bluuebunny/crossref_metadata_2025_split huggingface dataset:
The advancing age affects the frequency, functions and antibody repertoire of human B-1 cells, which secrete anti-tumor antibodies
Authors: Hernandez Ana Maria , Rodriguez-Zhurbenko Nely , Quach Tam D. , Hopkins Thomas J. , Rothstein Thomas L. 
Published: January 2023

Abstract
N-glycolylated (NeuGc) gangliosides are not naturally expressed in human tissues but are overexpressed in several tumors and have immunosuppressive capacity. The present work shows the existence of natural cytotoxic antibodies against NeuGcGM3, whose levels decrease with age. Furthermore, this reactivity was not detected in the sera of non-small cell lung cancer patients that matched in sex and age the assessed healthy donors. Aging decreases the efficiency of immune functions, which could be associated with an increased incidence of cancer. In order to identify and study the effect of aging on the B cell population involved in the response against NeuGcGM3 naïve, memory, plasma and B-1 cells were purified from healthy human donors PBMC and stimulated in vitro to produce immunoglobulins. Only B-1 cell supernatants recognized tumor cells expressing this ganglioside. Interestingly, B-1 cells from NSCLC patients had a reduced activation capacity. The percentage of B-1 cells and their capacity to spontaneously secrete IgM decreased with age in healthy donors. In fact, expression levels of Xbp1 and Blimp1 were significantly lower, while those of Pax5 were significantly higher, in elderly donors. Elderly donors showed a B-1 antibody repertoire with higher number of repeated clones and therefore a reduced variability in comparison with young individuals. It was interesting to observe in aged individuals an increased frequency of VH3-23 and VH4-34 usage, genes which have been linked to autoimmune diseases and hematopoietic malignancies, while there is a significant reduction of clones expressing the sequence VH3-33, frequently found on antibodies against PC, oxidized LDL, apoptotic cells and induced after vaccination with Pneumavax23.
Splenic microenvironment is important in the survival and growth of Chronic Lymphocytic Leukemia in mice
Authors: McKenna Mary Kathryn , Noothi Sunil K , Alhakeem Sara Samir , Byrd John C , Muthusamy Natarajan , Rangnekar Vivek , Bondada Subbarao 
Published: January 2023

Abstract
Chronic Lymphocytic Leukemia (CLL), the most common adult leukemia in the western world, is characterized by accumulation of clonally expanded CD5+CD19+ B lymphocytes in the peripheral blood and secondary lymphoid organs. CLL cell survival is dependent on B cell receptor signaling and cross-talk within the leukemia microenvironment. CLL presents with enlargement of the spleen, and lymph nodes due to accumulation of long-lived lymphocytes with impaired apoptotic mechanisms. The Eμ-Tcl1 mice, with a B cell specific overexpression of the oncogene, T cell Leukemia 1 (Tcl1), serve as an excellent model to study CLL as they develop a CLL like disease by 9–13 months of age. They exhibit splenomegaly, enlarged lymph nodes and a high peripheral blood white count. Adoptive transfer of primary CD5+CD19+ CLL cells from the Eμ-Tcl1 mice into syngeneic recipients leads to rapid disease development. Ultrasonography of recipient mice showed a dramatic enlargement of the spleen, which preceded detection of CLL cells in the blood. Unlike various genetic modifications, which can only delay the onset of CLL, splenectomy in these models, cures or significantly delays disease development from 30–35 days to 250–270 days. Interestingly, splenectomy did not delay CLL development as significantly in animals deficient for prostate apoptotic response-4 (Par-4) compared to C57BL/6 wild type mice. Par-4 is a known tumor suppressor protein that can be secreted and induce apoptosis selectively in cancer cells but not in normal cells. Par-4 appears to regulate a specific microenvironment required for CLL growth. Presently we are investigating the role of Par-4 in the microenvironment and the cell types that are critical for CLL growth within the splenic niche.
Modulation of signaling preference of NK cells by different forms of NKG2D ligands and its implications
Authors: Dhar Payal , Basher Fahmin , Zhang Jinyu , Wu Jennifer 
Published: January 2023

