bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cytoplasmic labile zinc regulates IL-1β production in human monocytes/macrophages via mTORC1-induced glycolysis
Authors: Kim Bonah , Lee Su Jeong , Kang Yeon Jun , Lee Ga Hye , Kim Hee Young , Lee Won-Woo 
Published: January 2023

Abstract
Zinc is an essential micronutrient that plays critical roles in multiple facets of biological processes, including growth, development, proliferation, and differentiation. Since dysregulated zinc homeostasis impairs overall immune function and resultantly, increases susceptibility to infection, much attention has been recently paid to immunomodulatory roles of zinc in immune cells. Rapid and efficient production of proinflammatory cytokines, such as IL-1β and TNF-α, is a primary effect function of monocytes/macrophages for host defense. However, the molecular mechanism involved with zinc-mediated modulation of cytokine production in monocytes/macrophages needs to be better understood. In present study, we provide evidence that cytoplasmic labile zinc is essential for modulation of IL-1β production in human monocytes/macrophages via mTORC1-induced glycolysis. We found that cytoplasmic level of labile zinc in human monocytes/macrophages was largely influenced by extracellular zinc concentrations, possibly via zinc importers such as ZIP1- and ZIP8-mediated influx. The phosphorylation of S6 kinase, a substrate of mTORC1, was significantly enhanced by zinc-mediated inhibition of PP2A, a phosphatase contracting S6K phosphorylation and consequently, IL-1β production was increased by activated mTORC1-induced glycolysis. These results provide new insight how labile zinc modulate a production of cytokines by metabolic reprograming in human monocytes/macrophages.
Bacterial Polyphosphates Modulate Macrophage Responses to Infection
Authors: Bosmann Markus , Roewe Julian , Stavrides Georgios , Marini Federico , Smith Stephanie A. , Reinhardt Christoph , Morrissey James H. 
Published: January 2023

Abstract
Polyphosphates are linear polymers of inorganic monophosphate residues (Pi) and these ancient metabolites exist in all living cells. In bacteria, polyphosphate kinases (PPK) and polyphosphatases (PPX) control the accumulation of polyphosphates from ATP as long chains (&gt;300–1000 Pi) for energy storage and prokaryotic homeostasis.
Here, we demonstrate that bacterial (long-chain) polyphosphates bound and were internalized by mouse macrophages. Long-chain polyphosphates modulated the expression of more than 1,800 LPS/TLR4-regulated genes in macrophages. This interference by polyphosphates included antagonism of hundreds of interferon-regulated genes as explained by reduced interferon-β release and responsiveness, reduced phosphorylation of STAT1 and diminished major histocompatibility complex II. Polyphosphates also prevented the LPS/TLR4-induced induction of NOS2 and M1 polarization. In C57BL/6J mice infected with live E. coli, PPK deficiency or polyphosphate neutralization using recombinant PPX resulted in better survival rates, amplified macrophage recruitment and improved pathogen clearance.
In conclusion, bacterial polyphosphate metabolites interfere with macrophage responses and the manipulation of such interactions could help to restore immunity to infection.
Myeloid cells expressing CK2α modulates resistance to systemic Listeria monocytogenes infection
Authors: Larson Sandy R , Crisler Cris , Lenz Laurel L 
Published: January 2023

Abstract
Reversable phosphorylation is a key regulatory mechanism of nearly all cellular processes. Kinase activity driving phosphorylation has implications in cell growth, survival, development, differentiation, and importantly, immune responses to pathogens. The protein kinase CK2 (casein kinase II) is a ubiquitously expressed and highly conserved haloenzyme with over 500 direct target substrates identified. Because of this promiscuous activity, CK2 is extensively involved in immune cell function, described in both innate and adaptive immune cells. However, functionality of CK2 is not clearly delineated, largely due to the lack of murine models available for study. Engineered mice deficient in either the catalytic α or α′, or the regulatory β subunits are either non-viable or non-fertile. Therefore, we have generated a conditional murine knockout model of CK2α by cre-mediated excision in lysozyme M (LysM) expressing cells. Progeny are viable and fertile, with no deficiencies of immune cell populations. Using this model system, we have identified a role of CK2α in negatively regulating monocyte and neutrophil infiltration and endosome acidification during the bacterial Listeria monocytogenes (Lm) infection, correlating with uncontrolled bacterial replication. Further, decreased inflammatory myeloid cells in infected tissues is a cell-intrinsic effect of CK2α. Future work is aimed to understand the mechanisms by which CK2α negatively regulates the accumulation of immune cells in infected tissues. Additionally, we now have a tool for investigating in vivo effects of CK2α that we can use to extend beyond these initial studies to evaluate its role in other bacterial/viral infections, autoimmunity and even cancer.
Mycobacterium tuberculosis ESX-1 induces eicosanoid mediated neutrophil influx to the lung during Tuberculosis
Authors: SAQIB MOHD , Mishra Bibhuti B 
Published: January 2023

