bluuebunny/crossref_metadata_2025_split huggingface dataset:
Neuroinflammatory Signatures of Complement Component 4 in the Subventricular Zone of Autism Spectrum Disorder and Schizophrenia
Authors: Mou Ta-Chung , Lane Malcolm , Ireland Derek D C , Clark Sarah M , Tonelli Leonardo H 
Published: January 2023

Abstract
Early inflammatory insults are the most recognized environmental risk factor associated with neurodevelopmental psychiatric disorders. The subventricular zone (SVZ) is a region heavily involved in neurodevelopment and neuroimmune interactions through the lifespan. Complement component 4 (C4), a classical molecule involved in inflammatory responses, has been associated with schizophrenia (SZ) by genetic studies. However, whether C4 is involved in other neurodevelopmental disorders, such as autism (ASD), is a matter of current studies. Moreover, while C4 in SZ has been studied in the context of synaptic pruning, little is known about its neuroinflammatory role. Using in situ hybridization with radioactive riboprobes and RNAscope, we identified robust astrocytic expression of C4 in the SVZ and in the septum pellucidum. C4 was also expressed in ependyma, neurons, and Ki67+ cells, but not microglia, with higher expression in SZ as compared to controls and ASD. Moreover, the number of Ki67+ cells was significantly higher in ASD compared to SZ and controls. Targeted transcriptomic analysis of inflammatory pathways revealed a strong association of complement system genes with SZ, and to a lesser extent ASD, as well as generalized inflammation without an association with known infectious pathways. Additional analysis of differentially expressed genes (DEGs) showed that ASD DEGs were enriched in adaptive immune system functions such as Th cell differentiation, while SZ DEGs were enriched in innate immune system functions, including NF-κB and toll like receptor signaling. Taken together, these results support a role for C4 into inflammatory-neuroimmune dysregulation observed in SZ and ASD pathology.
Supported by National Institutes of Mental Health grant P50 MH103222 (Silvio O. Conte Center for Translational Mental Health Research).
PD-1 restricts the development and effector function of tissue regulatory T cells in experimental autoimmune encephalomyelitis
Authors: Liang Dan , Judge Jennifer , Markson Samuel , Guinn Samantha , Collier Jenna Lynn , Shahid Osmaan , Manne Sasikanth , Kuchroo Juhi , Fung Megan , Pauken Kristen E , Vignali Dario A A , Wherry John E , Sharpe Arlene H 
Published: January 2023

Abstract
PD-1 restricts T cell effector functions, regulates T cell tolerance, and maintains immune homeostasis, thereby playing pivotal roles in cancer, autoimmune and infectious diseases. Our studies and others show that PD-1 can restrain Treg suppressive function in autoimmunity and cancer. Here, we investigate the role of Treg intrinsic PD-1 in regulating pathogenic and protective CD4+ T cell responses in experimental autoimmune encephalomyelitis (EAE) by inducing PD-1 deletion on FoxP3+ cells prior to disease development. This deletion of PD-1 only in Treg protects mice from severe disease, and both flow cytometry and transcriptional analysis of central nervous system (CNS) CD4+ T cells show enrichment of tissue effector Treg (eTreg) subsets at peak of disease. We find that PD-1 restrains the generation and suppressive function of these effector Treg. This tissue eTreg signature is also observed in human and mouse Treg subsets from the tumor microenvironment, and are potent suppressor populations. Further studies are underway to determine how PD-1 impacts TCR signaling and FoxP3 transcriptional co-activators in Treg cells. Our study provides deeper insights into how PD-1 regulates T cell tolerance, underscoring the important role of PD-1 in regulating Treg differentiation and suppressive function during autoimmune diseases, and provide mechanistic insights for PD-1 modulation in cancer and autoimmunity.
Supported by NIH P01AI108545
Regulation of myeloid-derived suppressor cell expansion by TRAF3 during chronic inflammation
Authors: Jung Jaeyong , Zhu Sining , Lalani Almin , Sant’Angelo Derek B , Covey Lori R , Ostrand-Rosenberg Suzanne , Xie Ping 
Published: January 2023

