bluuebunny/crossref_metadata_2025_split huggingface dataset:
TLR-dependent anti-commensal antibodies regulate intestinal immune homeostasis (MUC9P.750)
Authors: Koch Meghan , Reiner Gabrielle , Lugo Kyler , Barton Gregory 
Published: January 2023

Abstract
Maternal antibodies have been suggested to be important for establishing intestinal immune homeostasis in developing offspring, yet the mechanisms by which maternal antibodies achieve this remain unclear. We analyzed the gut immune response in mice born to antibody-deficient dams and found that maternal antibodies regulate the activation of mucosal CD4+ T cells. Interestingly, maternal IgA, the dominant mucosal antibody isotype, was not necessary for this regulation. As such, we developed an unbiased flow cytometric based assay to assess the microbiota-specific antibody response, thereby allowing us to investigate if other antibody isotypes are important for enforcing intestinal immune homeostasis. Unexpectedly, we found that mice generate a robust anti-commensal IgG2b- and IgG3- response. This commensal-specific IgG response occurs through a pathway independent of T cells, yet dependent on signaling through TLR2 and TLR4. These TLR-dependent anti-commensal IgG antibodies are transmitted from mothers to their offspring. There, they function to regulate T follicular helper responses and subsequently dampen germinal center B cell responses. This work reveals a feedback loop whereby T cell-independent antibody responses function to regulate mucosal CD4+ T cell responses and thereby establish equilibrium between the host and its resident microbiota.
Impact of microbiota on ocular immunity (MUC9P.745)
Authors: Kugadas Abirami , Gadjeva Miheala 
Published: January 2023

Abstract
To evaluate the impact of microbiota on ocular immunity, the susceptibility of germ free mice (GFM) and conventional mice to ocular Pseudomonas aeruginosa infection was studied. We found that the GFM were significantly more susceptible to infection than the conventionally housed mice. This was exemplified by increased corneal bacterial presence and associated pathology. Quantitative LC-MS/MS analysis of the eyewashes showed that the elevated susceptibility of the GFM to infection correlated with alterations in the baseline levels of ocular immune effectors, suggesting that priming of ocular immunity with commensal microbiota strengthened the innate immune barrier function of the eye. Unlike the gut microbiota, not many bacteria are isolated from the conjunctiva, posing the question whether low levels of ocular commensals may be sufficient to prime local immunity. Colonization of GFM with ocular commensal Coagulase Negative Staphylococcus sp restored the resistance to infection. Consistently, similar corneal P. aeruginosa burden was detected in the challenged monocolonized GFM and conventional mice. Notably, the expression levels of innate immune effectors such as complement C3 was restored in the monocolonized mice. This increase in the lacrimal gland-derived immune effectors was IL-6 driven as neutralization of IL-6 increased the susceptibility to infection. In summary, these data show that tonic signals by local commensals are sufficient to promote innate immune responses.
Enhanced detection of eosinophils simultaneously with other leukocyte subsets using flow cytometry (TECH2P.902)
Authors: Guenther Garret , Kang Jia , Nunez Rene 
Published: January 2023

Abstract
Eosinophils differentiate in the bone marrow prior to migration to the vascular periphery and eventual residency in tissues. While they can either be a harmful or helpful component of the immune system, they help aid in the allergic response as well as modulate inflammation. Detection of eosinophils in blood samples was typically performed using H&E staining methods, however flow cytometric analysis of whole blood samples allows researchers to identify multiple cell types in a single sample that couldn’t be done before. Here, we were able to identify a distinct eosinophil population amongst clearly separated lymphocytes, monocytes, and granulocytes comparing FSC vs SSC using an ACEA NovoCyte flow cytometer. When compared to another flow cytometer, we were unable to clearly resolve other leukocyte subsets simultaneously with eosinophils; rather additional adjustments had to be made to resolve the eosinophil population at the expense of viewing the other leukocyte subsets. Eosinophils were also analyzed using their inherent auto-fluorescence in the FITC and PE channels, then back-gated to FSC vs SSC to confirm their identity. This shows that in order to identify multiple leukocyte subsets simultaneously using FSC vs SSC, a flow cytometer with a large dynamic range is essential for this type of analysis.
Enabling platforms for multiplex diagnostics (TECH2P.905)
Authors: Dufresne Claude 
Published: January 2023

