bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD4+ resident memory T cells dominate immunosurveillance and orchestrate local recall responses
Authors: Beura Lalit K , Fares-Frederickson Nancy J , Steinert Elizabeth Marie , Scott Milcah C , Thompson Emily A , Fraser Kathryn A , Schenkel Jason M , Vezys Vaiva , Masopust David 
Published: January 2023

Abstract
Migratory dynamics of T cells influence outcomes of infections and cancer. This study addresses how CD4+ T cell immunosurveillance strategies compare to CD8+ T cells by analyzing migration, transcriptome, phenotype, and function. After acute viral infection, memory CD4+ T cells predominantly utilized residence to survey nonlymphoid tissues, albeit not as stringently as observed for CD8+ T cells. In contrast, memory CD4+ T cells were more likely to be resident within lymphoid organs than CD8+ T cells. Migration properties of memory-phenotype CD4+ T cells in non-SPF parabionts were similar, generalizing these results to diverse infections and conditions. CD4+ and CD8+ resident memory T cells (TRM) shared overlapping transcriptional signatures and location-specific features, such as granzyme B expression in the small intestine, revealing tissue-specific and migration property-specific, in addition to lineage-specific, differentiation programs. Functionally, mucosal CD4+ TRM reactivation locally triggered both chemokine expression and broad immune cell activation. Thus, residence provides a dominant mechanism for regionalizing CD4+ T cell immunity, and location enforces shared transcriptional, phenotypic, and functional properties with CD8+ T cells.
Single-cell mRNA analysis of colon phagocyte heterogeneity identifies two major macrophage developmental pathways
Authors: Kang Byunghyun , Alvarado Luigi J. , Lehman Michael , Kim Bong-Hyun , Li Peng , Cho Hyeseon , He Jianping , Larochelle Andre , Kelsall Brian L 
Published: January 2023

Abstract
Tissue macrophages (MPs) develop distinctive functions due to unique transcriptional programs driven by local environmental ques. In the intestine, it is known that MPs are heterogeneous cells that differentiate from blood monocytes and embryonic precursors, and are essential for homeostasis with the vast microbiome through their ability to kill invading microbes, clear apoptotic cells, and produce regulatory cytokines. However, the full extent of MP heterogeneity, their developmental relationships, and how the intestinal microbiota affects MP generation is not known. Here, we performed single cell RNA sequencing of colonic myeloid cells from specific pathogen free (SPF) and germ free (GF) C57BL/6 mice and found extensive heterogeneity of colon MP, with at least six different identifiable populations, with less heterogeneity of dendritic cells (DCs). Unsupervised modeling of developmental pathways predicted two major trajectories from common CCR2+precursors resulting in CD11c+CD9+MMRintCD121b+and CD11c−CD9intMMRhiCD121b−MP populations. The generation of both mature populations of colon MPs was impaired in germ-free mice, which coincided with the appearance of a dominant unique population of MPs, whereas the generation of DC populations in GF mice were unaffected or increased. Furthermore, the two major MP populations were clearly distinct from other populations regarding their gene expression profile, localization within the lamina propria, and ability to phagocytose macromolecules from the blood. These data uncover the diversity of intestinal myeloid cell populations, highlight the importance of microbiota on the unique developmental as well as anatomical and functional fates of colon MPs.
Examining the Immune Composition of the Intestinal Mucosa by Mass Cytometry
Authors: Tyler Christopher J , Lundborg Luke , Yeasmin Shaila , Rivera-Nieves Jesus 
Published: January 2023

Abstract

BACKGROUND
The inflammatory bowel diseases (IBD) can affect various segments of the gastrointestinal tract, and the biology behind regional differences in disease remains poorly understood, in part due to a lack of understanding of regional variation of the particular cell subsets present. By utilizing mass cytometry, our objective was to identify regional differences in the immune composition of the digestive tract in healthy individuals.

METHOD
Intestinal pinch biopsies were obtained from the following sites of healthy individuals (n=10); duodenum, terminal ileum, ascending colon, transverse colon, descending colon, and rectum. Immune cells were isolated from biopsies and stained with a panel of 37 markers for analysis using mass cytometry.

