bluuebunny/crossref_metadata_2025_split huggingface dataset:
Altered B cell Development and Anergy in the Absence of Foxp3 (143.20)
Authors: Leonardo Steven , Josephson Jennifer , Hartog Nicholas , Gauld Steven 
Published: January 2023

Abstract
The importance of regulatory T cells in immune tolerance is illustrated by the human immune dysregulatory disorder IPEX (immune dysregulation, polyendocrinopathy, enteropathy, Xlinked), caused by a lack of regulatory T cells due to decreased or absent expression of Foxp3. Although the majority of work on regulatory T cells has focused on their ability to suppress T cell responses, the development of significant autoantibody titers in patients with IPEX suggests regulatory T cells also contribute to the suppression of autoreactive B cells. Although these observations suggest that regulatory T cells have an important role in B cell biology, no studies have directly determined whether an absence of regulatory T cells impacts B cell development or B cell tolerance. Using a murine model of IPEX, harboring a mutation in the Foxp3 gene derived from a human IPEX patient, we show that B cell development is significantly altered in the absence of regulatory T cells. Furthermore, we identify a loss of B cell anergy as a likely mechanism to explain the production of autoantibodies and loss of tolerance that occurs in the absence of regulatory T cells. Our results suggest that regulatory T cells, by either direct, or indirect mechanisms, modulate B cell development and anergy.
Breakdown of autoreactive tolerance in mice deficient in apoptotic clearance (143.24)
Authors: Shao Wenhai , Kuan Anita , Wang Charlie , Zhen Yuxuan , Eisenberg Robert , Cohen Philip 
Published: January 2023

Abstract
The Mer receptor tyrosine kinase (Mertk) helps in clearance of apoptotic cells through the bridging molecules Gas6 and/or Protein S. Mertk deficient (Mer-KO) mice develop lupus-like autoimmunity with gradual accumulation of apoptotic cells in spleen and thymus. Such Mer-KO mice have two-fold expanded numbers of splenic marginal zone (MZ) B cells, yet, they develop normal response to OVA comparing to B6 control mice. When challenged respectively with type I (NP-LPS) and type II (NP-Ficoll) T-independent antigens, Mer-KO developed greater antibody responses to NP-Ficoll than their B6 counterparts. We further explored the role of the Mertk in regulating autoreactive B cells. Mer-KO mice bearing a DNA-specific immunoglobulin heavy-chain transgene (3H9/Mer-KO) spontaneously produced anti-dsDNA antibodies at the age of 3-5 months. Allotype specific ELISA allowed us to identify the transgene as source of autoantibody in Mer-KO/3H9 mice. We further analyzed MZ and follicular (FO) B cells by ELISPOT. Antibody-secreting cell counts (ASC) of MZ B cells are 70 (3H9/Mer-KO), 40 (Mer-KO), and 20 (3H9), respectively, while ASC counts of FO B cells are all around the baseline level of 20 cells. Taken together, our data show that Mer has a major effect on the development of the marginal zone B-cell compartment. Mer is important in establishing DNA-specific B cell tolerance in 3H9 anti-DNA transgenic mice.
CD5-dependent CK2 activation pathway regulates T cell activation and threshold for anergy (143.27)
Authors: Sestero Christine , McGuire Donald , DeSarno Patrizia , Raman Chander 
Published: January 2023

Abstract
We have reported that a major biological activity of CD5 is to regulate the survival of activated T cells, a property mediated by the CD5-dependent CK2 activation pathway. In order to address the physiological role of the CD5-dependent CK2 activation pathway in T cell activation, the generation of Th effector cells, and autoimmunity, we generated by gene targeting a mouse (CD5ΔCK2BD) that expressed CD5 lacking the ability to bind and activate CK2. CD4 and CD8 T cells from CD5ΔCK2BD mice exhibited enhanced AICD and hyporeactivity to antigen receptor stimulation compared to T cells from CD5WT and CD5-/- mice. The ability to mobilize calcium and upregulate CD69 were greatly enhanced in CD5-/- T cells but not in CD5WT T cells or CD5ΔCK2BD T cells, indicating that the CD5-dependent CK2 activation pathway did not regulate TCR-proximal signaling pathways. CD5-/- mice developed attenuated experimental autoimmune encephalomyelitis (EAE) when immunized with 150 μg MOG, yet the disease in CD5ΔCK2BD mice was comparable to that seen in CD5WT mice. In contrast, immunization with tolerogenic doses of MOG resulted in attenuated EAE in CD5WT mice but not in CD5-/- and CD5ΔCK2BD mice. The results indicated that loss of the CD5-dependent CK2 pathway made T cells refractory to induction of anergy, a prediction that was recapitulated in in vitro recall assays. Our results reveal a novel role for CD5 in setting thresholds for anergy in T cells. Supported by grants from the NIH and NMSS.
Radioresistance of B-1 cells from (NZB/NZW) F1 mice: possible role of microRNAs (143.46)
Authors: Laurindo Maria , Salerno Erica , Popi Ana , Novaes e Brito Ronni , Yuan Yao , Kasar Siddha , Mariano Mario , Raveche Elizabeth 
Published: January 2023

