bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clinically relevant positive controls validate the use of serial FACS analysis, and video endoscopy in two murine adoptive transfer models of colitis (HUM1P.315)
Authors: Beal Dominic , Graham Sean , Lyng Gregory , Sonis Stephen 
Published: January 2023

Abstract
The adoptive transfer of naïve TH cells into immunodeficient mice is one of the best characterized immunological models of chronic colitis (CC). Traditional disease severity endpoints of weight loss and histopathology are often misleading since colitis may occur in the absence of weight loss, mice may not engraft, and weight loss and colitis may respond independently to established or experimental treatment. To overcome these limitations, we analyzed peripheral blood to track engraftment, and performed serial endoscopies to assess onset and severity of CC in two murine models of adoptive transfer-induced colitis: naive T cells from C57Bl/6J males sorted as either CD4+, CD45RBHIGH (FACS), or CD4+, CD44-, CD62L+ (negative magnetic separation) were injected IP into RAG2-/- males. Onset of colitis corresponded with the onset of weight loss, and day 14 peripheral blood CD4+ counts correlated well with both final weight loss (R2=0.74) and final endoscopy score (R2=0.79), indicating that day 14 CD4+ count was a reliable predictor of disease severity. Treatment with anti-TNFα had little effect on terminal weight loss, but significantly affected endoscopy score (2.0±0.15 vs. 1.1±0.14 at day 49 on a scale of 0-3). Together these observations suggest that endoscopy is a useful addition to weight change as a complete readout for assessing in-life disease severity in these models.
C/EBP homologous protein expression in alveolar macrophages promotes airway inflammation in an ovalbumin-based mouse model of asthma (HYP2P.329)
Authors: Thevenot Paul , Ghonim Mohamed , Boulares Hamid , Rodriguez Paulo 
Published: January 2023

Abstract
Lungs from asthmatic subjects are in a state of chronic Th2-driven inflammation with substantial shifts in myeloid cell function. We have recently shown that C/EBP homologous protein (Chop), a transcription cofactor linked with Integrated Stress Responses, is highly expressed in myeloid cells under chronic inflammatory processes. Therefore, we hypothesized that Chop plays a central role in the polarization of Th2 inflammation in a murine aerosol sensitization and challenge model. Increased expression of Chop was observed in alveolar macrophages (AMΦ) from ovalbumin (OVA)-challenged mice, compared to naïve controls. Chop expression appears to be mediated by Th2 cytokines as treatment of naïve AMΦ by a combination of IL-4 and IL-13 promoted a substantial increase in Chop expression. Interestingly, Chop deletion impaired AMΦ M2 polarization upon exposure to IL-4 and IL-13, suggesting an upstream role for Chop in promoting M2 responses. Using an OVA-based mouse model of asthma, we show that systemic Chop deletion blocked airway hyperresponsiveness, eosinophilia, and ovalbumin-specific IgE levels in mice following sensitization and challenge. Moreover, OVA-challenged Chop deficient mice exhibited decreased influx of F4/80+ AMΦ into the lungs accompanied by reduced accumulation of CD4+ T cells and increased numbers of CD8+ T cells. These results indicate a potential role for Chop in AMΦ in promoting Th2 inflammation in asthma.
Vasculotide can attenuate the immune inflammatory response and airway remodeling in chronic experimentally-induced airway inflammation in mice. (HYP2P.343)
Authors: Bourdeau Annie , Van Slyke Paul , Trop Sebastien , Dumont Daniel 
Published: January 2023

Abstract
Inflammation and airway remodeling are features associated with chronic asthmatic lungs. Previous reports have shown the benefits of the angiopoietin-1/Tie2 signaling axis on acute experimentally-induced asthma in mice. However, the more chronic effects as well as the interaction between the inflammatory and vascular components is poorly. Thus, we evaluated the impact of vasculotide, a synthetic Tie2 agonist in a chronic model of asthma. During the chronic allergic response in mice, we show that in addition to endothelial Tie2 receptor expression, there is an increased proportion of dendritic cells and monocytes expressing Tie2. Treatment of experimentally-induced chronic airway inflammation in mice with vasculotide correlated with a skewed CD4+ TH subsets and cytokine profile in the bronchoalveolar lavage of animals. This modified cytokine milieu promoted an altered endothelial cell response characterized by reduced expression of CD62E, CD54 and CD106 on endothelial cells, decreased cellular migration of inflammatory cells to the lung as well as reduced mucus accumulation. Analysis of lung tissues of animals treated with vasculotide revealed a decrease in collagen deposition and thinning of the smooth muscle cell layer in the airways of mice with chronic allergic airway inflammation. Our results demonstrate that vasculotide can impact the inflammatory and remodeling components in chronic asthma through its action on both cells of the immune response and on endothelial cells.
Salivary gland lymphocyte response to murine cytomegalovirus (INC1P.344)
Authors: Erick Timothy , Reilly Emma , Wands Jack , Brossay Laurent 
Published: January 2023

