bluuebunny/crossref_metadata_2025_split huggingface dataset:
Myeloid Cells in the Immune Response to Giardia
Authors: Fink Marc Y. , Maloney Jenny G. , Singer Steven M. 
Published: January 2023

Abstract
Infection with the protozoan parasite, Giardia, produces symptoms associated with intestinal distress, but does not induce intestinal inflammation. Long-term sequellae of this infection include malnutrition, Irritable Bowel Syndrome and Chronic Fatigue Syndrome. Previous work indicates that T-cells are critical for clearance of this parasite; however, it is unclear which cells are responsible for the activation of T-cells. As such, our goal was to examine the contribution of myeloid cells to Giardia immunity as possible T-cell activators, effector cells and/or immune regulators.
Small intestinal lamina propria cells were isolated from Giardia-infected wild-type or CCR2 deficient mice. Flow cytometry was used to determine cell phenotypes and to monitor cytokine response. EdU labeling was used to examine in situ proliferation of resident macrophages. Lastly, wild-type mice were treated with an anti-CSF1R monoclonal antibody to deplete macrophage populations.
Flow cytometry demonstrated that CD11b+, F4/80+, CD11cint, MHCII+, Ly6Cint, CX3CR1int cells accumulate in the small intestine and express ARG1 and NOS2 during infection. Intracellular cytokine staining revealed an increase in IL-10 with a concurrent decrease in TNF-α. Macrophage accumulation was the same in wild-type and CCR2-deficient mice, and EdU labeling suggested in situ proliferation of resident macrophages rather than recruitment of inflammatory monocytes. Mice treated with anti-CSF1R eliminated the parasites as well as control mice.
This study demonstrates that macrophages with a regulatory phenotype accumulate in the small intestine during Giardia infections and may explain why intestinal inflammation is not present during Giardia infections.
A malaria protein factor induces IL-4 expression by dendritic cells via TLR/MyD88-independent signaling, promoting Th2 development
Authors: Wu Xianzhu , Gowda Nagaraj M. , Gowda Channe D. 
Published: January 2023

Abstract
Dendritic cells (DCs) and cytokines produced by DCs play crucial roles in inducing and regulating pro/anti-inflammatory and Th1/Th2 responses. While DCs are known to produce Th1-promoting IL-12 in response to malaria and other pathogenic infections, it is widely thought that DCs do not produce Th2-promoting IL-4. Here, we show that a protein factor of malaria parasites induces IL-4 responses by CD11chiMHCIIhiCD3ɛ−CD49b−CD19− FcɛRI− DCs via MyD88-independent signaling. Malaria parasite-activated wild type DCs induced IL-4 responses by T cells. Interestingly, lethal parasites, Plasmodium falciparum and P. berghei ANKA, induced IL-4 response primarily by CD8α− DCs, whereas nonlethal P. yoelii induced IL-4 by both CD8α+ and CD8α− DCs. In both P. Berghei ANKA- and P. yoelii-infected mice, IL-4-expressing CD8α− DCs did not express IL-12 and a distinct population expressed IL-12. Further, in P berghei ANKA-infection, CD8α+ DCs expressed IL-12 but not IL-4, whereas in P. yoelii infection CD8α+ DCs expressed IL-4 but not IL-12. This differential IL-4 and IL-12 responses by DC subpopulations may contribute to differential Th1 and Th2 development and different clinical outcomes in lethal and nonlethal malaria. Our results for the first time demonstrated that a malaria protein factor induces IL-4 production by DCs, revealing the molecular mechanism that initiate the promotes Th2 development.
The transcription factor Bhlhe40 is a novel regulator of large peritoneal macrophages and type 2 immunity
Authors: Jarjour Nicholas N. , Bradstreet Tara R. , Schwarzkopf Elizabeth A. , Lin Chih-Chung , Cook Melissa E. , Huang Stanley Ching-Cheng , Taneja Reshma , Randolph Gwendalyn J. , Urban Joseph F. , Edelson Brian T. 
Published: January 2023

