bluuebunny/crossref_metadata_2025_split huggingface dataset:
2,3,7,8-Tetrachlorodibenzo-p-dioxin Induces Gimap5 Expression to Ameliorate Concanavalin-Induced T Cell-Mediated Liver Injury
Authors: Cannon Alkeiver S , Miranda Kathryn , Wilson Kiesha , Nagarkatti Prakash S. , Nagarkatti Mitzi 
Published: January 2023

Abstract
GTPase of the immune-associated protein 5 (Gimap5) has been implicated in immune cell maintenance and development. A mouse model comparable to autoimmune hepatitis in humans was used to study the alterations induced in the gene expression profile of immune cells upon administration of environmental toxin 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD). In this study, we tested the hypothesis that TCDD is capable of modulating gene expression to ameliorate autoimmune hepatitis severity. Mice were injected intravenously with 12.5 mg/kg concanavalin A (ConA) and treated intraperitoneally one hour after challenge with vehicle or 10 μg/kg TCDD. TCDD-treated mice showed lower levels of liver injury enzymes, alanine aminotransferase (ALT) and aspartate aminotransferase (AST), as compared to vehicle-treated mice. Histopathological analysis revealed a reduction in liver damage in the TCDD-treated group as compared to the vehicle-treated control. Furthermore, flow cytometric analysis of liver-infiltrating mononuclear cells demonstrated that following treatment with TCDD, there was an increase in FoxP3+CD4+ Tregs as well as a decrease in proinflammatory Tbet+ Th1 and RORγt+ Th17 T helper cell subtypes after induction of the disease. Single cell RNA-sequencing was conducted on liver mononuclear cells to further assess the changes induced by TCDD. The gene expression profile showed that TCDD treatment significantly upregulated the expression of vital genes such as Gimap5, a maintenance gene for normal liver function and survival of T cells. In summary, our data suggests that TCDD is capable of inducing Gimap5 expression in autoimmune hepatitis, which in turn modulates the progression of this disease.
IL-4 modulates reconstituted human T cells as a regulatory factor to mitigate GVHD in humanized mice model
Authors: Kuo Tzu-Jiun Tzu-Jiun , Sun Cheng-Pu , Tao Mi-Hua 
Published: January 2023

Abstract
Allogeneic hematopoietic cell transplantation is an essential treatment in hematological diseases. However, graft-versus-host disease (GVHD) is the leading cause of the mortality and treatment failures in transplant recipients. In humanized mice engrafted with human peripheral-blood mononuclear cells (PBMCs), the rapid development of lethal xenogeneic GVHD is also a major model limitation that hinders long-term and precise immune analysis. Therefore, the study aims to suppressed GVHD symptoms in human PBMCs-engrafted humanized mice model for the study of human immune responses. We investigated whether supplementary of human IL-4 that modulate naïve T cells maturing and differentiating can lead to GVHD amelioration. Adeno-associated virus (AAV) vectors were used to express human cytokines in humanized mice. We first demonstrated that rAAV vectors allow sustained expression of human IL-4 in vivo. After injection of PBMCs, GVHD symptoms observed in NSG mice were significantly suppressed in human IL-4 expressing recipients, with the survival of originally less than two months to more than 7 months without body weight loss. Long-term engraftment of human T cells were also observed. Among the reconstituted human cells, CD4+ T cells were the dominant expanding subpopulation, comparing to CD8+ dominated cell proliferation in mice with GVHD development. In addition, significantly high levels of human IgG in humanized mice sera (1 mg/ml, comparable to human’s) were detected 5 months after PBMC transplantation, suggesting the potential use for long-term study of human humoral immune responses. In conclusion, human IL-4 expression by AAV vectors is an efficient strategy to ameliorate GVHD in PBMC-engrafted humanized mice model.
Senescence-associated β-galactosidase activity and other markers of senescence are present in human peripheral blood mononuclear cells during healthy aging
Authors: Dewald Hannah K , Martínez-Zamudio Ricardo Iván , Vasilopoulos Themistoklis , Herbig Utz , Fitzgerald-Bocarsly Patricia 
Published: January 2023

