.03% for IE1 and 0.07% for gB, and for CD4 T cells was 0.12% for pp65, 0.06% for IE1 and 0.05% for gB. These data demonstrate that this novel alphavirus replicon particle vaccine for CMV elicited polyfunctional CD4 and CD8 T cells to all 3 CMV immunogens.
IgG1, but not IgG2a/c, is pathogenic in Leishmania mexicana infection (133.18)
Authors: Buxbaum Laurence U. , Thomas Bolaji N. 
Published: January 2023

Abstract
FcγRs and IL-10 are both required for chronic disease caused by L. mexicana infection of C57BL/6 mice. We recently showed that IgG1 and IgG2a/c can both induce IL-10 from macrophages and dendritic cells when bound to Leishmania amastigotes, but through different FcγR types. IgG1 induces IL-10 through FcγRIII, whereas IgG2a/c induces IL-10 primarily through FcγRI. Given that FcγRIII KO mice are resistant to L. mexicana infection and because IgG1 responses occur much earlier than IgG2a/c responses, we examined whether IgG1 is specifically required for chronic L. mexicana disease. In fact, mice lacking IgG1 are more resistant to L. mexicana infection. Furthermore, we found that IgG1 KO mice had an earlier and stronger IgG2a/c response to L. mexicana infection. This demonstrates that IgG1 is pathogenic and that IgG2a/c is not, and may in fact be protective. Thus IgG2a/c does not appear to induce a suppressive IL-10 response in vivo. This opens up the question of whether macrophages are the source of suppressive IL-10 responsible for chronic disease in vivo, or whether another cell type is responsible. In addition it has important implications for vaccines for leishmaniasis and potentially diseases caused by other pathogens that share this FcγR/IL-10 mechanism, as vaccines must be engineered to minimize IgG1 responses to amastigote surface epitopes.
Funded by VA Merit and the University of Pennsylvania.
TL1A, a TNF-like molecule, regulates the differentiation of human TH17 cells (133.21)
Authors: Michelsen Kathrin , Yu Qi , Ko Brian , Thomas Lisa , Takedatsu Hidetoshi , Landers Carol , Targan Stephan 
Published: January 2023

Abstract
TL1A is a member of the TNF superfamily that mediates a strong co-stimulation of TH1 cells by enhancing IFN-γ production by CD4+ T cells. The expression of TL1A is increased in inflamed mucosa of Crohn's Disease patients and in murine models of ileitis. We have shown that TL1A is induced in human antigen-presenting cells (APCs) by Fc γ Receptor but not TLR signaling. Furthermore, neutralizing TL1A antibodies attenuate colitis by attenuating TH1 and TH17 responses in a chronic model of colitis. However, it remains to be elucidated if TL1A enhances de novo differentiation of TH17 cells. Here we demonstrate that FcγR signaling in APCs leads to the concomitant induction of TL1A, IL-6, TGF-β, and IL-23. TL1A, in combination with TGF-β and IL-6, promotes the differentiation of human TH17 cells from naive CD4+ T cells and leads to a unique cell population of IL-17/IFN-γ producing TH17 cells. Furthermore, TL1A enhances the IL-17 production by committed CD45RO+CCR6+ TH17 cells. TL1A also enhances the induction of RORA mRNA but not RORC mRNA. However, the enhanced RORA mRNA levels corresponded to an increase in IL-17 mRNA under the same conditions suggesting that enhancement of RORA mRNA by TL1A is sufficient to enhance production of IL-17. Our findings establish an important role of TL1A in promoting human TH17 cell differentiation and function and may provide a potential target for therapeutic intervention in TH17 driven diseases.
The major Th2 polarizing component in schistosome eggs is a T2 ribonuclease (omega-1) that inhibits dendritic-T cell interaction (133.23)
Authors: Jankovic Dragana , Andersen John , Steinfelder Svenja , Cannons Jennifer L. , Schwartzberg Pamela L. , Sher Alan 
Published: January 2023