Abstract
Myeloid-derived suppressor cells (MDSCs) expand under pathological conditions and orchestrate the immunosuppressive and inflammatory network, and thus are recognized as a prime therapeutic target of human cancers and inflammatory diseases. In this study, we identified the tumor suppressor TRAF3 as a critical regulator of MDSC expansion. We found that myeloid cell-specific Traf3-deficient (M-Traf3−/−) mice exhibited MDSC hyperexpansion during chronic inflammation. Using mixed bone marrow chimeras, we demonstrated that TRAF3-mediated regulation of MDSC expansion involves cell-extrinsic mechanisms. To delineate the environmental factors that induce the aberrant expansion of MDSCs in M-Traf3−/− mice, we performed cytokine protein array analyses and in vivo neutralization experiments. Our results revealed that G-CSF is a major driver of MDSC hyperexpansion in M-Traf3−/− mice during chronic inflammation. Furthermore, we found that G-CSF was mainly produced by CCR4+ inflammatory monocytes and macrophages in mice during chronic inflammation. Taken together, our findings provide novel insights into the cellular and molecular mechanisms underlying TRAF3-mediated regulation of MDSC expansion, and open up new avenues for the development of MDSC-targeting therapeutics in disease conditions.
This study was supported by the National Institutes of Health grant R21 AI128264 (P. Xie), a New Jersey Commission on Cancer Research (NJCCR) grant DCHS19CRF005 (P. Xie), a Pilot Award of Cancer Institute of New Jersey through Grant Number P30CA072720 from the National Cancer Institute (P. Xie), and a Busch Biomedical Grant (P. Xie).
Setting the pace: CD4 T cell-intrinsic Arginase 1 orchestrates Th1 induction and contraction
Authors: West Erin E , Freeley Simon , Kaminski Marcin M , Merle Nicolas S , Veenbergen Sharon , Lee Duck-Yeon , St. John-Williams Lisa , Thompson J Will , Green Douglas R , Scholl-Buergi Sabine , Karall Daniela , Huemer Martina , Kemper Claudia 
Published: January 2023

Abstract
CD4 T cell-intrinsic engagement of the complement receptor CD46 controls nutrient influx and the metabolic reprogramming events that are essential for both the initiation and contraction of human Th1 responses, characterized by IFN-g and IL-10 production respectively. Here, we demonstrate that CD46 also orchestrates T cell arginine metabolism by upregulating the arginine transporter CAT-1 and, unexpectedly, Arginase 1 (Arg1). Arg1 has been well characterized in macrophages where it is associated with the IL-10 secreting M2 type, but its expression or function in T cells has not been described. Surprisingly, in CD4 T cell, Arg1 seems to restrain IL-10 production and contraction: T cells isolated from four patients with rare Arg1 deficiency mount strong Th1 responses but display significantly increased IL-10 switching and early contraction when compared to healthy control cells. Similarly, Arg1fl/fl CD4-cre+ mice infected with influenza virus are characterized by an enhanced Th1 response that contracts more rapidly, resulting in viral control and significantly reduced lung pathology. Unexpectedly, both Arg1-deficient mouse and human T cells produce normal levels of nitric oxide (NO) and polyamines. Metabolic profiling rather revealed that T cells lacking Arg1 have enhanced glycolysis, reduced TCA-cycle intermediates, and engage an “alternative” glutamine pathway often utilized by cancer cells. Normalization of these metabolic perturbations, through the targeting of specific metabolic enzymes, restored typical Th1 induction/contraction. Overall, these data unveil an unexpected intrinsic role for Arginase 1 as a pacemaker of the Th1 lifecycle, which could be harnessed for the amelioration Th1-driven pathologies.
Characterization of urine-derived immune cells from bladder cancer patients and comparison to tumor and peripheral blood
Authors: Tran Michelle A , Farkas Adam , Beaumont Kristin , O’Donnell Timothy , Mehrazin Reza , Wiklund Peter , Horowitz Amir , Galsky Matthew , Sfakianos John , Bhardwaj Nina 
Published: January 2023

Abstract

Background
Although PD-1/PD-L1 immune checkpoint blockade (ICB) therapy has achieved durable clinical responses in a subset of bladder cancer patients (15–25%), the majority do not respond. This has led to a need to identify clinically predictive biomarkers. Urine is an accessible material that may reflect cellular and/or genetic signatures related to ICB response. It has been shown that bladder cancer patient urine contains immune cells in addition to tumor cells. To study the concordance between urinary immune cells and the tumor immune microenvironment (TIME), we un-biasedly characterized bladder cancer patient urine and compared it to tumor and peripheral blood mononuclear cells (PBMC).

Methods
Matched tumor, urine, and PBMC from 8 bladder cancer patients were dissociated for single cell RNA sequencing (scRNAseq) and Cellular Indexing of Transcriptomes and Epitopes by Sequencing (CITE-seq) using 10X Genomics.

