bluuebunny/crossref_metadata_2025_split huggingface dataset:
TOLLIP Deficiency is Associated with Increased Mycobacterium tuberculosis-specific Anti-Microbial Monocyte Responses and Protection from Pediatric Tuberculosis in South Africa
Authors: Shah Javeed A , Musvosvi Munyaradzi , Shey Muki , Wells Richard D. , Peterson Glenna J. , Cox Jeffrey S. , Daya Michelle , Hoal Eileen , Gottardo Raphael , Hanekom Willem A. , Scriba Thomas J. , Hatherill Mark , Hawn Thomas 
Published: January 2023

Abstract
Pediatric tuberculosis (TB) is a significant cause of death in children and has a different pathogenesis than adult TB, but BCG-induced immunity is incomplete. Although genetic factors regulate BCG-induced immunity and TB susceptibility, these genes are mostly unknown and have largely been unexamined in pediatric TB. We hypothesized that TOLLIP, a TLR suppressor, orchestrates immune responses to M. tuberculosis (MTb) and susceptibility to pediatric TB. We discovered that a polymorphism of TOLLIP, rs5743854, regulated signaling in a promoter assay and was associated with TOLLIP mRNA deficiency in infant peripheral blood monocytes (p = 0.008, Kruskal-Wallis). The rs5743854 G allele was associated with protection from pediatric TB in a South African cohort (N=226 cases and 626 controls, dominant model, p = 0.002, odds ratio (OR) 0.63 unadjusted analysis; p = 0.011, OR = 0.58 adjusted for ethnicity by ancestry informative markers). The G allele was also associated with decreased frequency of BCG-specific CD4+IL-2 responses (p = 0.003, Kruskal-Wallis test) and CD4+ T-cell proliferation (p = 0.03, generalized linear model) in 174 infants vaccinated with BCG at birth and re-stimulated at 10 weeks of age. To examine mechanisms of protection, we created a TOLLIP-deficient THP-1 cell line. TOLLIP-deficient monocytes had increased MTb-induced TNF and reactive nitrogen secretion and reduced MTb replication compared to control cells. Together, these data suggest that TOLLIP deficiency is associated with enhanced anti-microbial monocyte responses that control MTb replication and provide protection against pediatric TB disease. Suppressing TOLLIP levels by host-directed therapeutics may provide adjunctive therapy for pediatric TB.
Activated B cells from BENTA patients display intrinsic differentiation defects and resistance to apoptosis
Authors: Arjunaraja Swadhinya , Snow Andrew L 
Published: January 2023

Abstract
We recently characterized a novel, B-cell specific congenital lymphoproliferative disorder termed BENTA (B cell Expansion with NF-κB and T-cell Anergy). BENTA disease is caused by heterozygous, germline gain-of-function mutations in the lymphocyte-specific signaling protein CARD11, triggering constitutive NF-κB signaling. BENTA patients present not only with splenomegaly and remarkable polyclonal immature/naive B cell expansion, but also display signs of primary immunodeficiency, including reduced percentages of class-switched/memory B cells, low serum IgM and poor antibody responses to specific vaccines. Using purified naïve B cells from several BENTA patients, we discovered profound intrinsic defects in plasma cell differentiation and antibody secretion in vitro in response to various stimuli. Although BENTA B cells proliferated slightly more than healthy donor B cells in vitro, they failed to differentiate into short-lived IgDloCD38hi plasmablasts or CD138+ long-lived plasma cells. Consistent with these defects, BENTA B cells exhibited low IgG secretion, and reduced induction of key proteins involved in class switching (IRF4) and late plasma cell commitment (XBP1). On the other hand, activated BENTA B cells exhibited an overwhelming survival advantage in culture, reflecting apoptosis resistance that may be linked to a CARD11-dependent, NF-κB-independent pathway. Together, these results provide new mechanistic insights into our understanding of both B cell lymphocytosis and humoral immunodeficiency in BENTA disease, which may emerge from impairments in cell death and differentiation programs normally triggered by B cell activation.
ASSOCIATION BETWEEN DISEASE ACTIVITY AND IL-17A/F, IL-23 AND IL-12/23 (p40) PRODUCTION BY PERIPHERAL BLOOD T LYMPHOCYTES IN BEHCET’S DISEASE
Authors: Yucel Aysegul Atak , Sonmez Cemile , Yesil Turan Hilmi , Kucuk Hamit , Mercan Ridvan , Yucel Eftal Ahmet , Goker Berna , Tufan Muge Aydin , Demirel Gulderen Yanikkaya , Sezgin Berna 
Published: January 2023

