bluuebunny/crossref_metadata_2025_split huggingface dataset:
A heterozygous mutation in the coiled-coil domain of STAT5B is associated with immune deficiency and dysfunction
Authors: Fritz Jill Marie , Majri Sonia , Villarino Alejandro , Zheng Lixin , Lenardo Michael J 
Published: January 2023

Abstract
The signal transducer and activator of transcription (STAT) family of transcription factors play a critical role in regulating gene expression required for immune development and function. In a patient diagnosed with primary immunodeficiency characterized by chronic lymphadenopathy, splenomegaly, immune thrombocyotopenia and necrotizing granulomas, we discovered a heterozygous missense mutation in STAT5B (c.617A&gt;G, p.Q206R). Cotransfection of Q206R and wild-type STAT5B in HEK293 cells reduced STAT5B-mediated transcriptional activity compared to wild-type STAT5B alone, suggesting that Q206R STATB dominantly-interfered with wild-type STAT5B function. Patient T cell blasts exhibited reduced phosphorylation of STAT5B in response to IL-2, IL-7 and IL-15 compared to healthy controls. Further, IL-2/STAT5-dependent gene transcription was impaired in patient T cell blasts. The patient exhibited increased lymphocytes (as well as monocytes and neutrophils) in the peripheral blood associated with a higher frequency and number of CD4+ T effector memory cells (TEMs). Further, CD4+ TEMs isolated from the patient were resistant to restimulation-induced cell death compared to healthy control TEMs in vitro. Collectively, these data suggest that the Q206R STAT5B variant reduced IL-2/STAT5B-mediated gene expression in human T cells, and defects in this signaling pathway are associated with increased survival of TEMs, disrupted immune homeostasis and disease.
ST8Sia6 deficiency is insufficient to induce complement-mediated attack on recent thymic emigrants
Authors: Belmonte Paul , Hsu Fan-Chi , Dash Barsha , Chung Ji Young , Arocha Sinibaldo Romero , Shapiro Virginia 
Published: January 2023

Abstract
T cells require additional developmental steps after positive selection before they are functionally competent and can enter the long-lived peripheral T cell pool. During maturation, T cells gain the ability to produce cytokines upon stimulation, and the ability to protect themselves from TNF and complement-mediated cell death. In three different mouse models with defects in T cell maturation (CD4-cre NKAP conditional knockout mice, CD4-cre HDAC3 conditional knockout mice and CD4-cre RUNX1 conditional knockout mice), recent thymic emigrant (RTE) numbers are greatly reduced due to complement-mediated attack initiated by the binding of IgM and activation of the classical pathway. These models have deficiencies in the addition of sialic acid modifications, which is normally increased during the later stages of T cell development. As stripping cell surface sialic acids by neuraminidase on peripheral T cells leads to binding of natural IgM and complement activation, it was hypothesized complement-mediated attack is due to a defect in sialylation. Transcriptionally, the sialic acid transferase ST8Sia6 is upregulated concurrently with T cell maturation after positive selection. To understand the role of ST8Sia6 in T cell maturation, we generated ST8Sia6 knockout mice. These mice are healthy and fertile. RTEs and mature T cells are present in the periphery in normal numbers, and do not show evidence of complement deposition. Therefore, loss of ST8Sia6 alone is insufficient to induce complement-mediated attack on RTEs and other co-expressed sialyltransferases may also be required to protect from complement deposition.
The Specific and Non-Specific Action of Rabbit Blood Serum in the Complement Fixation Test
Authors: Huddleson I. F. 
Published: January 2023

Summary
There is not sufficient evidence to condemn naturally infected milk as dangerous to rabbits by ingestion since it has shown no pathogenic action and no antigenic action.
A non-specific absorption of complement takes place in the presence of inactivated rabbit blood serum and a bacterial antigen of Bact. abortus.
Identification of immunological pathways altered by a pro-autoimmune genetic risk variant, PTPN22 1858C&gt;T, that enhances the prevalence of Type 1 diabetes
Authors: Schmiel Shirdi E. , Marquardt Kristi , Lin Xiaotian , Sherman Linda A. 
Published: January 2023

