bluuebunny/crossref_metadata_2025_split huggingface dataset:
Interferon gamma is involved in Borrelia-induced arthritis in mice lacking interleukin-17 (MPF7P.702)
Authors: Kuo Joseph , Nardelli Dean , Schell Ronald 
Published: January 2023

Abstract
Interleukin-17 (IL-17) has been shown to participate in the development of Lyme arthritis. Neutralization of IL-17 with antibodies inhibits the progression of arthritis in immunocompetent mice. Our study is the first to examine if Borrelia-induced arthritis occurs with IL-17-deficiency. We hypothesized that mice lacking IL-17 would have low to no Borrelia-induced arthritic symptoms. IL-17 knockout C57BL/6 mice were vaccinated with heat-inactivated Borrelia and/or infected with live spirochetes. Hind paw swelling, histopathology of ankle joints, serum cytokine levels, and T cell populations in tissues were measured. We found insignificant hind paw swelling and pathological changes in mice that were only vaccinated or infected when compared to control naïve mice, due to the Borrelia-resistant C57BL/6 background. However, mice that were both vaccinated and infected had more severe swelling and pathological changes, eliciting a robust immune response disregarding resistance. Also, elevated serum interferon gamma (IFNγ) levels and increasing frequencies of IFNγ-expressing lymphocytes with CD4 or CD8 were also observed in Borrelia-vaccinated and -infected mice. Therefore in these mice, IFNγ is directly involved in Borrelia-induced arthritis without IL-17. Future studies include identifying the role of IFNγ and other understudied IFNγ-expressing immune cells that induce Lyme arthritis in the absence of IL-17.
EphA2 promotes the development of experimental cerebral malaria. (MPF7P.707)
Authors: King Thayer , Mimche Patrice , Bray Christian , Brady Lauren , Aneke Janessa , Meyer Esmeralda , Galinski Mary , Lamb Tracey 
Published: January 2023

Abstract
Cerebral malaria is a severe complication of infection with the Plasmodium falciparum parasite, yet the etiology of the disease is not fully understood. In the well-established Plasmodium berghei ANKA (PbANKA) mouse model of experimental cerebral malaria (ECM), sequestration of parasitized red blood cells (pRBCs) and CD8+ T cells in the brain microvasculature is required for ECM development. However, our understanding of the receptors involved in CD8+ T cell adherence to the brain endothelium during ECM is currently incomplete. We hypothesize that EphA2, a member of the Eph family of receptor tyrosine kinases, is involved in the sequestration of CD8+ T cells in the brain during ECM through its interactions with membrane-bound ephrin-A ligands on CD8+ T cells. Using the PbANKA model of ECM, we show that EphA2 is upregulated in the brains of mice six days post-infection at the onset of cerebral symptoms in a lymphotoxin-α dependent manner. Ephrin-A ligands are also upregulated on splenic T cells of mice infected with PbANKA three days post-infection prior to potential trafficking to the brain. Importantly, EphA2-/- mice show significant resistance to death from ECM compared to EphA2+/+ mice. Similar numbers of pRBCs accumulate in the brains of EphA2-/- and EphA2+/+ mice, but EphA2-/- mice have less CD8+ T cells in the brain six days post-infection correlating with improved survival. Our data demonstrates a novel potential role for the EphA2 receptor in the pathogenesis of ECM.
Host vitamin D status is dependent on the microbiota. (MUC9P.747)
Authors: Bora Stephanie , Smith Philip , Patterson Andrew , Cantorna Margherita 
Published: January 2023