Abstract
The significance of ligand-induced activation of NK cell activating receptor NKG2D has been well established in controlling tumor growth in various experimental animal models. Amongst different types of NKG2D ligands, MHC I chain related molecule MICA/B is expressed most prevalently in various human cancers. In particular, proteolytically shed form of MICA/B (sMIC) has been shown to suppress the immune system and found to correlate with advanced disease stages in cancer patients. Previous studies in our lab using two humanized bi-transgenic mice, one expressing native ligand (TRAMP/MICB) and the other expressing the shedding-resistant mutant (TRAMP/MICB.A2) revealed opposite roles of these ligands in regulating tumor immunity. MICB expressing mice exhibited increased tumor progression, whereas MICB.A2 expressing mice had tumor free survival. These findings raised the intriguing question of why do these structurally similar forms of MIC have vastly different effects on tumor immunity. To address this question, we investigated the signaling events and functional outcomes in NK cells upon stimulation by the two forms of ligands. In vitro co-culture studies of NK cells with tumor cell lines expressing sMICB and MICB.A2 revealed elevated pro-inflammatory and immunosuppressive cytokine production by NK cells upon stimulation with sMICB. In contrast, NK cells stimulated with MICB.A2 displayed enhanced expression of cytotoxicity mediators and signaling molecules of cytotoxicity pathway. This suggests that signaling through sMICB may polarize the NKG2D signaling pathways with preferential activation of inflammatory cytokine pathways. Our data has uncovered a new potential mechanism whereby sMIC promotes tumor progression.
The role of TFEB in tumor-associated macrophages
Authors: Hodge Johnie O’Neil , Fang Liang , Fan Daping 
Published: January 2023

Abstract
Alternatively activated (M2-like) tumor-associated macrophages (TAMs) have been shown to promote immune suppression, invasion, angiogenesis, the establishment of distant metastases, and maintain the breast cancer stem cell population. However, the mechanisms governing TAM polarization are not fully understood. Transcription factor EB (TFEB) has been recently investigated due to its role as the master regulator of autophagy, but little is known regarding its role in governing macrophage activation or other immune response pathways. We show here that TFEB transcription and activation are reduced in TAMs, and TGF-β-induced activation of ERK signaling is the driving force behind this phenomenon. siRNA knockdown of TFEB in macrophages significantly enhances their alternative activation. Co-inoculation of these macrophages with breast tumor cells results in an increase of both tumor growth and infiltration of M2-like TAMs. Further mechanistic studies reveal that TFEB knockdown regulates M2 polarization by decreasing activation of SOCS3, which acts as an inhibitor of STAT3. These findings led us to further investigate whether increasing TFEB activation in TAMs could decrease their alternative activation and their tumor promoting properties. We found that activation of macrophage TFEB suppresses alternative activation, and introduction of these macrophages at the time of tumor initiation decreases tumor growth in multiple murine models. Therefore, our data demonstrate that the downregulation of TFEB is an integral part of immune editing in the tumor microenvironment, and provides a rationale for a new cancer treatment strategy involving the modulation of TAM polarization through activating TFEB.
Analysis of granulocytes and macrophages infiltrating a primary tumor site within the first 72 hours
Authors: Kurt Robert A , Zhang Chenyu , Newsom-Stewart Catherine M , Ptasinski Anna M , Greis William H 
Published: January 2023

Abstract
While a great deal of work has been reported on innate and adaptive white blood cells infiltrating solid tumors, as well as the pre-metastatic niche, there is a paucity of information concerning the early innate immune response to primary tumors. This is particularly evident in the very early stages; before a solid tumor is palpable. Here we used a gelatin sponge model to investigate white blood cells infiltrating a tumor site 12–72 hours following injection. Initial studies focused on identifying the total number of infiltrating cells as well as the number of lymphocytes, macrophages and granulocytes. While 12 hours after injection there was no difference in the total number of cells infiltrating the tumor site versus a control site injected with saline, at 24 and 48 hours there was a small, yet significant increase in the number of cells at the tumor site. The majority of the cells to arrive at the tumor and saline injected sites within the first 12 hours were Gr1+F4/80+ (60%), and this population of cells decreased to 36% by 72 hours at both sites. These data contrasted with the F4/80+Gr1− cells which increased over time from about 10% at 12 hours to 25% at 72 hours. Moreover, two significant differences were noted between the tumor and saline sites. At 24 hours there were significantly more F4/80+Gr1-cells at the tumor site, and the number of Gr1+F4/80− cells remained higher at the tumor site over time. These data indicate early macrophage recruitment and neutrophil retention at the primary tumor site. Gene expression analysis by quantitative RT-PCR is underway to investigate additional differences in these cell populations.
Control of gut bacteria-associated sepsis by an inhibitor of group 2 innate lymphoid cells in severely burned mice
Authors: Ito Ichiaki , Kobayashi Makiko , Finnerty Celeste C , Herndon David N , Suzuki Fujio 
Published: January 2023