Abstract
Protein tyrosine phosphatase nonreceptor type 22 (PTPN22) encodes lymphoid tyrosine phosphatase (Lyp) adaptor molecule that is involved in numerous biochemical pathways in hematopoietic cells. Studies have shown an association with the minor allelic variant of PTPN22, R620W, and increased incidence of autoimmune disorders such as type 1 diabetes (T1D). Approximately 10% of the North American population carry the R620W minor allele, which increases the risk of T1D by as much as 2–4-fold. The exact mechanisms by which this pro-autoimmune risk allele enhances the incidence of T1D is unknown. To address this gap of knowledge our lab introduced the murine ortholog of the human R620W allele, R619W, into the endogenous locus of Non-obese diabetic (NOD) mice that develop spontaneous T1D. NOD mice harboring this mutation (NOD 619W/W) demonstrated a similar phenotype seen in affected humans with accelerated onset of T1D and enhanced anti-islet autoantibodies (IAA). Clinical manifestation of T1D in humans is preceded by IAA in the serum. IAA affinity can often identify individuals at risk of developing diabetes. Germinal centers (GC) are lymphoid structures where affinity maturation occurs with the help of follicular helper T cells (Tfh) cells. An increase in circulating Tfh cells has been reported in newly diagnosed and pre-diabetic individuals, thus suggesting a role for GC reactivity in T1D. Similar to human observations, NOD 619W/W mice also show enhanced IAA and increased Tfh and GC B cell numbers before onset of T1D therefore, our lab is currently examining the immunological pathways altered by this pro-autoimmune risk alleles and their contributions to T1D.
Aire-dependent thymic FoxP3+ regulatory T cells in Type 1 Diabetes
Authors: Bridge Jennifer Anne , Yuen Benjamin , Dong Shen , Ye Jimmy , Kappler John W. , Bluestone Jeffrey A. , Anderson Mark S. 
Published: January 2023

Abstract
Aire expression within the specialized mTECs has been found to be important for enforcing negative selection of T cells, however, mounting evidence has demonstrated the importance of Aire in Treg differentiation and development. Autoimmune diseases like T1D arise from the failure to restrain immune responses against self. Utilizing a novel mouse model, where the Aire locus drives the selected expression of the high affinity DT receptor, revealed a significant reduction in the number of FoxP3+ Tregs within the thymus following treatment. Thus, we hypothesize that alterations in the thymic self-antigen expression and presentation via ablation of Aire-expressing mTECs, will affect the repertoire and function of Aire-dependent FoxP3+ Tregs. New high-throughput platforms and single-cell DNA barcoding technology will allow us to assess several thousand individual Tregs that have undergone Aire-dependent selection in the thymus. The identification of these unique clones and gene signatures will be further interrogated retrogenically, allowing for qualitative assessment of their function within the model of T1D. The antigen specificities of the Aire-dependent TCRs will be assessed via screening against peptide libraries using MHCII molecule strongly associated with NOD mouse T1D. We anticipate identification of novel sequences that are recognized by Tregs, which may lead to discovery of new self-peptides that contribute to T1D. Our proposed study aims to expand our knowledge on TCR repertoire of Aire-dependent FoxP3+ Tregs and the antigen-specificity in T1D, improving our understanding of the respective function of thymic-derived antigen-specific Treg suppression in peripheral tissues associated with autoimmune T1D.
IL-27 Protects from Autoimmunity by Suppressing IL-17 Production and Promoting Treg Suppressor Functions
Authors: Bockerstett Kevin A. , Wong Chun Fung , Ford Eric L. , DiPaolo Richard J. 
Published: January 2023