Abstract
The (NZB/NZW)F1 (B/W) mouse spontaneously develops an autoimmune disease comparable with systemic lupus erythematosus in humans. B-1 cells may be an important cell type in the development of autoimmunity and B-1 cells are distinguished from conventional B cells by developmental origin, surface marker expression and functions, including the production of autoantibodies. Here, we present findings that B-1 cells (but not conventional B cells) from the peritoneal cavity of B/W mice are resistant to 9 Gy of radiation in contrast to B-1 cells derived from C57BL/6 mice. Similar to NZB mice which have a point mutation near the mir-15a/16 loci on chromosome 14 associated with decreased expression of microRNAs miR-15a and miR-16, the B/W are hemizygous for this mutation and also have decreased miR-15a/16. We hypothesized that increased expression of targets of miR-16 in B-1 cells from the B/W mice (such as Bcl-2) could be responsible for the radioresistance observed. Transfection of peritoneal cells from B/W with miR-16 sensitizes the B-1 cells to irradiation. Moreover, 1 hour after irradiation, peritoneal cells from the radiosensitive C57BL/6 also demonstrated an increase in miR-16 expression. Our data indicate a possible role of microRNAs in contributing to the radiation response, with decreased miR-16 associated with radioresistance in B-1 cells derived from the B/W strain. The possible implications of these findings in the pathogenesis of autoimmune disease are under investigation.
Thymic self-reactivity selects for natural Th17 cells which can be distinguished by unique markers (143.49)
Authors: Nowyhed Heba , Marks Benjamin , Choi Jin-Young , Craft Joseph 
Published: January 2023

Abstract
CD4 T helper (Th) cells that secrete interleukin (IL)-17 (Th17) complement the function of Th1 and Th2 cells as a third effector lineage, with a distinct role in neutrophil recruitment necessary for extracellular pathogen clearance and the promotion of certain autoimmune conditions. We have previously demonstrated that natural Th17 (nTh17) cells develop within the thymus in a manner similar to that of natural T regulatory (nTreg) cells (Marks et al., Nat. Immunol. 2009), with activation in response to an increase of self-antigen due to tissue destruction and subsequent host protection via IL-22 secretion. In this study, we report microarray analysis of lymph node nTh17 cells, revealing specific patterns of gene expression compared to naïve, Th1, and nTreg cells. We identified 317 genes uniquely over- or under-expressed in nTh17 cells compared to controls. These results give new insights into the differences among naïve, nTreg, nTh17, and Th1 cells. We further identified two markers, IL-17RE and IL-17RC, that are expressed in nTh17 cells and not in the other subsets or in induced Th17 cells. Neither has been previously reported to be expressed in Th17 cells. IL-17RC appears to be functional in nTh17 cells, in that it offers a way for them to regulate the levels of secretion of IL-17A and IL-22. IL-17RC and IL-17RE could potentially be used as specific markers for natural Th17 cells.
Estrogen upregulates IL-17-secreting cells and IL-17 levels in normal and autoimmune mice: IL-27 and IFNγ downregulate IL-17 (143.50)
Authors: Khan Deena , Dai Rujuan , Karpuzoglu Ebru , Ahmed S. Ansar 
Published: January 2023

