bluuebunny/crossref_metadata_2025_split huggingface dataset:
The novel IgG-receptor FCRL5 is able to distinguish IgG2 disulfide isomers and displays increased affinity for heat-stressed IgG2
Authors: Alabi Oyeleye A , Dement-Brown Jessica , Tolnay Mate 
Published: January 2023

Abstract
Human Fc receptor-like 5 (FCRL5) is a novel IgG receptor that senses the whole IgG molecule, including the Fc and F(ab′)2 portions. Using surface plasmon resonance (Biacore) technology, we established that FCRL5 binds human IgG via a complex mechanism, where IgG isotype is just one determinant of the interaction. In particular, individual IgG2 samples were found to display a thousand-fold range of affinities as well as distinct kinetics, indicating that additional factors beyond the isotype affect binding. We further investigated two potential factors that may affect the interaction of IgG2 with FCRL5; structural heterogeneity related to IgG2 disulfide isoforms and charge variants mostly due to deamidation. We generated IgG2 samples enriched for either of the disulfide isoforms, IgG2-A and IgG2-B, and assessed their binding to FCRL5 using Biacore. We found that the IgG2-A isoform bound FCRL5 with up to 80-fold higher affinity than the IgG2-B isoform. IgG2 samples were also kept at 40°C for up to four weeks, a condition known to promote deamidation. We found that FCRL5 displayed higher affinity for heat-stressed IgG2, suggesting that the interaction is sensitive to deamidation of the IgG2. We conclude that FCRL5 is able to discriminate two independent changes in the IgG2 molecule, the disulfide isoform structure as well as charge variants likely related to deamidation. Of note, both conversion of disulfide isoforms and deamidation of the molecule are known to occur in vivo, on time scales comparable to the half-life of the IgG2 molecule. Therefore, our studies raise the possibility that FCRL5 discerns IgG2 variants produced in vivo in days or weeks, providing insight into the nature of the physiological FCRL5 ligand.
Discovery of a novel transcriptional regulator of sustained TLR-mediated inflammatory responses
Authors: LIN BIN , Dutta Bhaskar , Oh Kyu-Seon , Fraser Iain D.C. 
Published: January 2023

Abstract
Toll-like receptor (TLR) pathways have been intensively investigated for almost two decades. However, since most studies have focused on the initial signal transduction and transcription events, the mechanisms of late-sustained gene expression are poorly understood. To address this issue, we conducted an RNAi screen for regulators of TLR-induced IL-6 secretion in RAW264.7 macrophages, and identified a novel putative regulator of this process (Hit4). Hit4 was specifically required to sustain the late expression of genes including Il6, Il1a, Nos2 and Lcn2, but did not affect induction of transiently expressed genes such as Nfkbiz, Egr1, and Egr2. This effect of Hit4 was confirmed by CRISPR/Cas9 gene editing and by RNAi knockdown in primary macrophages. Most interestingly, Hit4 functioned specifically in sustaining rather than initiating transcription, as Hit4 depletion showed no effect on the early mRNA transcription before 2 hr for all gene classes. Further mechanistic investigation revealed that Hit4 functioned at the transcriptional level rather than post-transcriptional level, as Hit4 enhanced the level of nascent transcripts at later time points. Moreover, Hit4 KO RAW264.7 macrophages failed to sustain the response to persistent LPS stimulation, showing a response similar to that observed for transient LPS stimulation. This suggests a role for Hit4 in discriminating brief spurious pulses of infectious signal from a more sustained real infection signal. In summary, we have identified a novel transcriptional regulator that functions to sustain transcription of late inflammatory genes, providing important new insight to the control of TLR-induced gene program.(Supported by the Intramural Research Program of NIAID, NIH)
Characterization of the role of NLRP10 in dendritic cells
Authors: Vacca Maurizio , Mortellaro Alessandra 
Published: January 2023

