bluuebunny/crossref_metadata_2025_split huggingface dataset:
Maternal control of dietary and gut microbial antigen exposure promotes enhanced tolerance and prevents allergic responses
Authors: Knoop Kathryn A , McDonald Keely , Newberry Rodney D 
Published: January 2023

Abstract
The incidence of allergic disorders is rapidly increasing in children in Western societies, and is linked to allergen avoidance diets and decreased exposure to microbes and microbial antigens in early life. Exposure to dietary antigens and the microbiota via the gastrointestinal tract early in life is associated with a reduced risk of allergic disorders, however the optimal time to introduce these antigens, where the immune system encounters these antigens, and what controls the immune systems exposure to define this time in early life is largely unknown. We identified a window of time in early life in which luminal antigens are shunted to the colonic lamina propria (LP) immune system to induce enhanced tolerogenic responses. Delivery of luminal antigens to the colonic immune system during this time in life was mediated by the formation of goblet cell-associated antigen passages (GAPs), which were regulated by goblet cell (GC) intrinsic sensing of breast milk derived epidermal growth factor (EGF) and the blooming gut microbiota. Disruption of GAP formation and antigen delivery during this time in early life abrogated tolerance to dietary antigens, decreased regulatory T cells, and induced a skewing of the immune system toward systemic Th2 responses. We propose that maternal control of antigen delivery during infancy allows for the proper education of the immune system against dietary and microbial antigens, and induction of regulatory T cells necessary to suppress inappropriate Th2 responses later in life.
A unique population of IFNγ-producing innate lymphoid cells promotes Campylobacter-induced colitis
Authors: Muraoka Wayne T , Xia Qingqing , Halperin Sydney , Koroleva Ekaterina , Tumanov Alexei 
Published: January 2023

Abstract
Inflammatory bowel diseases (IBD) are thought to arise from an overly aggressive immune response to the intestinal microbiota. Campylobacter jejuni is a human enteric pathogen that is a major cause of gastroenteritis worldwide. C. jejuni infection also increases the risk of developing IBD-like sequelae. Prior studies showed IL-10 deficient mice develop fulminant colitis when infected by C. jejuni. However, the roles of adaptive and innate immune cells in promoting colitis remain unresolved. Here, we show that T cells are not necessary for C. jejuni-induced colitis. In vivo depletion of Thy1+ cells or neutralization of IFNγ in T cell deficient mice significantly ameliorated C. jejuni-induced colitis, indicating a pathogenic role for IFNγ-producing innate lymphoid cells (ILC1s). In mice, ILC1s are currently identified as NK1.1+NKp46+ cells and prior studies reported that depleting NK1.1+ cells ameliorated ILC1-driven colitis. Unexpectedly, despite effective depletion of NK1.1+ cells in mice lacking T cells, this treatment did not impact IFNγ production or colon pathology after C. jejuni infection. Phenotyping analysis revealed IFNγ-producing ILCs (Thy1+Lin−Tbet+Eomes−GnzA−RORγtlow/−) that accumulated in the colon during C. jejuni-induced colitis did not express NK1.1 or NKp46. Further, mice deficient for the transcription factor RORγt possessed markedly fewer NK1.1−NKp46− ILC1s in the intestine and exhibited reduced severity of C. jejuni-induced colitis. Based on the lack of NK and natural cytotoxicity receptors (NCR) and in accord with the current ILC nomenclature, we propose the tentative designation of NCR−ILC1. Together, these results identified a unique population of ILC1s that promote colitis.
Generation of colonic T-cell mediated protective immunity after rectal infection with LCMV
Authors: Trapecar Martin , Cohen Benjamin , Khan Shahzada , Woodruff Erik , Luong Julie , Sanjabi Shomyseh 
Published: January 2023

