bluuebunny/crossref_metadata_2025_split huggingface dataset:
Tumour eradication and induction of memory against murine mesothelioma by combined immunotherapy (53.22)
Authors: Ireland Demelza , Kissick Haydn , Krishnan Shruti , Madondo Mutsa , Beilharz Manfred 
Published: January 2023

Abstract
Numerous immunotherapy treatments for cancer are undergoing clinical trials or are already approved for use. One particular area of interest is targeting mechanisms of immune tolerance. Using a murine model of mesothelioma, we investigated the roles of regulatory T-cells (Tregs), intra-tumoural transforming growth factor (TGF)-β and the immunosuppressive cytotoxic T lymphocyte-associated (CTLA)-4 molecule in immune tolerance to tumours. It was found that treatments targeting a single immunosuppressive mechanism were not as effective against tumours as targeting multiple mechanisms simultaneously. Combined Treg depletion and intra-tumoural TGF-β neutralisation resulted in the generation of an anti-tumour response however; this treatment regime was limited by the eventual return of the Tregs around 7-10 days following depletion. In this study we report that combining CTLA-4 blockade with our previously published combined Treg depletion/TGF-β neutralisation regime not only permanently cleared established tumours in ~50% of treated mice. Most importantly, it was found that a combined triple treatment of anti-CD25 mAb, anti-CTLA-4 mAb and TGF-β soluble receptor resulted in prolonged depletion of Tregs, long term clearance of tumours and memory against tumour rechallenge. These data suggest that clinical application of immunotherapies against tumours may be improved by targeting multiple immune suppression mechanisms simultaneously.
TLR11-dependent and -independent recognition of Toxoplasma gondii enhances innate immunity to bacterial infection (53.3)
Authors: Neal Lori , Knoll Laura 
Published: January 2023

Abstract
Previous studies have shown that mice infected with the obligate intracellular parasite Toxoplasma gondii are resistant to infection with a variety of bacterial, viral, and fungal pathogens, but the factors and mechanisms mediating these phenomena are unknown. This study identifies the TLR11 ligand profilin (TgPRF) as one T. gondii factor sufficient to confer resistance to bacterial infection using a lethal Listeria monocytogenes challenge model. Mice stimulated with a single injection of purified TgPRF had a 3- to 4-log reduction in bacterial burden in the spleen and liver compared to control animals at day 3 post infection, did not develop clinical signs of disease, and survived for over 30 days. TgPRF stimulation was associated with a rapid increase in the number of neutrophils in the spleen and with production of CCL2, TNF-α, IL-12 and IFN-γ. TgPRF stimulation reduced bacterial burden in mice deficient in T cells, NK cells, IL-12 or IFN-γ, but did not provide any benefit in animals treated with neutrophil depleting antibody, suggesting its protective effect was primarily dependent on neutrophil recruitment. T. gondii-infected TLR11-deficient animals showed similar resistance to L. monocytogenes challenge, indicating that other T. gondii factors may function in similar, but TLR11-independent manners. Together these data show that TgPRF and other T. gondii factors may be used as therapeutics to stimulate innate immunity against bacterial infections.
The Role of MyD88 in the Adjuvant Activity of Neisseria meningitidis PorB (53.5)
Authors: Lohsen Mark , Liu Xiuping , Toussi Deana , Massari Paola , Wetzler Lee 
Published: January 2023