Abstract
Estrogen, an important immunomodulator, plays critical role in gender related immune disorders including autoimmune diseases by regulating Th1, Th2 and Tregs homeostasis. Despite extensive research, it is still not known whether estrogen modulates recently identified Th17 cell, and its key effector cytokine IL-17A (IL-17). To address this critical gap in literature, in the present study, we determined whether estrogen regulates potent proinflammatory IL-17 in wildtype (C57BL/6) and autoimmune lupus prone (NZB/W) mice. By comprehensive analysis, we report for the first time that estrogen upregulates not only IL-17 levels in the supernatants of activated splenocytes, but also IL-17-specific transcription factor, retinoic acid-related orphan receptor gamma T (RORγT). Addition of IL-17 suppressive, IL-27 or IFNγ, at the time of initiation of culture inhibit IL-17 induction significantly and expression of RORγT+ cells. However, delay in addition of IL-27 or IFNγ decreased IL-17 (albeit less profoundly) but not RORγT suggesting that once the cell is committed to IL-17 secreting cell then the magnitude of inhibitory effect of IL-27 and IFNγ is lowered. These novel studies may have far-reaching implications that could clarify our understanding of the mechanism of estrogen-mediated proinflammatory responses. Future studies are being conducted to study in detail the signaling events, which favor IL-17 induction in estrogen treated mice and inhibition by IL-27 and IFNγ.
Dendritic cells propagate early autoimmunity in the absence of functional Tregs (143.55)
Authors: Hoyer Katrina , Isakson Sara , Abbas Abul 
Published: January 2023

Abstract
BALB/c IL-2-KO mice develop systemic autoimmunity, dying by 4 wks from complications of autoimmune anemia. Disease in these mice is Th1-mediated, and IFNγ production is required for early disease (Blood 2008). We asked whether APC are required for IFNγ production by T cells in the IL-2-KO mouse. Our data show that APC:T cell interactions through B7, but not via B cells, are necessary for IFNγ production in the absence of IL-2, suggesting that dendritic cells (DC) mediate this alteration. Disease in IL-2-KO mice is marked by DC accumulation, activation and elevated cytokine production, including IL-12. Surprising, in the absence of IL-12 T cell activation and disease is unchanged, thus, IL-12 is dispensable for IFNγ induction as a part of the underlying kinetics of autoimmunity. This led us to ask whether DC are necessary for IFNγ induction and autoimmunity in these mice. The depletion of either CD11c-high-DC or plasmacytoid DC significantly augmented the survival of IL-2-KO mice, demonstrating that DC contribute to the propagation of autoimmunity. IL-2-KO DC induced heightened proliferation and cytokine production by naïve T cells compared to WT DC. Also, DC from mice without overt T cell activation still increased the proliferation of naïve T cells, suggesting that enhanced DC function is not dependent on T cell activation. Overall, our data suggest that DC are directly activated in the absence of functional Tregs, and are critical regulators of autoimmune development.
The initiative to Link Animal Models to Human DIsease (LAMHDI) (144.12)
Authors: Anderson David , Buccigrossi Robert , Ellisman Mark , Eppig Janan , Evans-Kavaldjian Haig , Grethe Jeffrey , Hamel Raymond , Kemnitz Joseph , Martone Maryann , Pohland Kimberly , Preuss Nina , Sullivan Maureen , Turner Judith , Wagner Keith , Westerfield Monte , Yurt Hakan 
Published: January 2023

Abstract
We report a new Web-based resource (LAMHDI) integrating information about animal models for human disease research. This NCRR supported resource provides a simple interface to existing animal model information for the biomedical research community. It provides more efficient access to animal model information, better categorizes and organizes existing data, facilitates interactions between researchers and resource providers, and promotes broader and more applicable use of animal models for human disease research. Approach Phase 1: A first-generation prototype is available at www.LAMHDI.org. This portal provides searchable access to existing mouse and zebrafish databases through an integrated search engine, as well as linked access to information about a variety of other species including nonhuman primates and rats. The portal also allows researchers to contribute data on animal models, thereby extending the impact of existing information through direct posting by the research community. Phase 2: The second generation will add extensive new information for additional animal model species and will increase the depth and connectivity of the data through development of semantic datastores. A sophisticated software architecture will support intelligent relational searches across diverse datasets to highlight associations between diseases, animal models, and biologic pathways.
New Protocol for the Enrichment of Untouched Human Dendritic Cells (144.14)
Authors: Cheng Qi , deMedeiros Ricardo , Hoium Brian , Resnik Ernesto , Erickson Christian , Ni Jessie 
Published: January 2023

