bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mincle activation and type-1 proinflammatory response during scrub typhus: new insight into innate immune recognition of Orientia tsutsugamushi
Authors: Fisher James Robert , Card Galen , Liang Yuejin , Trent Brandon , Soong Lynn 
Published: February 2023

Abstract
O. tsutsugamushi is an obligate intracellular bacterium and etiologic agent of scrub typhus. The lungs harbor the greatest burden of infection, displaying overzealous, type 1-skewed proinflammatory responses. Here, we investigated whether the C-type lectin receptor (CLR) Mincle contributes to immune recognition and dysregulation. Following lethal infection in C57BL/6 mice, we analyzed pulmonary differential gene expression via the NanoString technology. Of 671 genes analyzed, we found 312 significantly induced genes (adj. p &lt; 0.05) at the terminal phase of disease, among which Mincle (Clec4e) was the 4th greatest up-regulated gene (36 fold compared with mock). In addition, Mincle signaling partners (Fcgrs), Cxcr3, Ccr5, and their ligands, as well as Il27, were all highly up-regulated. To validate a role of Mincle in immune recognition, we exposed murine bone marrow-derived macrophages (MΦ) to live or heat-killed O. tsutsugamushi and analyzed a panel of CLRs and proinflammatory markers via qRT-PCR. While heat-killed bacteria transiently stimulated Mincle transcription that dissipated by 24 hpi, only live bacteria generated a sustained response; infection had limited impact on other tested CLRs. Mincle activation was validated by indirect immunofluorescence and western blot. Live, but not heat-killed, bacteria also generated sustained proinflammatory gene expression in MΦ (Cxcl9, Ccl2, Ccl5, Nos2, Il27). Our study provides the first evidence for selective activation of Mincle in sensing O. tsutsugamushi and a role of Mincle- and IL-27-related pathways in severe infection. Since there are no effective vaccines for scrub typhus, our study helps understand innate immune recognition of this poorly-studied bacterium.
Proliferation of Ly6C+ monocytes/macrophages contributes to their accumulation and inflammation in mouse skin wound healing
Authors: Pang Jingbo , Koh Timothy J. 
Published: February 2023

Abstract
Monocytes and macrophages (Mo/Mφ) play critical roles in all phases of normal skin wound healing. However, persistent accumulation of pro-inflammatory Mo/Mφ contributes to impaired healing in diabetes. The purpose of this study was to determine whether proliferation contributes to Mo/Mφ accumulation in skin wounds. First, proliferating Mo/Mφ (F4/80+Ki67+) were observed in wound cryosections from non-diabetic mice. Next, cell cycle analysis indicated skin wounding increased the number and percentage of Ly6C+F4/80lo/-Mo/Mφ in the S/G2/M phases peaking on day 6 post-wounding with significantly more proliferating cells in wounds from diabetic mice compared to non-diabetic mice. Moreover, unbiased scRNAseq analysis revealed that a cluster dominantly composed of wound Mo/Mφ from diabetic mice exhibited upregulation of genes associated with progression and regulation of cell cycle. Finally, a screen of potential regulators of wound Mo/Mφ proliferation showed increased wound CCL2 levels in diabetic versus non-diabetic mice and that local treatment with CCL2 protein increased proliferation of wound Ly6C+F4/80lo/-Mo/Mφ in non-diabetic mice. Importantly, adoptive transfer of mixtures of CCR2−/− and CCR2+/+ Ly6Chi Mo suggested a critical role for CCL2/CCR2 signaling in promoting Mo/Mφ proliferation in the wound environment. Together, these data demonstrate 1) Ly6C+/F4/80lo/-Mo/Mφ proliferate during skin wound healing with higher rates in wounds from diabetic mice than non-diabetic mice and, 2) a novel role for the CCL2/CCR2 signaling pathway in promoting skin Mo/Mφ proliferation, contributing to persistent accumulation of Mo/Mφ and impaired healing in diabetic mice.
Macrophages activate a MLK3/MEK signaling axis to trigger Annexin mediated membrane repair after cholesterol-dependent cytolysin damage
Authors: Ray Sucharit , Keyel Peter A 
Published: February 2023