Abstract
Tuberculosis disease (TB) is characterized by excessive inflammation and irreversible tissue damage. To determine the bacterial factors responsible for regulating inflammation, different TB susceptible mouse strains were infected with a pool of transposon mutants of M. tuberculosis (Mtb) that revealed transposon insertions into ESX1 locus impaired the ability to cause disease in susceptible mice and elicit inflammatory response comparable to wild type Mtb. However, the underlying mechanisms of ESX1 mediated tissue damage in vivo is obscure. This study aims at elucidating the mechanism by which Mtb ESX1 regulate tissue damaging inflammation in vivo. Mtb ESX-1 secretion function was required for inducing neutrophil infiltration to the lungs, of TB susceptible mice (iNOS−/−, IL1R1−/−, C3HeB/FeJ). Deletion of ESX-1 from virulent Mtb (ΔESX-1) had little impact on T cells, but significantly reduced the neutrophil/T lymphocyte ratio that favored efficient bacterial control. Co-infection of these mice with wild-type and ΔESX-1, rescued the defective neutrophil influx seen in mice infected with the ΔESX-1 alone. Further, the neutrophil influx elicited by wild type Mtb follows the loss of pro-surfactant C expressing type II alveolar epithelial cells that coincided with the generation of lipoxygenase derived eicosanoids in the lungs; indicating involvement of these chemotactic lipid mediators in initiating neutrophil recruitment in an ESX1 dependent fashion. Taken together, this study provides evidence for the involvement of ESX1 in regulating tissue damaging neutrophilic inflammation during TB, expanding the current understanding of the pleiotropic functions of type VII secretion apparatus.
Vitamin D modulates innate immunity in pulmonary tuberculosis
Authors: Singh Archana , Panda Sudhasini , Tiwari Ambrish , Luthra Kalpana , Sharma Surinder Kumar 
Published: January 2023

Abstract
Vitamin D and vitamin D receptor (VDR) may play role in altering host defence against Mtb via cathelicidin production and regulating nitric oxide (NO) production through inducible nitric oxide synthase (iNOS). 100 active pulmonary tuberculosis (PTB) patients, 75 household contacts and 70 healthy controls were recruited. Serum Vitamin D and NO levels were measured using CLIA and colorimetry based kit respectively. VDR and iNOS mRNA was studied in monocytes using qPCR. Serum VDR, cathelicidin and iNOS levels were measured using ELISA. Lower serum levels of vitamin D was found in PTB group compared to household contacts and healthy controls (P&lt;0.001). VDR (both mRNA and protein) and iNOS mRNA levels were also lower in PTB group compared to household contacts and healthy controls (P&lt;0.005)probably due to decreased levels of vitamin D. However, despite low serum vitamin D, serum cathelicidin levels were higher in PTB group compared to household contacts and healthy controls (p&lt;0.01) which could be attributed to direct trigger via toll like receptors where Mtb is multiplying actively. Serum iNOS and NO levels were higher in PTB group compared to household contacts and healthy controls (p&lt; 0.05), despite decreased iNOS expression which needs further evaluation. Increased VDR and iNOS expression in household contacts compared to both PTB and healthy controls could be due to optimum levels of vitamin D along with some exposure to Mtb as evident by increased levels of cathelicidin and NO compared to healthy controls. Adequate levels of vitamin D, VDR and iNOS that in turn promote production of antimicrobial cathelicidin and NO might have protective role against TB among household contacts and needs further investigation for its therapeutic potential.
Genetic mechanisms contributing to IFNβ regulation of Lyme Arthritis: involvement of the Borrelia burgdorferi arthritis associated locus 1 (Bbaa1)
Authors: Li Jinze , Ma Ying , Paquette Jackie , Teuscher Cory , Weis Janis J 
Published: January 2023