Abstract
The effect of SR3335 on infectious complications caused by enterococcal translocation was studied in severely burned mice. SR3335 is an RORα selective inverse agonist for group 2 innate lymphoid cells (ILC2). Sepsis caused by Enterococcus faecalis oral infection (106 CFU/mouse) was developed in decontaminated and proton pump inhibitor treated mice subjected to burn injuries (25%TBSA burn), as previously described. These mice were treated i.p. with 100 μg/mouse of SR3335 once daily for 3 days started 1 h after burn injury. SR3335 inhibits the maturation of ILC2 through the suppression of the RORα transcriptional activity. ILC2 were isolated form the intestines and analyzed by flow cytometry. The severity of infection was evaluated by mortality and bacterial growth in organs of these mice, as previously described. In the results, all severely burned mice died within 6 days of E. faecalis oral infection, while 100% of the same mice treated with SR3335 survived more than 10 days after the infection. E. faecalis grew seriously in the organs of burned mice after the infection, but the pathogen did not grow in burned mice treated with SR3335. The increased number of ILC2 and their cytokine products (IL-5 and IL-13) were detected in the lamina propria of mice 1 to 7 days after severe burn injury. However, the number of ILC2 was not increased in the lamina propria of burned mice treated with SR3335. SCIDbg mice resisted E. faecalis oral infection, while the same mice adoptively transferred with ILC2 were shown to be susceptible to the infection. These results indicate that ILC2 play a role in the severity of gut bacteria-associated sepsis, and the ILC2 inhibitor (SR3335) is suggested to be protective against these infections in severely burned mice.
Pulmonary innate type 2 inflammatory responses protect against Staphylococcus aureus-induced sepsis
Authors: Krishack Paulette , Louviere Tyler J , Sperling Anne I , Verhoef Philip A 
Published: January 2023

Abstract
Sepsis is defined as life-threatening organ dysfunction caused by the body’s response to infection. Sepsis is associated with overwhelming type 1 or type 17 immunologic inflammation, but the role of type 2 immune responses in sepsis remains unknown. To test the role of type 2 immune responses in sepsis, we treated mice with intratracheal IL-33 to induce an innate pulmonary type 2 response, followed by intravenous infection with S. aureus to induce sepsis. Surprisingly, type 2 responses were beneficial during acute infection, as IL-33 treatment protected mice from S. aureus-induced death. An increase in the ratio of lung eosinophils to neutrophils was revealed in IL-33 treated, S. aureus infected mice, suggesting that type 2 responses suppress lung neutrophilia. Further, no difference was found in splenic granulocytes or lymphocytes, indicating that activation of pulmonary immune responses alone was sufficient for protection. Infection of PLZF null mice, which have defective NKT and ILC2 cells normally required for type 2 innate responses after IL-33 treatment, resulted in accelerated mortality compared with wild type littermate controls. Moreover, IL-33 did not rescue PLZF null mice from death. However, infection of CD1d null mice, which lack only NKT cells but have intact ILC2s, demonstrated no difference in mortality. Thus, pulmonary type 2 innate immunity provided by ILC2s is critical for protection against S. aureus-mediated death.
Glutathione reductase facilates immune defense against fungal infections
Authors: Liu Yusen , Kim Victoria Y. , Jin Yi , Meng Xiaomei , Zhang Jian , Rogers Lynette K. , Nelin Leif D. 
Published: January 2023