Abstract
The innate immune system is critical to promoting host survival during bacterial necrotizing soft tissue infections (NSTIs). NSTI causing pathogens like Streptococcus pyogenes and Clostridium perfringens use pore-forming toxins to kill innate immune cells like macrophages. Toxins cause rapid influx of Ca2+, which triggers repair responses. Ca2+ sensitive proteins like Annexins repair via clogging the injured site and are shed with toxin pores on microvesicles during repair. However, the molecular mechanism orchestrating this repair is unknown. Since Mitogen Activated Protein Kinase (MAPK) pathways initiate long-term inflammatory responses, short-term signaling could drive membrane repair. We hypothesized that Mixed Lineage Kinase 3 (MLK3) activates a prosurvival kinase cascade (MEK/ERK) to activate membrane repair by microvesicle shedding in macrophages. We tested cell viability and repair responses to toxins after blocking proteins in the MAPK pathways using inhibitors or RNAi in primary murine macrophages and cell lines. We found MLK3 or MEK inhibition significantly reduced toxin resistance. Protective MLK3/MEK activity was downstream of Ca2+ and accounted for ~ 60% of all Ca2+ dependent repair. MEK inhibition led to increased cytosolic Ca2+ flux after toxin damage and reduced ANXA2, but not ANXA1 or ANXA6, translocation to damaged site. MEK blockade reduced shedding of toxin pores. Overall, these findings suggest that a Ca2+ dependent MLK3/MEK axis protects macrophages and other cells from pore-forming toxins by recruiting ANXA2 to the cell membrane to promote repair. Modulation of this axis could promote resistance to NSTIs.
Genetically Determined Strength of Natural Killer Cells is Enhanced by Adaptive Admixture of HLA class I Allotypes in East &lt;a&gt;Asians&lt;/a&gt;
Authors: Harrison Genelle F , Deng Zhihui , Zhen Jianxin , Zhang Guobin , Chen Rui , Sun Ge , Yu Qiong , Nemat-Gorgani Neda , Guethlein Lisbeth Ann , He Liumei , Tang Mingzhong , Gao Xiaojiang , Cai Siqi , Palmer William , Shortt Jonathan , Gignoux Chris , Carrington Mary , Zhou Hongyan , Parham Peter , Hong Wenxu , Norman Paul 
Published: February 2023

Abstract
Human natural killer (NK) cells are essential for controlling infection, cancer and fetal development. NK cell functions are modulated by interactions between polymorphic inhibitory killer cell immunoglobulin-like receptors (KIR) and polymorphic HLA-A, -B and -C ligands expressed on tissue cells. All HLA-C alleles encode a KIR ligand and contribute to reproduction and immunity. In contrast, only some HLA-A and -B alleles encode KIR ligands and they focus on immunity. By high-resolution analysis of KIR and HLA-A, -B and -C genes, we show that the Chinese Southern Han are significantly enriched for interactions between inhibitory KIR and HLA-A and -B. This enrichment has had substantial input through population admixture with neighboring populations, who contributed HLA class I haplotypes expressing the KIR ligands B*46:01 and B*58:01, which subsequently rose to high frequency by natural selection. Consequently, over 80% of Southern Han HLA haplotypes encode more than one KIR ligand. Complementing the high number of KIR ligands, the Chinese Southern Han KIR locus combines a high frequency of genes expressing potent inhibitory KIR, with a low frequency of those expressing activating KIR. The Southern Han centromeric KIR region encodes strong, conserved, inhibitory HLA-C specific receptors, and the telomeric region provides a high number and diversity of inhibitory HLA-A and -B specific receptors. In all these characteristics, the Southern Han represent other East Asians, whose NK cell repertoires are thus enhanced in quantity, diversity and effector strength, likely through natural selection for resistance to endemic viral infections.
SARS-CoV-2 mediated monocytes dysregulation may contribute to increase risk of cardiovascular risk in severe COVID-19 patients.
Authors: Gunasena Manuja Gayashan Bandara , Wijewantha Yasasvi , Bowman Emily , Gabriel Janelle , Kumar Amrendra , Kettelhut Aaren , Ruwanpathirana Anushka , Weragalaarachchi Krishanthi , Kasturiratna Dhanuja , Vilgelm Anna , Bednash Joseph , Pannu Sonal , Demberg Thorsten , Funderburg Nicholas , Liyanage Namal 
Published: February 2023