Abstract
Tissue-resident macrophages occupy key roles in immunity and physiology within organ microenvironments. Many tissue macrophages derive from embryonic progenitors and self-maintain locally as unique tissue-specific populations. Resident macrophages can also expand in response to type 2 stimuli including parasites and contribute to control of infection via the alternative activation program. However, the transcriptional basis for this capacity to proliferate in situ is poorly understood. We have observed that the transcription factor basic helix-loop-helix, member e40 (Bhlhe40) is highly expressed in a subset of hematopoietic cell types, including large peritoneal macrophages (LPMs). Based on this data, we hypothesized that Bhlhe40 is a key part of the LPM transcriptional network. We have found that peritoneal macrophages are selectively reduced in Bhlhe40−/− mice, in contrast to other resident macrophages. Mixed bone marrow chimeras, conditional knockout mice, and other approaches demonstrated a specific, intrinsic defect in mature, Bhlhe40-deficient LPMs. Bhlhe40−/− LPMs exhibited an alternative activation-like profile and impaired self-renewal. These functional perturbations were correlated with altered expression of gene sets pertaining to the endoplasmic reticulum (ER) and protein homeostasis, as well as ER morphology changes in Bhlhe40−/− LPMs. Using models of peritoneal type 2 immunity, we observed near-total loss of expansion of Bhlhe40-deficient LPMs, correlated with impaired control of an intestinal helminth. Our findings demonstrate critical roles for Bhlhe40 as a tissue-specific regulator of resident macrophage self-renewal and expansion during type 2 immunity.
Malnutrition Decreases both Effector and Memory CD4 T cells in Malaria Infection
Authors: Pilotos Jennifer , Smith Margaret Rose , Johnson Corey M. , Campbell Logan , Ibitokou Samad , Stephens Robin , Opata Michael M. 
Published: January 2023

Abstract
Malaria and other chronic infections generate effector and effector memory T cells, but the infection is not completely controlled. While malnutrition has long been suggested to be protective against malaria, recent studies have shown that malnourished children are more susceptible to malaria infections, with increased mortality rates. Current vaccination protocols result in immunity that decays, but the influence of malnutrition in the target population has not been explored. To better understand how malnutrition affects the development of protective T cells in this infection, we used a diet of moderate malnutrition to determine the activation of effector (Teff), central memory (Tcm) and effector memory (Tem) T cells. C57Bl/6 mice were fed on either protein sufficient (16%) or moderately malnourished diets (3% protein, and deficient in both Iron and Zinc) for 5 weeks. Upon infection, we observed a significant decrease in the number of effector cells at day 7 post-infection. This was accompanied by a significant drop in weight among the malnourished mice. Surprisingly, there was no significant deference in parasitemia by day 7p.i. At the memory timepoints (d60 p.i), we observed that the spleens of the malnourished mice were much smaller than the well-nourished control mice. There was a significant drop in the number of CD4 T cells and Tcm but the proportion of Tem increased in the malnourished mice and a lower cell number due to the few total CD4 T cells in the malnourished group. Our data indicate that the nutritional status of the host affect the pool of memory CD4 T cells generated in malaria infection. This could be one mechanism as to why T cell immunity decays in malaria patients as the disease is prevalent in resource-limited nations.
Epithelial TSC1-mTOR controls RIPK3-dependent necroptosis and susceptibility to IBD by sensing dietary and microbial metabolites
Authors: Xiao Hui 
Published: January 2023

Abstract
Intestinal epithelial cells (IECs) are responsible for the sensing and uptake of dietary and microbial metabolites, yet how dietary and microbial components exert on intestinal homeostasis and Inflammatory Bowel Diseases (IBDs) has yet to be better understood. Here we firstly identified epithelial necroptosis as a key process influenced by nutrients protein and fatty acid, as feeding high-protein or high-fat diet to mice led to mTOR activation and aberrant necrosis in IECs. Notably, genetic ablation of TSC1 in the IECs mimicked the effect of high-protein/fat feeding, resulting in mTOR dysregulation, epithelial necrosis and hyper-susceptibility to colitis. Mechanistically, our data demonstrated that aberrant activation of mTOR could result in the overexpression of RIPK3 in enterocytes, whereby rendering epithelial necrosis and ensuing colitis, a process could be reversed by microbiota-depletion or rapamycin treatment. Remarkably, this mTOR-RIP3-necrosis axis seemed to underlie the pathogenesis of human IBDs as well. Furthermore, we found that mTOR-regulated selective autophagy might have contributed to the degradation of RIPK3 in IECs, revealing an intricate association between autophagy and necroptosis. Together, our data unravel an unsuspected link between metabolism and necroptosis, and highlight a crucial role for TSC1-mTOR in the maintenance of intestinal homeostasis and prevention of colonic inflammation through coordinating epithelial metabolism, autophagy and necroptosis.
Protective effect of IFN-γ during experimental autoimmune encephalomyelitis is associated with the induction of inducible nitric oxide synthase.
Authors: Dietz Alina Kay , Robinson Rachel R. , Forsthuber Thomas 
Published: January 2023

