bluuebunny/crossref_metadata_2025_split huggingface dataset:
Estradiol Augments the Production and Actions of Polymorphonuclear Cells to Promote Lymphangioleiomyomatosis (LAM) Progression
Authors: Minor Briaunna M. N. , Hammes Stephen R 
Published: February 2023

Abstract
Predominately affecting women of reproductive age, lymphangioleiomyomatosis (LAM) is a rare lung disease characterized by slowly growing, estrogen-sensitive metastatic smooth muscle cell-like adenomas that result in cystic lung changes and loss of pulmonary function. We have previously reported that estrogen is required to maintain LAM-like tumor progression in a uterine-specific Tsc2-knockout murine model. Interestingly, the observed estrogen sensitivity in vivo is more markedly pronounced than that of our estrogen receptor-positive TSC2-null cells when stimulated with estradiol (E2) in vitro, suggesting that estradiol may act elsewhere to promote LAM progression. We hypothesized that, in addition to direct effects of estrogen on tumor cells, estrogen might also stimulate tumor growth by promoting polymorphonuclear cell (PMN) production in the bone marrow and actions in the tumor microenvironment. Here, using bone marrow cultures, we demonstrate that estradiol is a potent inducer of PMN production. This effect occurs equally with both male and female bone marrow. Employing both pharmacologic agents and bone marrow from ER_ knockout mice, we showed that ER_ is necessary for promoting a PMN fate for myeloid progenitors. While we see that the migratory and invasive capacities of TSC2-null cell lines are not augmented when stimulated with E2 directly, evidence shows estrogen promotes the pro-tumorigenic function of PMNs co-cultured with TSC2-null cell lines. These data provide insight into the robust effect of in vivo estrogen stimulation as estradiol may be a dual effector in LAM tumor progression and great target for anti-LAM therapeutic strategy.
Intravital imaging experiments track CD8 T cells searching for Plasmodium liver stages, but do the imaging frequency and duration impact the detection of CD8 T cell bias?
Authors: Zenkov Viktor S , Ganusov Vitaly V , Cockburn Ian A , O’Connor James , McNamara Hayley 
Published: February 2023

Abstract
Vaccine-induced CD8 T cells can provide sterilizing protection by killing Plasmodium sporozoites in mice, but how do they find those parasites? We recently showed that most liver-localized CD8 T cells search for infected hepatocytes randomly, with some cells displaying bias toward the infection site only when there is already a CD8 T cell cluster present. Our initial intravital microscopy experiments minimized tissue damage from laser exposure by using low imaging frequency (volume/1.5–2min) and were fairly lengthy (1–2h). In these experiments, activated CD8 T cells displayed Brownian-like turns and slow average calculated speeds of 1–3 um/min. Recently, we performed new experiments in which we recorded the movement of CD8 T cells at a higher frequency (volume/0.2min) at the cost of shorter experiments (30min). Interestingly, observed speeds ranged between 5 and 18 um/min, suggesting that the calculated speed of CD8 T cells depends on the details of the experiment. Further testing found that imaging frequency impacts the estimated average speed per cell: reducing the imaging frequency from 0.2min to 2min can reduce the average calculated cell speed 5-fold, partially explaining the discrepancy between our datasets’ speeds. Nonetheless, the new data still suggest that some CD8 T cells display attraction towards the infection site only when there is a cluster of CD8 T cells already present. Taken together, our results highlight the impact of details of intravital imaging experiments on inferred parameters of T cell movement in complex environments.
Skewed escape from X-inactivation underlies female bias in Sjögren’s syndrome
Authors: Shaw Teressa , Zhang Wei , McCoy Sara S. , Qiu Xueer , Pagenkopf Adam , Galipeau Jacques , Liang Yun 
Published: February 2023