Abstract
Adjuvants enhance the response to poorly immunogenic antigens in vaccines. Our laboratory has shown by ELISA that Neisseria meningitidis outer membrane protein PorB acts as an adjuvant by eliciting a greater serum concentration of antigen-specific IgG when compared to immunization with antigen alone, with Ovalbumin as the antigen. PorB is a TLR2 ligand and signals through the adaptor protein MyD88. We compared the adjuvant effect of PorB to other adjuvants (Alum, MPL & CpG) in WT & MyD88 KO mice. PorB adjuvant activity was ablated in MyD88 KO mice and consistent with previous investigations: MPL & CpG adjuvant activity was also MyD88-dependent while Alum acted independently of MyD88. In addition, we previously observed PorB had residual adjuvant activity in TLR2 KO mice. One hypothesis was that PorB could stimulate the Nalp3 inflammasome and induce IL-1β production without a second signal e.g. ATP. Therefore, bone marrow-derived macrophages were stimulated with PorB ± ATP. The IL-1β production was analyzed by ELISA and compared to IL-1β production from stimulation with other TLR ligands. We found that PorB required ATP in order to sufficiently produce IL-1β, disproving our hypothesis. Overall, our results show an importance for MyD88 in both the adjuvant pathway of PorB & other TLR ligand-based adjuvants. One future direction for this work is MyD88 deletion in an antigen-presenting cell-specific manner to examine the role of each cell type in the adjuvant activity of PorB.
Novel hemocyanin from the Fissurella latimarginata exhibits an outstanding immunogenicity and non-specific immunomodulatory effects in a melanoma model (53.6)
Authors: Becker María , Arancibia Sergio , Cecilia Espinoza , Salazar Fabián , Del Campo Miguel , Born Raimundo , Ferreira Jorge , Manubens Augusto , De Ioannes Alfredo 
Published: January 2023

Abstract
The versatile properties of mollusk hemocyanins in biomedical applications, including non-specific immunostimulant during the therapy of bladder cancer and carrier/adjuvant in cutting-edge therapeutic cancer vaccines, has increased the interest in finding new hemocyanins with better immunomodulatory properties. Here, we evaluate the physicochemical and immunological properties of the hemocyanin from Fissurella latimarginata (FLH). This protein shares with keyhole limpet hemocyanin (KLH) and Concholepas hemocyanin (CCH) the typical hollow cylinder structure and it is also made by two subunits (~350 KDa). ConA lectin staining of FLH, demonstrated the presence of mannosylated carbohydrate structures as well as in KLH and CCH. The humoral responses in mice demonstrated that antibody titers induced by FLH itself were significantly higher than KLH and CCH. However, it performance as a carrier protein to induce antibodies to the hapten DNFB, was similar to the other carriers using Freund’s adjuvant as a repository. More interestingly, FLH exhibits an outstanding antitumor activity prolonging mice survival, when was evaluated and compared with KLH and CCH in the B16F10 mouse melanoma model. In addition, flow cytometry analysis showed that FLH induced a significant number of tumor-infiltrating CD4+ cells and also, IFN-δ secretion. Together, these findings introduce a novel hemocyanin, with intrinsically more immunogenic and immunomodulatory capacities for antitumor therapy.
Protective role of R-spondin3 in mesenteric ischemia/reperfusion induced tissue injury (54.11)
Authors: Kannan Lakshmi , Kis-Toth Katalin , Yoshiya Kazuhisa , Thai To-Ha , Tsokos George 
Published: January 2023

Abstract
Ischemia reperfusion (I/R) injury represents a leading cause of morbidity and mortality. R-spondins, acknowledged as ligands of Wnt pathways, demonstrate their physiological role as mitogens, morphogens, stem cell factors, and wound healing agents in various development and disease models. We asked whether these proteins could prevent inflammation-associated tissue damage in ischemic disorders. C57BL/6J mice were pretreated with vehicle or R-spondin3 (R-spo3) before inducing mesenteric I/R for 30/180 min. At the end of reperfusion, intestinal tissues were harvested for different experimental analysis. Treatment with R-spo3 significantly decreased local intestinal injury. mRNA and protein analyses revealed that this effect was achieved through the modulation of the expression of pro-inflammatory cytokines (IL-6, TNFα and COX2) and deposition of IgM and complement. The decreased levels of IgM on tissues correlated with the reduced vascular leakage in the intestine of R-spo3 treated mice. Further we determined the effect of R-spo3 on endothelial barrier function in vitro using HUVECs and FITC-dextran (500kDa). R-spo3 pretreated groups showed significant reduction in histamine-enhanced FITC-dextran permeability of endothelial monolayer. Our results demonstrate that R-spo3 mitigates I/R-induced local tissue injury by preventing vascular leakage and decreasing the local inflammatory response. These findings confer the protective effects of R-spondin3 in I/R-related tissue damage.
The role of Syndecan-4 in the development of rheumatoid arthritis (54.23)
Authors: Endo Tsutomu , Morimoto Junko , Ito Koyu , Kanayama Masashi , Iwasaki Norimasa , Minami Akio , Uede Toshimitsu 
Published: January 2023