Abstract
The ability of the immune system to recognize a large variety of microbial-derived molecules relies on diverse families of pattern-recognition receptors (PRRs). One important family of PRRs is the nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs). Among all the NLRs, NLRP10 has unique features, as it is the only member lacking the ligand-binding leucine-rich-repeat (LRR) domain. NLRP10 protein is poorly characterized and its functions are still unknown. Nlrp10 is expressed almost in immune and non-immune tissues. In this study, we characterized the role of the murine Nlrp10 in host immune response. We firstly assessed the role of Nlrp10 in DCs by examining Nlrp10−/− DC ability to present protein antigens and thereby activate T cells. We found that ovalbumin-specific CD4+ T cells when co-cultured with Nlrp10−/− DCs produce less IFNγ compared to T cells co-cultured with WT DCs. We then measured the expression of some co-stimulatory molecules (CD80, CD86), as well as MHC class I and II in immature and LPS-matured Nlrp10−/− DCs. Expression levels of these molecules are similar in WT and Nlrp10−/− DCs. Furthermore, we evaluated the release of specific cytokines responsible for T cell activation in DC/T cell co-cultures. Preliminary data suggest that Nlrp10−/− DCs have a defect in IL-12 release as a result of impaired NFkB signaling. Nlrp10 contribution in the pathways regulating IL-12 production is currently under investigation. Since associations between NLRP10 and inflammatory disorders are emerging, a better understanding of the underlying mechanisms regulated by NLRP10 in immune response may help us to identify potential new targets for anti-inflammatory therapies.
Post-transcriptional elements within NLRP3 mRNA set the activation threshold of the inflammasome.
Authors: Varela Stephanie Nichole , Schwerk Johannes , Forero Adriana , Hong MeeAe , Arroyo Nicole , Chou Bryan , Savan Ram 
Published: January 2023

Abstract
NOD-like receptor protein 3 (NLRP3) is an intracellular sensor and a major component of the canonical inflammasome pathway, which causes inflammation and cell death when activated by two temporally discrete signals. While novel putative activators of the NLRP3 inflammasome have been identified, the mechanisms that regulate expression of NLRP3 itself are less understood. Alternative polyadenylation (AP) is a mechanism by which cells modulate expression through varying the length of the 3′ untranslated region (3′UTR). Through AP, the longer forms may provide unique regulatory elements, that allow for differential regulation of mRNA stability and translation. Here we sought to thoroughly characterize the features of the NLRP3 3′UTR and hypothesized that AP may significantly control the threshold of inflammasome activation. We examined AP in THP-1 monocytes and found that they express both long and short forms, with the long form demonstrating decreased stability relative to the short. We have attributed these differences to the presence of AU-rich elements (ARE), as ablation of AREs results in increased stability of the long NLRP3 mRNA. Additionally, we have demonstrated that a known post-transcriptional regulator of NLRP3, miR-223, preferentially regulates the long transcript as well as augments the kinetics of pyroptotic cell death. These studies demonstrate that alternative polyadenylation of the 3′UTR of NLRP3 has a marked impact on the threshold of inflammasome activation.
Inflammatory ILC2: An IL-25-activated circulating ILC population with a protective role during helminthic infection
Authors: Huang Yuefeng , Mao Kairui , Chen Xi , Kawabe Takeshi , Li Weizhe , Zhu Jinfang (Jeff) , Urban Joseph F. , Germain Ronald N , Paul William E 
Published: January 2023

Abstract
Innate lymphoid cells (ILCs) play important roles in host defense, tissue repair, metabolic homeostasis and inflammatory diseases. While ILCs are generally considered to mediate these functions in a tissue-resident manner, whether ILCs can move between sites when infection demands has not been carefully explored. We have reported the existence of an ILC population, inflammatory ILC2 (iILC2) cells, which are not present in the steady state but appear in high numbers in lung, liver, mesenteric lymph nodes and spleen upon IL-25 or Nippostongylus brasiliensis infection. iILC2 cells develop into natural ILC2-like cells and contribute to the expulsion of N. brasiliensis worms. They can also acquire IL-17-producing ability and provide protection against Candida albicans, indicating their multipotentiality or plasticity. Our recent data reveals that iILC2 cells are circulating cells, exchanging to a much greater extent between parabiotic mice than other ILC populations. iILC2 cells that appear after IL-25 administration or helminthic infection in peripheral, non-gut tissues such as the lung arise from distant resting ILC2 cells residing in the intestinal lamina propria. IL-25 induces rapid proliferation of these intestinal ILC2 cells and a change in their sensitivity to S1P-mediated chemotaxis, leading to lymphatic entry, blood circulation, and accumulation in many non-gut sites. These gut-derived cells are critical for anti-helminth immunity and tissue repair in the bowel and lung. Thus, inflammatory ILC2 cells are a circulating ILC population that is derived from the intestine but plays important roles in peripheral tissue host defense.
This work was supported by the Intramural Research Program of NIAID, NIH and by the USDA.
Analysis of caspase-1 regulated transcriptome in various tissues lead to identification of novel IL-1β, IL-18 and sirtuin-1 intendent pathways
Authors: Nanayakkara Gayani K , Li Ya-feng , Yu Sun , Choi Eric T , Wang Hong , Yang Xiao-Feng 
Published: January 2023