Abstract
Vitamin D deficiency is linked to susceptibility to inflammatory bowel disease (IBD). Mice deficient in vitamin D are more susceptible to experimental colitis than supplemented mice. Vitamin D alters the microbiota suggesting some of the effects of vitamin D on IBD are due to changes in the microbiota. Here we explored how the microbiota affect vitamin D metabolism using germ-free (GF) mice. Vitamin D is converted to 25D3. 25D3 is converted primarily in the kidney to 1,25D3, by 1α-OHase (Cyp27B1), or to 24,25D3 for degradation. Expression of Cyp27B1 was significantly higher in conventional (CV) versus GF kidneys. GF mice were given vehicle or 5μg vitamin D3 for 14 days and then conventionalized while maintaining their respective treatments for another 14 days. Liquid chromatography mass spectrometry was used to measure plasma vitamin D3, 25D3, 24,25D3, and 1,25D3 from GF mice before and after conventionalization. Vitamin D supplementation was less effective in GF mice. Conventionalized mice absorbed more vitamin D and had higher 25D3, 1,25D3, and 24,25D3. Furthermore, after correcting for differences in absorption between GF and CV mice, 1,25D3 was still significantly decreased in GF mice. This indicates GF mice neither absorb nor metabolize vitamin D normally. Differences in vitamin D metabolism between GF and CV mice suggest the microbiota play a role in regulating host vitamin D status, which could affect host immune responses and susceptibility to diseases like IBD.
Aryl hydrocarbon receptor activation enhances total but not antigen-specific IgA secretion in Leishmania major-infected mice (MUC9P.748)
Authors: Metten Maddie 
Published: January 2023

Abstract
Secretory IgA plays a critical role in neutralizing enteric toxins and protecting against some mucosal pathogens. In mice, activation of the aryl hydrocarbon receptor (AhR) with a single dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) has been shown to transiently decrease total fecal IgA levels but increase antigen-specific IgA levels after mucosal antigen challenge. The goal of this study was to determine if TCDD exposure would alter secretion of antigen-specific IgA levels after systemic pathogen challenge. Female C57Bl/6 mice were administered TCDD at 0 or 40 μg/Kg (po) one day prior to infection with Leishmania major promastigotes. Three weeks later, IgA levels were measured by ELISA. L. major-specific IgA levels in serum were readily detected but unchanged by TCDD treatment. L. major-specific lgA was undetected in the feces of any animal. In contrast, total fecal IgA levels (OD) were significantly increased by 34% in TCDD treated animals (p &lt; 0.05). Total spleen and bone marrow cell numbers were reduced significantly in TCDD-treated mice. The expression of IgM heavy chain, IgJ chain, or kappa light chain in these cells (measured by RT-RT-PCR) was unchanged by TCDD treatment. In contrast, the expression of IgA heavy chain was significantly reduced by approximately 50% in TCDD-treated animals. These results suggest that AhR activation enhances bulk IgA secretion but does not enhance secretion of antigen-specific IgA following systemic pathogen challenge.
Dynamic vaginal microbiota in pigtail macaques is associated with menstrual cycle and inflammation (MUC9P.753)
Authors: Klatt Nichole , Srinivasan Sujatha , Gile Jillian , Hensley-McBain Tiffany , Fiedler Tina , Fredricks David 
Published: January 2023

Abstract
A challenge in prevention of mucosal HIV transmission is incomplete understanding of correlates of vaginal transmission, including mucosal inflammation. Vaginal SIV/SHIV transmission in pigtail macaques (Macaca nemestrina) is an excellent model for HIV transmission, however little is known about how genital microbes may influence inflammation and transmission. Broad-range 16S rRNA gene PCR and pyrosequencing was performed on vaginal swabs to assess vaginal microbiome in healthy pigtail macaques. Bacteria were identified to the species or genus level. Luminex was used to measure cytokine and chemokine levels in vaginal swabs. Pigtail macaques had diverse bacterial communities at all phases of the menstrual cycle, with a range of dominant bacteria. Several species were identified that are commonly found in bacterial vaginosis (BV) in humans, and three animals exhibitied Lactobacillus-dominance. Longitudinal sampling demonstrated that the vaginal microbiome is dynamic and possibly influenced by the menstrual cycle. Animals with Lactobacillus-dominant vaginal microbiota had decreased IL-8 (p=0.009) and IL-1Ra (p=0.044). In conclusion, the macaque vaginal microbiota is diverse, dynamic, and can resemble the human vaginal microbiota, including BV-associated bacteria as well as Lactobacillus spp, which is associated with decreased innate inflammation. These data provide a foundation for understanding how the vaginal microbiome may impact risk of HIV/SIV transmission.
Gut commensal dependence of autoreactivity and Th17 cells in systemic autoimmunity (MUC9P.740)
Authors: Manfredo Vieira SILVIO , Ruff William , Hiltensperger Michael , Yu Andrew , Goodman Andrew , Kriegel Martin 
Published: January 2023

