bluuebunny/crossref_metadata_2025_split huggingface dataset:
HSV-2 infection induces TLRs and NF-κB-dependent cytokines in human cervical epithelial cells (133.41)
Authors: Li Hui , Li Xiaoling , Wei Yun , Tan Yun , Wu Xinxing 
Published: January 2023

Abstract
Herpes Simplex Virus type 2 (HSV-2) is one of the most common sexually transmitted pathogen worldwide. The host immune response induced by viral infection is cell-type specific. Little is known about the innate immune response to this virus in its natural host cells. In this study, we established an in vitro HSV-2 infection model with human cervical epithelial (HCE) cells. The viral infection was sufficient to induce expression of Toll-like receptors (TLRs), and western blot and reporter assays suggest that HSV-2 infection leads to dramatic activation of the NF-κB signaling pathway. The two critical cytokines,IL-6 and IFN-beta,were also induced after HSV-2 infection and maintained high levels during the course of experiment. The presence of NF-κB inhibitor, Isohelenin significantly blocked the production of both IL-6 and IFN-beta in response to HSV-2. Thus, our data provide direct evidence that the activation of NF-κB is required for the production of both IL-6 and IFN-beta induced by HSV-2 in HCE cells. Taken together, our results suggest the potential contributions of TLRs and a critical role of NF-κB in the innate immune response to HSV-2 infection in its natural host cells.
This work was supported by grants from the National Natural Science Foundation of China (No. 30500465,30810103052)
Functional Modulation of Dendritic Cells and Macrophages by Japanese Encephalitis Virus through MyD88 Adaptor Molecule-Dependent and Independent Pathways (133.42)
Authors: Eo Seong Kug , Han Young Woo , Rahman M. M. , Kim Seon Ju , Kim Seong Bum 
Published: January 2023

Abstract
Dendritic cells (DCs) are potent initiator of T cell-mediated immunity and undergo maturation during viral infections. However, few reports describing the interactions of DCs with Japanese encephalitis virus (JEV), which remains the most cause of acute and epidemic viral encephalitis, are available. In this study, we investigated the interaction of JEV with DCs and macrophages. JEV replicated its viral RNA in both cells with different efficiency, and we found that JEV infection of macrophages followed classical activation pathway of upregulation of costimulatory molecules and proinflammatory cytokine production (IL-6, TNF-¥á, and IL-12). On the contrary, JEV-infected DCs failed to upregulate costimulatory molecules (CD40, CD80, CD86, and MHC class II). Of more interest, along with production of proinflammatory cytokines, DCs infected by JEV released anti-inflammatory cytokine IL-10, which was not detected in macrophages. Moreover, signaling through MyD88 molecule, a pan-adaptor molecule of Toll-like receptors (TLRs), and p38 MAPK in JEV-infected DCs was found to have roles in production of cytokines and subversion of primary CD4+ and CD8+ T cell responses. We also found that IL-10 released from JEV-infected DCs mediated the reduced priming of CD8+ T cells but not CD4+ T cells. Our data suggest that JEV induce functional impairment of DCs through MyD88-dependent and independent pathways, which subsequently lead to poor CD4+ and CD8+ T cell responses, resulting in boosting viral survival and dissemination in the body.
Vesicular stomatitis virus infects primary murine microglia and astocytes and induces replication-dependent inflammatory cytokine production (133.47)
Authors: Marriott Ian , Chauhan Vinita S , Sterka David G , Grdzelishvili Valery Z 
Published: January 2023