Abstract
Many autoimmune diseases feature strikingly increased prevalence in females, such as systemic lupus erythematosus (female-to-male ratio 9:1), systemic sclerosis (female-to-male ratio 11:1) and Sjögren’s syndrome (female-to-male ratio 14:1). However, the molecular basis underlying female-biased autoimmunity remains elusive. To address this knowledge gap, we performed transcriptomic profiling of minor salivary gland-derived mesenchymal stromal cells (MSCs) from primary Sjögren’s syndrome (pSS) patients and control subjects. While autosomal gene expression profiles were largely comparable between pSS and control MSCs, we detected major differences in the regulation of X-linked genes. In control female MSCs, X-linked genes that escaped inactivation were expressed from both parental and maternal X chromosomes with a median paternal ratio of ~0.5. However, in pSS female MSCs, escapees exhibited preferential expression from one of the two X chromosomes. Concomitantly, pSS MSCs showed decrease in XIST levels and reorganization of H3K27me3+ foci in the nucleus, suggesting a global dysregulation of X-inactivation maintenance. The skewing in X escape in pSS MSCs was accompanied by mislocation of protein products encoded by the escapees. Given the hallmark features of inflammation and fibrosis in pSS salivary glands and growing evidence supporting MSCs contributing to tissue homeostasis, our data suggests that dysregulation of X-inactivation maintenance, a female-specific process, contributes to the female-bias in pSS susceptibility.
Promotion of Autoimmune Arthritis via Tryptophan Metabolism and Production of the Bacterial-Derived Tryptophan Metabolite Indole
Authors: Trent Brandon J , Jubair Widian , Fetchner Sabrina , Chriswell Meagan , Kuhn Kristine 
Published: February 2023

Abstract
Dysbiosis of gut bacterial communities in autoimmunity is a noted phenomenon in both murine models and human patients; however, the mechanisms of dysbiosis that promote disease pathogenesis remain unclear. In agreement with such studies, our lab has published that administration of antibiotics to deplete the microbiota late in the course of murine collagen-induced arthritis (CIA) significantly ameliorated disease. To understand the mechanisms by which microbiota depletion would significantly decrease CIA, we analyzed cecal metabolites by LC-MS during CIA and after antibiotic treatment and observed significant changes in various tryptophan metabolite levels. Interestingly, mice placed on a tryptophan-free diet had significantly decreased CIA development and increased in regulatory T cell (Treg) populations in the spleen. Further studies identified increases in indole, a bacterial-derived, tryptophan metabolite that strongly correlated with CIA progression. Direct administration of indole during CIA promoted disease and led to increases in T follicular cell (Tfh) and B cell populations in the peyer’s patches and mesenteric lymph nodes. Ex vivo stimulation of murine splenic B cells with indole resulted in significantly increased IgG production and a ≈10% increase in the frequency of CD23+ B cells. Our results suggest that gut dysbiosis due to CIA results in altered tryptophan metabolism and indole production, which promotes CIA pathogenesis via activation of T and B cell populations and antibody production. Precise understanding of which indole metabolites are involved and how they influence mucosal and systemic immune responses will help elucidate the role of intestinal dysbiosis in autoimmunity.
IL-17 dependent fibroblastic reticular cell training boosts host-protective immunity to mucosal infection
Authors: Wu Dongwen , Majumder Saikat , McGeachy Mandy J 
Published: February 2023

Abstract
Prior experience of inflammatory stimuli is thought to reduce morbidity and mortality to newly encountered infections through innate immune training that can be enhanced by childhood vaccination. Here we demonstrate that IL-17 signaling in lymph node fibroblastic reticular cells (FRC) is required for efficient clearance and protective antibody-mediated immunity to the gut bacterial pathogen Citrobacter rodentium. Furthermore, prior experience of gut inflammation led to an enhanced antibody response to C. rodentium without altering the kinetics of bacterial infection. However, colon inflammation was paradoxically reduced by previous inflammatory exposure, due to IL-10 production by B cells. Mechanistically, FRC-intrinsic IL-17 dependent activation mediated the trained response, which includes enhanced metabolic response of FRC to infection. These data identify a new pathway for immune training through FRC that modulates the adaptive response, enhancing protective antibody while reducing gut inflammation, to more efficiently preserve host health.
Stochastic pooling of IKK/NEMO complexes describes information transmission in the NF-KB pathway in response to IL-1 and TNF stimulation.
Authors: Cruz J. Agustin , Mokashi Chaitanya , Lee Robin E.C. 
Published: February 2023

Abstract
A myriad of inflammatory cytokines regulates signaling pathways for normal cellular physiology. The IKK complex is an integration hub for extracellular cytokines that govern NF-κB signaling. In response to inflammation, IKK is activated through recruitment to receptor-associated protein assemblies. How and what information IKK complexes transmit about the milieu are open questions. Here we track dynamics of IKK complexes and nuclear NF-κB to identify upstream signaling features that determine same-cell responses. Quantitative imaging experiments and computational modeling of single complexes reveals that their size, number, and timing relays cytokine-specific information with feedback control that is independent of transcription. Our results provide evidence for variable-gain stochastic pooling, a noise-reducing motif that enables parsimonious and cytokine-specific signaling. We argue that emergent properties of stochastic pooling motif are general principles of receptor signaling, and we present new modules of regulation of NF-kB signaling in response to cytokine stimulation.
FMS-like tyrosine kinase 3 ligand (FLT3L) administration alters rat classical and plasmacytoid dendritic cell phenotype in a tissue-specific manner
Authors: Carlson Kristin N , Verhagen Joshua , Verhoven Bret , Pavan-Guimaraes Juliana , Al-Adra David 
Published: February 2023