Abstract
Aging is associated with a decline in the immune system, termed “immunosenescence”. Cellular senescence is defined by persistent cellular growth arrest and overall loss of function. If immune cells undergo cellular senescence in vivo and whether cellular senescence has a role in immunosenesence remains controversial due to the lack of specific markers to reliably identify these cells in blood. Freshly isolated human peripheral blood mononuclear cells from younger and older donors were fluorescently labeled for senescence-associated β-galactosidase (SA-βGal). CD8+ T cells were sorted based on SA-βGal and analyzed by qRT-PCR, RNA sequencing, and microscopy. We found a higher percentage of cells with high SA-βGal in the older vs. younger cohort. This increase was apparent in NK cells, CD4+ and CD8+ T cells. CD8+ T cells with high SA-βGal displayed increased transcripts of p21, p16INK4a, and inflammatory cytokines. They also had greater DNA damage response foci and p16INK4a protein. SA-βGal increased as T cells matured, which was consistent regardless of age. Intriguingly, a portion of T effector cells did not exhibit a senescent phenotype and maintained low SA-βGal. RNA sequencing of CD8+ T cells revealed alterations in gene expression were associated with SA-βGal and cells with low, medium, and high SA-βGal shared similar profiles, independent of age. The presence of p16INK4a, DNA damage, and inflammatory cytokines demonstrate that SA-βGal is a strong indicator of cellular senescence. SA-βGal can be leveraged to further identify senescent immune cells and elucidate cellular senescence’s role in immunosenesence. Understanding this connection in healthy aging will further our understanding of premature aging seen chronic diseases.
Cellular Phenotypes in Cancer Patient Spleen and Peripheral Blood
Authors: Talmadge James E , Cole Kathryn , Ly Quan , Hollingsworth Tony , Cox Jesse , Stromnes Ingunn M , Padussis James , Foster Jason , Vargus Luciano 
Published: January 2023

Abstract
Adoptive cellular therapy (ACT) has progressed; however, it remains ineffective for most solid tumors. Further, neoadjuvant patients frequently lack tumor tissue sufficient to isolate and expand tumor infiltrating lymphocytes (TIL’s) that are the primary source of T-cells for ACT. However, patients with tumors near the splenic vasculature often undergo a splenectomy, providing a cellular resource for ACT. We document herein that spleens provide a high frequency and number of CD8+ T-cells, with low expression of exhaustion markers (program death ligand-1 (PD-1), T-cell immunoglobulin and mucin domain containing-3 (TIM-3), and lymphocyte activation gene-3 (LAG-3)) relative to the peripheral blood (PB). The memory phenotypes of these CD8+ T-cells (predominantly central and transitional) support their potential for expansion and their high frequency of PD-1 expression indicates tumor specificity as PD-1, is a surrogate for tumor specificity. Further, spleens have a low frequency of myeloid derived suppressor cells (MDSCs) subsets vs. patient PB. Our preliminary studies with spleens from 19 abdominal tumor patients revealed that the spleens, relative to the PB from the same patients, have a significantly higher frequency of CD8+ and PD-1+CD8+ T-cells, including central and transitional memory T-cells. Further, the CD8+PD-1+ T-cells in the spleen also have a low expression of TIM-3 and LAG-3 suggesting minimal exhaustion as supported by ELISPOT analysis of tumor specific CD8+ T-cells responses. Based on our studies we suggest that resected spleens from cancer patients including pancreatic ductal adenocarcinoma (PDAC) patients can provide a valuable source for ACT due to their high frequency of CD8+PD-1+ T-cells.
Development of regulatory IL-10-producing ILCs during type 2 inflammation
Authors: Nidetz Natalie F. , McGee Michael C , Limper Candice B , Ye Kaixiong , Islam Rezwanul , August Avery , Huang Weishan 
Published: January 2023