Results
scRNAseq of bladder cancer patient urine revealed CD4+ and CD8+ T, T regulatory (Treg), natural killer (NK), and B cells as well as neutrophils, dendritic cells, monocytes, and macrophages. The composition and transcriptional profiles of these cells were more similar to the tumor immune cells than to PBMC. Urine immune cells expressed hypoxia, anergy, and pro-inflammatory gene signatures that were more similar to tumor immune cells than PBMC.

Conclusions
Our work represents the first scRNAseq and CITE-seq of cancer patient urine. Our study shows several immune cells shed in bladder cancer patient urine and suggests they look phenotypically similar to the TIME. This has implications for future clinical applications as urine can be sampled non-invasively in scenarios when tumor resection may not be feasible.
Supported by the NIH grant: R01 CA249175-01.
Mechanisms by which Factor H protects Trypanosoma cruzi from the alternative pathway of complement.
Authors: Menon Smrithi Sugumaran , Ramirez-Toloza Galia , Wycoff Keith L , Shaughnessy Jutamas , Ram Sanjay , Ferreira Viviana P 
Published: January 2023

Abstract
Chagas disease is a neglected tropical disease caused by the protozoan Trypanosoma cruzi (T. cruzi). Complement (C) is a critical arm of the immune system. Various pathogens, including T. cruzi, hijack host regulators such as Factor H (FH) to inactivate C. FH regulates the alternative pathway (AP) of C in blood and on cell surfaces. FH contains 20 short consensus repeat (SCR) domains and binds to C activation products (i.e., C3b/C3d) or polyanionic host cell markers (e.g., sialic acid, glycosaminoglycans) on cell surfaces, and uses its 4 N-terminal SCRs to inactivate the AP. T. cruzi potentially binds FH by transferring host sialic acids onto its surface using its trans-sialidase enzyme. This study asked whether FH binds directly to T. cruzi in the absence of C3b and defines the FH domains involved. Only C-resistant infective trypomastigotes, but not C-sensitive noninfective epimastigotes, bound FH directly in a dose-dependent manner. Although SCR 19–20 was hypothesized as important for this interaction given it is the only known sialic acid-binding region, domain mapping indicated SCR 5–8 competitively inhibited FH binding by &gt;35% with lower contributions of other regions. FH related protein-5 (FHR-5), whose SCRs bear sequence homology to all known polyanion-binding regions (6–7, 10–14, 19–20), fully competitively inhibited FH binding to trypomastigotes. In addition, competitive survival experiments with serum and FHR-5 led to &gt;80% killing when the AP or all C pathways were active. This is the first study to show consequences of completely inhibiting FH protection on T. cruzi and to suggest multiple FH domains are involved in the interaction. These data may be harnessed for developing therapeutics and vaccine candidates against T. cruzi.
This work was supported by The University of Toledo Biomedical Research Innovation Program (VF), and The American Association of Immunologists Careers in Immunology Fellowship Program (VF and SSM).
T cell immunity to a classical strain of Klebsiella pneumoniae
Authors: Mackel Joseph J , Wasbotten Rachel , Dahler Aubree , Smith Catherine Morffy , Rosen David A 
Published: January 2023

Abstract
Klebsiella pneumoniae (Kp) is a versatile pathogen capable of causing numerous infections that are becoming increasingly difficult to treat due to increasing antibiotic resistance. Kp strains are categorized into two distinct pathotypes: classical and hypervirulent, which exhibit critical differences in pathogenesis. Despite causing the majority of clinical cases, adaptive immunity resulting from classical Kp infections is poorly understood. Our laboratory has recently developed a model of adaptive immunity to live infection with classical Kp in which wild-type mice surviving primary infection are protected from morbidity and mortality upon rechallenge. Passive transfer of convalescent serum from surviving mice to naïve recipients does not confer protection against cKp infection, suggesting antibody responses alone are inadequate to protect against cKp. CD69+CD4+ T cells and γδ T cells expand dramatically in the lung following primary cKp infection and mice deficient in T cells are not protected from reinfection, indicating a requirement for T cells in the protective response. Further analysis of mice deficient in either classical αβ T cells or γδ T cells indicates that either T cell subset is capable of mediating protective immunity against Kp reinfection. Production of IL-17A by both CD4 and γδ T cells is enhanced during secondary infection. Together these data indicate that a natural course of infection with classical Kp elicits populations of classical T cells and γδ T cells that mediate protection against reinfection, likely through the rapid production of IL-17A. These results will ultimately inform rational design of vaccines and immunotherapies to prevent and treat Kp infection.
Supported by AAI Careers in Immunology Fellowship
Longitudinal interactions between levels of serum cytokine and the microbiome from four body sites
Authors: Zhou Xin , Shen Xiaotao , Johnson Jethro , Spakowicz Daniel J , Zhu Chenchen , Zhou Wenyu , Zhou Yanjiao , Weinstock George , Snyder Michael 
Published: January 2023

