bluuebunny/crossref_metadata_2025_split huggingface dataset:
Administration of Melatonin Inhibits the development of DNFB-induced Dermatitis in NC/Nga Mice (36.9)
Authors: Kim Tae-Ho , Jung Jung-A , Cho Jeong-Je , Park Cheung Seog 
Published: January 2023

Abstract
Epicutaneously administered chemical haptens like 2, 4-dinitrofluorobenzene (DNFB) evoke an Atopic dermatitis (AD)-like dermatitis reaction in NC/Nga mice under Specific Pathogen Free (SPF) conditions. Melatonin (N-Acetye-5-methoxytryptamine, MA), synthesized by pineal gland, has different physiological functions, including control of seasonal reproduction, immnune system modulation, anti-aging effect, and function as a free radical scavenger. In the present study, we examined whether treatment with MA suppress AD-like skin lesions in NC/Nga mice treated with DNFB under SPF conditions. Intraperitonealy (i.p.) administration of MA to DNFB-treated NC/Nga mice was found to inhibit ear thickness increases and the skin lesions induced by DNFB. IL-4 and IFN-γ production by activated CD4+T cells from the lymph nodes of DNFB-treated NC/Nga mice was significantly inhibited by MA treatment. Moreover, total IgE level in serum was decreased by MA treatment. Our findings suggest that MA may suppress the development of AD-like dermatitis in DNFB-treated NC/Nga mice by reducing total IgE in serum, IL-4 and IFN-γ production of activated CD4+T cells.
Applying Immunological Methodology to Diagnosis (37.2)
Authors: English Rita 
Published: January 2023

Abstract
This is a high school unit investigating the role of proteins in our immune systems. Students are introduced to the basic structure of proteins and how that structure is related to antigen - antibody reactions. Immunohistochemistry, immunocytochemistry, enzyme-linked immunosorbent assay (ELISA), and immunoprecipitation techniques are discussed. Students work through a virtual ELISA lab to see its importance in medical diagnosis. Detection of antibodies is explained by introducing students to Western Blotting. Students perform gel electrophoresis to see how proteins are isolated in samples. Students research Western Blot procedure and apply that knowledge in a diagnostic exercise. Students presented their information and data to their peers in poster and lab report format.
Integrating Immunology into a Clinical Presentation-Based Medical School Curriculum (37.4)
Authors: Piskurich Janet F 
Published: January 2023

Abstract
Immunology can be a difficult discipline for first and second year medical students. As a teacher or learning facilitator it is sometimes hard to decide where to start. Even different text books adopt a variety of approaches. Clearly there is no one best way to begin to study the complex adaptive system that comprises our ability to respond to a wide variety of microorganisms, many of which have developed medically relevant means to evade the immune system. A further problem unique to the educational process of medical students is how to provide a relevant framework to enhance their retention of the concepts of basic immunology and their ability to apply these concepts later as they begin to practice medicine. Paul L. Foster School of Medicine is an emerging medical school that has adopted a clinical presentation-based curriculum. In this curriculum, students learn the basic science that is relevant to clinical science in the context of 120 specific common patient presentations. Novel approaches and the pros and cons of the integration of the basic science of immunology into a clinical-presentation based curriculum will be illustrated.
Indirect, Competitive Enzyme-Linked Immunosorbent Assay determination of Secretory Immunoglobulin A levels in saliva. (37.9)
Authors: Mwangi David 
Published: January 2023

Abstract
The amount of specific antibodies in the body can influence one's ability to fight or succumb to disease. Saliva provides the first line of defense against pathogens that enter the body through the mouth from the air or from ingested food. The primary antibody in saliva is salivary IgA. The students will complete a class activity that mimics the path taken by sIgA from its synthesis by plasma cells to its release from the mucosal membrane. In the first lab, students will construct a standard curve obtained from color intensity readings of serially diluted soft drink. This curve will be used to determine the concentration of soft drinks purchased from various sources (movie theatres, restaurants etc). This lab introduces students to the concept of dilution series and the use of standard curves in determining concentration of unknown samples.
In the second lab, the students will use an indirect competitive ELISA technique to determine the level of sIgA in unknown samples. It is desirable to complete this lab in collaboration with a local immunology lab to give the students a feel of medical research and make use of materials and equipment that may not be available in the school. Students will calculate the flow rate and corrected sIgA levels and use this information to determine the sIgA quantity in saliva samples. Students will use a combination of their own data and individual research to complete a research paper.
Role of micro-RNA and AU-rich elements in post-transcriptional regulation of interferon gamma (38.1)
Authors: Savan Ram , Yalamanchili Rajesh , Hakim Shakeeb , Young Howard A. 
Published: January 2023