RESULTS
Marked variations in immune cell composition were observed along the digestive tract, in particular T and B lymphocytes. Increased abundance of T lymphocytes (&gt;30% of total CD45+ cells) was identified in the duodenum and terminal ileum, whereas B lymphocytes became more abundant (&gt;30% of total CD45+ cells) towards the distal end of the tract. In addition, intestinal homing molecules displayed variation at each site; integrin α4β7, a target for IBD therapeutic intervention, was increasingly expressed in the terminal ileum and ascending colon, compared with the descending colon (Mean 40% vs 25% abundance, n=10, p&lt;0.01).

CONCLUSIONS
Mass cytometry has the ability to identify and characterize numerous distinct immune subsets throughout the intestinal mucosa, allowing a deep understanding of regional differences in immune composition. This understanding may shed light on the pathogenesis of IBD and allow standardization of intestinal sampling during the evaluation of therapeutics.
Pulmonary resident effector CD4+ T cells fail to effectively utilize glycolysis
Authors: Roberts Lydia M , Evans Tyler J , Bosio Catharine M 
Published: January 2023

Abstract
The transition of T cells from naïve to effector cells has been reported to require a metabolic shift from oxidative phosphorylation to glycolysis for the production of effector molecules such as IFN-γ. This phenomenon is primarily restricted to study of splenic T cells and it is largely unknown if T cells in other organs undergo the same metabolic shift. Intranasal vaccination establishes a pool of lung tissue-resident, effector memory T cells (trTem) that are typically sufficient to control a secondary infection via production of cytokines. Therefore, we hypothesized trTem would be highly glycolytic upon activation. To test this hypothesis, we compared the metabolic activity of both trTem and circulating Tem (cTem) isolated from mice vaccinated with either Bordetella pertussis or Francisella tularensis. As expected, regardless of the vaccinating pathogen, CD4+ cTem readily shifted to glycolysis following activation with anti-CD3/CD28 beads. In contrast, CD4+ trTem cells did not shift to glycolysis following the same activation scheme, nor did they increase mitochondrial respiration as a mechanism to compensate for the absence of glycolysis. Consistent with an inability to shift to glycolysis, ex vivo stimulation of trTem with antigen also resulted in limited uptake of glucose as compared to cTem. These data suggested that, unlike previous reports, pulmonary CD4+ trTem do not rapidly shift to glycolysis even in the presence of antigen. Together these data highlight the unique environment of the pulmonary compartment and present critical information that will influence our understanding of pulmonary T cell responsiveness during infection which may shape future vaccination strategies.
Distinct Roles of TβRIII and LRP-1 in Lactoferrin-mediated Foxp3+ Inducible Tregs
Authors: Kim Sun-Myoung , Jang Young-Saeng , Kim Pyeung-Hyeun 
Published: January 2023

Abstract
We recently found that lactoferrin (LF) stimulated mouse naïve CD4+T cells to differentiate into inducible Foxp3+Tregs (iTregs). However, LF decreased cell proliferation, CD62L shedding, and CD44 expression. In the present study, we characterized these phenomena at the receptor level. Addition of excess amount of soluble TβRIII (betaglycan) virtually abrogated the LF-induced Foxp3 expression but little affected LF-suppressed cell proliferation/CD62L shedding/CD44 expression. On the other hand, treatment of anti-LRP-1 Ab slightly enhanced LF-induced Foxp3 expression while substantially restored LF-suppressed cell proliferation/CD62L shedding/CD44 expression. These results indicate that LF enhances Foxp3 expression mainly through TβRIII and simultaneously suppresses cell proliferation/CD62L shedding/CD44 expression mainly through LRP-1. Thus, it is assumed that LF renders activated CD4+T cells to become central memory-like Tregs (CD4+Foxp3+CD62L+CD44lo).
BATF is an essential regulator of migratory ILC2 cell fate and function
Authors: Miller Mindy M , Patel Preeyam , Bao Katherine , Danhorn Thomas , O’Connor Brian , Reinhardt R Lee 
Published: January 2023