Abstract
Exact pathogenesis of Behcet’s Disease (BD) hasn’t been known but recent studies suggest it’s a genetic-based and immune-related disease. We aimed to investigate the association between disease activity and Th1, Th17 and Treg cells in BD patients via IL-17A/F, IL-23, IL-12/23p40, IL-35 produced by these cells. We also compared our findings with those of SLE patients to investigate their specificity for immunopathogenesis of BD. 15 active and 15 inactive BD patients, 12 active and 12 inactive SLE patients and 12 sex & age-matched healthy controls were involved. Peripheral blood lymphocyte cultures were done. Supernatants from PHA-stimulated and -unstimulated cultures were measured for IL-17A/F, IL-12/23p40, IL-23 and IL-35 levels in time-dependent manner (48 and 72 hrs) by ELISA. For disease activity International Study Group Diagnostic Criteria and SLEDAI index were used for BD and SLE, respectively. IL-17 and IL-23 increased parallelly in both BD and SLE; IL-17 was higher but another Treg cytokine IL-35 was lower in active BD and active SLE compared to healthy controls. After 48 and 72 hrs of PHA stimulation the highest levels of IL-17A/F, IL-22723p40 and IL-23 were measured in active BD followed by active SLE. Our data suggest that IL-17, IL-23 and IL-12/23p40; Th17 and Th1 responses play role in pathogenesis of BD. Finding IL-35 to be lower in active BD compared to inactive BD patients and healthy controls made us think that there can be a plasticity between Th17 and Treg cells depending on disease activity. This is the first time such plasticity is suggested for BD in literature. Our findings also show that IL-17/23 axis contributes to pathogenesis of SLE (another inflammatory rheumatic disease) in addition to other mechanisms like in BD.
Human liver contains two populations of NK cells based on CD49e expression
Authors: Yeang Han Xian Aw , Lin Yiing , Malkova Olga N , Yang Liping , Piersma Sytse J , Miner Cathrine , Oh Stephen T , Chapman William , Yokoyama Wayne M 
Published: January 2023

Abstract
Natural killer cells are a subset of innate immune lymphocytes that can kill transformed and infected cells, and produce cytokines that modulate the host immune response. Previous studies on human NK cells have primarily characterized NK cells isolated from peripheral blood. However, studies in mice from our lab and others indicate an additional tissue-resident NK (trNK) population that does not re-circulate in contrast to conventional NK (cNK). When the mouse liver was excised and flushed with saline, trNK cells were detected, indicating that the trNK cells reside in the liver sinusoidal space. Recent studies to identify trNK cells in the human liver have concentrated on the differential expression of CD56-dim/bright populations of NK cells and also CD49a, a marker we previously defined for mouse trNK cells in the liver. Herein we obtained samples from liver transplantation cadaveric donors. Matching donor venous blood and serial liver saline flushes were studied. Our preliminary data identifies a subset of putative liver-trNK cells based on lack of CD49e expression which appeared in the terminal liver flush aliquot and was not present in earlier liver flushes nor peripheral blood whereas CD49a expression was not discriminatory. Mass cytometry experiments with cytometry by time-of-flight instrument and a panel of 42 antibodies corroborated these findings. Interestingly, the liver-resident CD49e− cells have elevated EOMES but lower Tbet expression which contrasts the CD49a+ trNK cell populations in human and mouse liver. Finally, the putative trNK cells express different killer immunoglobulin-like receptor (KIR) repertoire than cNK cells thus, a unique population of trNK cells that reside in the sinusoidal space of the human liver.
Immunophenotypic Characterization of Hemorrhagic Shock and Poly-trauma in Non-Human Primates
Authors: Paredes Ruth Madelaine , Sooter Amanda , Vernon Philip Joseph , Glaser Jacob , Sheppard Forest 
Published: January 2023

Abstract

Background
Severe hemorrhage and trauma initiates an inflammatory cascade that promotes innate and adaptive immune-dysregulation. The kinetics of immune-cell subtype expansion, recruitment and retention, remain to be elucidated. We sought to immunophenotypically characterize the innate and adaptive cell response to hemorrhage and poly-trauma in non-human primates (NHPs).

Methods
Rhesus macaques (n=32) were randomized into four injury groups: 30min hemorrhagic shock (30HS), 60 min HS (60HS), 60HS + laparotomy, and 60HS + laparotomy + femur fracture and repair. After injury, animals were resuscitated and monitored for24 hours prior to euthanasia. Whole blood was collected at baseline (BSLN), end of shock (EOS), end of resuscitation (EOR), t=360 min and t=1440 min. Flow cytometric analysis was performed to identify T-cells, B-cells, Natural Killer (NK) cells and monocytes/macrophages/granulocytes. Results reported as mean±SEM, statistical analysis: two-way ANOVA, with p&lt;0.05 significant.