Abstract
Innate lymphoid cells (ILCs) are important for mucosal homeostasis and host defense against infectious pathogens. ILC subsets ILC1, 2, and 3, classified similar to T helper cells Th1, Th2, and Th17, are well characterized and known to promote both protective and harmful inflammatory responses. However, the immunosuppressive roles of ILCs are less understood. Regulatory ILCs that produce the immunomodulatory cytokine IL-10 have been recently defined, but the mechanisms regulating their development and function are not fully understood. Similar to other ILCs, regulatory ILCs were thought to be a pre-existing population, however unlike ILC1, 2, and 3 which all differentiate from ILC precursor cells (ILCPs), regulatory ILCs were thought to differentiate exclusively from upstream precursors, common helper-like innate lymphoid precursors (CHILPs) and common lymphoid cell precursors (CLPs). In our animal facilities, we found no evidence of pre-existing tissue resident IL-10+ ILCs in our investigations. In contrast, we find that co-stimulation of IL-33 and a γ-chain cytokine (IL-2, IL-4, or IL-7) robustly induces IL-10+ILCs that exhibit immunosuppressive functions in vitro and in vivo. Further, in addition to CLPs and CHILPs, we find that downstream ILC2Ps can develop into functionally suppressive IL-10+ ILCs following cytokine stimulation. Together, our data support an alternative pathway of IL-10-producing regulatory ILC development and regulation of inflammatory responses.
Dysregulated TGF-β Signaling Leads to Cell-Intrinsic Changes in T cell Development that Promote Allergic Inflammation
Authors: Haque Tamara , Weissler Kate , Frischmeyer-Guerrerio Pamela 
Published: January 2023

Abstract
Variants in genes in the Transforming Growth Factor Beta (TGFb) signaling pathway are associated with allergic diseases suggesting that this cytokine may be a central player in allergic disease. The role of TGFb in preserving T cell mediated self-tolerance is well-appreciated but how disruption in this pathway promotes allergic inflammation is not clear. TGFb signals through its receptor TGFβRI and II, leading to phosphorylation and nuclear translocation of Smad proteins 2 and 3. We previously demonstrated that patients with Loeys-Dietz Syndrome (LDS), an autosomal dominant disorder caused by mutations in TGFBR1 and TGFBR2 are highly predisposed to developing allergic disease. Both LDS patients and mice harboring a knock-in allele (Tgfbr1mut) of an LDS mutation known to cause severe disease in humans exhibit increased levels of total and food-specific IgE, consistent with their allergic tendencies. These serologic changes were associated with increased numbers of T follicular helper (Tfh) cells which promote memory B cell and plasma cell formation, and decreased T follicular regulatory (Tfr) cells, which suppress humoral immune responses, in LDS patients and mice. Using a mixed bone marrow chimera model we demonstrated that these alterations in T cell development are cell-intrinsic. Furthermore, OVA specific OTII cells harboring the Tgfbr1mut were more likely to accumulate in the peyer’s patch, differentiate into Tfh cells, and less likely to upregulate Foxp3 in response to orally ingested OVA. Tgfbr1mut OTII cells also produced more IL-4 and less IL-10 in response to OVA. These findings suggest that TGF-β signaling plays an important role in T cell development and function that, when disrupted, promotes allergic inflammation.
CD8 T cell fate is programmed by a miRNA-mediated mechanism
Authors: Pobezinsky Leonid 
Published: January 2023

Abstract
CD8 T cells are the cytotoxic effectors of the adaptive immune response, clearing virally infected and cancerous cells within the host. CD8 T cells acquire their cytotoxic function by differentiating into cytotoxic T lymphocytes (CTLs). Once the CTLs have cleared the antigen, the majority of responding cells will die during contraction, while a small population of the antigen-specific responders will remain, differentiating into long-lived memory cells that provide potent protection to the host upon re-encounter with the antigen. However, during chronic infection and cancer, CD8 T cells are often diverted into the so-called exhausted state, in which CTLs are losing their function. As such, the ability to program the differentiation of CD8 T cells into a particular lineage is an important therapeutic strategy. We have identified a global post-transcriptional mechanism that directs the fate of differentiating CD8 T cells. Specifically, we have found that let-7 miRNAs are downregulated upon T cell activation, allowing T cells to differentiate into CTLs. Moreover, we have found that prolonged expression of these miRNAs programs CD8 T cells to become memory cells, while complete loss induces exhaustion. Thus, these findings may have important implications for the improvement of current immunotherapies targeting CD8 T cell fate.
Type I IFN induces vascular permeability delivering iron binding proteins to the site of Cryptococcosis gattii infection resulting in local nutritional immunity and contained infection
Authors: Davis Michael J , Pinheiro Giovana , Hoke Liz , Chang Yun , Kwon-Chung June 
Published: January 2023