Abstract
A recently identified subset of group-2 innate lymphoid cells (ILC2s) migrates to mucosal tissues early during type-2 inflammation and is characterized by responsiveness to interleukin-25 and high expression of KLRG1. This study identifies the major transcriptional regulator required for their generation and function. RNA-sequencing and unbiased pathway analysis revealed the AP-1 transcription factor BATF as a potential modulator of ILC2 cell fate. Indeed, infection of BATF-deficient mice with the helminth N. brasiliensis showed a selective defect in IL-25-induced, ILC2-mediated worm clearance as well as a corresponding loss of IL17RB+ (IL-25 receptor) KLRG1high ILC2s in the lung. Moreover, these BATF-dependent migratory ILC2s serve as the major early source of IL-4 and IL-13 after infection. Interestingly, while the presence of KLRG1high ILC2s in the small intestine are unaffected by BATF-deficiency, their ability to produce IL-13 is significantly impacted. Defects in IL-13 expression by intestinal KLRG1high ILC2s corresponded with decreased goblet and tuft cell hyperplasia in BATF-deficient mice, supporting the critical role of these migratory ILC2s as early sentinels of mucosal barrier integrity.
Bivalent anti-CD3 Fab dimers drive T cells into a highly efficient fratricide with potential clinical application
Authors: Pages Diana Gil , Nelson Alfreda D , White Tommi A , Schrum Adam G 
Published: January 2023

Abstract
Non-covalent anti-CD3 Fab dimers (Bi-Fabs) bind to the TCR/CD3 complex and activate T cells. In the presence of proper co-stimulation, anti-CD3 Bi-Fabs promote T cell cycling and acquisition of effector functions on CD4 and CD8 T cells more robustly than other bivalent antibody species like F(ab′)2 and mAb with the same specificity. Surprisingly, the net effect of anti-CD3 Bi-Fab treatment on T cells in vitro is contraction of CD4 and CD8 T cells, via AICD. This is in contrast with the net T cell proliferation observed with soluble anti-CD3 mAbs that are stimulatory in the presence of proper co-stimulation. Bi-Fab binding to the TCR/CD3 complex, and direct cellular contact with Bi-Fab engaged T cells, are required to observe the net contraction of treated T cells. Using perforin-deficient T cells, and Fas-FasL blockade, we show this net contraction results from T cell apoptosis via Fas-FasL and CTL degranulation. Structural studies on Bi-Fabs suggest a rigid rod like structure. We hypothesize that the rigid rod allows for a potent crosslinking structure with both paratopes from the anti-CD3 Fabs capable to engage effector T cells on fratricide. In vivo, mice injected with anti-CD3 Bi-Fabs present depletion of T cells from blood, and absence of any adverse effects derived from T cell activation like cytokine storm. We have begun exploring the therapeutic potential of T cell depletion in mouse models of autoimmune disease with promising results.
Genetic and pharmacological validation of TAK1 inhibition in macrophages as a therapeutic strategy to effectively inhibit TNF secretion
Authors: Scarneo Scott A , Eibschutz Liesl , Hughes Phillip , Haystead Timothy 
Published: January 2023

Abstract
Immune challenge of invading macrophages at sites of infection is associated with release of TNF, which triggers a local cytokine storm as part of the normal inflammatory response. Whereas this response maybe beneficial in fighting off infections, similar responses triggered in autoimmune diseases contribute significantly to the underlying damaging pathology associated with these diseases. Here we show that Takinib, a highly discriminatory inhibitor of transforming growth factor Beta-activated kinase 1 (TAK1), selectively and potently reduces TNF production in pro-inflammatory THP-1 macrophages. A complete survey of 110 cytokines, showed robust loss of proinflammatory cytokine responsiveness to lipopolysaccharide (LPS) and interferon gamma (IFNg) challenge in response to Takinib. The mechanisms of action of Takinib was recapitulated in TAK1 KO macrophages. TAK1 KO cells showed significant loss of TNF production as well as release of IL-6 in response to LPS challenge. Furthermore, Takinib blocked the ability of exogenously added LPS to promote phosphorylation of, c-Jun, p38 protein kinases as well as downstream transcription factors regulated by nuclear factor κ-light-chain-enhancer of activated B cells (NFκB). In a mouse LPS challenge model, Takinib significantly reduced TNF serum levels. Our findings demonstrate that Takinib has utility in the treatment inflammatory disease by locally suppressing TNF production from invading macrophages.
Controlling the Residual Inflammatory Risk in Type 1 Diabetic Atherosclerosis
Authors: Yussman Martin , Wagner David H , Vaitaitis Gisela , Waid Dan 
Published: January 2023