Results
Ineach injury model, the proportion of lymphocytes decreased (p&lt;0.05), and myeloid population frequency increased (p&lt;0.05) from BSLN over the course of the experiment. Frequencies of lymphocytes decreased (p&lt;0.05); where as, frequencies of myeloid cells increased (p&lt;0.05). No significant differences were found between models.

Conclusion
The data demonstrates a shift in lymphoid to myeloid predominance following hemorrhage and injury, consistent with that reported in humans. Interestingly, severity of injury did not alter the magnitude of this response, implying an early trigger. Further investigations into post-traumatic immunomodulation should target myeloid cells.
Quantitative assay of human antibody-dependent cell-mediated cytotoxicity (ADCC) using peripheral blood mononuclear cells (PBMCs)
Authors: Sung Alexander P , Tang Jia Jie , Guglielmo Michael J. , Redelman Douglas D , Silvestre Isabel Barao , Gagen Julie Smith , Hudig Dorothy 
Published: January 2023

Abstract
ADCC can be essential for anti-viral immunity and supports many tumor immunotherapies. It is a challenge to quantify and one wants a fast assay that uses few lymphocytes when patient blood is limited. Three considerations affected our new assay design: 1) normal B cells within PBMCs introduce ‘cold target’ competition of radiolabeled tumor B cells if anti-B cell antibodies are present in the ADCC assays; 2) type 1 antibodies to CD20 B cell epitopes are internalized and cleared from the cell surface while type 2 anti-CD20 antibodies remain external; and 3) target cells with MHC I initiate KIR-regulation of lysis that will vary among donors. To address these issues, we used Daudi B leukemia cells that lack MHC I and pretreated the Daudi cells with the humanized nonfucosylated/glycoengineered monoclonal type 2 anti-CD20 antibody obinutuzumab (GA101 G2). Target cell pretreatment prevented ‘cold target’ competition. We used 4 hr 51Cr-release for its sensitivity and varied the PBMC to target (E:T) cell ratios to obtain lytic units. Flow cytometric counts of CD16A Fc-receptor positive cells with TrueCountR beads determined the exact number of ADCC effectors within the unfractionated PBMCs. There was low NK activity to Daudi cells, which required subtraction of simultaneous controls without antibody. We plotted ADCC activity as a log function of the E:T ratios and observed that ADCC was dependent upon interactions of multiple CD16A-positive effectors per target cell, which was expressed as lytic units per 106 CD16A cells, that varied 50 fold among donors. In addition, the linear slopes of CD16A cell lytic cooperativity varied 8 fold. In summary, we have established a fast quantitative assay for comparison of human ADCC that uses as few as 8 million PBMCs.
Sepsis compromises primary B cell-mediated responses
Authors: Griffith Thomas S , Condotta Stephanie A , Tygrett Lorraine T. , Rai Deepa , Yang Jessica A , Pape Kathryn A , Murphy Katherine A , Cabrera-Perez Javier , Waldschmidt Thomas J , Badovinac Vladimir P 
Published: January 2023

Abstract
Sepsis is the leading cause of death in non-coronary ICU’s, and one hallmark of sepsis patients is sustained immune suppression. Septic patients exhibit compromised innate and adaptive immunity, making them highly susceptible to secondary infections. Using the cecal ligation and puncture (CLP) model of sepsis, we recently showed the recovery of CD4 T cells from CLP-induced lymphopenia is accompanied by alterations in the composition and function of the Ag-specific CD4 T cell repertoire. These data led us to hypothesize that CD4 T cell-dependent B cell responses would also be impaired after CLP. Using the same CLP model, we immunized sham (control) and CLP mice with PE-conjugated 2W1S56 peptide 30 d after surgery, and then we quantitated the number of 2W1S-specific CD4 T cells and PE-specific B cells 7 d later. Compared to sham mice, CLP-treated mice immunized with PE-2W1S56 had a sustained reduction in number of 2W1S-specific Tfh CD4 T cells and naïve/memory PE-specific B cells after CLP. Next, we immunized sham and CLP mice with TNP-KLH plus either CpG or alum early (2 d) or late (30 d) after surgery to induce Th1 or Th2 Ab responses, respectively. We observed compromised production of anti-TNP Ab in mice immunized with either TNP-KLH/CpG or alum early (2 d) after CLP. Importantly, Ab production remained reduced in mice immunized with TNP-KLH/alum late (30 d) after CLP. Together, our data show CLP-induced sepsis impacts humoral immunity by affecting the number and function of Ag-specific B cells and CD4 Tfh cells. Moreover, these data suggest potential for defects in humoral immunity to newly introduced Ag (e.g., vaccines) in sepsis survivors long after the septic event has resolved.
Short-lived myeloid cell lifespan is regulated by a long non-coding RNA
Authors: Kotzin Jonathan J , Spencer Sean P , McCright Sam J , Uthaya Kumar Dinesh B. , Mowel Walter K , Makiya Michelle , Klion Amy D. , Williams Adam , Flavell Richard A , Henao-Mejia Jorge 
Published: January 2023