Abstract
Infectious triggers have been implicated in transient antiphospholipid antibody production but the cause of chronic antiphospholipid syndrome (APS) is unknown. We disrupted the gut microbiota composition in the (NZWxBXSB)F1 model of APS using antibiotics after maturation of the immune system. Targeting the microbiota with broad-spectrum antibiotics (ABX), vancomycin or ampicillin, but not neomycin or metronidazole, lowered pathogenic autoantibodies and mortality (p=0.014). On histology, manifestations of the autoimmune clotting disorder, i.e. myocardial infarctions, pulmonary emboli and strokes, were prevented. 16S rDNA sequencing on the MiSeq platform revealed preliminarily depletion of similar gram-positive anaerobic communities with vancomycin and ampicillin as were enriched in longitudinally collected human APS samples compared to controls. Systemic Th17 cells were detectable in our cohort in the absence of Th17-inducing segmented filamentous bacteria (SFB) but were reduced with ABX. IL-17 and Th17-differentiating factors IL-1b and IL-21 were diminished in serum whereas regulatory IL-10 was increased. Autoantigen- but not global anti-CD3-induced splenocyte proliferation was blunted after ABX (p=0.0012) supporting antigen-specific effects of the gut microbiota in systemic autoimmunity. In summary, gram-positive anaerobic gut microbiota are fundamentally involved in the pathogenesis of murine APS and systemic Th17 responses with broad implications for systemic autoimmunity.
Commensal flagellated A4 bacteria inhibit intestinal Th2 responses through induction of innate cell TGFβ production (MUC9P.742)
Authors: Wu Wei , Liu Houpu , Cao Anthony , Yao Suxia , Chen Feidi , Liu Han , Evans-Marin Heather , Duck Wayne , Elson Charles , Cong Yingzi 
Published: January 2023

Abstract
Interaction between host and microbiota maintains intestinal immune homeostasis. While commensal bacteria promotes Th1, Th17, and Tregs cells in lamina propria (LP) in the steady condition, it suppresses mucosal Th2 cells. Some specific commensal organisms have defined effects on mucosal immune function, e.g., SFB preferably promotes intestinal Th17 while certain Clostridium spp induce Tregs. However, it is still unclear whether there are specific commensal organisms down-regulating Th2 responses, and the mechanism involved. In this report, we showed that there were more LP Th2 cells in germ-free mice compared to the mice under SPF conditions. A4 bacteria, a commensal isolated from mouse cecum and produces immunodominant microbiota antigen, CBir1 flagellin, presented in intestinal lumen. In CBir1 TCR Tg mice, Th1, Th17 and Treg but not Th2 cells were present in the intestinal LP. When stimulated with CBir1 flagellin, naïve CBir1 Tg T cells differentiated into Th1, Th17 and Tregs but resisted Th2 differentiation. Aaddition of A4 bacteria, as well as pretreatment of APC with A4 bacteria, inhibited Th2 differentiation of OTII T cells stimulated with OVA and of wild-type T cells stimulated with anti-CD3 under Th2 polarizing conditions. A4 bacteria stimulated DC production of TGFβ, blockade of TGFβ abrogated A4 inhibition of Th2 cell development. Collectively, our data reveal that A4 bacteria inhibit intestinal Th2 responses through induction of innate cell production of TGFβ.
Chronic ingestion of low doses of cadmium alters the gut microbiome and immune homeostasis to enhance allergic sensitization (MUC9P.743)
Authors: Kim Eunsoo , Xu Xiaohua , Steiner Haley , Ahmer Brian , Cormet-Boyaka Estelle , Boyaka Prosper 
Published: January 2023

Abstract
Low doses of cadmium can be ingested due to its presence in contaminated water and its accumulation in leafy vegetables, fish and grains. Pollutants are believe to contribute to the increased incidence of allergy diseases. We addressed whether chronic ingestion of low doses of cadmium could impact allergic sensitization and thus, favor the development of allergic diseases. Conventional C57BL/6 mice given low doses of cadmium in the drinking water for 28 days exhibited a significant reduction of bacterial diversity, and an alteration of the Firmicutes to Bacteriodetes ratio. This treatment also activated both the canonical and the non-canonical NF-κB pathway and promoted proinflammatory cytokine and antimicrobial responses in the gut. The effects of cadmium were at least partially independent of the gut microbiome since germ-free C57BL/6 mice subjected to the same treatment developed the same profile of responses although at a lower degree. Finally, conventional mice chronically treated with low doses of cadmium developed higher antigen-specific IgE responses upon oral sensitization with OVA and cholera toxin as adjuvant. Furthermore, upon nasal antigen, cadmium-treated mice developed higher airway allergic response, which were characterized by the increased levels of IL-17 and Th1 responses control mice. In summary, the environmental pollutant cadmium can be a major regulator of gut immune homeostasis and a cause for increased allergic responses at distant mucosal sites.
Immuno-spin trapping to detect immune-derived free radicals in type 1 diabetes (TECH2P.903)
Authors: Tse Hubert , Anderson Brian , Ganini Douglas , Mason Ron 
Published: January 2023