Abstract
Prevention trials with statins reduce the relative risk of cardiovascular (CV) events by 10 to 40%. This leaves a ‘residual risk’ of 60–90% for which the CANTOS trial (IL1β inhibition) provided proof of concept that targeting inflammation reduces CV event rates; unfortunately, it did not change rates of type 1 diabetes (T1D) and infections in T1D patients were higher. A major molecular driver of auto-inflammation in both diseases is the CD40/CD154 inflammatory dyad with unique T cell subset, CD3+CD4+CD40+ (TH40 cell) known to be significantly expanded in T1D subjects. Normalization of the aberrant contact dependent interaction of this dyad has shown efficacy in animal models using antibody to CD154, reversing T1D and rate of atherosclerosis, but unfortunately has deleterious side effects when given in humans.
We measured TH40 cells in T1D subjects with high calcium scores (a measure of atherosclerotic burden) and those with low calcium scores. Additional to the higher percentage of TH40 cells compared to controls, we have found a statistically significant elevation in in INFg production in the high calcium score cohort, indicating a more pathogenic state of these cells.
We designed peptides capable of binding directly to the CD40 receptor, which have been shown to reverse T1D in animals. Peptide was administered to ApoE−/− mice, a model of atherosclerosis. Immunohistochemical staining demonstrated significant reduction in plaque as well as smooth muscle and collagen content. In-vitro analysis of TH40 mouse splenic cells showed modulation in IL2, INFγ and IL17, all potent cytokines of atherosclerosis.
This new information relays promise for a more directed diagnostic and therapeutic target for T1D related atherosclerosis.
Modeling graft-vs-host disease in humanized mice using different hematopoietic stem cells sources reveals inherent differences in Tc and myeloid cell biology
Authors: Hess Nicholas J , Hematti Peiman , Gumperz Jenny E. 
Published: January 2023

Abstract
Cancers and autoimmune diseases, which affect 1.6% and 4.9% of the U.S. population respectively, are often treated with chemotherapy and immunosuppressive drugs with variable response rates. One alternative treatment option are hematopoietic stem cell (HSC) transplantations. Unfortunately, the biology behind these transplants remains unclear despite being in practice for over 60 years. Our research aims to use a humanized mouse model to understand the different molecular mechanisms that lead to acute GVHD, hematopoietic recovery/rejection and GVL. Here, we will focus on our data recapitulating clinical GVHD in different graft sources. Injecting equivalent numbers (2E7) of bone marrow (BM) mononuclear cells and G-CSF mobilized (MB) mononuclear cells into non-irradiated NSG mice results in 50% & 0% penetrance of GVHD respectively at 12 weeks. Concurrently, we have measured the Tc proliferation and IFNγ secretion in these mice over the course of 12 weeks that reveals an elevated activation status in BM Tc’s from an early stage. Interestingly, stimulating the Tc’s directly in vitro results in equivalent activation potentials suggesting a prominent role of myeloid cells in the induction of GVHD and not Tc’s. The following investigations have shown an increased expression of the B7 complex on BM myeloid cells and an increase in their IL-6 secretion. These early observations suggest that the “accessory cells” injected from the different HSC grafts are vastly important for driving clinical outcomes. Understanding the molecular cross-talk between the “accessory” cells and the underlying cellular differences attributed to each HSC source will better inform clinicians on the best treatment practices for patients.
Defining the role of graft-infiltrating B cells in the acute rejection of kidney allografts in mice
Authors: JAIN DHARMENDRA , Yin Dengping , Chong Anita S 
Published: January 2023