Abstract
Neutrophils, eosinophils and “classical” monocytes collectively account for ~70% of blood leukocytes and are among the shortest-lived cells in the body. Precise regulation of the short lifespan of these myeloid cells is critical to maintain effective host responses to pathogens while minimizing the deleterious consequences of prolonged inflammation. However, how the lifespan of these cells is strictly controlled remains largely unknown. We identified a novel long non-coding RNA (lncRNA) that is highly and specifically expressed by neutrophils, eosinophils and “classical” monocytes in response to pro-survival cytokines. Ablation of this lncRNA in mice leads to a dramatic reduction in the survival of these myeloid cells, rendering these animals highly susceptible to bacterial infection and resistant to the development of immunopathology. To control the lifespan of these cells, this lncRNA epigenetically regulates the transcription of its neighboring pro-apoptotic gene in an allele-specific manner. Thus, in these highly inflammatory cells, changes in levels of this lncRNA provide a locus-specific regulatory mechanism that allows for rapid and dynamic control of apoptosis in response to extracellular signals. In concordance, expression of this lncRNA is significantly elevated in eosinophils from human patients with hypereosinophilic disorders, which are characterized by high plasma levels of the pro-survival cytokine IL-5. Our results indicate that locus-specific regulation of gene expression by lncRNAs is a powerful mechanism by which environmental cues are integrated to achieve optimal immune cell homeostasis. Moreover, this lncRNA may represent a potential therapeutic target for several inflammatory disorders.
Single cell variability in pro-inflammatory and antiviral gene responses in dendritic cells
Authors: Fribourg Casajuana Miguel L , Hartmann Boris M. , Tabbaa Omar P. , Ramos Irene , Zaslavsky Elena , Nudelman German , Albrecht Randy A. , Merad Miriam , Hayot Fernand , Jayaprakash Ciriyam , Kleinstein Steven H. , Garcia-Sastre Adolfo , Sealfon Stuart C. 
Published: January 2023

Abstract
Type I interferons (IFNs) can have both protective and deleterious effects in chronic infection and autoimmune disease. Cellular responses to type I IFNs are mediated by IFN-stimulated genes (ISGs), are cell type-dependent, and likely cell specific. Varying responses of individual cells to immune signals, such as type I IFNs, may be critical in orchestrating context and microenvironment appropriate immune responses. Simulations of an agent-based mathematical model of viral infection suggested that within each cell, the levels of pro-inflammatory ISGs induced by STAT dimers and the antiviral ISGs induced by the ISGF-3 heterotrimer can show low correlation. We used RNAseq to study global single-cell correlation of ISG groups in human peripheral blood CD1c+ dendritic cells infected with an influenza A H1N1 virus. Consonant with the simulations, distinct groups of ISGs were identified that showed high levels of single cell expression correlation within, but low to negative correlations across ISG subgroups. Using multiple probe FISH we find low single cell correlation of the pro-inflammatory ISG IL6 and the antiviral ISG RIG-I induction in cells stimulated with type I IFN. The single cell correlations in the induction of the two genes were 0.56, 0.55 and 0.19 in human peripheral blood plasmacytoid dendritic cells, monocytes, and monocyte-derived dendritic cells respectively. The generation of uncorrelated pro-inflammatory and antiviral ISG single cell responses within a dendritic cell population can increase the breadth of the immune response to noxious stimuli.
Ly49 Pro1 activity is associated with gene activation, not gene expression: Pro1 transcripts are not present in mature Ly49-expressing NK cells.
Authors: Anderson Stephen , McCullen Matthew V , Sen Shurjo , McVicar Daniel W 
Published: January 2023