Abstract
Caspase-1 is prominently involved in bridging disease risk factors/ danger signals to inflammatory response via its downstream targets such as IL (interleukin)-1β, IL-18, and Sirt-1 (Sirtuin-1). The microarray datasets derived from caspase-1 knockout tissues indicated that caspase-1 function can significantly impact the transcriptome. However, it is not known whether all effects exerted by caspase-1 on transcriptome are mediated only by its well-known substrates. Therefore, we hypothesized that the effects of caspase-1 on transcriptome may be partially independent from IL-1β, IL-18, and Sirt-1. To determine new global and tissue-specific gene regulatory effects of caspase-1, we took novel microarray data analysis approaches including Venn analysis, cooperation analysis, and meta-analysis. We made the following important findings: (1) Caspase-1 exerts its regulatory effects on majority of genes in a tissue-specific manner; (2) Caspase-1 regulatory genes partially cooperates with genes regulated by Sirt-1 during organ injury and inflammation in adipose tissue but not in liver; (3) Caspase-1 cooperates with IL-1β in regulating less than half of the genes involved in organismal injury, and cancer in mouse liver; (4) The meta-analysis identified 40 caspase-1 globally regulated genes across tissues, suggesting that caspase-1 globally regulates many novel pathways; and (5) The meta-analysis identified new cooperatively and non-cooperatively regulated genes in caspase-1, IL-1β, IL-18, and Sirt-1 pathways. Our findings suggest that caspase-1 regulates many new signaling pathways potentially via its known substrates and also via transcription factors and other proteins that are yet to be identified.
Assessment of the dual mechanism of inflammatory inhibition by the novel JODI series of compounds
Authors: Szollosi Doreen , Manzoor Mohammed , Lucero Victoria , Dhuguru Jyothi , Edafiogho Ivan , Ghoneim Ola 
Published: January 2023

Abstract
Previous experiments in our laboratory have screened several novel compounds for anti-inflammatory activity, including enaminone E121 and the related JODI series. These studies have concluded that these compounds decrease the release of pro-inflammatory cytokines TNF-α and IL-6 after stimulation with bacterial lipopolysaccharide (LPS). Additionally, the JODI compounds, which have an N-arylpiperazine ring added to the enaminone pharmacophore appear to also have chemokine inhibition activity. These studies in our laboratory suggest that the JODI compounds not only have a drug target in the Toll-like receptor pathway (after stimulation with LPS) but also a drug target in the MCP-1/CCR2 pathway. Ongoing studies are aimed at evaluating these compounds for their specific drug targets by assessing kinases including p38 which appears to be decreased after JODI treatment, as well as NF-kB translocation. Taken together, the JODI compounds represent a potentially novel series of anti-inflammatory compounds for inflammatory disorders in which efficacious treatments are urgently needed.
The nuclear receptor REV-ERBα modulates Th17 cell-mediated autoimmune disease
Authors: Loo Chin San , Chang Christina , Zheng Ye 
Published: January 2023

Abstract
T helper 17 cells (Th17) play an important role in controlling extracellular bacteria and yeast infections; however, dysregulated Th17 response can lead to detrimental autoimmune diseases such as multiple sclerosis. Th17 cell differentiation relies on IL6/TGF-β-induced expression of retinoic acid receptor-related orphan nuclear receptor RORγt, which is able to induce Th17 effector cytokines such as IL17A. Therefore, antagonizing RORγt activity may prove to be one of the therapeutic strategies for ameliorating Th17-mediated inflammation. Here we identify REV-ERBα (encoded by Nr1d1), a member of the nuclear hormone receptor family, as a transcriptional repressor that antagonizes RORγt-induced Il17a expression. Both RORγt and REV-ERBα bind to the same ROR response element (RORE) in Il17a promoter region. Elevated expression of REV-ERBα prevents RORγt binding to RORE and reduces Il17a expression. Furthermore, treatment with a synthetic REV-ERBα agonist, SR9009, significantly delays the onset and impedes the progression of experimental autoimmune encephalomyelitis (EAE), a Th17-mediated autoimmune disease. These results suggest that modulating REV-ERBα activity holds therapeutic potential for treatment of Th17 cell-mediated autoimmune diseases.
Characterization of varicella zoster virus-specific monoclonal antibodies
Authors: Sullivan Nicole L , Monslow Morgan , Sei Janet , Durr Eberhard , Chang Cathy , Mccausland Megan , Rouphael Nadine , Pulendran Bali , Ahmed Rafi , Vora Kalpit A 
Published: January 2023