Abstract
The human body is co-habituated with trillions of microbes, which are actively interacting with the human immune system. Our previous study demonstrated that a subset of individuals, characterized by a chronic absence of serum Interleukin (IL)-17 and IL-22, is more likely to be resistant to insulin compared with individuals with detectable serum IL-17/IL-22. Additional analysis pointed out that such an absence of IL-17 and IL-22 is associated with the low abundance of the gut microbiome that belongs to the class of Clostridia.
To better understand the interactions between serum cytokines and microbiome, we curated a dataset from the iHMP that contains 86 individuals with a study duration from 5 months to 6 years, whose microbiome has been characterized from four body sites include: stool (n=927), skin (n=1128), nasal (922), oral (n=1001). Their serum cytokines had been characterized using 62-plex Luminex antibody-conjugated bead capture assay. The longitudinal interaction between microbiome abundance and serum cytokine are modeled by Bayesian Mixed-Effects Model, and the interaction between microbiome richness and serum cytokine are modeled by Linear Mixed Models.
We found the interactions are strongest in the gut compared to the other three body sites, but it is highly territory specific. For example, Moraxella from the skin is negatively correlated with multiple cytokines, but not Moraxella from the nasal. Among all cytokines, IL-1 signaling is the strongest modulator of microbiome abundance in all four body sites, but LEPTIN and GMCFS are more potent when focused on richness. Our results indicate that the immune regulation of the microbiome is active on multiple body sites, and such interaction is genera and cytokine specific.
This work was supported by the National Institutes of Health Common Fund Human Microbiome Project (HMP) (1U54DE02378901). Dr. Xin Zhou received support from the Stanford Aging and Ethnogeriatrics (SAGE) Research Center under NIH/NIA grant P30AG059307. The SAGE Center is part of the Resource Centers for Minority Aging Research (RCMAR) Program led by the National Institute on Aging (NIA) at the National Institutes of Health (NIH). Its contents are solely the responsibility of the authors and does not necessarily represent the official views of the NIA or the NIH.
Aberrant survival of uterine natural killer subsets in uterus transplant recipients
Authors: Porrett Paige , Gonzalez Michael V , Garifallou James , Wright Emma D , Lucander Aaron C K , Bell Mar’Kayla Joy , Tyson Kennedy , Smiler Jacqueline , Mafra Fernanda , Da Silva Renata Pellegrino , Johnston Sarah , Naziruddin Bashoo , Testa Giuliano , Johannesson Liza , George James , Freud Aharon , O’Neill Kathleen 
Published: January 2023

Abstract
Uterine natural killer cells (uNKs) are critical mediators of pregnancy success, but how aberrancies in uNK survival or differentiation underpin pregnancy complications is unknown. To improve our understanding of normal and abnormal uNK biology, we studied 1) uNKs in uterus transplant (UTx) recipients at high risk for pregnancy complications and 2) uNK survival after exposure to pharmacologic immunosuppression in vitro. To address the first question, we analyzed uNKs isolated from endometrial biopsies of healthy controls (n=3) or UTx recipients (n=5) using scRNA-seq. In healthy controls, CD103-expressing uNK3 cells were the dominant uNK subset (30% of all uNK cells). In contrast, uNK1 cells were the most frequent (30%) in the majority of UTx recipients. This dominance of uNK1 cells in UTx recipients did not appear to arise only from loss of uNK3 cells, as the uNK1 transcriptional signature was abnormally upregulated in immature proliferating uNKs. Next, we used multiparameter flow cytometry to study single-cell suspensions of whole endometrium from deceased organ donors (n=4) that were cultured for one week with or without the calcineurin inhibitor FK506. Although FK506 significantly impaired the survival of CD103+ uNK3 cells in vitro, there was no evidence of uNK1 enhancement. Instead, FK506 appeared to selectively deplete CD39+ cells – a marker currently used to identify human decidual NK1 cells. Altogether, these results demonstrate that the distribution of uNK subsets is often altered in UTx recipients, and that FK506 can impact the survival of specific uNK subsets. Identification of the additional factors which impact uNK differentiation and survival will be necessary to understand the genesis of pregnancy complications.
Supported by NIH/NIAID (R01 AI 145905) University of Pennsylvania Institute for Immunology
ILC3 Expansion in Acute Myeloid Leukemia
Authors: Dinh Thanh Thanh Trinh , Lordo Matthew R , Shilo Nikolas , Altynova Ekaterina , Kronen Parker , Broughton Megan , Sellers Victoria , Collins Patrick , Freud Aharon G , Mundy-Bosse Bethany L 
Published: January 2023