Abstract
How viral-specific immunity is generated in the colon is not well understood, leaving a major gap in our knowledge of defining the parameters needed to develop vaccines against rectally transmitted pathogens. Clearance of most intracellular pathogens requires CD8 T cells, and since unlike the small intestine, the colon lacks organized lymphoid aggregates such as Peyer’s patches, generating protective immunity requires a series of orchestrated events to mobilize CD8 T cells to this mucosal organ. Complicating the generation of protective immunity is the fact that the colon naturally serves as an inductive site for tolerance against many commensal organisms and food antigens. Our central hypothesis, supported by our data, is that the tolerogenic nature of the colon limits induction of robust inflammatory responses needed to activate CD8 T cells for their efficient and timely recruitment to the colonic mucosa. We are using our newly established lymphocytic choriomeningitis virus (LCMV) rectal infection model to determine why CD8 T-cell priming is sub-optimal after rectal viral transmission of LCMV and why the activated CD8 T cells are unable to control the virus from spreading systemically. We are employing several genetic tools to ablate various monocytic and dendritic cell populations to determine their specific role in promoting CD8 T cell priming, activation, differentiation, and migration after rectal LCMV infection. This knowledge is required to break down barriers that have prevented development of successful T-cell-mediated vaccines against sexually transmitted RNA viruses, such as HIV, and will ultimately support the development of vaccines against sexually transmitted infections.
Enhanced innate CD161+ T cell responses in mucosal tissues of nonhuman primates
Authors: Rout Namita , Woods Adrienne 
Published: January 2023

Abstract
Innate T cells recognize non-peptide antigens presented by non-classical MHC molecules, such as CD1 and MR1. The three major populations within the innate T cell group, namely, γδ T cells, MAIT cells, and NKT cells express high levels of the C-type lectin molecule CD161. To understand the significance of CD161 expressing innate T cells in mucosal immunity, we investigated the phenotype and functions of these cells in mucosal tissues, lymph nodes and blood of rhesus macaques. The frequency of CD161+ T cells was higher in the lungs and colonic mucosa (9.4%±3.8 and 6.2%±1.9) in comparison to peripheral blood (2.9%±1) and mesenteric lymph nodes (0.5%±0.3). Regardless of the tissue compartment, 30 to 80% of the CD161+ T cells comprised of γδ T cells and TCR Vα 7.2+ MAIT cells in approximately equal proportions. However, only 1 to 6% consisted of CD1d tetramer+ NKT cells. Moreover, the mucosal tissue CD161+ T cells displayed a highly activated phenotype based on CD69 and HLA-DR expression in comparison to blood and lymph node cells. HMBPP stimulated γδ T cells from lungs and colonic mucosa produced significantly higher amounts of IFN-γ, Perforin and IL-17 than those in blood. Similarly, E.coli stimulated MAIT cells from mucosal tissues produced significantly higher amounts of IFN-γ and IL-17 than those in blood. The enhanced Th1 and Th17 type effector functions of mucosal innate T cells along with their cytotoxic potential suggests that they can mount early immune responses to mucosal pathogens like HIV and Mtb prior to the development of adaptive immune responses and thereby play a role in the initial control of pathogen burden/dissemination.
Loss of intestinal epithelial autophagy leads to catastrophic susceptibility to acute T. gondii-mediated inflammation
Authors: Burger Elise , Lopez-Yglesias Americo H , Yarovinsky Felix 
Published: January 2023

Abstract
The protozoan parasite Toxoplasma gondii triggers severe small intestinal immunopathology characterized by inflammation, Paneth cell loss and bacterial dysbiosis. Paneth cells are the predominant secretory epithelial cells which reside at the base of the crypt and release antimicrobial peptides that maintain intestinal homeostasis. T. gondii-triggered IFN-γ is a major inducer of Paneth cell death. However, mechanisms of IFN-γ-mediated Paneth cell death are unknown. We recently observed that under steady state conditions Paneth cells undergo active autophagy, which is both microbiota- and IFN-γ-dependent. To study autophagy’s role in T. gondii–induced Paneth cell loss we used mice deficient in the critical autophagy gene Atg5 in the intestinal epithelium (Villin-Cre × ATGfl/fl mice, E-ATG5−/−). Infected E-ATG5−/− mice exhibited catastrophic inflammation, characterized by the complete destruction of the intestinal crypts, establishing ATG5’s protective role during intestinal inflammation. To determine ATG5’s protective mechanism we generated ATG5-deficient intestinal organoids and observed that lack of ATG5 increased sensitivity to the TNF-induced cell death. Organoid death was further exacerbated when TNF stimulation occurred in combination with IFN-γ. Reexamining our in vivo model showed significantly elevated levels of TNF receptor 2 in infected E-ATG5−/− mice, indicating these mice are prone to TNF-triggered small intestinal immunopathology. Our results reveal that ATG5 expression within Paneth cells plays a highly protective role against cytokine-mediated immunopathology in the intestine during acute T. gondii infection.
The proteolytic activity of Porphyromonas gingivalis is a critical virulence factor in the murine model of periodontitis
Authors: Choi Youngnim , Baek Keum Jin , Choi Yun Sik 
Published: January 2023

