bluuebunny/crossref_metadata_2025_split huggingface dataset:
Dynamics of antimicrobial protein secretion in mouse milk
Authors: Abokor Ahmed , Saha Piu , Yeoh Beng San , Golonka Rachel , Vijay-Kumar Matam 
Published: January 2023

Abstract
Breast milk is the preferred food for the newborns and considered as complete ‘edible immune system’. In addition, breast milk is a source of innate and adaptive proteins which not only protects the naïve infant gastrointestinal tract from enteropathogens but also aid in the appropriate initial colonization of gut microbiota. In this study, we analyzed antimicrobial immune proteins in the milk from C57BL/6 dams at several time points, and observed distinctive patterns in the immune protein levels throughout the lactation period. The presence of innate immune proteins serum amyloid A (SAA), CD14, and notably lipocalin-2 (Lcn2) were observed in high quantities with Lcn2 and SAA present at microgram levels suggesting a potential niche for these proteins during neonatal immune development. Moreover, low dose LPS administration to dams significantly increased some of these proteins in the milk. In addition, adaptive immune proteins immunoglobulins (Ig) A and G with IgG were present in milk at higher quantities compared to IgA at day 5 post-delivery. Interestingly, class-switching occurs by day 15 indicating a selective adaptive immune preference towards IgA, which could be due to gut colonization of newborns with more complex gut microbiota with aging and IgA being a major gut homeostatic factor. Additionally, milk from IgA-deficient dams allowed for robust proliferation of E. coli compared to wild-type milk further signifying the potential involvement of IgA during the early colonization of microbes in the neonatal gut. Collectively, our findings provide insight in the various immune proteins with antimicrobial activity may play a major role in the initial microbial colonization in the gut, which has long-lasting consequences on the host.
Supported by NIH R01-CA219144, NCI Diversity Supplement (CA219144-05S1)
E-Syt1 ubiquitination by Nedd4 limits the caspase-11-mediated non-canonical inflammasome and endotoxemia
Authors: Ma Yilei , Lin Guoxin , Guo Hui , Yu Jianyu , Zhang Jian 
Published: January 2023

Abstract
Sepsis caused by gram-negative bacteria is thought to be largely due to the host’s response to LPS or endotoxin. Upon sensing LPS by its cytosolic receptor, caspase-11 (Casp-11) assembles a higher order structure that enables the activation of Casp-11 protease function, leading to gasdermin D cleavage and pyroptosis. However, the mechanisms for negatively regulating the activation of the Casp-11-dependent non-canonical inflammasome during sepsis are largely unknown. Nedd4 is a HECT-type E3 ubiquitin ligase that positively regulate T cell activation. However, its role in innate immunity is largely unknown. In this study, we found that although macrophages lacking Nedd4 produce comparable amounts of TNF-α and IL-6 in response to different TLR ligands, they produce more IL-1β upon LPS priming and stimulation with cholera toxin B (CTB) and EHEC but not ATP, nigericin, or other inflammasome stimuli. These data suggest that Nedd4 specifically regulates non-canonical inflammasomes. In support of this notion, mice lacking Nedd4, or lacking Nedd4 in myeloid cells, succumb to endotoxemia, which is rescued by deficiency for Casp-11 but not NLRP3. Mechanistically, Nedd4 appears to specifically bind to extended synaptotagmin 1 (E-Syt1), and targets E-Syt1 for ubiquitination, and loss of Nedd4 stabilize E-Syt1 protein which potentiates Casp-11 oligomerization and activation. Therefore, our data unveil a novel mechanism by which Nedd4 puts a brake on the Casp-11-dependent non-canonical inflammasome, thus preventing over-activation of innate immune responses during sepsis. The molecular insights gained in this study provide novel targets for modulating Casp-11-dependent inflammasome-mediated sepsis in patients.
Examining the relationship between epidermal gamma delta T cells and neuropeptide Y
Authors: Almaguer Berenice , Jameson Julie 
Published: January 2023