Abstract

Background:
Characterization of tissue-resident dendritic cell (DC) subsets, including classical DCs (cDCs) and plasmacytoid DCs (pDCs), is complicated experimentally by their rare nature. Many investigators have artificially increased DC yields through administration of the cytokine FMS-like tyrosine kinase 3 ligand (FLT3L). Despite its previous use experimentally, no study has yet described the effects of in vivo FLT3L administration on rat tissue-resident DCs. This study provides an advanced characterization of rat tissue-resident cDCs and pDCs following FLT3L injection.

Methods:
Lewis rats were injected once daily for 10 days with PBS vehicle or with 10, 50, or 100μg of FLT3L. Animals were sacrificed on the 11th day and dissected to obtain liver, spleen, and mesenteric lymph nodes. Cells were stained for 13-color flow cytometry and data was acquired on the Cytek Aurora spectral cytometer.

Results:
cDC and pDC enumeration increased in a dose-dependent manner following FLT3L administration. Expression levels of MHC I, MHC II, CD40, CD80, CD86, and PD-L1 were evaluated on liver, spleen, and mesenteric lymph node-resident cells. MHC II, CD80, and CD86 levels were significantly decreased on liver-resident cDCs but remained unchanged or increased on spleen and lymph node cDCs. Changes in MHC I/II and costimulatory molecule expression levels on pDCs varied by tissue and marker.

Conclusions:
FLT3L administration in vivo increased cDC and pDC enumeration, generated immature cDCs in the liver, and increased cDC maturation in the spleen and mesenteric lymph nodes. This comprehensive characterization of rat liver, spleen, and lymph node-resident DCs is key to understanding the effects of FLT3L on cDC and pDC phenotype.
Lineage tracing and transplantation studies indicate endothelial origin of B-1a cells in the mouse embryo
Authors: Yoshimoto Momoko , Nishida Chika , Cornelius Samuel , Valiente Noemi , Kobayashi Michihiro 
Published: February 2023

Abstract
Innate-immune like B-1a cells are unique in its functions and developmental origin compared to conventional adaptive immune B-cells (B-2 cells). Especially, their origin and developmental pathway have been argued over four decades between two proposed models: the lineage model and the selection model. Although many transplantation studies support the lineage model in which B-1a cells are derived from fetal progenitors, contradictory results of fetal liver transplantation studies raise a question as to whether fetal liver hematopoietic stem cells (HSCs) are the primary source of the peritoneal B-1a cells. In order to address this question, we took two approaches using transplantation assays and lineage tracing mouse models that enable us to trace Tomato+ progeny of endothelial cells (ECs, Cdh5CreERT mice) and HSCs (Fgd5CreERT2 mice) at a time of Tamoxifen (Tam) injection, respectively.

Results:
First, we transplanted HSC-precursors (pre-HSCs) from the aorta-gonad-mesonephros (AGM) region of the mouse embryo at embryonic (E) 11.5 into irradiated NSG neonates. Interestingly, we found more B-1a repopulating capacity than multi-lineage in the pre-HSC population. The lineage tracing of hemogenic ECs revealed that B-1a progenitors and HSCs were simultaneously produced from ECs during E9.5 to 10.5, but B-1a production is diminished after E11.5 while HSC were still labeled at E11.5. The HSC-tracing model demonstrated the minimal contribution of FL HSCs to the postnatal B-1a cell pool.