Abstract
Although helminth parasites are the most potent infectious stimuli for the induction of Th2 cells, the identity of their Th2-inducing components has remained elusive. Schistosoma mansoni eggs contain factors that trigger potent Th2 responses in vivo and condition murine dendritic cells (DC) to promote Th2 lymphocyte differentiation. We have performed a classical biochemical purification of the DC-dependent Th2-polarizing molecule(s) in both secreted egg products and in a water-soluble egg extract (SEA) and identified the active Th2-inducing component as a 31 kDa molecule identical to the previously described S. mansoni ribonuclease T2, omega-1. The Th2-promoting activity of this glycoprotein is sensitive to ribonuclease inhibition and does not require MyD88 or TRIF signaling in DC. In common with unfractioned SEA the purified native protein suppresses LPS-induced DC activation, but unlike SEA fails to trigger IL-4 production from basophils. We further demonstrate that SEA as well as purified omega-1 directly affect both DC morphology and the ability of these APC to interact physically with CD4 T lymphocytes. Based on this evidence, we propose that SEA and its omega-1 bioactive component promote Th2 polarization by lowering the strength of the activation signal delivered by DC.
This work was supported by the Intramural Research Program of the NIAID.
Immune Responses during Respiratory Syncytial Virus Pathogenesis in a Neonatal Ovine Model (133.46)
Authors: Sow Fatoumata Ba , Gallup Jack M , Olivier Alicia , Krishnan Subramaniam , Suzich JoAnn , Ackermann Mark R 
Published: January 2023

Abstract
Respiratory Syncytial Virus (RSV) is the leading cause of lower respiratory tract infection in infants and young children worldwide, causing severe pneumonia and bronchiolitis. The newborn ovine lung is well suited for comparison to the human perinatal lung, due to similarities in alveolar development, immune responses, and RSV susceptibility. To understand the role of innate and adaptive immunity in RSV pathogenesis, we inoculated lambs intratracheally at 3 days of age with human RSV (hRSV), and lung tissue was collected from the right cranial lobe at 3, 6, and 14 days post-inoculation for histopathology, viral load, cellular infiltration and cytokine production. hRSV antigen was present at low levels at day 3 of infection, increased 29.8-fold at day 6, and decreased to baseline levels at day 14. The presence of hRSV in the lung correlated positively with an increased accumulation of CD11b+ cells. Infection was associated with a decreased expression of IFN-β, TNF-α, TGF-β, IL-10, RANTES and PD-1, but with an increased expression of IL-8, MCP-1, MIP-1α, PD-L1, and IFN-γ. Laser capture microdissection analysis of macrophage-enriched populations from lung sections determined that MCP-1 was highly up-regulated in those cells, but that alveolar macrophages were not the source of increased IL-8 mRNA. The modulation of host immune responses by RSV may contribute to the enhanced susceptibility and severity of disease.
This project is funded by MedImmune LLC and by NIH grant # RAI062787A
Urushiol regulates the expression of TLR2, HBD and TSLP via NF-κB pathway in Staphylococcus aureus-treated HaCaT cells (134.50)
Authors: PARK JU YOUN , Chang Jae-Ho , Kim Myong-Jo , Kim Soo-Ki 
Published: January 2023

Abstract
TLR2, HBDs(beta-defensins) and TSLP(thymic stromal lymphopoietin) expressed from keratinocytes play key roles in controlling the progression of atopic dermatitis. These molecules might account for the accelerated colonization with S. aureus into eczematous skin barrier. In the course of screening natural substance in regulating TLR2, HBD and TSLP expression by using the inflamed keratinocytes, we found urushiol, major active ingredient of Rhus verniciflua Stokes which has been used as barrier protective in traditional medicine. The aim of this study was to clarify how urushiol regulates innate immune molecules, focused on TLR2, HBD and TSLP, in our in vitro infectious keratinocyte model, S. aureus-treated HaCaT cells. To the end, the expression of TLRs, cytokines and chemokines were assessed by by siRNA, RT-PCR, ELISA and Western blots. We found that urushiol treatment on HaCaT cells induced the expression of TLR2 and HBD2 but not that of TNF-α and TSLP. In S. aureus-treated HaCaT cells, urushiol pretreatment enhanced the expression of TLR2 and HBD2, whereas TNF-α and TSLP expression were in decrement. Further to explore the mechanism, inhibition of NF-κB decreased the expression of TLR2, HBD2 and TSLP, albeit increased TNF-α release. Cumulatively, our data indicate that urushiol regulates the expression of TLR2, HBD and TSLP via NF-κB pathway in S. aureus-treated HaCaT cells.
Demonstration of a severe complement defect in FcRγ-deficient mice (134.63)
Authors: Strait Richard T , Khodoun Marat , Mahler Ashley , Potter Crystal , Köhl Jörg , Finkelman Fred D 
Published: January 2023