Abstract
We have developed a new negative selection protocol for the enrichment of untouched human dendritic cells (DCs) from human blood. The protocol can be carried out in less than one hour, after PBMC enrichment, and is characterized as a two-step negative selection procedure, using two different cocktails of biotinylated antibodies that target non-DC lineage markers. DCs enriched with this protocol performed better in T cell proliferation assays, as compared to DCs enriched with a commercial kit. Flow cytometric analysis of the isolated cells show that 90% of the enriched cells are CD14-/CD19-/BDCA1+/BDCA2+/BDCA3+ (total DCs), as compared to less than 10% before cell enrichment. Within these isolated DCs 2-8% are plasmacytoid DCs (BDCA2+/CD45+) and 82-88% are myeloid DCs (20-40% BDCA1+/CD11c+, 20-40% BDCA3+/CD11c+, 15-20% CD16+/CD11c+ and 20-25% CD56+/MHCII+). On average 1 million DCs were isolated per 100 million PBMCs. The advantages of this protocol include: a) better performance in T cell proliferation assays, compared to DCs enriched with a commercially available kit, and b) higher myeloid DC heterogeneity in the enriched DC fraction. The above observations could be the result of the fact that the isolated DCs are not bound to antibodies that could initiate activation cascades, and that CD16+ and CD56+ DCs were absent from the cell population isolated with the commercial kit.
Identification of biologically significant genes using Gene Ontology (GO) and pathways analysis (144.16)
Authors: Golovan Serguei , Husain Mainul 
Published: January 2023

Abstract
Due to the small sample size and high dimensionality, it is not always possible to identify all of the biologically relevant genes from microarray analysis by simply selecting an arbitrary fold change and p-value thresholds. Such concerns are especially relevant in the analysis of immune response. It has been long known that large variability exist in immune responsiveness with the coefficient of variation (CV) as high as 230% reported for antibody response. Similarly, immune related genes were shown to have CV in excess of 100%. As a consequence, some genes involved in immune response will never be detected as differentially expressed as the number of replications required to detect statistically significant effects may be unobtainable even in well-designed experiments. To better understand which biological pathways were affected by allergenic response to ovomucoid treatment in mouse spleen, two gene sets for GO analysis were selected using different selection criteria. First set included differentially expressed genes that were biologically and statistically significant between treatments (&gt;1.5 fold, P&lt;0.05). Genes for the second gene set were selected based only on their biological significance (&gt;1.5 fold). It is demonstrated that not only new biologically significant genes were identified in previously detected GO and pathway categories (p&lt;0.05) but completely new categories (p&lt;0.05) were detected using second gene set.
Characterization of antibodies generated toward small hydrophobic environmental pollutants (144.17)
Authors: Spier Candace , Harris Thomas , Unger Michael , Kaattari Stephen 
Published: January 2023

Abstract
Antibodies are currently being exploited for their use as analytical reagents for the detection of environmental pollutants. They exhibit exquisite sensitivity and specificity such that they can be used to detect very low concentrations of targeted pollutants. One important class of pollutants for which antibodies are being developed is the polycyclic aromatic hydrocarbons (PAHs). Besides being small, PAHs are extremely hydrophobic, which increases the challenge in developing antibodies to these molecules. We have synthesized derivatives of PAHs and heterocycles (dibenzothiophene, benzothiophene, biphenyl, and fluorene) to serve as immunizing and screening haptens. By making modifications to our sera screening methods, some interesting aspects of antibody affinity to small hydrophobic haptens have been brought to our attention. Namely, the overall titer to these molecules is low (i.e., &lt;10,000 L/mol), the site of attachment greatly influences antibody specificity, and the choice of screening hapten can influence the characterization of the antibody. Knowing these outcomes promotes the need to expand screening and immunizing routines, which will increase the success of developing antibodies to other small hydrophobic pollutants. This research has been supported by a Cooperative Institute for Coastal and Estuarine Environmental Technology grant.
An Alternative Method for Intrathymic Injection in Mice (144.8)
Authors: Handon Robin , Mueller Kristen , Schwatzberg Pamela , Hoogstraten-Miller Shelley 
Published: January 2023