Abstract
Interferon gamma (IFN-g) is an important immunomodulatory cytokine that plays a significant role in innate and adaptive immune responses of the host. The tightly-regulated expression of this pleiotropic cytokine provides robust host protection from various infections and tumors while limiting the pathological damage to the host from the inflammatory response. Similar to many inflammatory cytokines, the IFN-g 3'UTR harbors AU-rich element (ARE), that is subjected to ARE-mediated decay (AMD). The destabilization of the mRNA by AMD results in a rapid decrease in IFN-g levels. MicroRNAs (miRNAs) are another class of RNA which have regulatory potential to impact post-transcriptional regulation of gene expression. Our study has identified a miRNA-29 binding site (BS) located at the 3' end of the 3'UTR of IFN-g. Several 3' IFN-g-UTR-mutant reporter constructs have been generated by mutating the ARE and miRNA-BS, revealing competing effects between the ARE and miRNA affecting the stability of the message. Although ARE and miRNA-BS individually inhibit translation of IFN-g, mutations of both the ARE and miRNA-BS does not result in any synergistic effect. From these results, we propose that the AMD and RISC pathways competitively interact in the post-transcriptional control of IFN-g gene expression.
Externalization of the leaderless cytokine IL-1F6 occurs in response to LPS/ATP activation of transduced bone marrow derived macrophages (38.10)
Authors: Martin Unja , Gurgel Jesse , Scholler John , Renshaw Blair , Gabel Chris 
Published: January 2023

Abstract
Interleukin (IL)-1F6, like many members of the IL-1 cytokine family, lacks a signal sequence. Moreover, IL-1F6 does not contain an obvious prodomain or a caspase cleavage site but is somehow released to act on target cells. The mechanism of cellular export of IL-1F6 was compared to that employed by IL-1β. In the case of IL-1β, release of the mature processed cytokine is induced by treating lipopolysaccharide (LPS)-primed monocytes/macrophages with an agonist of the P2X7 receptor, such as adenosine 5'-triphosphate (ATP). Mouse bone marrow derived macrophages (BMDMs) were selected as the model system because they express the P2X7 receptor and because they can be engineered via retroviral transduction to express genes of interest. Stable populations of BMDMs transduced with retroviral vectors encoding full length IL-1F6 or green fluorescent protein (GFP) constitutively expressed IL-1F6 and GFP but did not constitutively release these polypeptides to the medium. Evidence is presented that IL-1F6 can be externalized rapidly in response to sequential LPS and ATP activation via a non-selective process. Therefore, externalization of IL-1F6 from cells engaged in the synthesis of this cytokine proceeds in vitro via a stimulus-coupled mechanism similar to that employed by IL-1β.
Characterization of IFNλ1 (IL-29) gene regulation and analysis of inter-individual variation of IFNλ1 levels (38.18)
Authors: Siegel Rachael , Eskdale Joyce , Gallagher Grant 
Published: January 2023

Abstract
Type III interferons are a relatively new class of anti-viral cytokines comprised of three members: IFNλ1, 2 and 3. Through an uncharacterized mechanism, levels of IFNλ1 are reduced in bronchial epithelial cells of rhinovirus-infected asthmatics. In this study, we have examined the regulation of IFNλ1 transcription by utilizing BEAS-2B bronchial epithelial cells as a model system. We have identified the IFNλ1 minimal promoter region that is responsive to viral infection. Using fluorescent DNA footprinting of epithelial cells and purified peripheral blood mononuclear cells (PBMCs) stimulated with various mitogens, we are mapping the occupancy of transcription factor binding sites up to 4kb from the IFNλ1 transcription start site. Furthermore, we have identified key regulators such as NFκ-B that are involved in control of IFNλ1 expression in epithelial cells. Data obtained from a sample set of PBMCs stimulated with HSV has shown inter-individual variation in IFNλ1, both at the level of mRNA and protein production. These findings have been correlated with analysis of selected SNP haplotypes within the IFN-λ1 promoter. Genotyping of the IFNλ1promoter has led to identification of three major haplotypes, which may relate to variation in IFNλ1 levels. The data presented in this study represent an initial characterization of IFNλ1 gene regulation that can now be applied to understanding pathogenic regulation of IFNλ1.
Generation of IL-17A reporter mice to track IL-17A expression in vivo (38.19)
Authors: Price April E , Liang Hong-Erh , Locksley Richard M 
Published: January 2023