Abstract
Neuropeptide Y (NPY) is one of the most abundant peptides present in the central nervous system and is responsible for regulating food intake, energy metabolism, and emotional expression. Emerging evidence indicates that NPY could potentially play a role in serving as a transmitter between the nervous system and immune system and even within the immune system itself. Epidermal gamma delta (γδ) T cells play a role in governing wound repair and homeostasis within the skin by producing growth factors, cytokines, and performing cytolysis. Wound repair involves the transmission of pain signals and immune functions simultaneously, suggesting interaction and collaboration between the nervous and immune system. We isolated and cultured epidermal γδ T cells from wild type mice and used flow cytometry to determine the expression of NPY1R on epidermal γδ T cells prior to and after activation. The data indicates that NPY1R is present on epidermal γδ T cells and that expression increases when upon activation. In addition, we performed immunofluorescence microscopy to acquire images of skin sections and epidermal sheets to determine expression of NPY1R on epidermal γδ T cells during skin homeostasis. This data shows that NPY1R is present on select populations of epidermal γδ T cells in the skin and future directions will examine how this impacts the role of epidermal γδ T cells in wound repair and homeostasis in the skin.
Supported by the Arnold & Mabel Beckman Foundation (Beckman Scholars Program)
The neurotrophic factor Neuritin regulates T cell anergy and T regulatory cell function
Authors: Yu Hong , Nishio Hiroshi , Barbi Joseph , Mitchell-Flack Marisa , Vignali Paolo , Zheng Ying , Lebid Andriana , Chang Kwang-Yu , Fu Juan , Blosser Lee , Tam Ada , Pardoll Drew 
Published: January 2023

Abstract
T cell activation and tolerance are tightly regulated to ensure effective elimination of foreign antigen while maintaining immune tolerance to self-antigens. Development of T cell anergy and regulatory T cell (Treg) mediated suppression both contribute to the establishment of immune tolerance. Here, we show that neuritin (Nrn1), a conserved GPI-anchored surface molecule important for the development, survival and function of neurons, is highly expressed in anergic and Treg cells. Nrn1 deficient CD4 cells are resistant to Treg cell mediated suppression, display defective anergy induction, and have reduced peripheral Treg generation. Nrn1 deficient Foxp3+ Treg cells exhibit reduced control of inflammatory colitis. Moreover, upon induction of experimental autoimmune encephalomyelitis (EAE), Nrn1 deficient mice develop non-remitting disease and have increased spinal cord inflammatory infiltrates. These in vivo findings underscore the importance of Nrn1 in immune tolerance. Recently, Nrn1 was identified as an accessory component of the ionotropic AMPA receptor (AMPAR) complex in neurons. AMPARs mediate glutamate dependent cation flux and regulate cell membrane potential. Cell membrane potential can impact nutrient uptake, calcium influx, cell size, proliferation and survival. In vitro analysis reveals that Nrn1 deficient Treg cells exhibit reduced proliferation and survival, associated with higher membrane potential, reduced nutrient sensitivity, reduced glycolysis and mTOR activation. AMPAR blockade can correct proliferation defect in Nrn1 deficient Treg cells. These findings reveal Nrn1 as an important regulator of immune tolerance functioning through the modulation of glutamate activated AMPAR.
Transcription coactivator OCA-B/Pou2af1 is necessary and sufficient to promote T cell-intrinsic CD4 memory
Authors: Tantin Dean R , Sun Wenxiang , Kim Heejoo , Perovanovic Jelena , Ibarra Andrea , Hale J Scott , Williams Matthew A 
Published: January 2023

Abstract
The establishment of durable immunological memory remains a poorly understood aspect of lymphocyte biology and a limiting factor in vaccine development. Using a T cell-conditional OCA-B (Pou2af1) knockout model and a mouse viral pathogen, we show that expression of the transcription cofactor OCA-B within T cells is necessary for proper CD4+ memory T cell formation. We also show that ectopic OCA-B expression is sufficient to drive T cells towards a memory fate while having no effect on primary antiviral effector response. Bulk and single-cell gene expression profiling comparing cells transduced with OCA-B and empty vector at peak primary viral response identifies changes in gene expression consistent with later memory formation. Intersecting differentially expressed genes in bulk RNA-seq with prior ChIP-seq identifies direct targets increased (Tbx21, Zeb2, Gadd45b, Socs2) and decreased (Zbtb16, Ccr1) by ectopic OCA-B expression. Single-cell RNA-seq reveals expansion of short-lived effector T cell compartments with increased expression of Gadd45b and Socs2, as well as increased expression of memory-associated genes such as Slamf6, Il7r and Tcf1 in smaller clusters of effector cells with memory potential. We also describe a new OCA-B-mCherry reporter mouse that efficiently labels B and T lymphocytes and that shows high expression in CD4+ TCM cells. Cumulatively, the results demonstrate that OCA-B expression in T cells is necessary and sufficient to promote CD4 T cell memory in vivo.
Supported by grants from NIH (R01 AI100873)
The extracellular ATP receptor P2RX7 imprints a pro-memory transcriptional signature in effector CD8+ T cells
Authors: Van Dijk Sarah , Vardam-Kaur Trupti , da Silva Henrique Borges 
Published: January 2023