Abstract
Severe Acute Respiratory Syndrome Coronavirus 2 (SARS□CoV□ 2) is a highly pathogenic corona virus which causes COVID-19 and resulted in millions of deaths and led to a global public health emergency. An effective and appropriate immune response is essential to control and eliminate viral infections, however, dysregulated immune responses may lead to immunopathology in viral infections. Clinical data showed that COVID-19 may promote the development of cardiovascular disorders (CVDs). However, exact mechanism associated with CVDs in COVID-19 patients is currently unknown. Monocytes are one of the major immune cell subsets that contribute to host defense against many infections but also associated with immunopathology and development of CVDs. Thus, in this study we investigated the role of monocyte subsets in COVID-19 associated CVDs. By using high dimensional flowcytometry, immune biomarker assays and multi variant data analysis in healthy donors (n=17), severe-COVID-19 patients (n=20) (ICU) and COVID-19 recovered individuals (n=30), we found elevated intermediate monocytes and Monocyte chemoattractant protein-1, one of the most important chemokines that regulates migration and infiltration of monocytes to the subendothelial space, where they may become foam cells. Spearman correlation network analysis showed that strong correlation between increase intermediate monocytes in the whole blood and plasma cardimetabolic biomarkers such as fatty acid binding protein 4, C-reactive protein, soluble CD14 and LPS binding protein in ICU patients. Our data show for the first time that the possible role of monocytes in the development of CVDs in ICU patients.
Perforin-2 deficiency delays wound reepithelialization
Authors: O’Neill Katelyn Elizabeth , Padula Laura , Fisher Eva , Head Cheyanne , Wikramanayake Tongyu , Pastar Irena , Tomic-Canic Marjana , Strbo Natasa 
Published: February 2023

Abstract
Wound repair declines with age and contributes to a variety of health complications; however, the cellular and molecular mechanisms governing this decline are not well understood. We observed that the novel pore-forming antimicrobial protein Perforin-2 (P-2) is suppressed in the epidermis of diabetic foot ulcers (DFUs), a major diabetes complication that affects the elderly. Therefore, we sought to investigate the role of P-2 in inducing reepithelialization in wounded skin, particularly focusing on contributions of aging.
We induced wounds on the dorsal skin of 2 or 10-month-old C57/BL6 WT and P-2 deficient (−/−) mice using an 8 mm biopsy punch and assessed healing at day 3 and 6 post wounding. We analyzed reepithelialization and keratinocyte activation using H & E stained and Keratin-6 and Caveolin-1 antibody stained wound tissues, respectively.
We found that wound reepithelialization was delayed in 2-month old P-2 −/− mice on day 3 after wounding in comparison to WT, but there was no difference in reepithelialization by day 6. Interestingly, in 10-month old mice, wound reepithelialization was significantly delayed at day 6 in P-2 −/− mice compared to WT. This delay was accompanied by significantly decreased Keratin-6 expression at the wound edge. Immunofluorescence staining and flow cytometric analysis revealed significantly fewer GD T cells in the wound bed in aged P-2 −/− mice. These P2 −/− GD T cells also displayed significantly lower expression of activation markers such as NKG2D.
Our data demonstrate impaired keratinocyte activation and GD T cell infiltration in the absence of P-2 that is more pronounced in aged mice, indicating a potential role for this protein in inducing the inflammatory response upon wounding.
p38 MAPK activation by Toxoplasma gondii secretory effector protein GRA24: Role in cytokine regulation and induction of protective immunity
Authors: Mercer Heather Lynn , Snyder Lindsay M. , Menard Kayla L. , Doherty Claire M. , Bzik David J. , Fox Barbara A. , Denkers Eric Y. 
Published: February 2023

Abstract
The apicomplexan Toxoplasma gondii induces strong protective immunity in part dependent upon recognition by Toll-like receptors (TLR) 11 and 12 operating in conjunction with MyD88 in the murine host. However, these TLR are not present in humans, prompting us to investigate MyD88-independent signaling pathways of resistance. We show here that parasite dense granule protein GRA24 induces p38 MAPK activation and subsequent IL-12 production in bone marrow derived mouse macrophages (BMDM). Furthermore, Toxoplasma triggers two waves of p38 MAPK phosphorylation in MyD88+/+ and MyD88−/− BMDM. Using the uracil auxotrophic Type I T. gondii strain cps1-1, we demonstrate that the late response does not require active parasite proliferation, but strictly depends upon GRA24. By i. p. inoculation with cps1-1 and cps1-1:Δgra24, we identified unique subsets of chemokines and cytokines that were up and down regulated by GRA24. We demonstrate that cps1-1 triggers a strong host-protective GRA24-dependent Th1 response in the absence of MyD88. To gain deeper insight into genes regulated by GRA24, we employed qPCR to evaluate expression of transcripts for 84 cytokine and chemokine genes in BMDM infected with GRA24-positive compared to GRA24-negative parasites. Our results identify a specific signature of genes regulated by this dense granule protein. In conjunction, using qPCR we evaluated the impact of GRA24 on a panel of macrophage long noncoding RNA (lncRNA) regulated by Toxoplasma infection. Taken together, our data identify GRA24 as a major mediator of p38 MAPK activation, induction of IL-12 and other immune mediators, as well as protective immunity that operates independently of the TLR/MyD88 cascade.
Development of a quantitative COVID-19 Multiplex Assay and its use for serological surveillance in a low SARS-CoV-2 incidence community
Authors: Guarino Cassandra M , Larson Elisabeth , Babasyan Susanna , Rollins Alicia , Joshi Lok , Laverack Melissa , Parrilla Lara , Plocharczyk Elizabeth , Diel Diego , Wagner Bettina 
Published: February 2023