Abstract
IL-17A plays a crucial role in host defense against a variety of pathogens and also contributes to the pathogenesis of autoimmune diseases. Various studies have suggested that IL-17A can be produced by a multiple cell types, including CD4+ T cells, CD8+ T cells, γδ T cells, NKT cells, dendritic cells, macrophages and granulocytes. However, since most studies rely on ex-vivo restimulation to assay IL-17A production, it is unclear which cell types are the predominant sources of IL-17A during the actual course of an immune response in vivo. To address this issue, we generated IL-17A reporter mice in which IL-17A-expressing cells are marked by the presence of a surface marker. We verified the ability of these reporter mice to mark the activation of IL-17A-producing Th17 cells in vitro. We then used these mice to gain a better understanding of the temporal and cell-type specific patterns of IL-17A expression in the mouse model of experimental autoimmune encephalomyelitis (EAE). We demonstrated that a small subset of CD4+ T cells and a larger fraction of γδ T cells express IL-17A in the CNS of mice with EAE, and that IL-17A expression levels peak in the CNS before the onset of paralysis. We have thus far been unable to detect IL-17A expression during infection with bacterial or fungal pathogens, which raises questions as to the actual contribution of IL-17A in these models.
IRS-2 phosphorylation and association with p85 and Grb2 after engagement of type I IL-4 receptor (IL-4R). (38.2)
Authors: Heller Nicola M , Qi Xiulan , Junttila Ilkka S , Shirey Kari Ann , Vogel Stefanie N , Paul William E , Keegan Achsah D 
Published: January 2023

Abstract
We previously showed that IL-4 elicited stronger tyrosine phosphorylation of insulin receptor substrate (IRS)-2 than IL-13. This IRS-2 phosphorylation difference was observed even at high concentrations of IL-13 and was due to the exclusive ability of IL-4 to bind to type I IL-4 Rs (IL-4Rα + γC). Furthermore, IL-4 but not IL-13, significantly augmented expression of a subset of alternatively-activated macrophage (AAM) genes, arginase I, found in inflammatory zone (FIZZ)1, and Ym1 in WT bone marrow-derived macrophages (BMM). In this study, we further analyzed signaling differences between IL-4 and IL-13 downstream of the IRS-2 pathway. IL-4 stimulation resulted in enhanced association of the p85 subunit of PI-3' kinase with IRS-2 as compared to IL-13, while only IL-4 induced the association of Grb2 with IRS-2 in WT BMM. Since IRS-2 activation leads to PI-3' K activation, we investigated the effect of wortmannin on induction of the AAM genes. IL-4-induced expression of FIZZ1 was suppressed by wortmannin to the levels induced by IL-13. Arginase I and Ym1 expression was unaffected by wortmannin. These data suggest that the type I IL-4R can direct site-specific phosphorylation of IRS-2, resulting in enhanced recruitment of adaptor molecules and augmented expression of a subset of AAM genes. In addition, they suggest these genes are differentially affected by the IRS-2/PI-3' K pathway. (PHS AI38985)
Development of an Enzyme-Linked Immunosorbent Assay for Quantitative Determination of TSLP in Cell Culture Medium and Human Serum (38.20)
Authors: Conklin John D , Sun Bingang , Leetola Catherine , Kato Atsushi , Ji Shaoquan 
Published: January 2023