Abstract
The β-herpes virus MCMV, a homologue of HCMV, is a well-characterized animal model of viral infection that results in a non-replicative, chronic infection of an immune-competent animal. In most organs, MCMV is cleared within days by cytotoxic lymphocytes. However, the virus persists in the submandibular salivary glands (SMG) for several weeks, before establishing latency for the life of the host. NK cells are crucial for the early containment of MCMV before T cells can mount an effector response. Recent work from our laboratory has shown that SMG NK cells are hyporesponsive during MCMV infection. Here we show that SMG NK cells regain normal effector functions against MCMV when adoptively transferred into different tissue environments. This indicates that the hyporesponsive phenotype observed is tissue specific, and not cell-intrinsic. In addition, using E4BP4 deficient animals, we determined that two CD3-NK1.1+ subsets reside within the salivary glands. One subset is derived from the conventional NK cell population and relies on E4BP4 for development, while the other is E4BP4 independent. When the E4BP4 independent NK cells are transferred into different tissue environments, they produce IFN-γ in response to MCMV infection. The SMGs of E4BP4-/- mice also contain a novel population of CD3+NK1.1+NKp46+ cells that produce IFN-γ during MCMV infection. Altogether, our data show that the SMG tissue environment shapes a unique repertoire of NK1.1+ cells with distinctive phenotypes.
Skin inflammatory response to complex hydrocarbon mixtures (INC1P.357)
Authors: Gallucci Randle , Chastain Lerin , Kemp Jesse , Calhoun Kaitlin 
Published: January 2023

Abstract
Petroleum whether as crude oil or as various refined products is known to variably cause skin irritation. For instance, spill remediation workers that are exposed to fresh and weathered oil appeared to suffer a myriad of health effects, including contact dermatitis. However, the relationship between the form of petroleum and its ability to induce skin inflammation has not been examined. To investigate this, a mouse model of seven-day exposure to crude oil (weathered and fresh), JP8 jet fuel (refined) or acetone (control) was utilized, employing C57BL/6 and Balb/c mice. Immunohistochemistry, multiplex protein analysis, and real-time PCR were then conducted on isolated skin samples. Inflammatory cytokine expression showed that oil exposure significantly increased IL-1b, IL-6, CXCL2, CXCL10, CCL2, CCL3, CCL4, and CCL11 levels, the extent of which differed based on the form of petroleum as well as mouse strain. Histopathology from all forms of petroleum showed obvious inflammation in exposed skin, primarily characterized by epidermal thickening and dermal inflammatory cell infiltration which was most pronounced from jet fuel exposure. Classically activated macrophages dominated M2-like as indicated by CD86 versus CD206 expression respectively for all petroleum types. These results suggest that petroleum is indeed a skin irritant, and its form whether fresh, weathered, or refined may influence the severity of the skin inflammation.
Effect of various types of sea salt spa spring water on cytokine expression in primary human keratinocyte under TLR stimulation (INC1P.404)
Authors: Lee Kyung Ho , Kim Su-Jin , Woo So-Youn , Kim Jin-Wou 
Published: January 2023