Abstract
Intrathymic injection has traditionally been performed using surgical procedures to open the chest cavity for direct visualization and injection into mice thymus. This technique subjects the mice to significant pain and distress, which can alter T cell development and potentially increase mortality. Intrathymic injection can also be performed by blind injection into the thoracic cavity. However, the success rate for this technique is usually low. In order to improve success rates with non-surgical intrathymic injection, our lab has preformed intrathymic injections in isoflurane-anesthetized mice using ultrasound (US)-mediated guidance. The success rate using this technique has been high as the needle and injectate can both be visualized by US as they enter the thymus. Using this technique, we have been able to label thymocytes using either FITC or Biotin to trace their fates. Complications have been very rare and were limited to the accidental laceration of an intrathoracic vessel that required in the euthanasia of one mouse out of approximately 85 injected to date. Moreover, it is important to note, that we were able to see the immediate hemorrhage on US and euthanized the mouse while it was still anesthetized, thereby avoiding any pain or distress. Our results suggest that use of US-mediated guidance is a refinement that can help reduce morbidity, distress and animal usage in experiment involving intrathymic injection.
Promotion of engraftment by donor NK cell infusion and IL-15 in nonmyeloablative allogeneic bone marrow transplantation (145.18)
Authors: Liu Haiyan , Hu Bo , Lin DanDan , Bao Guangming , Liang Yong , He Yan , Wu Yan , Zhao Lixiang , Murphy William 
Published: January 2023

Abstract
Donor NK cells have been shown to be able to promote engraftment during allogeneic bone marrow transplantation (allo-BMT). The biological activity of NK cells can be significantly enhanced by IL-15. We carried out experiments to determine the effects of combining donor NK cell infusion and IL-15 administration in a murine nonmyeloablative allo-BMT model. Mice infused with donor NK cells and treated with IL-15 during BMT resulted in increased donor engraftment compared with either treatment alone, although single treatment could also significantly promote donor engraftment. The counts of donor derived cell subsets were all increased in the spleens of the recipient mice with combinational treatment compared with either treatment alone. The alloreactivity to donor type antigens was significantly reduced in the recipient mice with combinational treatment, which was manifested by decreased proliferation and IL-2 secretion of splenocytes from recipient mice in response to donor type antigens in mixed lymphocyte responses and decreased capacity of the splenocytes killing donor type tumor targets. In conclusion, the combination of donor NK cell infusion and IL-15 administration could promote donor engraftment and development of all donor derived cell subsets, as well as suppress the host allo-responses in a nonmyeloablative allo-BMT model. The results provide a promising way to promote donor engraftment without involving systemic and nonspecific suppression of the immune system.
Pancreatic islets engineered to display on their surface a novel form of FasL protein establish localized tolerance via Treg cells (145.2)
Authors: Zhao Hong , Yolcu Esma , Bandura-Morgan Laura , Shirwan Haval 
Published: January 2023

Abstract
Objective: Islets engineered to display on their surface a chimeric FasL (SA-FasL) protein induce localized tolerance in allogeneic recipients. This study tests if CD4+CD25+FoxP3+ Tregs play a role in the initiation and maintenance of tolerance. Methods: C57BL/6 mice were transplanted with SA-FasL-engineered BALB/c islets. The frequency of Tregs in various tissues were analyzed using flow cytometry and confocal microscopy. The role of Tregs in localized tolerance was tested by; i) in vivo depletion early or late post-Tx or ii) in an adoptive transfer model using sorted Treg cells from naïve, rejected, or tolerant animals with CD4+ T cells from naïve mice. Results: We observed increased numbers of Treg cells only within the islet grafts on day 5 post-Tx and long-term (&gt;100 days) SA-FasL-islet grafts as compared to controls, but not in peripheral lymphoid organs. In vivo depletion of Treg cells early (day 14) or late (day &gt;100) post-Tx resulted in acute graft rejection. Also, cotransfer of sorted 2-5x103 Tregs from LT (n=6; &gt;100 days) graft survivors, but not rejectors (n=4; MST=36±2.3 days) with 1x106 CD4+ T effector cells from naïve C57BL/6 prevented rejection of unmanipulated BALB/c islets in an adoptive transfer model. Importantly, Tregs from long-term graft survivors expressed higher levels of FoxP3 as compared to Tregs from naïve or rejectors mice. Conclusions: Localized tolerance achieved by SA-FasL-engineered islet grafts is initiated and maintained by Treg cells