Abstract
B-cell infiltrates has been observed in kidney biopsies undergoing clinical cellular or mixed rejection. Some studies suggest poorer graft survival is associated with the presence of B cell infiltrates in biopsies diagnosed with cellular rejection, whereas others have not. We have developed a mouse kidney transplant model to study the role of donor-specific B cells infiltrate kidney allografts undergoing acute rejection. To this end, C57BL/6 female mice were subjected to left nephrectomy and were transplanted with kidneys from BALB/c mice expressing 2W-OVA, transgene. Donor-specific B and T cells were identified with donor MHC Class I & II tetramers. All graft were rejected by D28 post-transplant and showed an average 36-fold increase in donor specific antibodies (DSA). At D14, ~5% of the donor specific B cells had a Fas+GL7+ germinal center phenotype in both SLO’s (pooled spleen and peripheral lymphnodes) and kidney allografts. In addition, about 47% of donor-specific T cells in both the SLO and allograft had a CD4+ICOS+ expression, suggesting Tfh phenotype. ~4000 and ~900 Anti Kd-IgG secreting B cells (ASC’s) were detected in both SLO’s and in kidney allografts, respectively. Our data raise the possibility that in situ donor-specific T and B cell interactions are occurring in SLOs and also in the rejecting allografts. We plan to use this transplant model to further investigate whether graft infiltrating B cells serve as antigen-presenting cells to alloreactive T cells mediating acute rejection, whether these B cells differentiate into ASC in situ, and to define the relationship between the intrarenal B cells and ASCs in the allograft and those expanded in the SLO.
The Th17 lineage in heart transplant rejection: Examining plasticity and pathogenicity
Authors: Williams Linus Clark , Iacomini John 
Published: January 2023

Abstract
Heart transplants are the only treatment for end-stage heart failure. Over 3000 heart transplants were performed in the United States in 2016, making heart transplant the most common cadaveric organ transplant. Long-term survival of heart transplants remains a significant clinical concern, and median survival rates after the first year have not significantly improved in 25 years. We have previously shown that pro-inflammatory co-morbidities such as hyperlipidemia contribute to accelerated IL-17 dependent rejection of the transplanted heart. Hyperlipidemia, an increased level of triglycerides and lipids in the blood, is extremely prevalent in the transplant patient population, with nearly 90% of heart transplant recipients becoming hyperlipidemic after their transplant, despite universal use of statins in the transplant patient population. The work described in this poster aims to further explore the mechanisms by which hyperlipidemia promotes Th17-mediated graft rejection, especially the potential interplay between the hyperlipidemic state and CD4 T cell plasticity. By using a MHC-II mismatched heart graft model, immunocytochemistry staining, a Th17 lineage trace, and genetic and chemical ablation of inflammatory cytokines and lineages, we aim to elucidate the differences in inflammatory T-cell infiltrate between graft-destructive accelerated hyperlipidemic rejection and chronic non-hyperlipidemic transplant rejection. This work may lead to more precise targeting of immunosuppression in heart transplant patients and improved survival and quality of life in this patient population
The enigmatic impact of donor T cell subsets on the therapeutic efficacy of tolerogenic protocols for transplant rejection
Authors: Lozano Marcos Iglesias , Bibicheff Darrel , Chicco Maria , Brandacher Gerald , Raimondi Giorgio 
Published: January 2023

Abstract
Costimulation blockade (CoB) regimens are a promising immunomodulatory strategy to prevent transplant rejection, especially when combined with antigen specific therapy. However their efficacy is affected by multiple factors. Recent reports have highlighted the unexpected capacity of passenger donor lymphocytes to enhance the recipient’s anti-graft response.
In this study we assess if T lymphocytes contained in the donor specific infusion (DSI) used in combination with anti-CD154 (MR1), a powerful CoB regimen, impact the regulation of mouse skin transplant rejection. Our data indicated donor T cells persist for at least 7d after infusion. When T cells were depleted from the DSI, transplant survival was significantly increased (MST=103 vs 68). Unexpectedly, the removal of CD4 T cells did not improve survival (MST=66). Concordantly, the addition of CD8+CD44hi memory T cells to T-depleted DSI restored the limited survival to that of full DSI (MST=71). ELISPot and flow cytometry results showed a greater anti-donor memory response in post-rejection spleen samples. Surprisingly, the removal of donor CD8 T cells in the DSI inoculum induced a survival even greater than that achieved with total T cell depletion (MST=148). The addition of CD4+CD44hi memory T cells to T-depleted DSI (inoculum still depleted in CD8 T cells), matched the survival of T-depleted DSI (MST=113) and was not associated to any increment in anti-donor memory response post-rejection.
Overall, these data reveal a novel role for donor lymphocytes in the modulation of recipient’s alloimmunity by DST+MR1. Identification of the mechanisms and specificities underlying this effect will be pivotal for the optimization of clinically effective tolerogenic therapies.