Abstract
Syndecan-4 (Syn4), a cell-surface heparan sulfate proteoglycan has been implicated as a critical mediator for inflammatory responses. Recent report has demonstrated that dendritic cell-associated heparan sulfate proteoglycan-dependent integrin ligand (DC-HIL) binds to Syn4 on activated T cells, resulting in the inhibition of T cell responses. Here, we have investigated the role of Syn4 in the development of Rheumatoid Arthritis (RA). We have used two RA models; T cell-dependent collagen-induced arthritis (CIA) model and T cell-independent collagen antibodies-induced arthritis (CAIA) model. In CAIA model, Syn4-wild-type and Syn4-deficient (Syn4 KO) mice showed similar levels of clinical severity. In contrast, in CIA model, Syn4 KO mice revealed attenuated RA development. Syn4 expression was detectable on activated CD4+ T cells and B cells in the draining LN (dLN). Syn4 KO mice showed a tendency to generate reduced numbers of Th1 and Th17 cells in the dLN after immunization, and helper T cells generated in Syn4 KO mice revealed different expression patterns of chemokine receptors. Although both groups showed comparable numbers of Tfh and GC (germinal center) B cells, Syn4 KO mice produced lower level of type-II collagen-specific antibody, as compared with WT mice. These results suggest that Syn4 may regulate the development of RA by modulating T and B cell responses. We are currently investigating if Syn4-deficiency affects the functions of Tfh and GC B cells.
Manzamine A (MZA) has differential effects on osteoarthritis (OA) fibroblast expression of MCP-1 and IL-6 (54.26)
Authors: Klosowska Karolina , Mc Dermott Sarah , Zanotti Brian , Volin Michael , Walczak Brian , Woods James 
Published: January 2023

Abstract
MZA is a complex alkaloid compound with anti-inflammatory properties that has recently been synthesized. OA has been characterized to have an inflammatory component involving increased cytokine release by synovial tissue fibroblasts. We hypothesized that MZA treatment of OA fibroblasts could decrease their production of pro-inflammatory cytokines at non-toxic concentrations. OA fibroblasts were isolated with IRB approval, treated with MZA or vehicle, and viability was assessed using multiple methods. OA fibroblast condition media (CM) was collected at various time points after MZA or control treatment, screened by antibody microarrays, and verified by ELISAs. Toxicity studies on OA fibroblasts suggested that concentrations up to 2.5 µM MZA were not toxic using a sensitive flow cytometry kit (results varied in a method-, time-, and concentration-dependent manner). Antibody microarray performed on CM pooled from 5 primary OA fibroblast lines treated with 10 µM MZA for 24 hrs suggested that monocyte chemoattractant protein (MCP)-1 and interleukin (IL)-6 were two of the most prominent proteins detected, and MZA appeared to decrease MCP-1 production. ELISAs suggested that concentrations of 0.5 µM MZA and higher significantly decreased production of MCP-1 (n=5; p&lt;0.05). In contrast, 1.25 µM MZA appeared to increase IL-6 production, while other concentrations inhibited it. Our data suggest that MZA could alter cytokine production at non-toxic concentrations.
Knockdown of Ste20-like proline/alanine-rich kinase (SPAK) attenuates intestinal inflammation in mouse (54.9)
Authors: Yan Yutao 
Published: January 2023