Abstract
Porphyromonas gingivalis, a periodontal pathogen, has an inter-strain variability in virulence. We recently reported that the cell invasion ability, but not cytokine proteolytic activities, of P. gingivalis has strong positive correlations with clinical parameters of subjects who harbored the isolates. To determine the contribution of the invasion ability and proteolytic activity to the virulence of P. gingivalis in the murine model of periodontitis, three clinical isolates KUMC-P1 (P1: low cell invasion ability and low proteolytic activity), KUMC-P4 (P4: low cell invasion ability but high proteolytic activity), and KUMC-P8 (P8: high cell invasion ability but low proteolytic activity) were orally inoculated in Balb/c mice. The net alveolar bone loss induced by three strains was P4 &gt; P8 &gt; P1, where the difference between P8 and P1 was not significant. The amounts of P. gingivalis within the gingival tissues examined by in-situ hybridization using a P. gingivalis-specific probe were increased by all three strains, which had a significant positive correlation with alveolar bone loss. Whereas P1 and P8 induced a significant IgA antibody response, P4 induced a slight but not significant IgG1 antibody response against P. gingivalis. In vitro, P4 had the greater effect than other strains on the epithelial barrier disruption evidenced from the reduction in the levels of tight junction proteins and transepithelial resistance. In conclusion, the proteolytic activity of P. gingivalis was a critical virulence factor in the murine model of periodontitis, which may be attributed to the paracellular pathway-dependent bacterial invasion of the gingival tissue in mice.
Fast and efficient enrichment of functional ILC2 from human whole blood
Authors: Valdez Yanet , Kyei Stephen K. , Poon Grace F.T. , Takei Fumio , Peters Carrie , Woodside Steve M. , Eaves Allen C. , Thomas Terry E. 
Published: January 2023

Abstract
Group 2 innate lymphoid cells (ILC2) are a functionally distinct subset of recently identified immune cells with important roles in type-2 immunopathologies such as allergies, asthma, helminth infections and other metabolic diseases. Studying these rare cells is challenging due to a lack of specific surface markers, and currently multicolor flow cytometric analysis and cell sorting are the only methods to characterize and isolate ILC2s. However, the scarcity of these cells makes flow cytometry time-consuming, expensive and often results in low purities and recoveries. Thus, better approaches for effective identification and isolation are essential to further understanding of ILC2 biology and function. We have developed a rapid and efficient method for enrichment of human ILC2 from whole blood. In brief, non-ILC2 cells in whole blood were crosslinked to red blood cells already present in the sample using RosetteSep™. The sample was then layered over Lymphoprep in a SepMate™ tube, spun at 1200 x g for 10 minutes (min), and the untouched, desired cells simply poured off. Cells were washed once and were then ready for subsequent analysis. Starting with only 0.01 – 0.07% in whole blood, ILC2 were enriched 350 ± 220 fold to 8.2 ± 6.8% in 35 min (means ± SD, n=17). Subsequent cell sorting from these pre-enriched samples was faster and yielded higher purity ILC2 than sorting from non-enriched controls (n=3, p&lt;0.05 paired t test). Sorted ILC2, both pre-enriched and non-enriched controls, were cultured and stimulated, and secreted similar high levels of IL-13 as assessed by ELISA, indicating that these cells are functional. In summary, ILC2 pre-enrichment improves sorting efficiency, increases ILC2 purity, and maintains ILC2 functionality.
Immunomagnetic enrichment of regulatory T cells directly from whole blood without the need for density gradient centrifugation.
Authors: Richter Anne , Steinbrück Philipp , Schenk Tobias , König Sindy , Siemer Ramona , Bacher Petra , Hohnstein Thordis , Scheffold Alexander , Dose Christian 
Published: January 2023