Abstract
While Cryptococcus neoformans primarily infect AIDS patients, Cryptococcus gattii (Cg) causes deadly mycosis in mostly non-HIV patients even in Cg endemic areas with high rates of AIDS. Previous work found that type I IFN (t1IFN), which is induced by HIV infection, protects mice from Cg. Thus, we sought to examine the mechanisms mediating t1IFN protection from Cg. Previous data ruled out cellular immunity, so we examined soluble immune factors. We found that induction of t1IFN by stabilized poly(I:C) (pICLC) induced the expression of serum proteins into the lung airspace fluids. PICLC induced the leak of intravenously injected FITC-dextran into lung airspace fluids suggesting that serum proteins reach the lung airspace via t1IFN-induced vascular permeability. The pICLC-mediated expression of serum proteins into the lung airspace was dependent on t1IFN but not IL-6, IL-1 or TNF-α indicating that this effect is mediated by t1IFN and not by other cytokines characterized to affect vascular permeability. Transferrin and ferritin were among the leaked serum proteins found in the lung airspace and were directly inhibitory to cryptococcus in vitro so we hypothesized that the presence of these proteins in the lung airspace may result in local iron deficiency. Iron supplementation reversed pICLC-protection while iron chelation mediated similar protection as pICLC. Together these data suggest that t1IFN induces vascular permeability in the lung which delivers iron binding proteins to the Cg niche. These host proteins restrict iron to Cg and limit infection. In addition to explaining the pICLC-induced protection from Cg, these studies illustrate a novel connection between t1IFN and vascular permeability that may be significant to other systems.
Mda5/MAVS are essential for host resistance against Aspergillus fumigatus
Authors: Wang Xi , Caffrey Alayna Katherine , Espinosa Vanessa , Cunha Cristina , Croteau Walburga , Lacerda João , Campos António , Lagrou Katrien , Maertens Johan , Rivera Amariliz , Carvohlo Agostinho , Cramer Robert A , Obar Joshua J 
Published: January 2023

Abstract
Type I and III interferons act as important activators of antifungal neutrophil response in the lungs. RIG-I like receptors (RLR) are cytosolic RNA sensors that signal through the MAVS adaptor in order to activate interferon responses against viruses. Whether this pattern-recognition receptor family has broader effects on host immunity against other pathogen families remains to be fully explored. Herein we demonstrate that Mda5/MAVS signaling was essential for host resistance against pulmonary Aspergillus fumigatus challenge through the regulation of antifungal leukocyte responses. Interesting, induction of type I interferons after A. fumigatus challenge was only partially dependent on Mda5/MAVS signaling, while type III interferon expression was entirely dependent on Mda5/MAVS signaling. Ultimately, type I and III interferon signaling drove the expression of CXCL10, which is critical in resistance against invasive aspergillosis in transplant patients. Importantly, we found that polymorphisms in Ifih1 and Mavs were associated with the incidence of invasive pulmonary aspergillosis in a human HSCT cohort. Moreover, polymorphisms in the Ifih1 gene alter the inflammatory response induced in patients with invasive pulmonary aspergillosis, which include interferon-responsive chemokines. In conclusion, our data broaden the role of the RLR family in the regulating innate immunity during infection to include a role in immunity against Aspergillus fumigatus in both mice and humans.
Innate immune responses are manipulated by a streptococcal communication system
Authors: Rahbari Kate , Federle Michael 
Published: January 2023