Abstract
IL-27 has been implicated in the regulation of several autoimmune diseases including multiple sclerosis, inflammatory bowel disease, and rheumatoid arthritis. However, a better mechanistic understanding of how IL-27 regulates individual autoimmune diseases is needed. Autoimmune gastritis involves chronic inflammation and tissue damage caused by self-reactive T cells and autoantibodies. Serious consequences of autoimmune gastritis include pernicious anemia and a significant increase in the risk of developing gastric cancer. We used a TCR transgenic mouse model (TxA23) in which mice develop autoimmune gastritis due to CD4+ T cells that express a transgenic TCR specific for a peptide expressed by parietal cells in the gastric mucosa. Disease in this model mimics many aspects of human disease including the development of autoantibodies and precancerous lesions. We recently discovered a critical role for IL-27 in regulating autoimmune gastritis. IL-27 deficient mice have more severe inflammation and tissue damage, and develop precancerous lesions much faster compared to control mice. Further analyses revealed that IL-27 expression suppressed IL-17A secretion by immune cells in vivo. In vitro we observed that IL-27 acts directly on autoreactive CD4+ T cells and reduced IL-17A secretion. Finally, IL-27 expression increased inhibitory receptors (LAG3, CD39, CD73, GITR, CD25) on CD4+FoxP3+ regulatory T cells (Tregs) and supported their ability to suppress autoreactive CD4+ T cells in vitro. These data elucidate an important role for IL-27 in protecting from autoimmune gastritis by suppressing IL-17A secretion and promoting suppressor functions of Tregs.
Antigen discovery for regulatory T cells (Tregs) in type-1 diabetes (T1D)
Authors: Dong Shen , Xu Daqi , Bridge Jennifer Anne , Purtha Whitney , Spence Allyson , Wells Danny , Wang Xiao hua , Tang Qizhi , Anderson Mark S. , Bluestone Jeffrey A. 
Published: January 2023

Abstract
Antigen specific Tregs are more efficient than polyclonal Tregs in their suppressive capacity, which makes them attractive tools for further targeted tolerogenic therapy. Autoreactive T cells specificity has been well characterized in type 1-diabetes but little is known about autoreactive Treg specificity. Whether Tregs recognize the same tissue-specific self-antigens as autoreactive T cell, or they recognize different self-antigens to regulate local immune responses via bystander suppression is unclear. Our goal is to identify antigen specificity from enriched islet infiltrated Treg by using previously validated peptide-major histocompatibility complex (pMHC) library displayed on yeast. We first performed single cell TCR sequencing on Tregs from the pancreatic islet of 12 weeks old NOD mice. Data show an abundance of the TCR allele TRAV5D4 usage in islet infiltrated Tregs that has been characterized to target a primary insulin peptide. Moreover, we also performed single cell RNAseq together with TCR sequencing on islet infiltrated CD4+ T cells from a T1D patient from the Network for Pancreatic Organ Donors with Diabetes. We were able to identify enriched TCRs with Treg gene expression profile that will be used to screen for antigen recognition on a DR4 pMHC yeast library. New antigen identification for pancreatic islet-infiltrated Tregs will allow us to improve our understanding of Treg deficiencies in T1D and may lead us to design new immunosuppressive therapies.
Dissecting the genetic networks that control regulatory T cell stability using pooled CRISPR screens
Authors: Cortez Jessica T. , Shifrut Eric , Lee Youjin , Mumbach Maxwell R. , Satpathy Ansuman T. , Granja Jeffrey , Subramaniam Meena , Roth Theodore , Simeonov Dimitre , Ye Chun J. , Chang Howard Y. , Van Gool Frederic , Marson Alexander 
Published: January 2023

Abstract
Regulatory T cells (Tregs) are a specialized subset of CD4+ T cells that suppress inflammation to maintain homeostasis and prevent autoimmunity. Treg development and function depend on expression of the master transcription factor Foxp3. While Tregs have been thought to be irreversibly committed to suppressive functions, lineage tracing studies have challenged this by revealing that Tregs can exhibit plasticity. Tregs that lose Foxp3 expression, termed ‘exTregs’, have been shown to acquire cytokine production capabilities of pro-inflammatory effector T cells and exacerbate autoimmunity. However, the gene regulatory programs that promote or disrupt Foxp3 stability in Tregs under various physiological conditions are not well understood. Here we have leveraged improved functional genetic tools, including pooled CRISPR screens, to identify nuclear factors that regulate Treg plasticity. A Foxp3 lineage-tracing reporter mouse model was used to confidently distinguish Foxp3− exTregs from contaminating Foxp3−T effectors. Using HiChIP, we identified Treg and exTreg-specific chromatin loops and systematically assessed their contribution to Foxp3 stability. Furthermore, we investigated transcriptional signatures of distinct exTreg populations residing in the peripheral organs using single-cell RNA sequencing. Collectively, this work helps us better understand the genetic networks that control stable expression of Foxp3 in Tregs as they encounter changing inflammatory environments.
CCR2 mediates transendothelial migration of human pathogenic Th17 cells
Authors: Parween Farhat , Kathuria Noshin , Zhang Hongwei H. , Farber Joshua M. 
Published: January 2023