Abstract
Development of central memory (Tcm) and resident memory (Trm) CD8+ T cells, which respectively promote immunity in the circulation and in barrier tissues, is not completely understood. Tcm and Trm cells may arise from common precursors, however their fate-inducing signals are elusive. We found that virus-specific effector CD8+ T cells display heterogeneous expression of the extracellular ATP sensor P2RX7. P2RX7-high expression is confined, at peak effector phase, to CD62L+ memory precursors which preferentially form Tcm cells. Among early effector CD8+ T cells, asymmetrical P2RX7 distribution correlated with distinct transcriptional signatures, with P2RX7-high cells enriched for memory and tissue-residency sets. P2RX7-high early effectors preferentially form both Tcm and Trm cells. Defective Tcm and Trm formation in P2RX7 deficiency is significantly reverted when the transcriptional repressor Zeb2 is ablated. Mechanistically, P2RX7 negatively regulates Zeb2 expression, at least partially through TGF-β sensing in early effector CD8+ T cells. Our study indicates that unequal P2RX7 upregulation in effector CD8+ T cells is a foundational element of the early Tcm/Trm fate.
Supported by grants from NIH (R00 AI139381, NIAID).
Maternally derived epidermal growth factor mediates protection in neonates from opportunistic intestinal pathogens.
Authors: Greenfield Kara G , Lothert Petra , Harlow Olivia , Knoop Kathryn 
Published: January 2023

Abstract
The intestinal epithelial barrier is vital in preventing translocating bacteria and subsequent systemic infections. This protection is provided by host factors, by the intestinal microbiota, and, during early life, by breast milk. While the beneficial functions of breast milk are numerous to the nursing infant, it has been observed that proteins such as antibodies and growth factors are secreted in higher concentrations early during lactation and decrease as the milk matures. One such growth factor, epidermal growth factor (EGF), regulates the development of colonic goblet cell-associated antigen passages (GAPs). Decrease of EGF through disruption of synchronous breastfeeding or deletion of the epidermal growth factor receptor from goblet cells alters the kinetics of GAP formation, which can serve as a passage for pathogenic bacteria. In a murine model of early GAP formation utilizing an inhibitor of EGF receptor, we observed a variety of gut-resident pathogens were able to utilize GAPs to disseminate systemically and cause disease, including Escherichia coli, Enterococcus faecalis, Enterobacter cloacae, and Pseudomonas aeruginosa. In human studies, we found a strong correlation between concentrations of EGF in the diet and stool of very low birth weight infants. Notably, formula-fed infants contained significantly less EGF in both diet and stool. Future work will assess EGF in donor milk, which we find has highly variable EGF concentration. This work has strong implications for how diet influences risk of developing infections from gut-resident pathogens and how maternal EGF present in breastmilk is synchronized to the newborn infant to provide appropriate protection in early life.
Supported by grants from NIH (AI144721, DK122187)
The cellular dynamics of early and transitional human breast milk
Authors: LeMaster Cas , Pierce Stephen , Geanes Eric S , Khanal Santosh , Elliott Staci , Scott Allison , Louiselle Daniel , McLennan Rebecca , Truog William , Maulik Devika , Lewis Tamorah , Pastinen Tomi , Bradley Todd 
Published: January 2023