Abstract
Recent studies of the Ly49 loci have challenged the traditional view of promoters, since transcriptional start sites for the variegated Ly49 genes were not found to be associated with regions possessing transcriptional activity in in vitro promoter assays. The upstream Pro1 bidirectional promoter previously associated with probabilistic activation of variegated Ly49 genes by immature bone marrow NK cells was shown to be active in mature NK cells and function as an enhancer element, suggesting that it may play a role in Ly49 transcript initiation in mature NK cells.
To directly assess if Pro1 transcripts are present in mature Ly49-expressing NK cells, RNA sequencing of the total transcript pool was performed on freshly isolated splenic NK cells sorted for expression of either Ly49G or Ly49I. No Pro1 transcripts were detected from the Ly49a, Ly49c or Ly49i genes in mature Ly49+ve NK cells that contained high levels of Pro2 or Pro3 transcripts. Ly49g Pro1 transcripts were found at low levels in both Ly49G+ve and Ly49G-ve populations, consistent with the presence of a small population of mature NK cells undergoing Ly49g gene activation, as previously demonstrated by culture of splenic NK cells in IL-2. Furthermore, Ly49 gene reporter constructs containing Pro1 failed to show any enhancer activity of Pro1 on the downstream Ly49 promoter active in mature Ly49-expressing NK cells. Taken together, the results are consistent with Pro1 transcription playing a role in gene activation in developing NK, and argue against a role for Pro1 in Ly49 gene transcription by mature NK cells.
The antiviral molecule Mx1 positively regulates the induction of type I IFN in response to influenza infection
Authors: Schattgen Stefan A , Oguin Thomas H , Thomas Paul G 
Published: January 2023

Abstract
Myxovirus resistance 1 (Mx1), a member of the dynamin-like GTPase family, has been long described to have an important function as a cell-intrinsic mechanism with broad antiviral activity against a wide range of viruses. How Mx1 imparts its antiviral activity, however, has been elusive and remains unknown. In this study we seek to understand the mechanism by which Mx1 inhibits Influenza A virus (IAV) infection in vitro. Here we found knock-down of Mx1 expression in A549 cells led to a significant reduction in the expression of interferon-stimulated genes (ISGs) following infection with IAV, along with increased viral replication. This observation is corroborated by decreased phosphorylation of interferon regulatory factor 3 (IRF3), a key transcription factor in the transcription of the ISGs, in cells depleted of Mx1. Though Mx1 is itself an ISG and its expression is upregulated in response to IFN receptor signaling, we found that it is present basally at low levels. A significant pool of Mx1 was found in the mitochondrial fraction in experiments examining its localization within the cell. Furthermore, immunoprecipitation of Mx1 in resting cells followed by mass spectrometry to investigate its potential interactions revealed a number of proteins known to be localized to the mitochondrial membrane, a location known to be an important platform for MAVS-dependent initiation of the interferon response to RNA viruses. These data taken together suggests a potential cooperative role for Mx1 in the recognition and initiation of the antiviral program during IAV infection.
Lactic acid inhibits IL-33-mediated mast cell inflammatory responses via HIF-1α suppressing miR-155 expression
Authors: Abebayehu Daniel , Spence Andrew , Qayum Amina Abdul , Taruselli Marcela T , McLeod Jamie J , Caslin Heather , Kolawole Elizabeth Motunrayo , Ndaw Victor S. , Barnstein Brian O , Oskeritzian Carole , Sell Scott , Ryan John J 
Published: January 2023

Abstract
Lactic acid (LA) is elevated in tumors, asthma, and wound healing, environments that also include mast cells and IL-33. However, how LA affects mast cell function is unknown. Therefore we evaluated how LA modifies the IL-33-mediated mast cell response. When bone marrow derived mast cells were cultured with LA, we noted reduced IL-33-mediated cytokine production, an effect that was both monocarboxylate transporter (MCT)1- and pH-dependent. LA selectively decreased IL-33-induced TAK1, JNK, ERK, and NFkB phosphorylation, but not p38 activation. Additionally, LA increased HIF-1α expression. Since HIF-1α has been shown to regulate the pro-inflammatory microRNA miR-155, we examined miR-155 expression. miR-155-5p was reduced by LA, an effect that was reversed by HIF-1α antagonism. More importantly, miR-155-5p overexpression abolished the suppressive effects of LA. Additionally, the negative regulator SOCS1, a known miR-155 target, was elevated by LA addition. This suggests that LA employs HIF-1a to suppress miR-155, indirectly increasing SOCS-1 and limiting cytokine production. These data were recapitulated in vivo, since C57BL/6 mice injected with LA showed less IL-33-induced plasma cytokine levels than control mice. Lastly, LA suppressed IL-33-mediated human skin mast cell activation, an effect that was also MCT1-dependent. Our data demonstrate that lactic acid, present in inflammatory and malignant microenvironments, alters mast cell function to suppress inflammation.