Abstract
Human thymic stromal lymphopoietin (TSLP) is a 24 kDa monomeric glycoprotein produced primarily by non-hematopoietic cells such as fibroblasts, epithelium, and stromal cells. TSLP induces myeloid cells to release T cell chemoattactents while enhancing maturation of CD11c+ dendritic cells that in turn promote a Th2 immune phenotype. We have developed a sandwich ELISA to quantitatively measure human TSLP concentrations in culture media and human serum. The capture and detection antibodies consist of antigen affinity-purified goat anti-human TSLP polyclonal antibodies generated from recombinant human TSLP as the immunogen. The assay has a linear detection range from 31 to 250 pg/ml, and a threshold of detection at approximately 2 pg/ml. Serum spike recovery averaged 92.6% for four human serums (2 male, 2 female) for spike concentrations from 31.25pg to 500pg/ml inclusive. Assay robustness was further demonstrated by acceptable linearity of sample dilutions, intra-assay and inter-assay precisions. When primary normal human bronchial epithelial cells were stimulated with dsRNA and/or IL-4, TSLP concentration in the culture supernatant of the double stimulated cells increased significantly (53pg/ml, sd 3.37) compared with that from dsRNA stimulated cells (22pg/ml, sd 1.5, p=0.001). Unstimulated cell media had no detectable TSLP. This assay will provide a valuable resource for the study of role of TSLP in normal and pathophysiological conditions.
Soluble Receptor for Advanced Glycation Endproducts (sRAGE) Induces Human Monocyte Survival and Differentiation (38.23)
Authors: Wang Yijie , Wang Hongmei , Piper-Hunter Melissa G , McMaken Sara , Mo Xiaokui , Opalek Judy , Schmidt Ann Marie , Marsh Clay B. 
Published: January 2023

Abstract
The receptor for advanced glycation end-products (RAGE) is a transmembrane receptor of the immunoglobulin super family or is the soluble portion of extracellular domain of RAGE (sRAGE). sRAGE is a competitive, negative regulator of transmembrane RAGE. RAGE and its ligands are overexpressed in a wide variety of diseases. However, the biological function of RAGE and sRAGE on human mononuclear phagocytes is unclear. Using flow cytometry, we found that sRAGE directly bound to human monocytes to mediate their survival and differentiation into macrophages, events that were dependent on cellular adhesion. In both monocytes and MDM, sRAGE treatment activated Akt, Erk and NFk-B signaling, and decreased caspase-3 activity. Interestingly, while sRAGE bound the surface of suspended pre-monocytic leukemic THP1 cells, it failed to stimulate intracellular signaling or cellular differentiation. Furthermore, we found sRAGE induced monocytes chemotaxis in vitro and recruited monocytes to the lung in vivo. Our data suggest a novel role for sRAGE in mononuclear phagocyte survival, differentiation, and cell recruitment of human monocytes. These observations suggest a potential role for sRAGE to modulate organ-based inflammation.
IL-17 elicits a neutrophil-attractant response to Neisseria gonorrhoeae infection (38.25)
Authors: Feinen Brandon , Gaffen Sarah L , Jerse Ann E , Russell Michael W 
Published: January 2023

Abstract
Infection of the genital tract with Neisseria gonorrhoeae is typically associated with an influx of neutrophils, but the mechanisms underlying this influx are not well understood. The importance of the newly defined lineage of Th17 cells in inflammatory immune responses to infection, including the recruitment of neutrophils, has become increasingly recognized. We have previously shown that N. gonorrhoeae elicits production of IL-17 and other Th17-associated cytokines in vitro and hypothesized that IL-17 plays a role in the induction of chemokines and cytokines which lead to the recruitment of neutrophils to the site of infection. We tested this using a mouse vaginal gonococcal infection model to compare neutrophil recruitment in wild type and IL-17 receptor (IL-17R) knockout mice. The IL-17RKO mice were severely deficient in recruitment of neutrophils to the genital tract. Mouse genital explants were incubated ex vivo with either N. gonorrhoeae or a control antigen and assayed for production of cytokines and chemokines. Genital explants from IL-17RKO mice incubated with N. gonorrhoeae showed decreased production of IL-6 and the neutrophil-attractant chemokines LIX and MIP2α compared to those from wild type mice. These results indicate that N. gonorrhoeae elicits a Th17 response which drives the production of neutrophil-attractant chemokines, resulting in the influx of neutrophils into the genital tract.
Neither the Peyer’s patches, mesenteric lymph nodes, nor the spleen are required for induction of chronic colitis in mice (166.16)
Authors: Koboziev Iurii , Takebayashi Koichi , Ostanin Dmitry , Gray Laura , Karlsson Fridrik , Jackson Sherry , Kosloski-Davidson Melissa , Burrows-Dooley Angela , Zhang Songlin , Grisham Matthew 
Published: January 2023