Abstract
Breast milk (BM) is a complex fluid containing factors essential for infant nutrition and immunity. Breastfeeding has been shown to be protective against infections and other immune-mediated diseases during the lactation period and beyond in later childhood. This suggests that BM also imprints the neonatal immune system and influences long-term health. BM also contains populations of maternal-derived cells. Which factors in BM that are important for neonatal health and how they change during lactation have not been well-defined. In this study, we used a single-cell transcriptomic approach to identify and define cell types of early and transitional milk. We collected BM samples from mothers of infants 2–5 days (early milk) and 8–12 days (transitional milk) after delivery. We applied single-cell RNA sequencing on over 154,000 BM-derived cells. We identified 25 transcriptionally distinct populations of cells in the BM. As expected, the most abundant cells in BM were mammary epithelial cells and macrophages. Monocytes, T cells, dendritic cells, and neutrophils were also present and had a higher frequency in week 2, suggesting that some immune cells may remain abundant in the early days of lactation and slowly decline as milk matures. We also detected a small number of stem and progenitor, natural killer and B cells in the BM at a higher frequency in week 1. This work provides an atlas of the cellular component in human milk at two timepoints of lactation. In addition to cell identity and frequencies, we have also uncovered unique molecular pathways that are activated in BM cells. This work will lay the foundation for future studies of how these cells influence neonatal health.
Supported by funding from Children’s Mercy Kansas City
Systemic and mucosal IgA responses are variably induced in response to SARS-CoV-2 mRNA vaccination and are associated with protection against subsequent infection
Authors: Sheikh-Mohamed Salma , Chao Gary , Isho Baweleta , Zuo Michelle , Cohen Carmit , Lustig Yaniv , Nahass Georgie , Salomon Rachel E , Blacker Grace , Fazel-Zarandi Mahya , Rathod Bhavisha , Colwill Karen , Jamal Alainna J , Li Zhijie , deLaunay Keelia Quinn , Takaoka Alyson , Garnham-Takaoka Julia , Patel Anjali , Fahim Christine , Patterson Aimee , Liu Angel , Haq Nazrana , Barati Shiva , Gilbert Lois , Green Karen , Mozafarihashjin Mohammad , Samaan Phillip , Budylowski Patrick , Siqueira Walter , Mubareka Samira , Ostrowski Mario , Rini James , Rojas Olga , Weissman Irving L , Tal Michal Caspi , McGeer Alison , Regev Gili , Straus Sharon , Gingras Anne-Claude , Gommerman Jennifer L 
Published: January 2023

Abstract
SARS-CoV-2 is a novel respiratory virus that has quickly spread across the globe. The virus uses a protein called Spike and its associated receptor binding domain (RBD) to interact with angiotensin converting enzyme-2 (ACE-2) on the surface of epithelial cells in the respiratory tract. Although a definite correlate of protection against COVID-19 has yet to emerge, many studies have quantified anti-Spike and anti-RBD IgG antibody (Ab) levels, as well as neutralizing Ab in the blood to ascertain immunity. This approach misses out on Ab that are produced in the upper respiratory tract (URT) mucosa – the site of viral encounter. Whether intramuscularly (i.m.) administered COVID-19 vaccines can promote immunity in the mucosa is not well understood. We recently completed a study where we showed that anti-Spike/RBD IgG could be detected in the saliva following i.m. vaccination with either two doses of mRNA vaccines (Pfizer or Moderna) or with a heterologous dosing of Astra Zeneca followed by an mRNA vaccine. Administration of a second dose of mRNA boosted the IgG but not IgA response, with only 30% of participants remaining positive for IgA at this timepoint. At 6 months post-dose 2, these participants had diminished anti-Spike/RBD IgG levels, although secretory component associated anti-Spike Ab were more stable. Examining two prospective cohorts we found that participants who experienced breakthrough infections with SARS-CoV-2 had lower levels of vaccine-induced serum anti-Spike/RBD IgA at 2–4 weeks post-dose 2 compared to participants who did not experience an infection, whereas IgG levels were comparable between groups. These data suggest that COVID-19 vaccines that elicit a durable IgA response may have utility in preventing infection.
We received funding support from CIHR (Fund #15992), a COVID-19 Immunity Task force grant, an “Ontario Together” province of Ontario grant, a CIHR team grant to CoVARR-Net, a Donation from the Royal Bank of Canada (RBC) and a donation from the Krembil Foundation to the Sinai Health System Foundation.
Accelerated tumorigenesis in a colorectal cancer model in Siglec-E knockout mice
Authors: Maniaci Brandon Lee , Friedman David , Crotts Sydney , Rajcula Matthew , Theodore Keith , Lee Hyun Se Kim , Shapiro Virginia 
Published: January 2023

Abstract
The lifetime risk for colorectal cancer in the United States is approximately 4%. Individuals with Inflammatory Bowel Disease, including Ulcerative Colitis and Crohn’s Disease, have a substantially increased risk of developing colorectal cancer. The mechanisms for accelerated tumorigenesis due to enhanced inflammation are not fully characterized. Siglecs (sialic acid immunoglobulin lectin-like proteins) are a family of inhibitory receptors that are negative regulators of the immune response. Siglec-E is an inhibitory receptor that is expressed by innate immune cells, including monocytes, macrophages, neutrophils and dendritic cells. Interestingly, mice deficient in Siglec-E have accelerated development of tumors and reduced survival in a spontaneous mouse model of colorectal cancer (TS4-cre LSL-KRasG12D APClox468/wt). While tumors develop at approximately 6 months in mice with Siglec-E, tumors develop at approximately two months in Siglec-E knockout mice. Initial results indicate that Siglec-E knockout mice also have accelerated gut inflammation using the DSS colitis model as compared to WT mice. Current studies are examining inflammation that develops during tumorigenesis in Siglec-E knockout mice.
Ovarian cancer immunogenicity is governed by a narrow subset of progenitor tissue-resident memory T-cells
Authors: Galindo Carmen Maria Anadon , Yu Xiaoqing , Hanggi Kay , Biswas Subir , Chaurio Ricardo , Mandal Gunjan , Martin Alexandra , Payne Kyle K , Innamarato Pasquale P , Harro Carly M , Mine Jessica , Sprenger Kimberly , Cortina Carla , Powers John J , Perez Bradford A , Gatenbee Chandler D , Prabhakaran Sandhya , Marchion Douglas , Heemskerk Mirjam H , Curiel Tyler J , Anderson Alexander R A , Wenham Robert M , Rodriguez Paulo C , Conejo-Garcia Jose R 
Published: January 2023