Abstract
Lyme disease shows a spectrum of clinical disorders in patients that are infected with the tick borne spirochete Borrelia burgdorferi. Studies have shown that C3H mice develop severe arthritis while C57BL/6 (B6) mice display mild arthritis when infected with the same isolate of B. burgdorferi. In order to study the role of the host genetics in regulating the severity of Lyme arthritis development, we used forward genetics and identified a genetic region, Bbaa1, that includes the Type I IFN cluster on Chr4. We further developed congenic mice isolating the C3H allele of Bbaa1 onto the background of B6 mice and determined that Bbaa1 regulates arthritis severity by upregulating Type I IFN. Antibody blocking revealed IFNβ, but not IFNα, as the key effector that drives severe Lyme arthritis. Because there are no SNPs between the C3H and B6 IFNβ genes, we hypothesized that other genetic factors in the Bbaa1 locus regulate the differential expression of IFNβ and Lyme arthritis severity. RNA silencing was used to identify candidate genes within Bbaa1 that regulate IFNβ expression in myeloid cells. Cdkn2a was identified as a strong regulator of IFNβ expression. Although Cdkn2a encodes two proteins, p16 and ARF, only the silencing of Arf resulted in the suppression of IFNβ expression. Additionally, the overexpression of ARF led to enhancement of IFNβ. We propose that ARF regulates IFNβ by two pathways involving the proteins p53 and BCL6. Indeed, blocking of BCL6 by a small molecule inhibitor induced IFNβ activation in the joint tissue of B6 mice and enhanced the severity of Lyme arthritis. Future work will focus on the impact of the two pathways on IFNβ expression and arthritis severity in mice.
Pathogenic T cell subset Th17 is regulated by chromatin remodeling dependent manner to switch Th1 phenotype
Authors: Hashimoto Kahoko , Kobayashi Daichi , Oda Toshiki , Hasegawa Akihiro , Nakayama Toshinori 
Published: January 2023

Abstract
In many autoimmune disease such as type 1 diabetes, rheumatoid arthritis, inflammatory bowel disease (IBD), it has been reported that pathogenic Th17 cells exhibit remarkable plasticity to classical Th1 phenotype at the site of autoimmune inflammation. This effecter Th17 cells is also called non-classical Th1 cells because they show the ability to produce IFNγ, similar to conventional Th1 cells, and also continue to produce IL-17 cytokine. The precise mechanism of transcriptional regulation in Th17/IFNγ cells is still not clear yet. By using the splenic naive CD4 T cells, cells were cultured under the Th17 condition including IL-6 and TGFβ for 3 days to 7 days and then cells were added IL-12 to provide Th1-like differentiation condition. In vitro induced Th17 cells were stimulated with anti TCR antibody and assessed the ratio of IL-17 protein production, RORγt and T-bet expression as protein level by flow cytometer. Then, induced Th17 cells were performed to the secondary culture under the several concentration of IL-12 and TGFβ to be able to detect the production of IFNγ, and T-bet. Also we analyzed the specific gene expressions of transcription factor by RT-PCR or quantitative PCR on each day. In this study, we focused on the plasticity of Th17 to Th1 and also compared to the effecter T cell function. We focused on the function of chromatin remodeling in Th17 cells producing both Il-17 and IFNγ by performing ChIP analysis of double producing Th subset and detected that this type plasticity is unique in pathogenic Th17 cell.
CTLA-4-dependent trogocytosis promotes the interactions between Tregs and antigen-presenting cells
Authors: TEKGUC MURAT , WING JAMES BADGER , OSAKI MOTONAO , LONG JIA , SAKAGUCHI SHIMON 
Published: January 2023