Abstract
A number of technologies have evolved that are very well suited for the creation and manufacturing of multiplex diagnostics tests. This presentation will illustrate how biological content can be used to create high quality diagnostics devices. Examples covering different platforms will include R&D and production aspects of lateral flow strips, microarrays in microtiterplates, and of custom biochips. While Lateral Flow test strips have been around for a long time, their multiplex version is just now emerging in the next wave of low cost diagnostics. Printing several smaller lines or multiple dots, instead of single analyte lines, brings about multiplex benefits, such as providing more information per tested sample. In central diagnostic labs, the use of microarrays in microtiter plates allows for a high degree of multiplexing together with a very high sample throughput. When combined with the right content, microtiter plates can form the basis for robust IVD tests. We will illustrate this with an example for Human Papilloma Viruses, and sexually transmitted diseases. Finally, a number of companies have developed unique diagnostic format platforms. Often based on custom fabricated biosensors, electrodes or other CMOS structures need to be loaded with biological substances in a way that does not damage surface features. Combining advanced printing technology with a label-free photonic biosensor-based immunoassay platform, is one example that will be presented
A novel approach to generate LIGHT mutants with selectivity to binding partners (TECH2P.925)
Authors: Liu Weifeng , Nathenson Stanley , Almo Steven 
Published: January 2023

Abstract
LIGHT is a member of TNF superfamily, which participates in interactions with many TNF receptors, including membrane-anchored HVEM and LTβR, as well as solely soluble DcR3. Interaction of LIGHT with HVEM and LTβR directly triggers intracellular signaling, which regulates a wide range of immune response, such as T cell proliferation, thymocyte differentiation and lymphocyte trafficking and migration. LIGHT:DcR3 interaction tends to attenuate the immune response. Besides LIGHT, DcR3 also neutralizes two critical inflammatory molecules TL1A and FasL. The mechanisms that support the broad specificity of LIGHT need to be defined and the functions of LIGHT interacting with different receptors await to be dissected. We have solved the crystal structures of LIGHT:HVEM and LIGHT:DcR3 complexes, revealing the critical residues involved in the interactions of LIGHT with HVEM and DcR3. We have also developed a novel approach to generate LIGHT mutants with altered binding selectivities to different receptors, which allow the further dissection of the functions of LIGHT. This novel method creates LIGHT mutants by transplanting the loops responsible for receptor recognition from TL1A to the corresponding positions of LIGHT. The generated LIGHT chimeras still possesses the wild-type like binding abilities with DcR3 and LTβR, but dramatically compromise the binding affinities with HVEM. Applications of the LIGHT chimeras reveal cooperative signaling of LIGHT through HVEM and LTβR.
Immune regulatory effects of Fingolimod (FTY720) on T cells (THER7P.959)
Authors: Gandhi Roopali , Mazzola Maria 
Published: January 2023

Abstract
FTY720 is the functional analogue of sphingosine and is the first oral treatment for relapsing remitting MS. We investigated the immune regulatory effects of Fingolimod (FTY720) on T cells in Multiple Sclerosis (MS) and controls. FTY720 function by blocking immune cell trafficking and prevents relapses in Multiple Sclerosis. We found that T cells activated in the presence of FTY-720 were less inflammatory with decreased proliferation and cytokines expression. FTY720 treated T cells have decreased expression of granzyme B and induced less neuronal cytotoxicity. We showed, FTY720 treated T cells have unique anti-inflammatory immune gene profile. The anti-inflammatory effect of FTY720 on T cells is dependent upon increased expression of a regulatory transcription factor TCF1 (Transcription factor 1). TCF1 has been previously linked to T- cell differentiation and lineage specification in mice. We show that the TCF1 could regulate human T cell differentiation. Knock down of TCF1 gene expression in human T cells switched anti-inflammatory T cells to pro-inflammatory T cell phenotype. In summary, we showed that FTY720 could regulate human T cells differentiation and its immune regulatory function is dependent upon the expression of regulatory transcription factor TCF1.
IL-23 promotes the development of hepatocellular carcinoma through the induction of IL-17-producing ILC3 (TUM10P.1044)
Authors: Liu Haiyan , Liu Yonghao , Song Yuan , Hu Bo 
Published: January 2023