Abstract
Leukocyte extravasation is a multistep process characterized by rolling and arrest on the endothelium followed by transendothelial migration (TEM). Arrest and TEM by T cells typically requires the activities of chemokines and their receptors. Previously, we showed that CCR2 identifies highly differentiated, readily activated CD4+ memory T cells in human blood with diverse effector capabilities, and we showed that CCR6 is the signature chemokine receptor for Th17 cells. Here we describe studies of the migratory capabilities and effector profiles of CD4+ CCR6+ CCR2+ as compared with CD4+ CCR6+ CCR2− and CD4+ CCR6− CCR2− T cells from human blood. Using flow chamber assays with TNFa-activated human umbilical vein endothelial cells, we found that among the CCR6+ cells, only the CCR2+ subset was able to cross the endothelial monolayer efficiently. Using antibodies against the CCR6 ligand, CCL20, and a specific inhibitor of CCR2, we found that CCR6 was important for arrest and that CCR2 was critical for subsequent TEM. Using intracellular staining of cells activated ex vivo, we found that although IL-17-expressing cells were found within all CCR6+ subsets, the CCR6+CCR2+ cells were distinguished by their production of IFNg, GM-CSF, and TNFa, thereby demonstrating the cytokine profile of pathogenic Th17 cells as identified in mouse models of autoimmune disease. Taken together, our studies suggest that chemokine receptors on memory Th cells play non-redundant roles in the egress from blood, that providing T cells with CCR6 and CCR2 could enhance their trafficking into tissue, and that, on the other hand, blocking CCR2 alone could prevent the extravasation of pathogenic Th17 cells and ameliorate tissue damage in Th17-cell driven disease.
Using Rexinoids to Program Effector T cells for Homing to Mucosal Sites of Virus Transmission
Authors: Manhas Kavita , Blattman Joseph N. , Marshall Pamela A. , Wagner Carl E. , Jurutka Peter W. 
Published: January 2023

Abstract
Vaccines are the most effective defense against infectious diseases. To be most successful, vaccines must target the induced immune response to chief sites of pathogen entry: epithelial and mucosal surfaces. We have previously shown that the biologically active form of Vitamin A, all-trans retinoic acid (ATRA), can directly modulate the expression of gut mucosal-homing receptors on T cells as binding of ATRA to the retinoic acid receptor (RAR), which partners with the retinoid X receptor (RXR), promotes transcription of specific genes. Rexinoids, synthetic ligands for the RXR, have potential to mimic or improve the immune effects of vitamin A by enhancing RAR-mediated transcription. Additionally, rexinoids have a greater potential for clinical use due to their improved solubility and stability, making them candidates to incorporate into vaccines. Here, we explored the possibility of different rexinoids to function as adjuvants during vaccination to program effector T cell expression of the gut-homing receptors CCR9, α4β7, and CD103. CD8 T cells obtained from transgenic mice were co-cultured with peptide and set concentration of each rexinoid, followed by flow cytometric analysis of receptor expression. We observed increased expression of gut-homing receptors during activation in the presence of certain rexinoids, indicating that they can mimic the effects of ATRA. Furthermore, we saw a cooperative relationship between other rexinoids and ATRA, as co-culturing with sub-optimal doses of each resulted in enhanced expression. Our findings lend support for the prospective use of these rexinoids as adjuvants during vaccination and in therapeutic approaches against diseases transmitted across mucosal surfaces.
Utility of recombinant Hedgehog proteins and RANKL to generate bone cell models in vitro
Authors: Soto Hortensia , Moyron-Quiroz Juan , Kim Daniel , Chattopadhyay Mohar , Tam Miguel , Yang Xifeng , Nguyen Dzung , Zhou Jie 
Published: January 2023