Abstract
Background: Balneotherapy is widely used for the treatment of inflammatory skin disease, but the mechanisms are only partly understood. We previously reported spa spring water affected the secretion of inflammatory cytokines from keratinocytes (KCs) in vitro and possibly affected the T cell differentiation. Objective: In this study, we evaluated effect on cytokine secretion in primary KCs of three types of sea salt spa spring water from HaeUnDae, Magumsan, and Sokmodo, selected by low, middle and high salt concentrations, respectively. Methods: We isolated mRNA from KCs and evaluated the expression of TLR1 to TLR10 by RT-PCR. Next, we investigated the immune modulatory effect of three sea salt type spring waters on the expression of cytokines (IL-8, TNF-α, IL-1β, IL-6, and GM-CSF) in the primary KCs under TLR1-6 stimulation by ELISA. Results: The treatment of three types of sea salt spa spring waters induced suppression of inflammatory cytokines under TLR stimulation in the primary KCs. Notably, cytokine secretion from primary human KCs to TLR3 ligand was prominent. Decrease of IL-8, TNF-α, IL-1β, IL-6, GM-CSF secretion under TLR stimulation were observed, depending on the type of TLR stimulations. Conclusion: Three different type of sea salt spa spring water treatment affected the secretion of inflammatory cytokines production from KCs under TLR stimulation. These immune parameters and protocols can be used to evaluate immune modulatory effect of spa spring water in Korea.
ATP protects against sepsis through macrophage P2X7 purinergic receptors by enhancing bacterial killing (INC2P.407)
Authors: Hasko George , Csoka Balazs , Németh Zoltan , Törö Gabor , Idzko Marco , Zech Andreas 
Published: January 2023

Abstract
Extracellular ATP binds to and signals through P2X7 receptors (P2X7R)s to modulate immune function in both inflammasome-dependent and independent manners. We show here using P2X7-/- mice as well as pharmacological receptor and channel ligands that ATP release through connexin/pannexin channels and subsequent P2X7R activation are crucial for the control of mortality, bacterial dissemination and inflammation following sepsis induced by cecal ligation and puncture. Our results with P2X7-/- bone-marrow chimeras, adoptive transfer of macrophages, and myeloid-specific P2X7-/- mice indicate that P2X7R signaling on macrophages is required for the protective effect of P2X7Rs. P2X7R signaling protects through enhancing bacterial killing by macrophages, but independently of the inflammasome. In summary, targeting P2X7Rs provides a new opportunity for harnessing an endogenous protective immune mechanism in the therapy of sepsis.
IL-21 is essential for the optimal control of Mycobacterium tuberculosis infection (INC2P.416)
Authors: Cheekatla Satyanarayana , Venkatasubramanian Sambasivan , Tripathi Deepak , Paidipally Padmaja , Welch Elwyn , Tvinnereim Amy , Vankayalapati Ramakrishna 
Published: January 2023

Abstract
IL-21 is a Th17 cytokine and is known for its anti-tumor and anti-viral effects. In the current study using IL-21receptor (IL-21r-/-) and IL-21 knockout (IL-21-/-) mice, we determined the role of IL-21 and IL-21r in Mycobacterium tuberculosis (M. tb) infection. Around 50% of M. tb H37Rv infected IL-21r-/- mice died in six months compared to no deaths in infected control mice. M. tb infected IL-21r-/- mice have enhanced bacterial burden (7.3 ± 1.0 x 106 vs. 0.7 ± 0.08 x 106,p&lt;0.001), reduced infiltration of CD4+ T-cells (0.9 ± 0.01 x 105 vs. 1.4 ± 0.03 x 105, p&lt;0.001), increased frequency of PD-1,2B4,CD160 & EOMES (transcription factor) and reduced frequency of IL-12Rβ positive T-cells in lungs compared to wild type M. tb infected mice. T-cells from M. tb infected IL-21-/- mice proliferated less, produced less IFN- γ in response γ-irradiated M. tb and expressed more EOMES compared to wild type M. tb infected mice, and this was reversed by recombinant IL-21. EOMES siRNA increased IFN- γ production (p=0.007) and IL-12Rβ expression by T-cells of M. tb infected wild type mice. We also found T-cells from M. tb infected IL-21-/- mice enhance production of IFN-α (p=0.01), IL-10 (p=0.01), reduce IL-12 (p=0.02) and IL-1β (p=0.04) expression by M. tb infected macrophages and were unable control bacterial growth in macrophages compared to T-cells from M. tb infected wild type mice. The sums of our findings suggest that IL-21 and IL-21r are essential for optimal control of M. tb in mice
Characterization of phagocytic cell responses to in vitro stimulation with two commensal fungi. (INC2P.422)
Authors: Limon-Tello Jose , Wolf Andrea , Leal Christian 
Published: January 2023