Abstract
Inflammatory bowel diseases (IBDs), principally Crohn’s disease and ulcerative colitis, are characterized by epithelial barrier disruption and alterations in immune regulation. Ste20-like proline/alanine-rich kinase (SPAK) plays a role in intestinal inflammation, but its underlying mechanisms need to be defined. SPAK KO mice exhibited significant increases of intestinal barrier function. Knock-down of SPAK significantly decreased paracellular intestinal permeability to FITC-dextran, and lowered significantly the sodium ion selectivity of tight junctions in intestinal epithelial cells. Further, the expression of junction proteins β-catenin, claudin-2 decreased. In contrast, expression of ZO-1, Occludin, E-cadeherin increased significantly, whereas the change of expressions of Claudin-4 and Claudin-1 was not noticeable. In vivo studies using mouse model of colitis induced by dextran sulfate sodium (DSS) and trinitrobenzene sulfuric acid (TNBS) showed that KO mice were more tolerant to DSS or TNBS treatment than wild-type animals, as demonstrated by clinical and histological characteristics and MPO enzymatic activities. KO mice also demonstrated ameliorated colon inflammation by mouse endoscopy compared to wild type mice. Consistent with this notion, we found that SPAK knockdown attenuated the increases of inflammatory cytokines production in vivo and also ameliorated bacterial translocation under DSS treatment, which together likely reduce intestinal epithelial permeability.
Elevated IL -1β levels are associated with resistance to Yesinia pestis in B10.T(6R) mice (55.11)
Authors: Sarkar Sanghita , Lambert Nathaniel , Nilles Matthew , Bradley David 
Published: January 2023

Abstract
We have previously demonstrated that inbred B10.T(6R) mice are resistant to Yesinia pestis (KIM5 strain) and resistance decreases with age. It has been shown that Y. pestis is cleared by neutrophil engulfment. We evaluate here the potential contribution of IL-1β, a proinflammatory cytokine important for neutrophil recruitment and activation. IL-1β levels were measured from spleens and sera collected 60 and 72 hour post infection from young (resistant) and middle-aged (susceptible) B10.T(6R) mice and control (susceptible) C57BL/6. Splenic IL-1β production was statistically higher in young B10.T(6R), compared with both middle-aged B10.T(6R) and C57BL/6 mice. Serum IL-1β levels were also moderately higher in young B10.T(6R) mice when compared to middle aged B10.T(6R) and C57BL/6 mice. IL-1β produced by Y. pestis-infected peritoneal-derived macrophages taken from uninfected young and middle-aged B10.T(6R) mice and C57BL/6 mice was measured at 2,4,8 and 24 hour. IL-1β levels were significantly elevated in peritoneal macrophages infected in vitro from young B10.T(6R) mice compared to decreased levels in both middle-aged B10.T(6R) and C57BL/6 derived peritoneal macrophages. These studies suggest a direct correlation between elevated IL-1β levels and resistance to Y. pestis infection observed in young B10.T(6R) mice.
Skin wound healing and cytokine response promoted by a quorum-sensing molecule from Pseudomonas aeruginosa (55.16)
Authors: Kanno Emi , Kawakami Kazuyoshi , Miyairi Shinichi , Tanno Hiromasa , Ishii Keiko , Hayashi Denso , Maruyama Ryoko , Tachi Masahiro 
Published: January 2023