Abstract
Vesicular stomatitis virus (VSV) is a rhabdovirus that closely resembles its deadly cousin, rabies virus. Intranasal inoculation of mice with VSV leads to infection of the central nervous system (CNS) and an acute encephalitis that results in a high rate of mortality similar to that observed during human rabies infections. Importantly, this encephalitis is accompanied by the rapid activation and proliferation of resident brain cells including microglia and astrocytes, suggesting a role for such cells in the initiation and/or progression of CNS inflammation. Our recent study showed that these glial cells express RIG-I and MDA5, cytosolic receptors for replicating RNA virus motifs. However, it is presently unclear whether microglia and astrocytes can harbor replicating VSV, or if such replication is required for the inflammatory immune responses of these resident CNS cells. In this study we demonstrate that primary murine microglia and astrocytes are permissive for VSV infection and replication. Importantly, active replication appears to be required for robust inflammatory mediator production as such responses were markedly lower in cells exposed to either inactivated or replication defective viral particles. Interestingly, this effect is not limited to neurotropic Mononegaviruses as the paramyxovirus, Sendai virus, can similarly infect glial cells and elicit inflammatory immune responses. Finally, we have confirmed that in vivo intranasal VSV administration results in viral infection and replication within mouse glial cells in situ. This study suggests that viral replication within resident glial cells could play a key role in the development of lethal CNS inflammation following neurotropic Mononegavirus infection.
NADPH oxidase-derived ROS mediates mast cell degranulation induced by secretory products secreted by Trichomonas vaginalis (133.5)
Authors: Min Deulle , Shin Myeong Heon 
Published: January 2023

Abstract
Trichomonas vaginalis is known to be a sexually transmitted protozoan parasite causing vaginitis in women. During infection in vivo, secretory products by T. vaginalis can disrupt the epithelial junctions so that these products gain access to, and alter the function of, underlying cells of the innate immune system. Mast cells are known to play a key role in mucosal immune responses. In this study, we investigated whether NADPH oxidase-derived reactive oxygen species (ROS) are closely associated with mast cell degranulation by Trichomonas-secreted products. When Trichomonas-secreted products were added to human mast cells (HMC-1 cells) in a transwell system (pore size 0.2 um), it caused extensive degranulation of mast cells, which was confirmed by ultrastructural change, increased expression of CD63 at the cell surface, and liberation of granular proteins such as histamine. In addition, mast cells were found to harbor different isoforms of NADPH oxidase. Stimulation with Trichomonas-secreted products induced intracellular accumulation of ROS in mast cells. Next, we investigated whether NADPH oxidase-derived ROS are one of the main signaling molecules responsible for Trichomonas-triggered mast cell degranulation. Pretreatment of cells with NADPH oxidase inhibitors such as DPI or apocyanin strongly inhibited ROS generation and mast cell degranulation induced by Trichomonas-secreted products. These results suggest that secretory products by Trichomonas vaginalis contain signaling molecules to trigger activation of NADPH oxidase in mast cells, which triggers degranulation leading to initiating and regulating mucosal inflammation.
HCV core/gC1q Receptor ligation transduce a signal across the plasma membrane possibly by beta1 integrin complex. (133.51)
Authors: Tosello-Trampont Annie , Tacke Robert , Hahn Young S. 
Published: January 2023

Abstract
Hepatitis C virus (HCV) infection in humans is associated with viral persistence, which leads to the development of liver diseases. HCV core protein has been reported to be detectable in bloodstream of HCV-infected patients, and contains the immunomodulatory function. We have previously demonstrated that the interaction of HCV core with the complement receptor gC1qR inhibits the inflammatory response, particularly the TLR4-mediated production of IL-12 in monocytes. The inhibition of IL-12 production depends on the activation of the PI-3 kinase pathway. However, it is still unknown how this receptor anchors at the cell surface to transduce intracellular signaling since it has no transmembrane or GPI domain. To determine the protein(s) involved in the gC1qR complex responsible for anchoring gC1qR, we examined the association of gC1qR with β1 integrin (a putative transmembrane partner). Here, we show by performing GST-gC1qR pull down assays on human cell lysats, that β1 integrin specifically interacts with gC1qR. Moreover, we examine the cell surface localization of gC1qR and β1 integrin by immunofluorescence microscopy after THP-1stimulation by HCV core. Our data demonstrate that HCV core enhance the formation of gC1qR/β1 integrin complexes on HCV core-treated monocytes. These results suggest that β1 integrin may play the role of anchoring protein to gC1qR on the cell surface.
NIH grant RO1AI057591
T cell receptors mediate docking of cytotoxic granules at the immunological synapse (133.7)
Authors: Qu Bin , Pattu Varsha , Junker Christian , Becherer Ute , Matti Ulf , Rieger Heiko , Schwarz Eva C , Rettig Jens 
Published: January 2023