Abstract
Over 2.3 million people are affected by multiple sclerosis (MS), the most common demyelinating autoimmune disease of the CNS. The relapsing-remitting form of MS is the most common initial diagnosis; however, the vast majority of patients will eventually develop secondary-progressive MS. The mechanisms that contribute to the progression of MS are largely unknown. IFN-γ, a pro-inflammatory cytokine, has long been implicated in the pathogenesis of MS and its murine model, experimental autoimmune encephalomyelitis (EAE); however, protective functions have also been demonstrated. Previous studies in our lab revealed mice lacking IFN-γ signaling developed severe and progressive EAE due to an increased presence of myelin debris and lipid peroxidation in the CNS. Recently, we identified a significant increase of inducible nitric oxide synthase (iNOS) production in the brains of WT as compared with IFN-γR−/− mice at peak of disease by qRT-PCR. iNOS is an inducible enzyme that produces nitric oxide (NO) upon activation by immunological stimuli such as IFN-γ and LPS. There are conflicting data on the role of iNOS in the pathogenesis of EAE. Our preliminary findings indicate macrophages are the primary contributors to iNOS expression during disease progression. To further explore the role of iNOS in disease progression, we are studying the underlying mechanisms, key molecules involved in this process, and the effects of treatment with iNOS inhibitors on disease course, general pathology and biochemical composition of the CNS. Our results will contribute to a better understanding of the underlying mechanisms and may lead to new treatments to prevent progression of MS.
The Effect of Helminth-Induced Immunomodulatory Therapy (HINT) in Relapsing Remitting Multiple Sclerosis
Authors: Hernandez Gianna L. , Ganansky Caleb , Sandor Matyas , Fleming John , Fabry Zsuzsanna 
Published: January 2023

Abstract
This study examines the immune response stimulated by Trichuris suis ova when used as a treatment for Multiple Sclerosis. The hygiene hypothesis posits that insufficient exposure to microorganisms due to modern sanitation can predispose susceptible individuals to immune dysregulation. A pilot HINT 1 study indicated that TSO could effectively regulate the immune system of relapsing-remitting MS patients. Here, we investigate this hypothesis further with a longer treatment period. The HINT phase 2 clinical trial consisted of 16 treatment-naïve patients with newly diagnosed RRMS. Patients were given TSO buffered with 15mL of Phosphate buffered saline every two weeks. Anti-TSO immune response was measured by IgG1, IgA and IgE ELISAs of patient serum and eosinophilia count. Changes in immune cell populations were monitored by flow cytometry of peripheral blood mononuclear cells. CD4+ T cells were further analyzed as Th1, Th2, Th17, TREG, and TFH cells. During TSO treatment, the average number of new Gd+ lesions in the HINT patients was reduced. In addition, there was a significant increase in the anti-TSO immune responses as measured by IgG1 and IgE ELISAs, and increased eosinophilia during the second month of treatment. Also, TSO administration led to a decrease in the percentage of activated CD4+ and CD8+ T cells during therapy accompanied by an increase in the presence of TREG cells during treatment, a reduction in the presence on TFH cells in circulation, and an increase in IL-4-expressing T cells. This data implies a shift from a Th1 inflammatory state to a Th2 anti-inflammatory state during HINT II TSO treatment and suggests that immunoregulatory therapy through probiotic helminth treatment could be a viable option as a treatment for MS.
Oral Therapy with Colonization Factor Antigen I (CFA/I) Activates Stable Regulatory T Cells (Tregs) in the Pancreatic Lymph Nodes (PaLNs) of Non-Obese Diabetic (NOD) Mice
Authors: Nelson Andrew Scott , Maddaloni Massimo , Hoffman Carol , Abbott Jeffrey , Pascual David 
Published: January 2023