Abstract
Quorum-sensing systems regulate virulence factors in Pseudomonas aeruginosa, which produces two autoinducers, N-3-oxododecanoyl or N-butanoyl-L-homoserine lactone (3-oxo-C12- or C4-HSL, respectively). Production of each HSL is regulated by the lasI or rhlI gene. Recently, HSLs are considered to play important roles in modulating immune responses. In this study, we addressed the effect of HSLs on the wound healing in skin. Wounds were created on the back of SD rats. P. aeruginosa PAO1 and its lasI or rhlI-deletion mutant (ΔlasI or ΔrhlI) were inoculated at the wound surface. Rats were injected intraperitoneally with anti-3-oxo-C12-HSL or -C4-HSL antiserum or administered each HSL at the wound surface. The wound tissues were harvested for analysis of re-epithelialization, accumulation of neutrophils and synthesis of TNF-α. Inoculation with PAO1 significantly accelerated the healing process, as evaluated by re-epithelialization, on day 3 post-wounding, which was associated with the local infiltration of neutrophils and synthesis of TNF-α. These responses were not observed when inoculated with ΔlasI or ΔrhlI in place of PAO1 and cancelled when co-treated with these anti HSL antisera, respectively, but not with control rabbit serum. By contrast, administration of either HSL in the absence of PAO1 significantly accelerated the re-epithelialization. These results suggest that 3-oxo-C12-HSL and C4-HSL may be deeply involved in the skin wound healing during P. aeruginosa infection.
Dectin-1 Signaling Recruits Elements of the Autophagic Machinery to Candida albicans-containing Phagosomes (55.2)
Authors: Tam Jenny , Tanne Antoine , Khan Nida , Mansour Michael , Becker Christine , Sokolovska Anna , Stuart Lynda , Xavier Ramnik , Vyas Jatin 
Published: January 2023

Abstract
A classic hallmark in the autophagic process is the reversible conjugation of LC3 to the autophagosomal membrane. Autophagy has been shown to target pathogens that reside in the cytosol or within autophagosomes for lysosomal degradation. Therefore, components of the autophagy pathway can participate in the effective elimination of viruses, bacteria, and parasites, though their role in fungal immunity is poorly understood. We show that activation of Dectin-1 by its ligand β-1,3-glucan leads to recruitment of LC3 to the phagosome and describe multiple intermediate signaling steps. The cytoplasmic tail of Dectin-1 contains an immunoreceptor tyrosine-based activation (ITAM)-like motif which is phosphorylated in response to binding of β-1,3-glucan, leading to relocation from the cell surface to the phagosome. We show that Dectin-1 activation by intact fungal pathogens or by chemically defined ligands leads to LC3 recruitment to the phagosomal membrane. Mutation of the tyrosine phosphorylation site in Dectin-1 abolished LC3 recruitment, suggesting the involvement of spleen tyrosine kinase (Syk) in this subcellular redistribution of LC3. Likewise, impairment of ROS generation by knockout of NADPH oxidase (NCF4) downstream of Syk inhibited LC3 recruitment to the phagosome, whereas knockout of Card9 did not affect LC3 recruitment. These observations suggest a previously unobserved signaling pathway connects Dectin-1 to the autophagic machinery via Syk signaling and ROS generation.
Role of IL-18 in the IFN-γ production by innate lymphcytes in response to contact-dependent interaction with dying bacterial-infected macrophages. (55.20)
Authors: Kubota Koichi 
Published: January 2023

Abstract
We have previously proposed that macrophage cell death caused by bacteria triggers initial IFN-γ production by innate lymphocytes at an early stage of bacterial infection (Scan. J. Immunol. 71:199-209, 2010; Front. Immun. 2:26, 2011). Here we explore the role of IL-18 in this phenomenon by using functional T cell hybridoma B6HO3, a hybrid between YACUT lymphoma and a relevant innate T cell. B6HO3 cells produced IFN-γ dependently of IL-18, but not IL-12p70, in response to cell-cell contact-dependent interaction with dying Listeria monocytogenes (LM)-infected macrophages. Despite the fact that B6HO3 cells required above 300 pg/ml of IL-18 to produce IFN-γ, the amounts of IL-18 secreted by dying LM-infected macrophages were far below this threshold level. IL-18 binding protein (IL-18BP) inhibited this IFN-γ production, but its ED50 was about 30 nM, far exceeding the physiological level of IL-18BP. LPS-primed macrophages, when stimulated with ATP, underwent cell death and secreted the similar amounts of IL-18, but their co-culture with B6HO3 cells did not result in producing IFN-γ. Inhibitors of caspase-1 and cathepsin B did not inhibit LM-infected macrophage cell death, indicating that neither pyroptosis nor pyronecrosis was involved. These results suggest that when macrophages are killed by bacteria, IL-18 plays a pivotal role in the production of IFN-γ by B6HO3 cells through an as yet undefined cell-cell interaction between dying bacterial-infected macrophages and B6HO3 cells.
Binding of soluble yeast β-1,3/1,6 glucans to complement receptors leads to immunomodulatory effects on both innate and adaptive human immune cells (55.8)
Authors: Grossman William , Chan Anissa , Gorden Keith , Bykowski-Jonas Adria , Walsh Richard , Ertelt Katie , Vasilakos John , Antonysamy Mary , Ottoson Nadine , Bose Nandita 
Published: January 2023