Abstract
IL-23 has been shown to play a pathogenic role in autoimmunity through the promotion of Th17 cells. However, its role in tumor development remains controversial. Some studies demonstrated that IL-23 could activate T cells and NK cells to suppress tumor metastasis and growth. However, it has also been shown to promote tumor growth in colon cancer, lung cancer, breast cancer and melanoma. In the current study, we established a murine hepatocellular carcinoma (HCC) model and found that IL-23 could promote HCC development. The in vitro study demonstrated that IL-23 could directly promote HCC cell proliferation but not affect its apoptosis. Furthermore, although IL-23 could significantly increase the percent of Tc17 (CD8+IL-17+), the majority of IL-17-producing cells induced by IL-23 in the liver was innate lymphoid cell type 3 (ILC3). IL-17 could further suppress anti-tumor immune responses through the recruitment and promotion of myeloid derived suppressor cells (MDSC). Our study, for the first time, demonstrated that IL-23 could enhance HCC development through direct promotion of tumor cell proliferation, as well as suppressing anti-tumor immune response by inducing IL-17-producng ILC3.
A role for IL-13 in the progression of prostate tumors (TUM10P.1046)
Authors: Gurusamy Devikala , Shoe Jennifer , Hocker James , Hurwitz Arthur 
Published: January 2023

Abstract
IL-13 is predominantly a Th2-derived cytokine that displays a variety of effects on inflammatory and stromal cells. Receptors for IL-13 are expressed by several cancers including prostate carcinoma. Although expression of IL-13 was previously positively correlated with human prostate cancer, there are no studies that validate the functional contribution of IL-13 during prostate cancer progression. In this study, we examined the importance of IL-13 during prostate tumor progression using Transgenic Adenocarcinoma of the Mouse Prostate [TRAMP] model of prostate cancer. Il13-deficient [Il13-/-] mice were bred onto the TRAMP background to generate TRAMP+Il13+/+,TRAMP+Il13+/-and TRAMP+Il13-/- genotypes. Mice were euthanized at 25, 30, 40 and 45 weeks of age and tumors were characterized for weight, leukocyte infiltration, and tumor histopathology. Preliminary data suggests IL-13 is important for prostate tumorigenesis, as tumors obtained from TRAMP+Il13-/- mice were smaller than those obtained from TRAMP+Il13+/- or TRAMP+Il13+/+mice. On-going studies continue to examine differences in histopathology, nature and function of immune infiltrates. Future studies are aimed at understanding distinct cellular sources of IL-13, as well as precise targets of IL-13 that contribute to tumor progression, focusing on both cells of hematopoietic lineage as well as epithelial and stromal cells. Findings from these studies may identify IL-13 as a novel target for treating prostate cancer.
IL-36γ promotes antitumor immune responses (TUM10P.1028)
Authors: zuo shuting , Wang Xuefeng , Lu Binfeng 
Published: January 2023

Abstract
Introduction of an appropriate inflammation to tumor should greatly increase tumor immunogenicity and thereby help breaking immune tolerance to tumor antigens and increase response rates of immunotherapy. Cytokines play a pivotal role in regulating tumor immunogenicity and antitumor immunity. IL-36γ is important for the IL-23/IL-17-dominated autoimmune diseases as well as anti-BCG Th1 immune responses. However, the impact of IL-36γ on tumor immunity is unknown. Here, we found IL-36γ stimulated IFN-γ production by CD8 T cells synergistically with TCR signaling and/or IL-12. Importantly, IL-36γ exerted profound antitumor effects in vivo and transformed the tumor microenvironment in favor of tumor eradication. Furthermore, IL-36γ strongly increased the efficacy of tumor vaccination. Our work also showed that IL-36γ expression in human cancers inversely correlated with cancer progression. Our study establishes a novel role of IL-36γ in promoting antitumor immune responses and suggests its potential clinical translation in cancer immunotherapy.
Combination therapy of radiation and interleukin 12 induced regression of large orthotopic liver cancers by modulation of tumor microenvironment (VAC12P.1111)
Authors: Tao Mi-Hua , Wu Chia-Jen , Tsai Yi-Ting , Wu Ping-Yi 
Published: January 2023