Abstract
Several studies have examined the relative roles of complement vs. Fc receptors (FcRs) in host protection and inflammatory disease. Many of these studies have concluded that the more important role is played by stimulatory Fc receptors. These conclusions were based, however, on the assumption that complement activation occurs normally in the absence of stimulatory Fc receptors. We have used FcRγ-deficient mice, which lack all stimulatory FcRs, to examine this assumption. FcRγ-sufficient and -deficient mice were found to have similar levels of C3 in blood. Also, as expected, similar amounts of IgG2a became associated with blood leukocytes from both mouse strains after intravenous injection of a complement fixing IgG2a mAb to MHC class I Ag. Surprisingly, however, &gt;10-fold more C3 became associated with the blood leukocytes from FcRγ-sufficient mice than those from FcRγ-deficient mice. Additionally, &gt;6-fold more C5a was generated in blood from FcRγ-sufficient than deficient mice. These results demonstrate that mice that lack all stimulatory FcRs have a severe abnormality in complement activation. This abnormality may well contribute to the decreased protective immunity and increased resistance to inflammatory disease characteristic of FcRγ-deficient mice that have previously been ascribed solely to direct effects of the absence of stimulatory FcRs.
A Meta Analysis of Relationship between C - Reactive Protein and Respiratory Tract Infection in Children (134.64)
Authors: Shen Chao-Bin , Gu Jun , Xia You-Jia 
Published: January 2023

Abstract
BACKGROUND: Although several studies have evaluated serum concentrations of C-reactive protein (CRP) as a predictor of bacterial respiratory tract infections or pneumonia in this patient population, the utility of this test remains unclear. OBJECTIVE: The purpose of this meta-analysis was to quantitatively define the utility of serum CRP as a predictor of bacterial respiratory tract infections or pneumonia in acutely ill children. METHODS: Multiple databases were searched, bibliographies reviewed,and 13 authorities in the field were queried.The quality of each included study was determined have 6 metrics: diagnostic criteria; study author; CRP agent; patient age; CRP assay of method; and CRP results before patients who received antibiotics. Data was extracted from each article; the primary outcome measure was the means of patients with bacterial or not bacterial infection and serum CRP concentrations exceeding 8 mg/L. RESULTS: Thirteen studies fulfilled inclusion criteria. Combining all of the studies demonstrated a pooled study population of 2760 patients.The level of serum CRP in the children with bacterial infection were higher 3.13 times than viral infection(Z=7.61,p£1/40.00001,95% confidence interval=2.33-3.94),and were higher 3.02 times than control group(Z=10.26,p£1/40.00001,95% confidence interval =2.44-3.60).The level of serum CRP in children with viral infection were higher 0.68 times than control group(Z=4.74,p£¼ 0.00001,95% confidence interval=0.40-0.96). Children with bacterial infection were significantly serum CRP concentrations than children with nonbacterial infections.CONCLUSIONS:The serum CRP level has value for early recognition of bacterial infections could guide treatments,reduce misuse of antibiotics.
Complement Factor H Regulation in the Central Nervous System (134.70)
Authors: Fraczek Laura , Martin Carol , Martin Brian 
Published: January 2023