Abstract
Despite repeated associations between T-cell infiltration and patient outcome, human ovarian cancer remains poorly responsive to immunotherapy. We report that hallmarks of tumor recognition in ovarian cancer-infiltrating T-cells are primarily restricted to tissue-resident memory (TRM) cells. In mouse models we found that TRM T-cells were better than the re-circulating counterpart at controlling tumor growth. Single-cell RNA/TCR/ATAC sequencing of 83,454 CD3+CD8+CD103+CD69+ TRM cells and 24,175 CD3+CD8+CD103− re-circulating TILs showed that progenitor (TCF1low) tissue-resident memory T-cells (TRMstem cells) arise from transitional recirculating T-cells, which depends on antigen affinity/persistence, resulting in oligoclonal, trogocytic, effector lymphocytes. This effector population develops into proliferative lymphocytes that eventually become exhausted TRMs. Immunohistochemistry of 122 high-grade serous ovarian cancer tissues showed that only TRMstem cells, but not re-circulating TCF1+ T-cells, predict ovarian cancer outcome. Therefore, ovarian cancer is indeed an immunogenic disease that depends on ~13% of CD8+ tumor-infiltrating T-cells (~3% of CD8+ clonotypes), which are primed against high-affinity antigens and maintain waves of effector TRM cells.
Support for Shared Resources was provided by Cancer Center Support Grant (CCSG) CA076292 to H. Lee Moffitt Cancer Center and by CCSG CA010815 to The Wistar Institute. This study was supported by grants from NIH (R01CA157664, R01CA124515, R01CA178687, R01CA211913 and U01CA232758 to JRCG; R01CA184185 and RO1CA262121 to PCR.)
Sexual dimorphism and the effect of therapeutic low-dose estrogen in head and neck squamous cell carcinoma
Authors: Knitz Michael W , Bickett Thomas E , Darragh Laurel B , Bhatia Shilpa , Van Court Benjamin , Piper Miles , Douglas Sarah , Karam Sana D 
Published: January 2023

Abstract
Differences between males and females have been identified in a variety of immune-related diseases. Generally, females show stronger immune responses, but also present a higher incidence of auto immune disease. Reasons for this have not been concretely identified but estrogen is believed to be a main driver of sex-dependent differences in biological behavior of disease and response to therapy. Using castration, oophorectomy, and estrogen rescue experiments in various preclinical head and neck squamous cell carcinoma (HNSCC) models, we identify estrogen as a regulator of immune response to treatment. The effect of estrogen is dose-dependent, as higher doses fail to demonstrate the beneficial effects of low dose estrogen. This response is dependent on regulatory T cells and dendritic cells, as knockout of these cell types eliminates the disparity between male and female responses. We also identified estrogen receptor beta agonists as a potential therapeutic strategy providing similar treatment benefit as low dose estrogen, reversing the effects of oophorectomy and stimulating cytotoxic T cell function. Survival analysis from HNSCC patients provided correlational validation of the potential benefit of estrogen as it demonstrates increased overall survival in younger women. Our data implicate low dose estrogen as a potential therapeutic strategy to enhance response to immunotherapy.
Sana D. Karam is funded by the NIDCR (R01 DE028529-01, R01 DE028282-01) and Colorado Head and Neck Cancer SPORE and receives funding from Astra Zeneca, Genentech, and Ionis. This study was partly supported by the National Institutes of Health P30CA046934 Cancer Center Support Grant, including the Bioinformatics and Biostatistics Shared Resource Core, the Cancer Center Flow Cytometry Shared Resource, and the Genomics and Microarray Shared Resource at the University of Colorado. This study was also partly supported by the Cancer League of Colorado.