Abstract
In autoimmune Type 1 diabetes (T1D), islet-infiltrating leukocytes synthesize noxious free radicals to collectively mediate b-cell lysis and enhance diabetogenic T cell responses as a pro-inflammatory third signal. We recently demonstrated a crucial role for NADPH oxidase (NOX)-dependent superoxide in T1D, as NOX-deficient Non-obese diabetic (NOD.Ncf1m1J) mice were highly resistant to autoimmune diabetes due to an enhanced alternatively-activated M2 macrophage phenotype, diminished pro-inflammatory cytokine and Type I interferon synthesis upon Toll-like receptor stimulation, dampened Th1 cytokine responses, and blunted CD8 T cell cytotoxicity. To further define the synergy between free radicals and T1D, we employed a 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) immuno-spin trapping technique to identify the generation of free radicals upon T cell activation and infection with Coxsackie B4 (CB4) virus, a putative diabetogenic enterovirus and hypothesized that NOD.Ncf1m1J immune cells would exhibit severe reductions in DMPO adducts. In the absence of superoxide, NOD.Ncf1m1J macrophages and CD4 T cells exhibited a significant decrease in DMPO adducts upon CB4 infection and autoantigen stimulation, respectively, in contrast to wild type cells. Future studies will utilize this technique to visualize the interface between free radicals and immune cells during spontaneous T1D progression and determine if immuno-spin trapping can be used as a biomarker during spontaneous T1D progression.
Ribosome profiling to investigate in vivo molecular mechanisms of immune function (TECH2P.906)
Authors: Jin Hyun Yong , Xiao Changchun 
Published: January 2023

Abstract
Genomic analyses of gene expression of immune cells have so far focused on their transcriptional program by quantifying mRNA abundance, measured by microarray or more recently by RNA-seq. The mRNA abundance is often presumed as a proxy of the gene expression, but the correlation between mRNA abundance and protein level, which is the true end point of gene expression, frequently shows lack of sufficient correlation due to the apparent regulation of gene expression at the layer of translation. Here, we present the applications of an emerging technique, Ribosome profiling (Ribo-seq), on primary immune cells derived from various genetic mutant mice developing immune diseases. This directly captured genome-wide maps of protein synthesis (translatome) by allowing quantification of ribosome footprint of each mRNA with high resolution and depth. Side-by-side comparison of transcriptome and translatome in the disease models revealed the signaling pathways that are predominantly regulated in translation levels are prevalent, highlighting the critical roles of translational control in immune function.
Novel single wall carbon nanotube electrode system capable of quantitative detection of CD4+ T cells (TECH2P.909)
Authors: Park Gayoung , Kim Joon Hyub , Min Nam Ki , Lee Kyung-Mi 
Published: January 2023

Abstract
We developed a fully-integrated SWCNT-based immunosensor capable of selective capture and linear quantification of CD4+ T cells by employing the two-step O2 plasma treatment processes. These processes overcome the current hurdle in the quantification of a specific subset of lymphocytes by removing the byproduct of spray-coated SWCNTs that hinders charge transfer and by creating large number of anchoring sites for anti-CD4 antibodies for stable cell binding. As a result, a linear range of electrochemical signal is generated by SWCNT electrode in proportion to the bound CD4+ T cells. Our CD4+ T cell-immunosensor provides a simple and low-cost platform which can fulfill the requirement for the development of point of care diagnostic technologies for HIV patients in the resource-limited countries.
Intranasal immunization with a chimeric influenza antigen delivered by detoxified anthrax toxin induces broad spectrum immunity against influenza viruses (TECH2P.756)
Authors: Diaz-Arevalo Diana , Arévalo Maria T. , Li Junwei , Navarro Ashley , Chen Yanping , Zeng Mingtao 
Published: January 2023