Abstract
The complement system is an important component of innate immunity that requires proper regulation to avoid inappropriate activation. This regulation is provided by many proteins including complement Factor H (CFH), a secreted protein of the alternative pathway. Despite its regulatory function, gene and promoter polymorphisms of CFH have been implicated in disease of many patients with age-related macular degeneration and atypical hemolytic uremic syndrome, indicating that a better understanding of this gene's regulation is valuable. Presently, little work has been done to address the role of CFH in the central nervous system (CNS). We have cloned the murine CFH promoter and have made truncation constructs to examine CFH gene regulation in cells of the CNS. Our studies show that CFH mRNA is present in CNS cell types, and that specific regions throughout the promoter contain possible enhancers and repressors. Database mining of these regions indicates that there are potential transcription factor binding sites conserved between many different species, which led us to investigate specific transcription factor binding interactions in these regions. Through these studies we hope to elucidate the transcriptional regulation of CFH in the CNS.
This work is supported by the National Multiple Sclerosis Society.
In Vivo Inhibition of Type I Interferon Transcription in Genital HSV-2 Lesions (134.72)
Authors: Peng Tao , Zhu Jia , Klock Alexis , Koelle David , Wald Anna , Corey Larry 
Published: January 2023

Abstract
We performed transcriptional analysis and immunocytochemistry (ICC) of sequential biopsies of lesional tissue of immunocompetent persons with recurrent HSV-2 infection. Histologic analysis of these biopsies indicated a massive infiltration of monocytes/macrophages along with a large amount of myeloid and a small number of plasmacytoid dendritic cells in the dermis of these lesional biopsies. IFNB1 and IFN-α were weakly expressed and IFNG potently induced during time periods in which there was abundant detection of HSV-2 antigens and gene expression. Transcriptional arrays of the same anatomic area over time (newly healed, 2 and 4 weeks post healing) in which no HSV DNA/RNA or antigen was detected continued to show that IFNB1 and IFN-α were barely detectable. IFNG persisted in lesional tissue, albeit at lower levels as compared with active lesions. Interferon-stimulated genes (ISGs) were also markedly up-regulated with expression patterns more clearly resembling those in primary human fibroblasts treated by IFNG than by IFNB1. The presence of a very large number of innate cells capable of sensing HSV-2 infection and synthesizing type I IFN by both histologic and transcriptional analyses associated with extremely low levels of IFN-α and IFNB1 even in the earliest lesional biopsies suggests a potent alteration in host defense during HSV-2 infection in vivo. This block of type I IFN by HSV-2 may be a major factor in allowing the virus to breakthrough host mucosal defenses.
Toll-Like Receptor 3 Signaling on Macrophages is Required for Survival Following Coxsackievirus B4 Infection (134.77)
Authors: Richer Martin Joseph , Lavallée Danielle J , Shanina Iryna , Horwitz Marc S. 
Published: January 2023

Abstract
Toll-like receptor 3 (TLR3) has been proposed to play a critical role in the innate detection of viruses. However, as several reports have demonstrated that TLR3 signaling is either dispensable or even harmful following certain viral infections, the role of TLR3 in anti-viral immunity remains controversial. Here, we asked whether TLR3 is required for the immune response to coxsackievirus B4 (CB4), a common human pathogen associated with pancreatitis, myocarditis and diabetes. We demonstrate that TLR3 signaling is required for survival following CB4 infection. TLR3 deficient mice produce lower levels pro-inflammatory mediators and are unable to control viral replication resulting in severe cardiac damage and increased mortality. We further demonstrate that the MyD88-dependent signaling pathways are dispensable in the response to this RNA virus and are unable to compensate for the loss of TLR3. Finally, we demonstrate that adoptive transfer of WT macrophages is sufficient to reestablish protection in a TLR3 deficient host by reducing cardiac pathology suggesting that TLR3 signaling on macrophages is critical for survival following CB4 infection. Our results demonstrate that TLR3 is not simply part of a redundant system of viral recognition, but rather TLR3 plays an essential role in recognizing the molecular signatures associated with specific viruses including CB4.
This work was supported by a CIHR grant to MSH.
Poly-T oligonucleotides enhance TLR7/8 agonist induced responses in glial cells through a TLR7-dependent mechanism (135.11)
Authors: Du Min , Butchi Niranjian , Peterson Karin E 
Published: January 2023