Abstract
The commensal community in the gut is not only comprised of a very diverse bacterial population but also of an equally diverse fungal community whose role in the gut ecosystem is only beginning to be explored. It is therefore important to be able to isolate and grow out these fungal commensal organisms to characterize host immune responses. By employing the use of selective media we successfully cultured two fungal organisms from the gastrointestinal tract of healthy mice; the basidiomycetous yeasts Rhodotorula mucilaginosa and Guehomyces pullulans. Yeast cells from both organisms stained positive for chitin with calcofluor white, and β-1,3 glucans with a mAb against β-1,3 glucan, or recombinant Dectin-1. Phagocytosis by bone marrow derived dendritic cells and macrophages was less efficient when compared to Candida albicans. Syk phosphorylation was visualized by microscopy after 30 minutes of stimulation. TNFα production was decreased in dendritic cells stimulated with both isolates versus Candida albicans however, cytokine production was not Dectin-1 dependent as Dectin-1 knock out cells produced similar levels of cytokine when compared to WT cells. These are preliminary results which indicate that Dectin-1 may be dispensable for the inflammatory response against these fungi and future work will investigate the role other immune receptors play in the host response to these commensal organisms.
Investigation of heat shock protein interaction with individual CD91 ligand binding domains (INM1P.436)
Authors: Sedlacek Abigail , Mizote Yu , Pappan Nikos , Binder Robert 
Published: January 2023

Abstract
Heat Shock Proteins (HSPs) are a structurally diverse class of proteins that respond to cellular stress. Many of these proteins naturally function to chaperone proteins and peptides throughout the cell. Upon release into the extracellular space, HSPs can act to potentiate immune responses against their chaperoned antigens. HSPs bind the receptor CD91on the surface of antigen presenting cells (APCs) which results in internalization of the HSP+antigen and downstream signaling resulting in APC activation. Interestingly, HSPs that bind CD91 do not share structural homology and result in different patterns of cytokine secretion by APCs following activation. CD91 is comprised of a 515kDa alpha chain which is responsible for ligand binding and an 85kDa transmembrane beta chain which is responsible for signal transduction. Within the alpha chain, 4 separate ligand binding domains have been identified. In these studies, we aim to determine which ligand binding domains are responsible for binding individual HSPs and if differences in binding domain usage result in the different patterns of APC activation. To accomplish this, we have constructed individual mini-genes comprised of a single ligand binding domain tethered to the full length beta chain. These mini-genes were then transfected into RAW309Cr.1 macrophage cells which does not express CD91. These cells will be used to assess HSP binding and downstream events to gain a better understanding of HSP-CD91 interactions.
RIG-I Signaling in the Innate Immune Response After Stroke (INM1P.445)
Authors: de Rivero Vaccari Juan Pablo , Brand Frank , de Rivero Vaccari Juan Carlos 
Published: January 2023

Abstract
Stroke is associated with an inflammatory response that contributes to deleterious outcomes. However, the molecular basis in the regulation of inflammation remains poorly understood. Here, we examine the activation of retinoic acid inducible gene-like (RIG) receptors (RLRs) and the involvement of RLR signaling in the regulation of type I IFNs following mid-cerebral artery occlusion (MCAO). In this study we show that in the rat hippocampus RIG-I and IFN-α expression are increased after MCAO. Similarly, we show that in astrocytes in culture, oxygen glucose deprivation (OGD) resulted in increased RIG-I and IFN-α expression. These data are consistent with immunohistochemical analysis of hippocampal sections, indicating that in glial fibrillary acidic protein (GFAP) positive cells there was an increase in RIG-I immunoreactivity after MCAO. Moreover, n-propyl gallate (n-PG) can be used to inhibit RIG-I/IFN-α signaling in astrocytes. In conclusion, these data indicate that RIG-I is involved in the inflammatory response after stroke.
Insulin resistance in macrophages results in a distinct metabolic profile promoting an M2-like phenotype affecting both acute and chronic innate immune responses (INM1P.430)
Authors: Ieronymaki Eleftheria , Theodorakis Emmanuel , Aznaourova Marina , Vaporidi Katerina , Margioris Andrew , Eliopoulos Aristides , Tsatsanis Christos 
Published: January 2023