Abstract
Group A Streptococcus (GAS) employs numerous systems to regulate genetic programs, and infection causes a spectrum of outcomes in humans ranging from commensalism to superficial disease (e.g. pharyngitis) or invasive disease (e.g. necrotizing fasciitis). Quorum sensing (QS) is a mechanism of cell-to-cell communication used by bacteria to synchronize genetic programs across populations. We previously determined that activation of the Rgg2/3 QS system in GAS results in bacterial surface alterations, leading us to hypothesize that QS could alter interactions with host cells. To test this, macrophages were infected with mutant strains of GAS containing either constitutively activated (QS-ON) or repressed (QS-OFF) Rgg2/3, and pro-inflammatory cytokine responses were measured. Macrophage responses to QS-ON GAS were attenuated, resulting in decreased NFκB activity and decreased production of TNFα, IL-6, and IFNβ compared to infection with QS-OFF GAS. Macrophage responses to mixed QS populations remained attenuated, suggesting the effect of QS-ON GAS is dominant. Furthermore, QS-ON GAS suppressed TNFα responses induced by various TLR agonists, suggesting an active mechanism of repression occurs downstream in these pathways. Together, these data suggest that GAS utilizes Rgg2/3 QS to manipulate innate immune responses. Disabling bacterial communication using a QS-inhibitor restored pro-inflammatory responses against the bacteria, highlighting its potential application as an alternative treatment strategy.
The Function of Brain Th1-like regulatory T cells in Suppressing Lethal Immune Pathology and Neurological Deterioration during Cryptococcus neoformans Meningoencephalitis
Authors: Xu Jintao , Zhao Jessica , Ivey Michael , Lopez Rafael , McClellan Brandon , Do Steven , Malepati Akanksha , Kroon Samantha , Mysel Alexa , Olszewski Michal A 
Published: January 2023

Abstract
Infection-induced inflammation in the brain frequently drives severe immunopathology, resulting in lasting neurological sequelae or high mortality. Cryptococcus neoformans meningoencephalitis (CM) is one of the leading causes of neurological death worldwide. The pathology of CM is increasingly recognized to be driven by brain inflammation, especially in immunocompetent or immune reconstituted patients. The function of FOXP3+ regulatory T (Treg) cells in maintaining immune balance under these conditions is still controversial.
Using a murine CM model, we found that Tregs massively accumulate in the mouse brain, suppressing lethal immune pathology and neurological deterioration during CM. Brain-infiltrating Tregs showed a CD44highCD62Llow activated status similar to effector T cells. Intriguingly, brain Tregs acquire a Th1-like phenotype, expressing transcription factor T-bet, IFN-γ, and chemokine receptor CXCR3. This Th1-like phenotype is critical for Treg accumulation in the brain during CM. Treg depletion using an anti-CD25 antibody intensifies CNS inflammation and results in deteriorated neurological symptoms and increased mortality. Mechanistically, we showed that CNS Tregs regulate activation of recruited monocyte-derived cells as well as cytokine production of T effector cells, both of which promote brain immunopathology during CM. Interestingly, Treg depletion only showed a minor effect on glia activation. Therapeutically, we showed that selectively enhancing Treg frequency through low-dose IL-2 treatment improves mice survival and neurological functions during CM. Our findings suggest that Tregs and their products may provide therapeutic opportunities for neuronal protection during CM.
Bacterial Extracellular Vesicle Mediated Host-Pathogen Interactions in Pneumococcal Infections
Authors: Yerneni Saigopalakrishna S , Werner Sarah , Azambuja Juliana H , Ludwig Nils , Aggarwal Surya , Eutsey Rory , Lucas Peter C , Campbell Phil G , Hiller Luisa 
Published: January 2023

Abstract
Extracellular vesicles (EVs) represent a highly sophisticated cell-to-cell mailing system across all biological kingdoms. EVs have long been characterized from many Gram-negative species and, recently from Gram-positive bacteria, including the major respiratory pathogen Streptococcus pneumoniae (pneumococcus).
Our studies reveal that pneumococcal-derived vesicles can be internalized by macrophages, T cells, and epithelial cells. In vitro, EVs induce cytokine signaling in macrophages, including dose-dependent NF-kB signaling in murine RAW 264.7 and human primary macrophages. When administered systemically into a mouse, pneumococcal EVs result in splenomegaly and induced a sepsis-like cytokine storm. When immobilized in a hydrogel implant for local administration into a mouse, pneumococcal EVs recruited lymphocytes and macrophages. Moreover, pneumococcal lipoproteins are major contributors to NF-kB signaling and inflammatory responses, as these phenotypes were substantially reduced with EVs from a lipoprotein deficient strain (Δlgt) as compared to EVs from the wildtype strain. Taken together, in vivo studies suggest that pneumococcal vesicles alone are sufficient to induce inflammatory responses and tissue damage in mammalian hosts.
Overall, our data suggest that pneumococcal EVs display potent immunomodulatory effects on host immune cells highlighting their pivotal role during the infectious process, either by manipulating host responses or by triggering host-defense systems. Thus, pneumococcal EVs are virulence determinants and may be effective tools for vaccine development.