Abstract
TCR enrichment and cytotoxic granules accumulation at the immunological synapse are necessary for cytotoxic T lymphocyte-mediated killing. Using immunocytochemistry, confocal microscopy, 4D fast live cell imaging and total internal reflection fluorescence microscopy, we analyzed the localization and transport of recycling TCR vesicles and cytotoxic granules in parallel. We found that recycling TCR vesicles are very often connected with cytotoxic granules. Both are co-transported in a paired manner to the IS where recycling TCR enhance the docking and release of cytotoxic granules. Quantitative analysis of vesicluar transport revealed that pairing enables CTL to kill within a few minutes after synapse formation. These data not only highlight an important function of recycling TCR, but also show that pairing of different vesicles is an unexpected mechanism to target vesicles to a defined area within CTL.
This project was funded by the Deutsche Forschungsgemeinschaft (Graduate Research School 1326, DFG grant RE 1092/6-1 to J.R and M.H.) and a HOMFOR grant from the Medical Faculty (to J.R. and M.H).
CD57 defines a functionally unique subset of NK cells in humans (134.1)
Authors: Lopez-Verges Sandra , Milush Jeffrey M , Pandey Suchitra , Norris Philip , Nixon Douglas , Lanier Lewis L 
Published: January 2023

Abstract
CD57 is a marker of terminal differentiation on human CD8+ T cells and possibly on human NK cells. Newborn and fetal NK cells do not express CD57, however the frequency of CD57-bearing NK cells increases with age. Utilizing PBMC obtained from healthy donors we are assessing the phenotypic and functional differences between CD57+ and CD57neg NK cells. CD57+ NK cells express different activation markers and a distinct repertoire of NK receptors suggestive of a more mature phenotype. Preliminary data also indicate that there is a higher frequency of IFNγ CD57+ NK cells compare to CD57neg NK cells when stimulated through their activating receptors, CD57+ NK cells are less sensitive to activation-induced apoptosis, and interestingly, proliferate less when co-cultured with target cells and/or cytokines. The combination of a more activated phenotype and a decreased capacity to proliferate suggests CD57+ NK cells are terminally differentiated. Studies are underway to assess the phenotypic and functional differences of CD57+ and CD57neg NK cells during infection with HIV and other viruses.
Funding: NIH. SLV is a Cancer Research Institute/Irvington Institute Fellow. LLL is an American Cancer Society Professor.
Myosin IIA associates with NK cell lytic granules to enable their interaction with F-actin and function at the immunological synapse (134.13)
Authors: Sanborn Keri B , Rak Gregory D , Maru Saumya Y , Difeo Analisa , Martignetti John A , Favier Remi , Banerjee Pinaki P , Orange Jordan S 
Published: January 2023

Abstract
NK cell cytotoxicity requires the formation of an actin-rich immunological synapse (IS) with a target cell and the polarization of perforin-containing lytic granules toward the IS. Following the polarization of lytic granules, they traverse through the actin-rich IS to join the NK cell membrane in order for the directed secretion of their contents to occur. We examined the role of myosin IIA in facilitating this pre-final step in lytic NK cell IS function. Lytic granules in and derived from a human NK cell line, or ex vivo human NK cells, were constitutively associated with myosin IIA. When isolated using density gradients, myosin IIA-associated NK cell lytic granules directly bound to F-actin and the interaction was sensitive to the presence of ATP under conditions of flow. In NK cells from patients with a nonsense mutation in myosin IIA, NK cell cytotoxicity, lytic granule penetration into F-actin at the IS, and interaction of isolated granules with F-actin were all decreased. Similarly, inhibition of myosin function also diminished the penetration of lytic granules into F-actin at the IS, as well as their final approach to and their dynamics at the IS. Thus, NK cell lytic granule-associated myosin IIA enables their interaction with actin and final transit through the actin-rich IS to the synaptic membrane, and can be defective in the context of naturally occurring human myosin IIA mutation.
Supported by NIH grants R01 AI067946 (JSO) and T32-GM07229 (KBS).
TNF driven activation of invariant NKT cells favours regulation of combined gut and joint inflammation. (134.22)
Authors: Elewaut Dirk , Jacques Peggy , Van Beneden Katrien , Apostolaki Maria , Lambrecht Bart , Kollias George 
Published: January 2023