Abstract
Bone remodeling is a lifelong process that begins during the embryonic stage and continues after birth. The two main bone cells involved in bone remodeling are osteoblast (bone formation) and osteoclast (bone resorption). The strict balance between bone formation and bone resorption is maintained by several cytokines and growth factors, including RANKL, the Hedgehog family of proteins (SHH, DHH, and IHH), and M-CSF secreted by bone cells, as well as systemic hormones. Any imbalance in bone remodeling might result in bone related diseases. Osteoblast differentiation was first studied using mouse mesenchymal cell line C3H10T1/2 cells. Recombinant SHH, DHH, and IHH were able to induce osteoblast differentiation of C3H10T1/2 cells. Bone morphogenetic protein 9 (BMP-9) enhanced the differentiation of C3H10T1/2 cells induced by Hedgehogs. Osteoclasts are multinuclear cells that are derived from hematopoietic precursors of the monocyte/macrophage lineage and their differentiation depends on osteoblast-produced RANKL and M-CSF. In order to study osteoclast differentiation, and the role of different cytokines in bone homeostasis; murine bone marrow derived macrophages (BMDM) were obtained and treated with mouse GM-CSF for 7 and 14 days. FACS analysis of cellular markers and functional assays confirmed the identity of macrophages. Subsequently, BMDM were differentiated to osteoclast with mouse M-CSF and RANKL. The multinucleated cells were identified by cell morphology and functional TRAP detection. Similar results were obtained with RAW264.7 cells, a mouse macrophage cell line. Our results confirm that BMDM and osteogenic cells lines are good models to study cytokines involved in cell differentiation and bone remodeling.
Bone marrow lympho-myeloid malfunction in obesity requires precursor cell-autonomous TLR4
Authors: Borghesi Lisa A. , Liu Ailing , Chen Minhui , Stefanovic-Racic Maja , Kumar Rashmi , O’Doherty Robert , Ding Ying , Jahnen-Dechent Willi 
Published: January 2023

Abstract
Obesity, a prevalent condition in adults and children, impairs bone marrow (BM) function. However, the underlying mechanisms are unclear. Here we show that obese mice exhibit poor emergency immune responses in a toll-like receptor 4 (TLR4)-dependent manner. Canonical myeloid genes (c-fms, pu.1, runx) are enhanced, and lymphoid genes (flt3, e47, ebf) are reduced. Using adoptive transfer and mixed BM chimera approaches we demonstrate that myeloid&gt;lymphoid bias arises after only six weeks of high fat diet and depends on precursor cell-autonomous TLR4. Further, lean mice exposed to the TLR4 ligand lipopolysaccharide (LPS) at doses similar to that detectable in obese serum recapitulates BM lympho-myeloid alterations. Together, these results establish a mechanistic contribution of BM cell-intrinsic TLR4 in obesity-driven BM malfunction and demonstrate the importance of LPS. Our findings raise important questions about the impact of maternal obesity and endotoxemia to fetal hematopoiesis since fetal immune precursors are also sensitive to TLR4 signals.
Common lymphoid progenitor derivation of a conserved dendritic cell subtype is mediated by Bcl11a
Authors: Dekker Joseph D. , Rhee Catherine , Hu Zicheng , Lee Bum-Kyu , Lee Jiwon , Iyer Viswanath R. , Ehrlich Lauren I. , Georgiou George , Ippolito Gregory C. , Tucker Haley O. 
Published: January 2023

Abstract
Plasmacytoid dendritic cells (pDCs) are a distinct subset of dendritic cell that specializes in the production of type I interferons (IFNs) upon engagement of pattern recognition receptors to promote antiviral immune responses. They have been implicated in the pathogenesis of autoimmune diseases that are characterized by a type I IFN signature, yet, pDC also can induce tolerogenic immune responses. Whereas an understanding of pDC physiology, immune function, and clinical significance has increased considerably, their development has not been unequivocally traced to either lymphoid or myeloid lineages nor has its transcriptional regulatory network been fully clarified. We observed that mb1-Cre mediated deletion of the Bcl11a transcription factor in common lymphoid progenitors (CLP) resulted in a ~15% reduction in pDC cellularity. Adoptive transfer of bone marrow from Bcl11aF/Fmb1-Cre mice recapitulated partial pDC loss. Mb1-Cre:YFP+ reporter mice contained ~85% YFP− and ~15% YFP+ pDCs, indicating a myeloid–lymphoid dichotomy of pDC generation. As compared with myeloid pDC, CLP-derived pDC (termed B-pDC) display a B cell-like gene expression profile, including hallmark targets directly transactivated by Bcl11a in B cells. B-pDCs demonstrate preferential homing to secondary lymphoid organs, are inherently activated compared to myeloid pDC, expand more rapidly upon TLR9 engagement, and are genetically and phenotypically homologous to a DC subtype recently discovered in human blood. We conclude that the pDC compartment comprises two cell types with overlapping but distinct functions that are conserved between mouse and man.