Abstract
β-glucans are considered as fungal pathogen-associated molecular patterns (PAMPs) that are recognized by cells of the human innate immune system. Few studies have focused on recognition and immunostimulatory functions of soluble β-glucan (sβG) on innate and adaptive human immune cells. In this study, we investigated recognition and immunomodulatory functions of an analytically well-characterized form of yeast-derived sβG by human neutrophils, monocytes, and B-cells. Binding studies demonstrated that the sβG bound to human neutrophils and monocytes via complement receptor 3 (CR3) and B-cells via CR2, and that receptor binding was complement-dependent. Opsonization of soluble β-glucan was detected by the presence of complement protein, iC3b on β-glucan-bound cells. Binding of sβG on human neutrophils resulted in enhanced directed migration of neutrophils towards IL-8. In the presence of other PAMPs (eg, LPS), binding of sβG was increased on both neutrophils and monocytes. Interestingly, when sβG was combined with either a TLR7/8 agonist (imidazoquinoline CL097) or a TLR9 agonist (CpG oligonucleotide, CpG 2006), enhanced activation of B-cells was observed as measured by B cell proliferation. These results demonstrate for the first time that complement opsonization of sβG makes it amenable not only for recognition by several complement receptors, but also modulation of immune functions of both innate and adaptive immune cells.
Hydrophobin RodA protein is essential for masking the host immune response and enhancing fungal survival of Aspergillus fumigatus during corneal infection (55.9)
Authors: de Jesus Carrion Steven , Leal Jr. Sixto M. , Paul-Latge Jean , Pearlman Eric 
Published: January 2023

Abstract
Fungal infections of the cornea (i.e keratitis) are caused mainly by filamentous fungi, with Fusarium and Aspergillus species being the most common agents. The surface layer of A.fumigatus spores is composed of hydrophobic RodA protein (RodAp) that coats the conidial surface, making it inert to the immune system. Thus, we proposed that RodA protein (RodAp) masks surface β-glucan immune recognition in the cornea during fungal keratitis. Chemical removal of RodA (using hydrofluoric acid (HFl)) and genetic removal (ΔrodA) lead to increased exposure of surface β-glucan in conidia, and decreased macrophage activation. Dectin-1-/- mice corneas infected with ΔrodA conidia for 6h exhibited decreased cytokine production, and neutrophil infiltration into the cornea. Dectin-1-/- macrophages exposed to ΔrodA and HFl treated conidia showed decreased cytokine production, NFkB activation, and phagocytosis. To study the role of RodAp in innate immune evasion, C57Bl/6 mice corneas infected with ΔrodA conidia for 96h exhibited less corneal opacity and enhanced ability to kill the fungus when compared to the corneas infected with the WT strain. Also, human and murine neutrophils showed increased fungal killing when exposed to ΔrodA hyphae in vitro, compared to the G10 strain. This data shows a role for RodAp in masking the host mediated β-glucan immune response to conidia and establishes a novel role for surface RodAp in enhancing fungal survival in the cornea during fungal keratitis.