Abstract
Invariant natural killer T (iNKT) cells are a subset of glycolipid reactive T cells, implicated in the protection against microbial pathogens and immune regulation. While the pathways leading to iNKT cell activation in the context of infectious diseases have been uncovered, much less is known how iNKT cells become activated in the context of chronic inflammatory diseases. Here, we unveil a novel mode of iNKT cell activation which occurs during chronic TNF driven inflammation through generation of inflammatory dendritic cells. This in turn, leads to a spontaneous activation of iNKT cells by promoting CD1d mediated presentation of endogenous glycolipids. This mode of activation is capable of attenuating combined Crohn's like ileitis and joint inflammation occurring in TNFΔARE mice, and thus may represent a natural counter regulatory mechanism to dampen TNF driven inflammation.
NK cell-mediated, Kupffer cell-dependent inhibition of liver injury attending biliary obstruction and cholestasis (134.25)
Authors: Cheng Chao-Wen , Wintermeyer Philip , Gregory Stephen H. 
Published: January 2023

Abstract
Natural killer (NK) cells are innate immune effector cells that participate in the first line of host defenses. In addition to expressing cytotoxic activity, NK cells produce cytokines and engage other cell populations to modulate adaptive immunity. Compared to other organs, the liver contains a large percentage of NK cells suggesting that NK cells play a unique role in the liver; the function of hepatic NK cells, however, is not well understood. Here we examined the contribution of NK cells to the tissue injury that occurs following bile duct ligation (BDL) in a mouse model of biliary obstruction. The number of activated hepatic NK cells was increased markedly at 18 hours following BDL. NK cells activation was suppressed in mice rendered Kupffer cell-depleted prior to ligation. Higher plasma alanine aminotransferase activity, an indictor of increased liver injury, occurred in NK cell-depleted mice. In contrast, IL-6 mRNA expression and protein production in the livers of BDL, NK cell-depleted mice were suppressed. Purified Kupffer cells derived from NK cell-depleted mice following BDL produced less IL-6 in culture than did Kupffer cells derived from non-depleted animals. Treatment with recombinant murine IL-6 reduced liver injury in BDL, NK cell-depleted mice. Taken together, these data suggest that hepatic NK cells suppress cholestatic liver injury by stimulating IL-6 production by Kupffer cells.
PI3-Kinase Links NKG2D Signaling to a CrkL Pathway Involved in Natural Killer Cell Adhesion, Polarity, and Granule Secretion (134.3)
Authors: Segovis Colin M , Schoon Renee A , Dick Christopher J , Nacusi Lucas P , Leibson Paul J , Billadeau Daniel D 
Published: January 2023

Abstract
The NK cell activating receptor NKG2D plays a critical role in the destruction of malignant cells, but many of the cell-signaling mechanisms governing NKG2D-mediated cellular cytotoxicity are unknown. We have identified an NKG2D-mediated signaling pathway that governs both conjugate formation and cytotoxic granule polarization. We demonstrate that an interaction between the regulatory subunit of PI3K, p85, and the adaptor protein CrkL is required for efficient NKG2D-mediated cellular cytotoxicity. We show decreased NK cell - target cell conjugate formation in NK cells treated with PI3K inhibitors or depleted of CrkL. Independent of adhesion, we find that microtubule organization center polarization toward target cells expressing the NKG2D ligand MICA or toward anti-NKG2D coated beads is impaired in the absence of CrkL. Antibody-stimulated granule release is also impaired in NK cells depleted of CrkL. Furthermore, our data indicate that the small Ras-family GTPase Rap1 is required for NK cell killing and is activated downstream of NKG2D engagement in a PI3K- and CrkL-dependent manner. Taken together, our data identify an NKG2D-activated signaling pathway that collectively orchestrates NK cell adhesion, cell polarization and granule release.
A simple new method for negative enrichment of monocytes from mouse blood and bone marrow (134.39)
Authors: Zaborowska Catherine , Fairhurst Maureen , Fairey Mark , Fogg Darin , Thomas Terry 
Published: January 2023