Abstract
The brain does not contain a resident dendritic cell population so resident cells such as microglia and astrocytes play an important role in initiating innate immune responses in the central nervous system (CNS) following virus infection. However, the activation of these cells following stimulation of virus-recognizing toll-like receptors (TLRs) such as TLR7 and TLR8 remains unclear. Poly T oligonucleotides (pT-ODNs) were recently shown to enhance signaling through TLR8, including murine TLR8 which was previously thought to be non-functional. In the current study, we analyzed the ability of pT-ODNs to enhance TLR7/8 activation of murine astrocytes and microglia. We found that the TLR7/8 agonist CL075 (3M002) induced the expression of glial cell activation markers and induced the production of multiple proinflammatory cytokines and chemokines. Addition of pT-ODNs enhanced the expression of proinflammatory cytokines and chemokines as well as the microglia/macrophage activation markers F4/80 and Iba1. Surprisingly, pT-ODNs enhancement of CL075-induced glial activation was abolished in TLR7 deficient mice, with the exception of Gfap (glial fibrillary acidic protein) mRNA upregulation. Thus in glial cells, pT-ODNs enhanced TLR7/8-induced proinflammatory responses through a TLR7-dependent pathway.
This research was supported by the Intramural Research Program of the NIH.
Desialylation of TLR4/MD2/CD14 enhances the response to LPS (135.16)
Authors: Feng Chiguang , Whitford Melissa , Zhang Lei , Rallabhandi Prasad , Vogel Stefanie N. , Stamatos Nicholas M. 
Published: January 2023

Abstract
Objective: Since bacterial neuraminidase (NANase) treatment of intact human PBMCs led to a synergistic increase in cytokine expression, we hypothesized that components of the TLR4 receptor complex were sialylated, and that removal of sialyl residues enhanced LPS-initiated signaling.
Methods: HEK293T cells were transfected with TLR4, MD2, CD14 and ELAM-Luc expression vectors. They were treated with NANase, stimulated with LPS, and assayed for NF-kB-driven luciferase expression. Sialylation was assayed by lectin blot after immunoprecipitation from lysed transfected cells.
Results: LPS stimulated NF-kB activation and IL-8 expression only in cells transfected with TLR4/MD2/CD14 or in cells transfected with TLR4/CD14 supplemented with supernatants (SNT) of MD2-transfected cells. NANase treatment of TLR4/CD14/MD2 cells led to enhanced NF-kB activation. While NANase treatment of either TLR4/CD14 cells or MD2 SNTs each enhanced the LPS response in the reconstituted culture, optimal responses occurred when both were treated. Lectin blots revealed MD2 to be sialylated with a α2,6 linkage since SNA but not Maackia bound to MD2.
Conclusions: We conclude that removal of sialyl residues from MD2 and perhaps from TLR4/CD14 enhances the response to LPS. Thus mobilization of endogenous sialidase in PBMCs could modulate the activity of the TLR4/CD14/MD2 complex and serve as a therapeutic target.
Support: NIH grant no. HL086933-01A1
Regulation of pro-inflammatory cytokines by interferon-gamma during endotoxin tolerance (135.38)
Authors: Chen Janice , Ivashkiv Lionel B. 
Published: January 2023

Abstract
Endotoxin tolerance is a phenomenon whereby previous exposure of macrophages to microbial products, such as LPS, induces a transient period of hyporesponsiveness upon subsequent LPS challenge and is characterized by diminished release of pro-inflammatory cytokines. Previous reports have shown that interferon-gamma (IFN-γ) can prevent and reverse endotoxin tolerance, and our study aims to define the mechanisms by which this occurs. We found that exposure of CD14+ human monocytes to 100 pg/mL LPS for 24 h was sufficient to blunt their ability to produce IL-6 and TNFα upon secondary LPS challenge. However, IFN-γ pretreatment abrogated tolerization of IL-6, and to a lesser extent, TNFα, induced by LPS as well as cross-tolerization induced by Pam3Cys and IL-1β. IFN-γ did not reverse the defects in LPS signaling induced by tolerization. Tolerized monocytes exhibited greatly reduced LPS-induced IL-6 gene expression, while IFN-γ-activated monocytes retained the capacity to upregulate IL-6 mRNA despite tolerizing signals. IFN-γ-mediated regulation of tolerance occurred at the level of transcription, as revealed by IL-6 primary transcript measurements and RNA polymerase II binding to the IL-6 promoter. Current efforts are directed at identifying key factors that might mediate the ability of IFN-γ to abrogate endotoxin tolerance. This work is supported by the CRI Predoctoral Fellowship and NIH T32 AIO7621.