Abstract
Regulatory T cells (Tregs) are responsible for the maintenance of immune homeostasis. They express the coinhibitory receptor CTLA-4 in high quantities, whose loss causes severe systemic autoinflammation in both mice and humans. CTLA-4 limits the expression of costimulatory CD80/CD86 proteins by antigen-presenting cells (APCs). However, the details of this mechanism are not fully understood. When we analyzed in vitro cocultures of CD80- or CD86-GFP expressing murine dendritic cells (DCs) with ex vivo sorted Tregs, we found that mutant Tregs, whose CTLA-4 lacked the cytoplasmic tail necessary for its endocytosis function, could still uptake CD80- or CD86-GFP from DCs via CTLA-4-dependent trogocytosis. Moreover, the specific CD80/86 uptake by CTLA-4, also facilitated the indirect transfer of membrane lipid particles and other surface proteins. Both flow cytometry and confocal microscopy revealed that immune synapse formation between Tregs and APCs was enhanced depending on CTLA-4 expression by Tregs. Confocal microscopy also demonstrated that uptaken CD80/CD86-GFP proteins colocalized with LFA-1 capping, an indicator of stable immune synapses. Furthermore, in vivo delivery of CTLA-4 WT and knockout (KO) Tregs into CD45.1+RAG2 KO mice showed that transfer of congenic CD45.1 protein from host cells to donor Tregs was significantly diminished in CTLA-4 KO Tregs compared to WT Tregs. The tail portion of CTLA-4 was also found to be dispensable for CD80/86 downregulation and suppressive functions of Tregs. In conclusion, CTLA-4-promoted immune synapse formation provides a stable platform between Tregs and APCs, which results in both depletion of CD80/CD86 via trogocytosis and indirect uptake of other surface proteins from APCs.
Differential transcriptional response of plasmacytoid dendritic cells in mice with lupus
Authors: Baeza Carlos Wong , Mora Albany Reséndiz , Pensado Monserrat Figueroa , Aveleida Giovanna Barrera , Ángeles Karen Sánchez , Cortés Cinthya Portas , Gómez Eréndira Molina , Saldívar Carla Landa , Baeza Isabel Wong , Baeza Isabel 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is an autoimmune, chronic and multi-systemic disease. SLE patients present high levels of type I IFN, which is mainly produced by plasmacytoid dendritic cells (pDCs). In this work, we analyzed the gene expression of pDCs from mice with lupus induced by non-bilayer phospholipid arrangements (NPA). An increase in type I (alpha and beta) IFN was detected in the serum of mice immunized with NPA-bearing liposomes, compared with mice that received smooth liposomes or saline solution. Furthermore, 20 days after the beginning of the administration of the NPA-bearing liposomes, a statistically significant increase in the absolute number of pDCs was found in the spleen and in the mesenteric lymph nodes of mice administered with these liposomes, compared with mice administered with saline solution or with smooth liposomes. When we compared the gene expression of pDCs from mice with lupus with pDCs from mice that received smooth liposomes, 9 of the 22 over-expressed genes corresponded to the innate response and 13 to the adaptive response, while 35 of the 47 under-expressed genes corresponded to the innate response and 12 to the adaptive response. After analyzing the gene expression patterns, we propose that there may be three subsets of pDCs in the spleens of mice with lupus: one that produces high levels of type I IFN; a second one that produces pro-inflammatory and immuno-regulatory cytokines and thus participates in both innate and adaptive responses; and a third one that recognizes NPA-bearing liposomes and processes and presents them through the CD1 molecule.
The secretory pathway-resident SNARE Sec22b regulates nitric oxide and cytokine production in dendritic cells
Authors: Descoteaux Albert , Dion Renaud , Descoteaux Julien , Fabié Aymeric , Stäger Simona , Duque Guillermo Arango 
Published: January 2023

Abstract
Soluble NSF attachment receptor (SNARE) proteins regulate the vesicle transport machinery in phagocytic cells. Within the secretory pathway, Sec22b is an ER-Golgi intermediate compartment (ERGIC)-resident SNARE that controls phagosome maturation and function in macrophages and dendritic cells. The ERGIC controls the release of cytokines and may impact that of nitric oxide (NO), the latter of which is synthesized by the Golgi-active inducible nitric oxide synthase (iNOS). Whether the ERGIC SNARE Sec22b controls NO and cytokine secretion, is unknown. Using bone marrow-derived dendritic cells (BMDC), we demonstrated that iNOS colocalizes with ERGIC/Golgi markers, notably Sec22b and its partner syntaxin-5 (Stx5), in the cytoplasm and at the phagosome. Pharmacological blockade of the secretory pathway hindered NO and cytokine release, and inhibited NFκB translocation to the nucleus. Importantly, RNAi-mediated silencing of Sec22b revealed that NO and cytokine production were abrogated at the protein and mRNA levels. This correlated with deregulated mitogen-activated protein kinase (MAPK) signalling and reduced nuclear translocation of NFκB. Collectively, our data unveiled a novel function for the ERGIC, and its resident SNARE Sec22b, in the release of inflammatory mediators.
PKC signaling plays an essential role in dendritic cell differentiation
Authors: Chavel Colin A , Lee Kelvin P 
Published: January 2023