Abstract
The mononuclear phagocyte system is comprised of tissue macrophages, dendritic cells, blood monocytes and their bone marrow (BM) progenitors. Transgenic mouse models have recently provided insight into the biology of monocytes in vivo. However a general method for isolating monocytes from mice is needed to better study their functions.
We describe a rapid and simple method for the enrichment of monocytes from mouse BM and peripheral blood that does not require a density gradient and yields high purity and recovery. BM was harvested from femurs and tibia by crushing the bones. Blood was collected with heparin and red blood cells were removed by ammonium chloride lysis. The monocytes were then enriched using immunomagnetic, column-free negative selection (EasySep®). Briefly, unwanted cells were specifically labeled with dextran-coated magnetic particles using a cocktail of bispecific tetrameric antibody complexes. The sample was placed in a magnet and the supernatant containing unlabeled monocytes was collected. The separation procedure can be automated with a pipetting robot (RoboSep®). Purity of CD11b+Ly6G- cells as assessed by flow cytometry ranged from 80-93% for BM and 92-98% for blood with recovery of 46 ±11 % (n=38) and 25 ±10 % (n=20) respectively. This protocol will provide easy access to monocytes, enriched from peripheral blood and BM, for further studies of immune and inflammatory responses.
Effect of glycyrrhizin (GL) on β-defensin production in tissues surrounding the burn area of thermally injured mice (134.44)
Authors: Yoshida Shohei , Shigematsu Kenji , Asai Akira , Kobayashi Makiko , Herndon David N , Suzuki Fujio 
Published: January 2023

Abstract
The decreased production of antimicrobial peptides in tissues surrounding the burn area has been demonstrated in patients and animals with severe burn injury. Previously, sepsis stemming from burn wound infections with Pseudomonas aeruginosa was effectively controlled in burn mice treated with GL. Therefore, in this study, effects of GL on β-defensin production in tissues surrounding the burn area were examined. BALB/c mice with 25% TBSA of 3rd degree flame burn were treated i.p. with GL (Minophagen Pharmaceutical Co., Ltd., Tokyo, Japan) at various schedules after burn injury. Eight punch biopsies (8-mm diameter) per mouse were obtained from skin tissues surrounding the burn area of these mice 24 hrs after burn injury. The homogenates of 8 biopsies were assayed for MBD-1 and MBD-3 by ELISA. In the results, normal mouse skin homogenates consistently contained 0.8 ± 0.4 μg/ml of MBD-1 and 6.9 ± 1.8 μg/ml of MBD-3. However, MBD-1 and MBD-3 were not detected in homogenates of tissues surrounding the burn area. When 10 mg/kg of GL was administrated i.p. to mice 2 and 12 hrs after burn injury, the amounts of MBD-1 and MBD-3 in the homogenates of tissues surrounding the burn area recovered to the level shown in normal mouse tissue specimens. MBD production in skin tissues of normal mice was not influenced by GL treatment. These results indicate that, after treatment with GL, the decreased production of β-defensins in tissues surrounding the burn area is improved in thermally injured mice. This study was supported by SHC NA #8610.
A critical role for calcium flux in phagocytes for apoptotic cell engulfment and the anti-inflammatory response (134.52)
Authors: Gronski Matthew Adam , Kinchen Jason M , Juncadella Ignacio , Franc Nathalie , Ravichandran Kodi S 
Published: January 2023

Abstract
Calcium plays a key role as a second messenger in many cell types, but a role for calcium signaling during apoptotic cell removal is unclear. Using studies in cell lines and in the context of a whole organism, we demonstrate that apoptotic cell recognition induces both an acute and sustained calcium flux within phagocytes and that such calcium flux is essential for engulfment. Furthermore, we provide evidence that both the release of calcium from the endoplasmic reticulum and the entry of extracellular calcium via CRAC channels into the phagocytes are important during engulfment. Moreover, knockdown in C. elegans of stim-1 and jph-1, two genes linked to the entry of extracellular calcium into cells, led to increased persistence of apoptotic cells in the nematode. Loss of these genes appeared to affect early signaling events, leading to decreased enrichment of actin adjacent to the apoptotic cell during corpse removal. We also show that calcium is crucial for the anti-inflammatory TGF-β response seen in phagocytes during engulfment of apoptotic cells. Taken together, these data point to a previously unappreciated and evolutionarily conserved role for calcium flux at two distinguishable steps: formation of the phagocytic cup and internalization of the apoptotic cell, and the anti-inflammatory signaling induced in phagocytes by contact with apoptotic cells.