Abstract
A serological COVID-19 Multiplex Assay was developed and validated using serum sample sets from convalescent patients and those collected prior to the onset of the 2020 pandemic. After initial testing of multiple potential antigens expressed in a mammalian system, the SARS-CoV-2 nucleoprotein (NP) and receptor-binding domain (RBD) of the spike protein were selected for the final human COVID-19 Multiplex Assay. A comparison of synthesized and mammalian expressed RBD proteins revealed clear advantages of the mammalian protein. Antibodies directed against NP strongly correlated with SARS-CoV-2 virus neutralization assay titers (rsp = 0.726), while the anti-RBD correlation was moderate (rsp = 0.436). Pan-Ig, IgG, IgA, and IgM against the NP and RBD antigens were evaluated on the validation sample sets. Detection of NP and RBD specific IgG and IgA had outstanding performance (AUC&gt;0.90) for distinguishing patients from controls, but the dynamic range of the IgG assay was substantially greater than for IgA. The COVID-19 Multiplex Assay was then utilized to identify seroprevalence to SARS-CoV-2 in people living in a low-incidence community in Ithaca NY. Samples were taken from a cohort of healthy volunteers (n=332) in early June 2020. Only two volunteers had a positive result on a COVID-19 PCR test performed prior to serum sampling. Serological testing revealed an exposure rate of at least 1.2% (NP) or as high as 5.1% (RBD), which was higher than the measured incidence rate of 0.16% in the county at that time. This highly sensitive and quantitative assay will be useful for continuing to monitor community exposure rates, duration of immune response following infection, and monitoring vaccination responses as vaccines become widely available.
IL-33 activates ILC2 expansion and modulates endometriosis
Authors: Miller Jessica E , Lingegowda Harshavardhan , Symons Lindsey K , Bougie Olga , Young Steven L , Lessey Bruce A , Koti Madhuri , Tayade Chandrakant 
Published: February 2023

Abstract
Introduction: Chronic inflammation drives endometriosis (EM) and its associated symptoms. Interleukin (IL)-33, has gained therapeutic interest as it is elevated in the plasma, peritoneal fluid (PF), and lesions of EM patients. However, how IL-33 contributes to EM is largely unknown. For the first time, we demonstrate, using both human samples and murine models, that IL-33 expands local group 2 innate lymphoid cells (ILC2s) and that IL-33 activated ILC2s modulate hallmark features of EM.

Methods:
We identified ILC2s (CD45+Lin−CRTH2+CD127+ST2+) in the PF of EM patients using flow cytometry. Using immunofluorescence, we determine whether IL-33 forms epithelial or stromal cell niches within the EM lesion. Then, using our well established EM model, we induced EM in female C57Bl/6 mice and treated with PBS or IL-33. Hallmark features of EM were assessed including inflammation (using multiplex cytokine analysis), immune cell profiling (using flow cytometry), and lesion architecture (using immunohistochemistry and Nanostring transcriptomic profiling). To establish cause and effect, we treated ILC2 deficient mice with PBS (n=5) or IL-33 (n=5). Unpaired t-tests and one-way ANOVAs were done when appropriate.

Results/Discussion:
In humans, ILC2s are present and elevated in EM patients (n=5) compared to healthy donors (n=3) and IL-33 colocalized with both epithelial and stromal cells within the EM lesion. Using murine models, IL-33 drove hallmark features of EM including increased inflammation, alterations in PF immune cells (including eosinophils, T cells, macrophages and ILC2s), as well as increased lesion proliferation, neurogenesis and fibrosis. Using ILC2 deficient mice, we show that IL-33 induced pathology is ILC2 dependent.
Adipocyte-specific PIK3C3/VPS34 controls adipogenesis, metabolically-triggered inflammation, insulin resistance, and adaptive thermogenesis in mice
Authors: Song Wenqiang , Yang Guan , Postoak J. Luke , Zhang Jianhua , Wu Lan , Van Kaer Luc 
Published: February 2023