Abstract
Herpes zoster (HZ or shingles) is the clinical manifestation of varicella zoster virus (VZV) reactivation. HZ typically develops as people age due to decreased cell mediated immunity. However, the relative importance of humoral immunity against VZV remains somewhat controversial. The goal of this study was to examine the breadth and functionality of VZV-specific antibodies. Direct enumeration of VZV-specific antibody secreting cells (ASC) via ELISPOT showed that Zostavax® vaccination can induce both IgG and IgA ASCs seven days after vaccination, but not IgM ASCs (IgG&gt;IgA≠IgM). The frequency of VZV-specific ASCs was from 33–55% of the total IgG ASCs. ASCs were also single-cell sorted from five subjects (&gt; 60 years old) and 25 VZV-specific monoclonal antibodies (mAbs) were successfully cloned and characterized. These mAbs had on average ~ 20 somatic hypermutations per VH gene, similar to the number of mutations seen in mAbs characterized after seasonal influenza vaccination (Li GM, PNAS, 2012). Fifteen of the 25 mAbs were gE-specific, whereas the remaining mAbs were gB, gH or gI-specific. The antibodies were then tested for in vitro neutralization and inhibition of cell-to-cell spread, which will be discussed. These data indicate that Zostavax® vaccination can induce a memory B cell recall response and gE is the predominant antibody target. However, the relative importance of these antibodies for protective immunity against VZV reactivation will be discussed.
Vaccine display on synthetic bacterial spores enhances protective efficacy against Staphylococcus aureus infection
Authors: Karauzum Hatice , Updegrove Taylor , Wu I-Lin , Datta Sandip K , Ramamurthi Kumaran 
Published: January 2023

Abstract

Background
Modification of bacterial spore surfaces with proteins has previously been proposed as vaccine display platforms, however, relied on viable spores.

Objective
In the present study we describe how synthetic spore husk-encased lipid bilayers (SSHELs) can be utilized as a display platform to present protein antigens like ovalbumin and mutant staphylococcal alpha toxin (HlaH35L) as vaccines. This platform is based on 2 self-assembling proteins SpoVM and SpoIVA from Bacillus subtilis.

Methods
C57BL/6 mice were immunized with SSHEL (with or without adjuvant), antigen or antigen covalently bound to SSHEL (SSHEL_antigen) thrice intramuscularly at two-week intervals and serum antibody titers against SpoIVA or antigens were determined 7 days after each immunization. In select experiments mice were challenged intravenously with a lethal dose of S. aureus USA300 2 weeks after last vaccination. Untreated mice served as controls.

Results
Immunization of mice with SSHEL induced a similar antibody response towards SpoIVA as SSHEL that was adjuvanted in CpG or MPL-A suggesting adjuvant properties of this synthetic lipid-bilayer construct. Post vaccination serum of mice immunized with ovalbumin covalently bound to SSHEL (SSHEL_OVA) or HlaH35L covalently bound to SSHEL (SSHEL_HlaH35L) contained SpoIVA specific as well as antigen-specific (OVA or HlaH35L) antibodies. Infection of mice with a lethal dose of USA300 resulted in 100% survival of mice immunized with SSHEL_HlaH35L compared to 20% survival of non-immunized mice (P=0.01), and 40% survival of mice immunized with HlaH35L (P=0.049).

In progress
In ongoing studies we are evaluating the mechanism of action of SSHEL_HlaH35L that lead to protection during infection.
miR-148a controls plasma cell differentiation and survival
Authors: Pracht Katharina , Meinzinger Julia , Daum Patrick , Côrte-Real Joana , Hauke Manuela , Schulz Sebastian , Roth Edith , Wittmann Jürgen , Jäck Hans-Martin 
Published: January 2023