Abstract
At homeostasis, type 3 innate lymphoid cells (ILC3s) are tissue-resident and have key roles in maintaining mucosal immunity. Recently, the expansion of ILC3s has been described in solid tumors; however, the mechanism driving this finding is not completely understood. In the setting of mouse and human acute myeloid leukemia (AML), we have observed an expansion of ILC3s. Using an AML murine model, we observed a 7-fold increase in ILC3s in the intestine (the normal ILC3 niche) of leukemic mice relative to controls and a 3-fold increase of ILC3s in the bone marrow (intestine: 0.208% +/− 0.04 AML vs 0.032% +/− 0.04 WT, n=5, p&lt;0.001; bone marrow: 1.312% +/− 0.20 AML vs 0.067% +/− 0.20 WT, n=5, p&lt;0.0001). We hypothesized that the ILC3 increase was due at least in part to increased ILC3 differentiation. To address this hypothesis, we isolated the innate lymphoid cell precursor (ILCP) from normal human donors and co-cultured with AML cells which resulted in increased ILC3 frequency (59% +/− 5% with AML vs 28% +/− 5% without AML, n=15, p &lt;0.001). This observation was also recapitulated in a human PDX model in NSG mice (3.22% +/− 7.8% without AML vs 23.91% +/− 7.8% with AML, n=10; p&lt;0.0001). Our published data have shown AML blasts secrete aryl hydrocarbon receptor (AHR) ligands, which can bind and activate the transcription factor AHR in ILC precursors and mature ILCs. By manipulating AHR activity ex vivo, we showed that ILC3s are expanded in AML in part due to AML-mediated AHR activation (60% +/− 5.5% with AML vs 27% +/− 5% AML+AHR inhibitor, n=15, p&lt;0.001). Collectively, these data support a model in which AML-mediated AHR activation promotes ILC3 expansion by skewing ILCP differentiation towards ILC3. Future studies will define the role of ILC3s in AML progression.
Supported by grant from NIH (1R01CA255860-01A1)
Combining Immunohistochemistry and In Situ Hybridization to Characterize Immunosuppression in the Tumor Microenvironment
Authors: Cartwright Emily , Oliver Ashley , Kalyuzhny Alex 
Published: January 2023

Abstract
While immune-based cancer treatments have been successfully deployed in the clinic for hematological malignancies, treatment of solid tumors has proven more challenging. Comprehensive understanding of the dynamic landscape of the tumor microenvironment (TME) will lead to new therapies and improved efficacy of current therapies. Immunohistochemistry (IHC) and in situ hybridization (ISH) are the two techniques at the forefront of TME characterization. Combining RNA and protein detection with ISH and IHC reveals cell-type specific gene expression and the cellular sources of secreted proteins. Traditional ISH-IHC workflows require harsh pre-treatment of tissues prior to detection with primary antibodies, destroying the target epitope for some antibody clones. Bio-Techne’s RNAscopeTM RNA protein Codetection Assay improves the likelihood of successful antibody staining by incubating with primary antibody first, before initiating the RNAScope pretreatment step. With minimal optimization time, we were able to visualize protein expression using IHC validated antibodies for important immunosuppression biomarkers such as HIF-1 alpha, FoxP3, PD-L1, and CD11b. Using a cancer tissue microarray (TMA), we show this workflow is compatible with many tissue types, including pancreas, uterus, liver, breast, lung, prostate, and colon tissue. In combination with RNAScope probes CD8A, CD4, PD-1, TGFB1, and VEGF we were able to identify immunosuppressive cells in the TME, including Regulatory T cells (Tregs) and tumor associated macrophages (TAMs). Adoption of this workflow will streamline profiling of the TME, facilitating a more comprehensive evaluation of solid tumors for biomarker discovery and immunotherapy development.