Abstract
Type 1 diabetes (T1D) is an autoimmune disease in which insulin-producing pancreatic β cells are destroyed by auto-aggressive T cells. Tregs in T1D have been shown to lose their suppressive function and/or become inflammatory, complicating Treg immunotherapy. Oral treatment with the adhesin from enterotoxigenic E. coli, CFA/I fimbriae, protects against autoimmunity. Although CFA/I fimbriae act in a bystander or in an antigen (Ag)-independent fashion, protection is ultimately dependent upon the induction and/or activation of auto-Ag-specific Tregs. Our initial findings showed that oral dosing with a Lactococcus lactis vector expressing CFA/I fimbriae (LL-CFA/I) reduces incidence of T1D in NOD mice by 45%. We hypothesized that dosing NOD mice more frequently would induce Foxp3+ Tregs in the pancreas and PaLNs to provide more robust protection. To test this, 4 wk-old NOD mice were orally dosed with LL-CFA/I varying the dosage and frequency of treatment. When dosed every 2 wks, the frequency of insulitis was reduced by more than half at 11 wks of age, and tetramer staining revealed a 2-fold reduction of insulin-specific CD4+ and CD8+ T cells within the PaLNs. Additional groups were followed until 16 wks of age, and examined for Treg and inflammatory T cell phenotypes. Treatment with LL-CFA/I reduced 2-fold IFN-γ-producing CD8+ T cells in the spleen and PaLNs. PaLN Foxp3+ T cells from the PBS and vector control groups, but not the LL-CFA/I-treated mice, co-expressed Tbet and IFN-γ, suggesting that Tregs from LL-CFA/I treated mice maintained their regulatory function at 16 wks. These data show that orally dosing with LL-CFA/I diminishes or halts T1D progression via activation of stably functional Foxp3+ Tregs. Work supported by AI121745.
Human alloreactive-regulatory T cell profile after ex vivo expansion using CD40L-stimulated B cells or monocyte-derived dendritic cells
Authors: Lee Linda M. , Zhang Hong , Liang Horace , Lee Karim , Thomson Angus W. , Tang Qizhi 
Published: January 2023

Abstract
Alloreactive regulatory T cells (arTregs) expanded with CD40L-stimulated B cells (sBcs) are being tested clinically to induce tolerance and control rejection in liver and kidney transplantation. In research settings, allogeneic monocyte-derived DCs (moDCs) have been shown to expand arTregs. We directly compared the efficacy of sBcs and cytokine-matured moDCs in expanding arTregs, and the phenotype of sBc-stimulated arTregs (sBc-arTregs) and moDC-stimulated arTregs (moDC-arTregs). arTregs expanded twice as much after stimulation with moDCs than with sBcs. Neither sBcs nor moDCs produced much TNFα, IL-1α, IL-1β, IL-6, IL-12P70, or IL-23 that can destabilize Tregs. sBc- and moDC-arTregs expressed similar percentages of Treg-defining markers - FOXP3, HELIOS, CD25, CD27, and CD62L. moDC-arTregs had slightly elevated FOXP3 and CD62L MFI. Compared to primary Tregs, sBc- and moDC-arTregs expressed similar or decreased levels of TBX21, GATA3, and RORC mRNA. However, greater percentages of moDC-arTregs produced IFNγ, IL-4, and IL-17A, and a greater percentage of sBc-arTregs produced IL-17A, but far less than those percentages produced by similarly stimulated conventional CD4+ T cells (Tconvs). The majority of cytokine-expressing arTregs were FOXP3+. Percentages of CXCR3 and CCR4, but not CCR6, increased after sBc and moDC stimulation. Using a 770-gene Nanostring panel and unsupervised clustering, sBc- and moDCs-arTregs were found to be mostly similar, but distinct from primary Tregs and further separated from Tconvs. Thus, arTregs expanded by sBcs or moDCs are phenotypically similar, likely stable, and had features that may enable them to traffic to inflammatory sites, e.g. transplanted organs undergoing alloimmune attack.
Prolonged cold ischemia storage of renal allografts shifts DSA specificity and enhances glomerular injury.
Authors: Gorbacheva Victoria Y. , Fan Ran , Baldwin William , Valujskikh Anna 
Published: January 2023

Abstract
Prolonged cold ischemia storage (CIS) of donor organs is a major risk factor for allograft injury. We tested how increased CIS time influences humoral alloimmunity in a mouse model of renal transplantation.
B6 (H-2b) mice received BALB/c (H-2d) renal allografts subjected to 0.5 h or 6 h CIS. At 14 d posttransplant, 6 h CIS recipients had increased titers of donor-MHC class II but not class I reactive IgG DSA, elevated frequencies of both class I and class II-reactive antibody secreting cells, higher frequencies of donor-reactive IFNγ-secreting T cells and higher proportion of graft glomeruli infiltrated with macrophages compared to 0.5 h CIS controls. B cell depletion inhibited DSA generation, intragraft C4d deposition and macrophage glomerular infiltration on d14.
Regardless of CIS duration, serum creatinine levels at d 60 posttransplant were comparable to non-transplanted mice. By d 60, 0.5 h CIS grafts had moderate segmental sclerosis in a limited number of glomeruli. In contrast, 6 h CIS grafts had extensive glomerular injury including thickened capillary loops, sclerosis, thrombotic microangiopathy, intracapillary fibrin thrombi, and mesangiolysis, and elevated intragraft levels of acute kidney injury markers NGAL and osteopontin. In addition, 6 h CIS recipients had increased serum levels of anti-class II but not anti-class I IgG DSA whereas the frequencies of IFNγ-producing donor-reactive T cells were low and comparable in both groups.
Our findings suggest that the augmented humoral rather than cellular immune responses account for the transplant glomerulopathy after prolonged CIS. Posttransplant inflammation spreads the specificity of DSA responses from class I to class II and may thus influence renal allograft pathology.
Vasoactive intestinal peptide synthesis in transplant recipients regulate donor T cell alloreactivity and graft-versus-host disease
Authors: Li Yiwen , Waller Edmund K. 
Published: January 2023