Conclusion:
Our data addressed the long-lasting controversy and showed HSC-independent B-1a cell development in the mouse embryo.
The Inhibitor of MyoD Family A Promotes Terminal Myeloid Differentiation
Authors: Houser Jeremy S , Patel Maulin , Onopiuk Marta , Tsiokas Leonidas , Humphrey Mary Beth 
Published: February 2023

Abstract
The differentiation of hematopoietic stem and progenitor cells is tightly regulated to maintain immune and blood homeostasis while also responding to injury and immunological insult. Myeloid lineage-committed progenitor cells are highly proliferative, but do not self-renew and must be continually replenished by the differentiation of hematopoietic stem cells. The process of terminal differentiation is controlled by various cell-intrinsic and - extrinsic factors, and the interactions between these factors remain not well understood. We hypothesize that the Inhibitor of MyoD Family A (I-mfa) protein is a novel regulator of myeloid lineage differentiation through its interaction with other cell-intrinsic factors.
To examine this, mice which lack I-mfa expression (I-mfanull) were compared to wild type (WT) 129Sv age- and sex-matched controls. I-mfanull animals had reduced spleen size (both in weight and total cellularity), but had preserved relative proportions of B, T, and myeloid cells as analyzed by flow cytometry when compared to WT. Analysis of the bone marrow (BM) myeloid lineage-committed progenitors showed an accumulation of common myeloid progenitors (CMP) and granulocyte-macrophage progenitors (GMP) in I-mfanull animals when compared to WT. In serial colony forming unit assays I-mfanull BM produced fewer mature colonies and continued to produce colonies after 5 passages, suggesting aberrant self-renewal.
Using myeloid cell lines lacking or overexpressing I-mfa, the molecular mechanism by which I-mfa acts is being elucidated. These results taken together suggest I-mfa acts to promote differentiation of myeloid cells, an effect which may be important in the treatment of myeloid neoplasms or bone marrow failure.
Microbial exposure in early life alters the ontogeny and function of CD8+ T cells
Authors: Tabilas Cybelle , Reynaldi Arnold , Daly Ciarán W.P. , Mon Kristel Yee , Wesnak Samantha , Smith Norah L. , Davenport Miles P. , Grimson Andrew W. , Rudd Brian D. 
Published: February 2023

Abstract
Evidence has shown that microbial exposure in early life can have long-lasting effects on the health status of an individual. However, the underlying basis of how microbes shape the immune system is not well understood. In this study, we utilized a novel pet shop mouse model and compared the immune development and function of laboratory mice raised in either a clean (SPF) or dirty (pet-shop) environment. When we challenged the two groups with an intracellular pathogen in adulthood, we found that mice raised in the dirty environment mounted a more robust CD8+ T cell response and were highly resistant to infection. Adoptive transfer experiments with CD8+ T cells from clean and dirty mice into T cell deficient recipients indicated that differences in immune susceptibility were linked to changes in the CD8+ T cell compartment. Moreover, the dirty mice had an increased accumulation of antigen-specific CD8+ T cells and produced more effector molecules. To understand how the ontogeny of the CD8+ T cell compartment was altered in the dirty mice, we used our fate-mapping timestamp mice and found a higher proportion of fast-acting fetal-derived CD8+ T cells and fewer slow-acting adult-derived CD8+ T cells present in the dirty mice. We found that immune susceptibility could be normalized between clean and dirty mice by depleting the fetal layer of CD8+ T cells. Collectively, these findings suggest that microbial exposure leads to permanent changes in the immune system by altering the developmental layering of the peripheral T cell compartment. We are currently using a combination of RNA- and ATAC-seq to gain a deeper understanding of how the microbial environment mediates changes to CD8+ T cells at the gene regulatory level.
The distinct co-stimulatory requirements for thymic MAIT and NKT cell development
Authors: Celli Sofia I , Watanabe Masashi , Alkhaleel Farrah , Hodes Richard 
Published: February 2023

Abstract
Mucosal-associated invariant T (MAIT) cells are a unique lineage of T cells that develop in the thymus. MAIT cells are positively selected by TCR recognition of microbial metabolite / MHC Class I-like molecule MR1 complexes presented on CD4/CD8 double-positive thymocytes. The developmental program of MAIT cells has been characterized as similar to that of invariant Natural Killer T (NKT) cells. Both MAIT and NKT cells acquire innate-like effector phenotypes during thymic development, and in each case differentiate into multiple subsets, with mature NKT cells categorized as either NKT1, NKT2 or NKT17, and mature MAIT cells as MAIT1 or MAIT17. We previously found distinct B7/CD28 and CD40L/CD40 co-stimulatory requirements for the development of thymic NKT subsets in BALB/c mice. In the absence of B7/CD28 co-stimulation, all NKT subsets were decreased, while only the NKT2 subset was specifically abrogated when lacking CD40L/CD40 co-stimulation. In the present study, we investigated the co-stimulatory requirements for MAIT cell development. Interestingly, MAIT17 cells were significantly increased when lacking either B7 or CD40 co-stimulation, while MAIT1 development was largely unaffected. These results revealed distinct co-stimulatory requirements for the thymic development of MAIT and NKT subsets. We are currently investigating the underlying mechanisms leading to the increase of MAIT17 cells in the absence of co-stimulatory signals.
Chemically-induced colitis is more severe in mice expressing TLR4 SNPs: a novel model for studying human Inflammatory Bowel Disease
Authors: Vlk Alexandra , Vogel Stefanie N 
Published: February 2023