Abstract
The objective of this study was to evaluate the role of PPs, MLNs and spleen in the pathogenesis of chronic colitis induced in mice. For the first series of studies, we transferred sorted, naïve (CD4+CD45RBhigh) T-cells (5x105 cells) into sham-operated RAG-1-/- mice or into RAG-1-/- mice that had their MLNs, spleen or both tissues surgically removed. We found that at 8 wks following T-cell transfer, all four of the surgically-manipulated RAG-1-/- groups developed moderate-severe colitis that was not different among the four groups. Flow cytometric analysis revealed large and similar numbers of IFN-γ producing T-cells within the colonic lamina propria (LP) of all four groups. For the second series of studies we intercrossed lymphotoxinβ-deficient (LTβ-/-) mice with RAG-1-/- animals to generate LTβ-/-xRAG-/- offspring that were devoid of Peyer’s patches and most peripheral lymph nodes. Surprisingly, and in contrast to what has been reported for LTβ-/- mice, we found no evidence of MLNs in the LTβ-/-xRAG-/- mice suggesting that lymphocytes may be required for MLN development in LTβ-/- mice. Adoptive transfer of CD45RBhigh T-cells into splenectomized LTβ-/-xRAG-/- mice or their littermate controls (LTβ+/+x RAG-/-) induced rapid onset of colitis suggesting that neither the PPs, MLNs nor the spleen are required for induction of disease. The mechanisms by which naïve T-cells are converted to colitogenic effector cells in this model are currently under investigation.
Dietary vitamin B9 is a survival factor of intestinal regulatory T cells (166.2)
Authors: Kunisawa Jun , Hashimoto Eri , Ishikawa Izumi , Kiyono Hiroshi 
Published: January 2023

Abstract
It is well known that dietary factors regulate versatile immunological functions, but the detail molecular and cellular mechanisms remain to be investigated. Here we show that vitamin B9 is a survival factor of regulatory T (Treg) cells in the intestine when it is supplied from diet. In vitamin B9-null condition, naïve T cells could differentiate into Treg cells, but differentiated Treg cells failed to survive. This effect was dose-dependent and associated with the decreased expression of anti-apoptotic Bcl-2, but independent from the IL-2- and CD28-mediated survival signals. Treg cells thus showed normal responses against these survival stimulations even in vitamin B9-null condition, suggesting that vitamin B9 is the third survival factor of Treg cells. Consistent with these findings, in vivo depletion of dietary vitamin B9 resulted in the reduction of intestinal, but not serum vitamin B9, which led to the specific decrease in the numbers of intestinal but not splenic Treg cells. These findings provide new links between dietary vitamin B9 and the gut immune system, which could be used for the development of new mucosal immunotherapy by controlling intestinal vitamin B9-mediated Treg cell network.
HDAC-mediated repression of IFN-γ responses is conserved in mouse and human trophoblast cells (166.20)
Authors: Murphy Shawn , Sullivan Danielle , Weise Paige , Choi Jason , Bushway Meghan 
Published: January 2023

Abstract
Successful pregnancy requires that the semi-allogeneic conceptus evades maternal immune-mediated rejection, while the maternal immune system simultaneously protects mother and fetus from infection. IFN-γ is a pro-inflammatory cytokine that activates innate and adaptive immunity, induces apoptosis and inhibits cell proliferation. During normal pregnancy, IFN-γ is secreted in both the human and mouse uterus. Placental trophoblast cells (TBCs) are the only cells derived from the fertilized egg that contact maternal blood, and they play important roles in controlling maternal immunity to the conceptus. Human and mouse TBCs are resistant to both IFN-γ-mediated apoptosis and activation of MHC class II expression. Moreover, while epithelial cells can clear Toxoplasma infections when exposed to IFN-γ, TBCs cannot. These collective results suggest that TBCs are resistant to IFN-γ. We recently demonstrated that IFN-γ-inducible gene expression is inhibited in mouse TBCs by histone deactylases (HDACs). Our current studies utilizing selective HDAC inhibitors and siRNA suggest that HDAC-2 and/or -3 are responsible for repressing IFN-γ-inducible gene expression in mouse TBCs. Importantly, HDAC-mediated repression of IFN-γ responses is conserved in Jar human TBC-derived choriocarcinoma cells. In contrast, HDACs activate these same genes in human cells of non-trophoblastic origin. These collective results suggest that HDACs function by a novel mechanism to repress IFN-γ-responses in TBCs.