Abstract
Glutathione reductase (Gsr) catalyzes the reduction of glutathione disulfide to glutathione, which plays an important role in Redox regulation. We expressed Gsr as a GST-fusion protein and produced a rabbit polyclonal antibody against Gsr. Using this antibody we examined Gsr expression levels in various mouse tissues. We found that Gsr was highly expressed in a variety of tissues, including lung, kidney, eyes, spleen, thymus, and bone marrow, while Gsr expression was very low in muscle and heart. Importantly, Gsr was not detected in any tissues of the Gsr hypomorphic mice. Because Gsr is expressed in lymphoid tissues and is implicated in phagocytic functions, we assessed the effect of Gsr deficiency on immune defense against a fungal pathogen, Candida albicans. We report in this study that Gsr-deficient mice exhibited substantially enhanced susceptibility to C. albicans challenge. Upon C. albicans infection, Gsr null mice exhibited dramatically increased fungal burdens in the kidneys, cytokine and chemokine storm, striking neutrophil infiltration and histological abnormalities in both the kidneys and hearts, and substantially elevated mortality. Large fungal foci surrounded by massive numbers of neutrophils were detected outside of glomerulus in the kidneys of Gsr null mice but not in wildtype mice. Examination of the bactericidal functions of the neutrophils from Gsr-deficient mice in vitro revealed normal phagocytosis and yet attenuated oxidative burst activity. Thus, Gsr-mediated redox regulation is crucial for fungal clearance during host defense against fungal challenge.
Immature myeloid cells (IMCs) increased in the peripheral blood of patients with alcohol use disorder (AUD) suppress the bactericidal activity of monocytes
Authors: Asai Akira , Yasuoka Hidetaka , Ohama Hideko , Tsuchimoto Yusuke , Fukunishi Shinya , Kobayashi Makiko , Higuchi Kazuhide , Suzuki Fujio 
Published: January 2023

Abstract
AUD patients have an increased risk of serious opportunistic infections. IMCs have been described as suppressor cells for host antibacterial defenses. In this study, IMCs isolated from AUD patients were examined for their inhibitory functions on the bactericidal activity of healthy donor (HD) monocytes. IMCs and monocytes were isolated from the peripheral blood of 10 AUD patients admitted to Osaka Medical Collage and 5 HD. HD monocytes (1 × 105 cells/ml) were stimulated with heat-killed Enterococcus faecalis, then cultured in media supplemented with the conditioned media of patient IMCs (24 hr-culture fluids, 10% v/v, 1 × 106 cells/ml). Twenty-four hrs after the cultivation, monocytes were harvested and tested for their bactericidal activities against E. faecalis. In some cases, patient IMCs, cultured with 5 μg/ml of retinoic acid (RA) for 24 hrs, were utilized for the experiments. Also, both groups of IMCs were analyzed for RA receptor (RAR) expression by flow cytometry. In the results, as compared to HD peripheral blood IMCs, the numbers of IMCs increased in AUD patients. HD monocytes were shown to be bactericidal against E. faecalis. However, the bactericidal activity of HD monocytes was not demonstrated when they were cultured with media supplemented with the conditioned media of patient IMCs. HD IMCs differentiated to monocytes or dendritic cells after cultivation with RA. However, AUD patient IMCs were shown to be not susceptible to RA treatment (patient IMCs, RAR−; HD IMCs, RAR+). These results suggest that, through the production of some soluble factors active against killing functions of monocytes, patient IMCs increase the susceptibility of AUD patients to opportunistic infections.
Protective role of ILC2 in sepsis-induced acute lung injury
Authors: Shu Qiang , Lai Dengming , Fan Jie 
Published: January 2023

Abstract

Objective
Group 2 innate lymphoid cells (ILC2), a recently identified population of innate immune cells, were suggested to be protective in infection. However, the underlying mechanism remains unknown. The current study aimed to determine the protective role of ILC2 in sepsis-induced acute lung inflammation and the underlying mechanism.

Methods
WT and IL-33 knockout mice were subjected to cecal ligation and puncture (CLP) to induce sepsis. The number of ILC2 and the concentration of cytokines including IL-4, IL-9 and IL-13 in the lung tissue were measured by flow cytometry and ELISA, respectively, at different time after CLP. In the in vitro studies, lung-derived ILC2 were co-cultured with mouse lung endothelial cells (LEC) and macrophages (MΦ), respectively, and challenged with LPS and/or TNF-α for up to 48 h. The LEC apoptosis and the ability of MΦ phagocytosis of bacteria were then detected by confocal fluorescence microscopy.

Results
Sepsis induced IL-33-dependent recruitment of ILC2 in the lung after CLP. The levels of ILC2-derived IL-9 and IL-13 significantly increased in the lung tissue of WT septic mice and in the supernatant of cultured ILC2 after LPS stimulation. IL-9 protects LPS/TNFα-induced LEC apoptosis, whereas, IL-13 decreased MΦ apoptosis in response to LPS/TNF-α. In addition, both IL-9 and IL-13 promote MΦ phagocytosis of bacteria.