Abstract
Obesity and insulin resistance affects innate immune responses. In this study, we evaluated the role of insulin resistance on macrophage activation and metabolism and how these changes may affect acute inflammatory responses and sustained responses such as those regulating the gut microbiome. Herein we demonstrate that primary macrophages from mice exposed to high insulin concentration, either in vitro or in vivo in the model of High Fat Diet as well as macrophages from Akt2-/- mice or from mice lacking IGF1R from macrophages, develop insulin resistance as determined by defective glucose uptake in response to insulin. These macrophages obtained M2-like phenotype and acquired a distinct metabolic profile characterized by decreased glycolytic and increased oxidation capacity. Mice bearing insulin resistant macrophages exhibited reduced acute lung injury in response to polymicrobial sepsis, as evaluated using the Cecal Ligation and Puncture model. The intestinal microbiome was similar in Akt2-/-, LysMCre+/IGF1Rfl/fl and HFD-induced obese mice and differed from this of WT mice, suggesting that insulin resistant macrophages impact microbial flora composition. These data suggest that insulin resistance in macrophages promotes an M2-like phenotype and a distinct metabolic program affecting acute inflammatory responses and shaping the gut microbiome irrespective of the diet.
Upregulation of aryl hydrocarbon receptor expression in CD4 T lymphocytes during an immune response in vivo. (INM1P.431)
Authors: Fechner John , Julliard Walker , O'Driscoll Chelsea , Mezrich Joshua 
Published: January 2023

Abstract
Activation of the aryl hydrocarbon receptor (AHR) can impact T-cell differentiation and the maintenance of tissue-resident T cells. AHR expression, measured by RT-PCR or Western blot has been shown to be low in most CD4 T-cell subsets with highest expression found after stimulation of naïve CD4 T cells under Th17 conditions in vitro. The current study measures AHR expression within CD4 T-cell subsets of naïve or skin-grafted mice using flow cytometry. Cells were freshly isolated from the spleen, lymph nodes or skin of either untreated mice or mice that underwent allogeneic skin transplant. AHR expression in cells from untreated mice was highest in IL-17A+ CD4 T cells, followed by FoxP3+ and IFNγ+ cells. Naïve, CD62L+ CD4 T cells had the lowest expression. AHR expression in splenic or lymph node CD4 T cells from skin-grafted mice was similar to naïve mice with 20 - 30 % of FoxP3+ and &lt;10% of FoxP3- being AHR+. However, CD4 T cells within the skin grafts showed elevated AHR expression compared to spleen or LN with 75 - 85 % of FoxP3+ and 40 - 70% of FoxP3- being AHR+. This is comparable to the &gt;90% AHR+ CD4 T cells found when splenocytes were stimulated in vitro with anti-CD3 antibody in the presence of TGFb and IL-6. We conclude that the up-regulation of AHR expression in CD4 T cells in vitro also happens during immune responses in vivo and is not restricted to a single T cell subset, and may be important in T cell differentiation, trafficking, and cytokine production.
Two dimensional affinity range correlates with phenotypic response to lipid antigens in iNKT cells (IRC4P.600)
Authors: Neeld Dennis , Rossjohn Jamie , Evavold Brian 
Published: January 2023

Abstract
Invariant natural killer T cells (iNKT) are a unique group of T cells that possess an invariant T cell receptor (TCR) alpha chain paired with a limited repertoire of TCR beta chains that recognize lipid antigens presented by the MHC-I like molecule, CD1d. The micropipette adhesion frequency assay was used to define 2D affinities of iNKT cells for a panel of lipid:CD1d antigens. Despite having a limited diversity of αβ TCRs, NKT cells possess approximately a 100-fold range of affinities for the lipids tested. Our results show that αGalCer, an INFγ inducing lipid, has an approximately 10-fold higher affinity than OCH, an IL-4 skewing lipid in B6 mice. Moreover, thymic B6 iNKT cells, which predominantly produce Th1 cytokines (IFNγ), possessed a 4-fold higher affinity as compared to iNKT cells from Balb/C mice that favor more of a Th2 (IL-4) response. iNKT cell phenotype is also affected by their location as cells from the liver of Balb/C mice, which are predominantly IFNγ producers have a 2-fold increase in affinity for αGalCer compared to IL-4 predominant thymic iNKT cells. In addition to affinity differences related to phenotype, TCR expression levels of thymic derived iNKT cells reveal that Balb/C mice express twice as much TCR as the B6 mice. Taken together, these results indicate that NKT2 cytokine producing iNKT cells possess lower 2D affinity and higher TCR density compared to their NKT1 producing counterparts.