Abstract
Differentiation of progenitor cells into dendritic cells (DC) is a complicated process that requires multiple signaling cascades. Tumor-derived factors can inhibit this process in part through repression of protein kinase C isoform β (PKCβ) gene expression mediated by STAT3 activity. How PKC signaling exerts this differentiation response has not been fully characterized. We found PKC required both the NFκB and ERK pathways to induce DC differentiation, as examined through T-cell proliferation assays where either pathway was blocked by chemical inhibition. We uncovered novel interactions for PKC dependent activation of both the NFκB and ERK during DC differentiation. The canonical NFκB pathway was activated by PKCβ through the CARD11 complex via novel mechanisms, including bypassing the established role for both TAK1 and MALT1 in downstream IκB phosphorylation and a newly characterized mechanism for IKK activation. The non-canonical NFκB pathway was also activated in a PKCβ dependent manner outside of the typical ubiquitin dependent degradation cascades activated by TRAF complexes. The ERK pathway was activated through cRaf by multiple PKCs in part through inhibition of RKIP activity. Multiple ERK targets were activated, including RSK and Jun-Fos complexes, as well as an ERK dependent Foxo3a depletion not yet characterized in DC differentiation. Furthermore, we found crosstalk mechanisms between the three pathways that resulted in early or late activation of the pathways and was seen at both the gene and protein levels. Taken together, these results indicate PKC as an important signaling node for DC differentiation.
miR-146a −/− deficiency in the DC compartment results in increased effector phenotype CD4+ and CD8+ T cells in response to LCMV infection
Authors: Flynn Patrick A , Lind Evan 
Published: January 2023

Abstract
miR-146a is a micro-RNA that acts on transcripts upregulated in response to TLR induced signaling. Activation of TLRs results in maturation and survival of dendritic cells (DCs) to prime T cells. Interestingly, little is known how miR-146a regulates DC phenotype/function and whether it has direct effects on the regulation of T cell function. To probe this question, we utilized a miR-146a conditional knockout mouse model to study both DCs and lymphocytes. Our in vitro studies of DCs show that miR-146a KO BMDCs have increased uptake and processing of DQ-OVA, suggesting increased antigen uptake and processing. Additionally, miR-146a KO BMDCs had increased gMFIs of both CD80 and CD86, suggesting increased co-stimulatory potential. To test for differences in T cell activation and priming, we used the LCMV infection model to analyze T cell responses in the context of miR-146a KO DCs in vivo (miR-146a cKO). We found that miR-146a cKO mice showed increased cross-presenting XCR1+ cDC1s and XCR1+ CD103+ DCs compared to miR-146a competent mice. T cells from miR-146a cKO mice had increased CD44 and PD-1 double-positive effector phenotype. Naïve miR-146a cKO mice had increased Tcm-like CD4+ T cells. Naïve miR-146a cKO mice had increased Tcm-like and Tscm-like CD8+ T cells. Our data indicate that the lack of miR-146a results in a skewing of DC subsets to favor XCR1+ DC1 and XCR1+ CD103+ populations who are known to excel at cross-presentation and migrate through tissues. Our data also suggests that miR-146a cKO mice activate more effector phenotype T cells and increase the proportion of Tcm and Tscm-like T cell populations which result in more robust adaptive T cell responses.
Essential role of NPAT in regulating T cell development and function
Authors: Cui Zejin , Zhao Fan , Lu Linrong 
Published: January 2023

Abstract
The T-cell development process is precisely controlled at different stages. For example, cell proliferation during T cell development is limited to immature double negative (DN) thymocytes. Here we want to investigate the mechanism of stage-specific regulation of cell proliferation in this process. Nuclear protein ataxia-telangiectasia (NPAT) is an in vivo substrate of cyclin E-Cdk2 kinase and plays a critical role in coordinating histone gene expression during the G1/S-phase transition and to ensure S-phase entry, thus play a regulatory role in determining the proliferation level of different cells.
By the using of conditional gene deletion in thymocytes, we found that ablation of NPAT resulted in severe thymocyte development defect including reduced thymocyte number and developmental arrest from ISP to DP, indicating a critical role of NPAT in thymocyte development. We have also constructed a mouse strain that ablates NPAT expression in peripheral mature T cells. NPAT deficiency in peripheral T cells only resulted in a mild decrease in the number of CD4 and CD8 cells, while maintained normal T-cell homeostasis in peripheral lymphoid organs. However, the proliferative capacity of naïve CD4 or CD8 T cells in NPAT cKO mice were impaired upon activation, which correlated with less severe symptoms in experimental autoimmune encephalomyelitis (EAE) disease upon antigen-immunization. We are currently exploring the regulation mechanism of NPAT function in different cell stages and will test the possibility of targeting NPAT in treat autoimmune diseases.