Abstract
Vasoactive intestinal polypeptide (VIP) is an immunosuppressive neuropeptide with pleiotropic effects. Activation of donor T cells leading to graft-versus-host disease (GvHD) in allogeneic bone marrow transplant (allo-BMT) causes secondary upregulation of immune co-inhibitory pathways, including VIP expression in T cells and dendritic cells. VIP signaling favorably regulates the activation status and the graft-versus-leukemia (GvL) activities of donor T cells (Li et al., Cancer Research 2016). We tested the hypothesis that absence of VIP expression by recipient cells would enhance donor T cell activation and increase GvHD in allo-BMT. VIP-KO mice transplanted with TCD BM alone had only 43.3% survival at day 75 compared with 73.3% survival among the WT recipients with no different in levels of donor chimerism. The addition of donor splenocytes to the graft resulted in significantly more GvHD-related mortality in VIP-KO recipients compared with WT recipients, with 80% vs 100% day 75 survival among recipients of 1×106 splenocytes (p=NS) and 0% vs 40% survival (p&lt;0.01) among recipients of 3×106 donor splenocytes. VIP-KO recipients transplanted with wildtype cells had similar co-inhibitory pathway expression (PD1, Lag3, Tim3) but elevated cytokine production of IL2, IL4, IL10, IFNγ and TNFa by donor CD4+ and CD8+T cells post allo-BMT compared to WT recipients. VIP is expressed by residual host leukocytes up to 10 days post-transplant confirmed by confocal microscopic imaging. Host cells express GFP transgene under the control of VIP promoter, consistent with the hypothesis that VIP production by host antigen presenting cells represents a critical pathway in regulatory immune activation of donor T cells.
Recipient mucosal-associated invariant T cells control graft-versus-host-disease within the colon.
Authors: Varelias Antiopi , Bunting Mark , Ormerod Kate , Koyama Motoko , Olver Stuart , Straube Jasmin , Kuns Rachel , Robb Renee , Henden Andrea , Cooper Leanne , Lachner Nancy , Gartlan Kate , Lantz Olivier J. , Kjer-Nielsen Lars , Mak Jeffrey , Fairlie David , Clouston Andrew , McCluskey James , Rossjohn Jamie , Lane Steven , Hugenholtz Phil , Hill Geoff 
Published: January 2023

Abstract
Mucosal-associated invariant T (MAIT) cells are a unique innate-like T-cell subset that responds to a wide array of bacteria and yeast through recognition of riboflavin metabolites presented by the MHC I-like molecule, MR1. Here we demonstrate using MR1 tetramers that recipient MAIT cells are present in small but definable numbers in graft-versus-host disease (GVHD) target organs and protect from acute GVHD in the colon following bone marrow transplantation (BMT). Consistent with their preferential juxtaposition to microbial signals in the colon, recipient MAIT cells generate large amounts of IL-17A, promote gastrointestinal tract integrity, and limit the donor alloantigen presentation that in turn drives donor Th1 and Th17 expansion specifically in the colon after BMT. Allogeneic BMT recipients deficient in IL-17A also develop accelerated GVHD, suggesting MAIT cells regulate GVHD, at least in part, by the generation of this cytokine. Indeed, analysis of stool microbiota and colon tissue from IL-17A−/− and MR1−/− mice identified analogous shifts in microbiome operational taxonomic units (OTU) and mediators of barrier integrity which represent pathways controlled by similar, IL-17A-dependent mechanisms. Thus, MAIT cells act to control intestinal microbiota and barrier function to attenuate pathogenic T-cell responses in the colon, and given their very high frequency in humans, likely represent an important population in clinical BMT.