Abstract
Influenza remains to be a significant public health problem worldwide, despite the heavy campaign for flu vaccination in many countries. The conventional influenza vaccines are based on stimulating immune responses against hemagglutinin by virus attenuation or inactivation. However, they must be reformulated annually because of antigenic shift and drift of circulating influenza viral strains. These seasonal vaccines do not always match the circulating strains, and there is the ever-present threat that avian influenza viruses may adapt to be transmitted in humans. Development of a broadly protective or universal influenza vaccine would have a significant impact on public health. We constructed a new chimeric influenza antigen, which is composed of the relatively conserved matrix protein 2 (M2) and stem region of hemagglutinin (HA2) of influenza A viruses in combination with a detoxified anthrax edema toxin delivery system. This vaccine candidate has been shown to elicit T cell and humoral responses against influenza antigens and cross-strain protective immunity against influenza viruses, when it is intranasally delivered to mice. In addition, the vaccination of CD4+ and CD8+ T cell knock out mice do not protect against influenza virus infection. Thus, this novel vaccine strategy induces the CD4+ and CD8+ memory T cell responses which play an important role in cross-strain protection against influenza viruses.
A model of somatic hypermutation targeting in mice based on high-throughput immunoglobulin sequencing data (TECH2P.910)
Authors: Cui Ang , Diniro Roberto , Briggs Adrian , Adams Kris , Vander Heiden Jason , O'Connor Kevin , Vigneault Francois , Shlomchik Mark , Kleinstein Steven 
Published: January 2023

Abstract
Analyses of somatic hypermutation (SHM) patterns in B-cell immunoglobulin (Ig) genes have important basic science and clinical applications, but are often confounded by the intrinsic biases of SHM targeting on DNA motifs (i.e., hot/cold spots). Modeling these biases has been hindered by the difficulty in identifying mutated Ig sequences in vivo in the absence of selection pressures, which skew the observed mutation patterns. To generate a large number of unselected mutations, we immunized heavy-chain transgenic mice with nitrophenyl (NP) to generate NP-specific λ+ B cells, and used next-generation sequencing to examine the unproductive κ light chains. Most of these sequences had out-of-frame junctions and were presumed uninfluenced by selection. Despite being non-functionally rearranged, they were targeted by SHM and displayed a higher mutation frequency than functional sequences. We identified 29,005 mutations and used them to construct a quantitative SHM targeting model, which was consistent with classic hot/cold-spots, but revealed additional highly mutable motifs. We observed comparable targeting for functional and non-functional sequences, suggesting similar biological processes operate at both loci. In contrast, the level of C to T transitions in mice was higher than in human κ chains, suggesting lower levels of DNA repair activities. In summary, our model of SHM targeting for mice provides insights into the SHM process and supports future analyses of mutation patterns.
Quantitative proteomics reveals a role for epigenetic reprogramming during human monocyte differentiation (TECH2P.757)
Authors: Zhang Kangling , Nicholas Dequina , Tang Hui , Zhang Qiongyi , Xu Feng , Langridge William 
Published: January 2023

Abstract
Epigenetic reprogramming is thought to play an important role during monocyte differentiation. Complementary to cell surface markers, the characterization of monocytic cell lineages by mass spectrometry based protein/histone expression profiling opens a new avenue for studying immune cell differentiation. Here, we report the application of mass spectrometry and bioinformatics to identify changes in human monocytes during their differentiation into macrophages and dendritic cells. Our data show that linker histone H1 proteins are significantly down-regulated during monocyte differentiation. While highly enriched H3K9-methyl/S10-phos/K14-acetyl tri-modification forms of histone H3 were identified in monocytes and macrophages, they were dramatically reduced in dendritic cells. In contrast, histone H4 K16 acetylation was found to be markedly higher in dendritic cells than in monocytes and macrophages. We also found that global hyperacetylation generated by the nonspecific histone deacetylase HDAC inhibitor Apicidin induces monocyte differentiation. Together, our data suggest that specific regulation of inter- and intra-histone modifications including H3 K9 methylation, H3 S10 phosphorylation, H3 K14 acetylation, and H4 K16 acetylation must occur in concert with chromatin remodeling by linker histones for cell cycle progression and differentiation of human myeloid cells into macrophages and dendritic cells.