Abstract
microRNAs (miRNAs) are critical regulators of central and Ag-driven B cell development. However, it is unknown whether miRNAs regulate the differentiation of B cells into plasma cells (PCs) and control the survival of long-lived PCs.
We recently showed that miR-148a, the most abundantly expressed miRNA in PCs, is upregulated in activated B cells and promotes in vitro plasmablast differentiation and survival by targeting Bach2, MiTF, PTEN and Bim.
To investigate if miR-148a supports PC differentiation in vivo, we established a conditional B cell-specific miR-148a knock-out mouse. Non-immunized mice showed a significant decrease in serum Ig, antibody-secreting cells (ASC) and splenic PCs. 28d after boost, TNP-KLH-immunized 148a-KO mice had lower serum TNP-IgG and decreased numbers of splenic ASCs and bone marrow (BM) PCs.
To ask whether miR-148a controls PC survival, we generated tamoxifen-inducible miR-148a deficient mice. Tamoxifen-mediated miR-148a deletion, 28 days after boost with TNP-KLH, resulted in reduced numbers of plasmablasts and IgG-ASCs in the spleen and of resting PCs and IgG-ASCs in BM. In parallel, increased numbers of resting PCs were found in the blood, indicating a possible egress from survival niches.
These findings support the hypothesis that miR-148a controls the differentiation of B cells into plasmablasts as well as the survival of long-lived PCs. We will now identify the underlying mechanism by a biosystems approach utilizing transcriptome and proteome profiles of fresh follicular B cells and BM PCs.
Proteome analysis was done in collaboration with the Warscheid lab at the University of Freiburg.
Affinity maturation of anti-lipid IgG antibodies produced via germinal center
Authors: Baeza Carlos Wong , Resendiz Albany , Landa Carla , Angeles Oscar , Tescucano Alonso , Wong Carlos , Flores-Romo Leopoldo , Baeza Isabel 
Published: January 2023

Abstract
Non-bilayer phospholipid arrangements (NPA) are lipid associations different to the bilayer and are found on cell membranes; they participate in different cellular functions and are transient, but when they are stabilized by drugs such as chlorpromazine, they become immunogenic and induce the formation of anti-NPA antibodies. Mice that receive stabilized NPA share several characteristics with patients with systemic lupus erythematosus, such as the presence of anti-cardiolipin, anti-histone and anti-nuclear antibodies, lupus band, malar exanthema and glomerulonephritis. Mice with this lupus model have been shown to develop the disease as a consequence of the production of anti-NPA antibodies; in addition, most of these antibodies are of the IgG class and are produced mainly by the germinal center pathway. In the case of protein antigens, germinal centers are the sites where class-switch recombination, somatic mutation, memory generation and affinity maturation occur. To determine if affinity maturation, a hallmark of the antibodies produced via germinal center reactions, occurs in the anti-NPA IgG antibodies produced in germinal centers, we performed antigen-specific ELISAs, both in the absence and in the presence of urea. We observed that the percentage of urea-resistant (high affinity) anti-NPA IgG antibodies in relation to the total anti-NPA IgG antibodies increases over time, and it correlates with the number of NPA-specific germinal center B cells.
Th17 inflammation in the lungs is sustained by multiple exposures to house dust and is dependent on TLR4 and conventional dendritic cells
Authors: Shalaby Karim H , Lyons-Cohen Miranda , Thomas Seddon Y , Whitehead Gregory S , Nakano Hideki , Cook Donald N 
Published: January 2023

Abstract
Microbial products can prime antigen-specific T helper 17 cell responses to inhaled allergens. However, once established, it is unclear how pathogenic lung Th17 responses are sustained. To study this, we sensitized mice to inhaled ovalbumin (OVA)together with various environmental adjuvants, including house dust extract(HDE). Animals were then chronically challenged with OVA alone, or OVA plus the adjuvant previously used during sensitization. Challenge with OVA plus HDE elicited greater airway neutrophilia, lymphocytosis and IL-17A than did challenge with OVA alone. HDE exposure with OVA also promoted greater expansion of adoptively transferred, OVA-specific, IL-17A-fate mapping Th17 cells. These effects were independent of enzymatic activity in HDE, but required TLR4 in recipient mice. Analysis of Tlr4 bone marrow chimeric mice and conditional TLR4 knockout mice indicated that Th17 proliferation is sustained by HDE via TLR4 signaling predominantly in hematopoietic cells, including CD11c+ cells, rather than in airway epithelial cells. IL-23 contributes to the maintenance of these lung Th17 cells. Use of genetically modified mice lacking distinct lung myeloid cell subsets, and co-culture of various lung antigen-presenting cell types with mature Th17 cells, revealed that conventional dendritic cells (either CD103+ or CD11b+), rather than lung macrophages or B cells, support the revival of Th17 clonal expansion and activation. Thus, maintenance of Th17-dependent inflammation in the lung requires persistent innate immune responses, TLR4, and the presence of conventional dendritic cells.