Abstract
Innate immune cells sense “danger” encountered in the environment using pattern recognition receptors (PRRs). The Toll-Like Receptors (TLRs) are a large family of PRRs, among which TLR4 is unique in the complexity of its downstream signaling. Single nucleotide polymorphisms (SNPs) expressed in the ectodomain of human TLR4 (D299G/T399I), have been shown to affect the functionality of this receptor. Population genetics studies have shown that these SNP frequencies vary geographically and among different ethnic backgrounds, with both SNPs co-segregating in ~10% of Caucasians. Humans expressing these TLR4 SNPs are LPS-hyporesponsive and exhibit increased susceptibility to certain bacterial infections and severe colitis. Our laboratory engineered a knock-in mouse strain that homozygously expresses homologous TLR4 SNPs (D298G/N397I). TLR4-SNP mice and their macrophages are LPS-hyporesponsive and exhibit altered susceptibility to bacterial and viral infections. To evaluate the mechanisms by which TLR4 SNPs contribute to Inflammatory Bowel Disease (IBD), we utilized a chemically-induced colonic inflammation and repair model to compare the responses of wild-type (WT) vs. TLR4-SNP mice. After 7 days of 3% dextran sodium sulfate (DSS) in the drinking water, followed by an additional 7 days of recovery, TLR4-SNP mice were found to be significantly more sensitive than WT mice to DSS-induced colitis (e.g., weight loss, fecal blood, colon length, histopathology, and a significant delay in recovery after DSS removal). Ongoing studies to determine if differences in the microbiota of TLR4-SNP mice vs. WT mice contribute to this phenotype are expected to provide insights into mechanisms underlying human IBD. Supported by NIH T32 AI095190.
Shared B cell but not T cell intratumoral immune repertoire among patients with low grade appendiceal mucinous neoplasms
Authors: Murthy Pranav , Weber Daniel , Sharma Sagar N. , Saini Pragosh , Pan Wenjing , Byrne-Steele Miranda , Han Jian , Pai Reetesh K. , Zureikat Amer H. , Choudry Haroon A. , Lotze Michael T. 
Published: February 2023

Abstract
Introduction: Low grade mucinous appendiceal neoplasms (LAMNs) are indolent tumors that present with widespread peritoneal metastases. Despite standard of care cytoreductive surgery and heated intraperitoneal chemotherapy (CRS-HIPEC), patients with LAMNs often recur within 1–3 years and respond poorly to chemotherapy.
Methods & Results: Histopathologic review identified clusters of lymphocytes associated with epithelial hypercellularity among larger pools of mucin. Utilizing dimer avoidance multiplex-PCR and next generation sequencing, we explored the adaptive immune response in patients with recurrent LAMNs (n=10). Bulk sequencing of tumor tissue following initial CRS-HIPEC revealed a predominant expression of B cell over T cell receptors (mean expression: 0.5% Vα, 1.4% Vβ, 0.01% Vδ, 12.6% IgH, 47.6% Igκ, 37.8% Igλ). Very interestingly, we observed a heightened IgE fraction (8.4%±1.1) in patients with LAMNs, greater than both healthy donor blood (0.57%±0.36, p&lt;0.0001, n=239) and pretreated high-grade tumors (n=66, 3.3%±2.4, p&lt;0.0001). Although the TCR repertoire was extremely private in patients, the BCR repertoire included several shared public and private clones: 11 IgH private CDR3s shared between 2 patients, 64 Igκ public CDR3s shared among &gt;7 patients, 261 private Igκ CDR3s shared among 3 patients, 20 public Igλ CDR3s shared among &gt;7 patients, and 137 private Igλ CDR3s shared between 2 patients, an intriguing and unexpected finding.

Conclusions:
This, we believe, is the first report of adaptive immunity in low grade mucinous neoplasms that identifies a skewed but diverse infiltration of T and B cells, that warrants study in additional patients.