Abstract
Regulatory T cells (Tregs) play a pivotal role in maintaining peripheral tolerance of unspecific or excessive immune responses. Therefore, the enrichment of these cells and their cultivation is of great interest for basic and translational research.
Accordingly, there is a considerable need for continually improved and reliable cell isolation protocols. We introduce here a rapid, easy, and convenient strategy for the isolation of regulatory T cells directly from anticoagulated whole blood without the need for density gradient centrifugation.
The new immunomagnetic separation protocol provides an easy and fast solution for the highly specific isolation of CD4+ CD25highCD127− regulatory T cells directly from whole blood within 30 minutes ready for downstream application such as flow cytometric phenotyping, gene expression studies, cytokine analysis, suppression assays, or cell expansion.
The procedure, leads to populations of CD4+ CD25highCD127− cells with high yields (2×104/mL whole blood), purities (&gt;90%) and viability (&gt;98%) as proven by flow cytometric analysis. In addition, enriched Tregs were capable for in vitro expansion for several weeks while remaining stable FoxP3 expression.
Generation of functional mini thymus-like units with self-assembling EAK16-II/EAKIIH6 hydrogel
Authors: Tajima Asako , Liu Wen , Pradhan Isha , Bertera Suzanne , Lakomy Robert , Rudert William A , Trucco Massimo , Meng Wilson S , Fan Yong 
Published: January 2023

Abstract
A functional thymus is crucial to produce mature, self-tolerant T cells. Crosstalk between the residing thymic stromal cells, especially the predominant population of thymic epithelial cells (TECs), and the developing thymocytes is essential for thymopoiesis. The survival and proliferation of TECs depend on a unique 3 dimensional (3-D) configuration of the thymus microenvironment. Disorganization of TECs results in a loss of gene expressions essential for TEC growth and survival, as well as inability to produce functional T cells. Recapitulating the property of thymus has been proven to be challenging.
In our study, we have incorporated EAK16-II, a low molecular weight peptide that self-assembles into beta-sheet fibrils, and its histidinylated analogue, to induce 3-D aggregation of TECs. When the adaptor complex comprised of anti-EpCAM IgG, anti-His and recombinant protein A/G molecules were mixed with TECs in the hydrogel, TECs were captured into small clusters. TECs cultured in this condition maintained their molecular properties and were viable up to 2 weeks. Furthermore, when the TEC clusters in the hydrogel were transplanted into athymic nude mice, T cell development was observed, and these newly generated T cells proliferated upon stimulation with allogenic cells. These results demonstrate that self-assembling EAK16-II/EAKIIH6 system with addition of tri-component adaptor complex may be a useful tool to organize TECs in a 3-D-like structure to support T cell development, suggesting a possibility to generate injectable mini thymus-like units to restore adaptive immune system.
A SMARTer approach to profiling the human T-cell receptor repertoire
Authors: Taylor Sarah , Yasuyama Nao , Farmer Andrew 
Published: January 2023

Abstract
Profiling T-cell receptor (TCR) repertoires involves characterizing the diversity of TCR nucleotide sequences in a sample, and is an increasingly popular approach for analyzing the composition of the adaptive immune system. While low-throughput approaches have yielded important insights concerning TCR repertoire dynamics, development of next-generation sequencing (NGS) technologies has dramatically expanded research prospects.
Using SMART® technology, we have developed an NGS library preparation kit for TCR profiling that employs a 5′ RACE-like approach to capture full-length variable regions of TCR-α and/or TCR-β subunits. With this approach, 10 ng–3 μg of input RNA or 50–10,000 purified T cells undergo reverse transcription using dT priming. Following first-strand cDNA synthesis, semi-nested PCR is used to amplify TCR-specific sequences and add Illumina® sequencing adapters.
Starting with peripheral blood RNA, libraries containing TCR-α and TCR-β sequences were generated and sequenced. For each replicate &gt;70% of reads were on-target, and the most highly represented clonotypes remained consistent across a range of input amounts. A separate sensitivity assay demonstrated that RNA corresponding to a single clonotype could be detected above background levels when spiked into input RNA at a relative concentration of 0.1 %.
Our approach enables the user to obtain sequencing-ready TCR libraries from RNA in ~2.5 hours of hands-on time. Whereas approaches that utilize genomic DNA as starting material require multiplexed PCR strategies, RNA based amplification of TCR sequences use single primer pairs for each subunit. In this way, our approach minimizes the likelihood of sample misrepresentation due to amplification biases.
Camelid nanobodies specific to human pancreatic glycoprotein 2
Authors: Hanack Katja , Schlör Anja , Holzlöhner Pamela , Listek Martin , Bauer Cindy , Butze Monique , Micheel Burkhard , Hentschel Christian , Sowa Mandy , Roggenbuck Dirk , Schierack Peter , Füner Jonas , Schliebs Erik , Goihl Alexander , Reinhold Dirk 
Published: January 2023