Abstract
Interactions between cells of the immune and metabolic systems play critical roles in controlling immune responsiveness and metabolic health. When these interactions are dysregulated, metabolic diseases such as obesity or lipodystrophy may develop. We have focused on cellular processes related to autophagy, a cellular self-degradation process, in controlling interactions between the immune and metabolic systems. In this study, we analyzed mice with an adipocyte-selective deficiency in PIK3C3/VPS34, a key early player in autophagy. These animals showed reduced white and brown adipose tissue (WAT and BAT) with enhanced macrophage infiltration, systemic immune activation, and spontaneous insulin-resistance. The phenotype of these animals bears a striking resemblance with the disease manifestations seen in individuals with lipodystrophy. Mechanistically, we found that conditional Pik3c3-deficiency inhibits adipogenesis in both WAT and BAT by downregulating glucose uptake and fatty acid metabolism, and impairs the thermogenic function of BAT by downregulating the expression of key regulators (Prdm16, Ucp1, and Cidea). Upon acute cold exposure, Pik3c3-deficient mice became cold-sensitive under fasting conditions due to insufficient thermogenesis in BAT. In contrast, these animals exhibited blunted beige fat thermogenesis in subcutaneous WAT in response to administration of a b3-adrenergic receptor agonist. Collectively, our data demonstrate that PIK3C3 is a vital regulator of adipose tissue development, metabolically-triggered inflammation, insulin resistance, and adaptive thermogenesis. Hence, we identify PIK3C3 as a potential therapeutic target for metabolic diseases such as type 2 diabetes.
Meningeal lymphatics near the cribriform plate undergo lymphangiogenesis and directly engage in leukocyte crosstalk during neuroinflammation
Authors: Hsu Martin , Madrid Andy , Laaker Collin , Choi Yun Hwa , Herbath Melinda , Sandor Matyas , Fabry Zsuzsanna 
Published: February 2023

Abstract
Meningeal lymphatics in the dural layer surrounding the dorsal regions of the brain, basal regions, and near the cribriform plate have all been implicated in the management of neuroinflammation and edema by facilitating the drainage of fluid, macromolecules, antigens, and cells. We have previously published that during neuroinflammation only the lymphatics near the cribriform plate are able to undergo extensive morphological changes through lymphangiogenesis in response to neuroinflammation during Experimental Autoimmune Encephalomyelitis (EAE), a mouse model of Multiple Sclerosis. In this study, we hypothesize that these cribriform plate lymphatics likely have other phenotypic changes as a consequence of neuroinflammation-induced lymphangiogenesis, and that identifying these phenotypic alterations may shed light on potentially novel roles of meningeal lymphatics during neuroinflammation. Single cell RNA sequencing reveals the upregulation of genes involved in leukocyte crosstalk and regulation such as chemotaxis/adhesion and antigen processing/presentation, suggesting that cribriform plate lymphatics may play a direct role in regulating adaptive immunity in addition to its canonical roles in facilitating drainage. We also demonstrate that cribriform plate lymphatics are able to capture CNS-derived antigens and functionally engage in crosstalk with dendritic cells and CD4 T cells. These data characterize cribriform plate lymphatics and demonstrate that these vessels become dynamic in response to neuroinflammation to facilitate excess drainage as well as directly modulate immunity through leukocyte crosstalk.
Sodium chloride generates anti-inflammatory Th17 cell responses but amplifies Th17 cell pathogenicity in pro-inflammatory cytokine microenvironments
Authors: Zielinski Christina , Matthias Julia 
Published: February 2023

Abstract
Ionic signals have recently been demonstrated to affect T cell polarization and function. Sodium chloride (NaCl) was proposed to accumulate in peripheral tissues upon dietary intake and to promote autoimmunity via the Th17 cell axis. We here demonstrate that together with amplification of human Th17 cell signature properties, NaCl gave rise to a stable anti-inflammatory Th17 cell fate that we found to be restricted to distinct pathogen specificities. The p38/MAPK pathway involving NFAT5 and SGK1regulated FoxP3 and IL-17 expression in high NaCl conditions. The NaCl induced acquisition of an anti-inflammatory Th17 cell fate was confirmed in vivo in the mouse model of experimental autoimmune encephalomyelitis (EAE), which demonstrated strongly reduced disease symptoms upon transfer of NaCl-treated encephalitogenic Th17 cells. However, NaCl was coopted to promote murine and human Th17 cell pathogenicity, if T cell stimulation occurred in a proinflammatory and TGF-b low cytokine microenvironment. This revealed a context dependent dichotomous role for NaCl in shaping the pathogenicity of Th17 cells. NaCl might therefore prove beneficial for the treatment of chronic inflammatory diseases in collaboration with cytokine blocking drugs.