Conclusions
IL-33-mediated ILC2 recruitment in the lung following sepsis protects lung injury through reducing apoptosis of LEC and MΦ and promotes bacteria clearance by MΦ. Increasing ILC2 and/or ILC2-derived cytokines in the lung may present a new therapeutic strategy for acute lung injury in sepsis.
GM-CSF controls the balance between effector and regulatory T cell lineage in experimental autoimmune encephalomyelitis (EAE)
Authors: Ghosh Debjani , Curtis Alan Dale , Wilkinson Daniel S , Mannie Mark D 
Published: January 2023

Abstract
Previous studies showed that GM-CSF-deficient (Csf2-deficient, Csf2−/−) mice exhibited profound resistance to experimental autoimmune encephalomyelitis (EAE). The present study focused on the myelin oligodendrocyte glycoprotein (MOG) 35–55-specific model of murine EAE in Csf2-deficient and wildtype C57BL/6 mice. The study addressed whether disease resistance of Csf2-deficient mice reflected a role for GM-CSF in controlling the functional balance between effector and regulatory T cells. The main finding was that treatment with the anti-CD25 mAb PC61 rendered Csf2-deficient mice fully susceptible to severe EAE, similar to that of C57BL/6 mice. When both donors and recipients were treated with PC61 in a passive model of EAE, adoptive transfer of myelin-specific Csf2−/− T cells into Csf2−/− recipients resulted in chronic severe EAE. Csf2-deficient T cell repertoire was characterized by elevated CD3+ T cell frequencies that reflected accumulations of naïve CD44null-low CD4+ and CD8+ T cells but normal frequencies of CD4+ CD25+ FOXP3+ T cells. Overall, this finding suggested that Csf2−/− mice had secondary deficiencies in T cell sensitization to environmental antigens. In accordance, MOG35-55/ Complete Freund’s Adjuvant -sensitized Csf2−/− mice exhibited deficient peripheral sensitization to MOG, whereas pretreatment of sensitized Csf2−/− mice with PC61 enabled the robust induction of MOG-specific T cell responses in the draining lymphatics. In conclusion, the EAE resistance of Csf2−/− mice, at least in part, reflects deficient induction of effector T cell function that cannot surmount normal Treg barriers. EAE effector responses in Csf2-deficient mice however are unleashed upon depletion of regulatory CD25+ T cells.
The Crystal Structure of the IL-33 Signaling Complex Reveals the Molecular Basis for Receptor Sharing in the IL-1 Family.
Authors: Guenther Sebastian , Bowers Amanda L. , Deredge Daniel , Bonsor Daniel A. , Beadenkopf Robert , Wintrode Patrick L. , Sundberg Eric J. 
Published: January 2023

Abstract
Cytokines of the IL-1 family are potent stimulators of the immune system. As seen in other cytokine families, cytokine redundancy due to receptor sharing is characteristic for the IL-1 family. Today five primary receptors have been described that each bind a subset of the 11 known cytokines within this family. Once an agonistic cytokine-receptor pair has formed, it can engage one of the two co-receptors and initiate signaling through MyD88. While IL-18Rβ is the unique co-receptor for the IL-18/IL-18Rα pair, IL-1RAcP is shared between the other four primary receptors, IL-1RI, IL-1RII, ST2 and IL-36R. Crystal structures of IL-1β bound to IL-1RI or IL-1RII in complex with IL-1RAcP previously revealed the general architecture of signaling complexes in the IL-1 family. IL-33 is the latest cytokine to be discovered in the IL-1 family. Since its first description in 2005 it has been recognized as a central driver of type 2 immunity. Here we present the crystal structure of IL-33 in complex with its receptors ST2 and IL-1RAcP. The structure reveals a remarkable conservation of the ternary architecture of signaling complexes in the IL-1 family. We observe common principles of co-receptor binding, also seen in the other two ternary structures containing IL-1RAcP. However, hydrogen/deuterium mass exchange spectroscopy and extensive alanine-scanning analysis combined with direct binding analysis of the IL-1β and IL-33 signaling complexes demonstrate that the binding energy is distributed quite differently within the two. Co-receptor binding of ST2/IL-33 is much less dependent on the cytokine than on the receptor, while for IL-1β/IL-1RI the cytokine plays a much more important role.