Abstract
Pancreatic secretory zymogen-granule membrane glycoprotein 2 (GP2) has been identified to be a major autoantigenic target in Crohn’s disease patients. It was discussed recently that a long and a short isoform of GP2 exists whereas the short isoform is often detected by GP2-specific autoantibodies. In the outcome of inflammatory bowel diseases, these GP2-specific autoantibodies are discussed as new serological markers for diagnosis and therapeutic monitoring. To investigate this further, camelid nanobodies were generated by phage display and selected against the short isoform of GP2 in order to isolate specific tools for the discrimination of both isoforms. Nanobodies are single domain antibodies derived from camelid heavy chain only antibodies and characterized by a high stability and solubility. The selected candidates were expressed, purified and validated regarding their binding properties in different enzyme-linked immunosorbent assays formats, immunofluorescence, immunohistochemistry and surface plasmon resonance spectroscopy. Four different nanobodies could be selected whereof three recognize the short isoform of GP2 very specifically and one nanobody showed a high binding capacity for both isoforms. The KD values measured for all nanobodies were between 1.3 nM and 2.3 pM indicating highly specific binders suitable for the application as diagnostic tool in inflammatory bowel disease.
Expression of Interferon-inducible Genes is Influenced by Sex Hormones in SLE
Authors: Singh Ram Pyare , Hahn Bevra , Hossain Awlad 
Published: January 2023

Abstract
SLE is a female-biased disease with female to male ratio of 9:1. Accumulating evidence suggests the existence of a nexus between inflammatory pathways and 17b-estradiol, resulting in increased interferon stimulated genes (ISGs), autoantibodies and dysregulation of immune cells in SLE. However, the mechanisms by which 17b-estradiol interacts with IFN and T cells, B cells and antigen presenting cells (APC) in SLE is poorly understood. Women may be more susceptible than men to SLE and other autoimmune diseases in part because many healthy women have higher interferon regulated genes. Similarly, female BWF1 lupus mice have increased expression of interferon genes compared to age and sex- matched males. We found that plasma estradiol levels positively correlated with levels of serum/plasma IL-6, and 21/IL-23 in SLE patients. Plasma 17b-estradiol levels are significantly increased in female SLE patients compared to healthy females (p&lt;0.01). Furthermore, we found that estradiol increases proinflammatory cytokines, and level of estradiol positively correlated with expression of miR21, 25, and 186 in SLE patients. Expression of miR21, miR25 and miR186 are positively correlated with SLEDAI score in SLE. Understanding the role of 17b-estradiol in the regulation of pro-inflammatory pathways, ISGs, and microRNAs will further advance our understanding of SLE disease pathology and may identify additional novel therapeutic targets.
Modification of soluble common gamma chain function controls collagen-induced arthritis
Authors: Ko Eunhee , Goh Taesik , Lee Byunghyuk , Hong Changwan 
Published: January 2023

Abstract
IL-2Rγ (γc) is an essential subunit in γc cytokine signaling which is very important in development and homeostasis of immune cells. Recently, we showed that the soluble form of γc (sγc) is generated by alternative splicing and their production is enhanced in activated T cells. Moreover, we found that sγc inhibits γc cytokine signaling and enhances Th17 differentiation, consequently exacerbates an inflammation. Since Th17 cells are known to play pivotal roles in the pathogenesis of autoimmune rheumatoid arthritis (RA) disease, we have questioned whether sγc affects RA pathogenesis. Consistent with EAE model, we found that sγc overexpressing mice with collagen induced arthritis (CIA) displayed accelerated clinical score of arthritis compared to WT with CIA. In addition, serum level of sγc was significantly increased in CIA mice compared to WT, indicating that sγc may contribute to exacerbate the pathogenesis of RA. It is conceivable that a blockade of sγc function potentially improves autoimmune diseases. Although sγc dimerization is more functional than monomer, the dimerization mechanism of sγc is not fully understood. Using a mutant sγc form which cysteine (Cys) is replaced to alanine (Ala), we confirmed that the generation of sγc homodimer is elicited by disulfide bond. This result suggests that Cys is critical to form the dimer structure. To further study, we are designing sγc inhibitor to block sγc functions and then testing inhibitory effect in RA animal model.