Abstract
Hepatocellular carcinoma (HCC) is one of the leading causes of cancer death worldwide with few therapeutic options. Radiation is one of the mainstay cancer therapies. We hypothesize that combination of radiation and local interleukin 12 (IL-12) immunotherapy may change the suppressive tumor microenvironment to a proinflammatory condition favorable in induction of antitumor immune responses. IL-12 has potent antitumor activities through activation of CD8+ T cells and natural killer cells, reprogramming several types of myeloid-derived suppressor cells to a phenotype that favors immune activation, and inhibiting new blood vessel formation within tumors. Our data showed that combination of radiation and intratumoral injection of an adenoviral vector encoding IL-12 (Ad/IL-12) led to tumor regression in the majority of animals bearing large orthotopic hepatocellular carcinoma and significantly prolonged their survival. Single treatment of radiation or Ad/IL-12 delayed tumor growth but did not result in tumor regression. Mechanistic studies revealed that combination therapy greatly increased the cell numbers and activities of tumor-infiltrating CD8+ T cells, while the cell numbers and suppressive functions of MDSC and tumor-associated macrophages (TAM) were significantly decreased. Our results strongly suggest that a combinatorial treatment of radiation and IL-12 may represent an alternative therapy for advanced HCC.
Rate of pESC plasmid loss in Saccharomyces cerevisiae in the absence of selective pressure (VAC13P.1126)
Authors: Jordan Jasmine , Nash Paul 
Published: January 2023

Abstract
Recombinant yeast has been proposed as a vaccine modality, especially for inducing CTL responses. The production of yeast expressing proteins of interest would be facilitated if they can be grown in less expensive non-selective medium. This would only be practical if the population of yeast retains high levels of plasmid over the duration of the growth. We tested the Saccharomyces cerevisiae multiple gene auxotroph with pESC-URA to determine the rate of plasmid loss in the population. The yeast maintain the plasmid much longer than anticipated, suggesting that growth in non-selective medium may be a viable method. Induction of protein expression increases the competitive advantage of plasmid loss. Growth rates of the yeast and of plasmid loss under these variable conditions was also determined.
A non-cytolytic role for NK cells during gp96-mediated immunotherapy (VAC13P.1129)
Authors: Sedlacek Abigail , Binder Robert 
Published: January 2023

Abstract
Immunogenic Heat Shock Proteins (HSPs) are capable of eliciting potent immune responses against their chaperoned antigens and are being investigated as a potential cancer immunotherapy. The current model of HSP mediated immune responses includes roles for Dendritic Cells (DCs) and T cells in priming and executing the HSP-mediated response. Additionally, NK cells are necessary to inhibit tumor growth following immunotherapy with HSPs. NK cells are innate lymphoid cells whose effector functions include cytokine production and target cell lysis. Despite their requirement, little is known about what role(s) NK cells play in HSP-mediated immune responses. In these studies, we aim to determine if NK cells are acting as helper cells or effector cells using the prototypical immunogenic HSP, gp96. Our results demonstrate that depletion of NK cells specifically during the effector phase (post-gp96 immunization) results in a loss of control of D122 tumor growth. Interestingly, NK cells isolated from mice immunized with gp96 do not exhibit increased killing ex vivo, suggesting NK cells may be acting as helper cells instead of directly killing tumor cells. Additionally, we determined that NK cells improve DC activation in response to gp96 in vitro, suggesting NK cells may also play a role in priming HSP-mediated T cell responses. Collectively, these data indicate that HSP-activated NK cells are capable of acting as helper cells, potentially during multiple stages of the immune response.
Trafficking of conventional dendritic cells into the central nervous system in response to SIV and Morphine creating viral reservoir in follicular DC within cervical lymph nodes of rhesus macaques (VIR9P.1146)
Authors: Jain Pooja , Nehra Artinder , Hollenbach Rebecca , Sagar Divya , Callen Shannon , Yao Honghong , Buch Shilpa , Khan Zafar , Byrareddy Siddappa 
Published: January 2023

Abstract
Neuroinvasion by HIV leads to neurocognitive diseases and alters the permeability of the BBB. Conventional dendritic cells (cDCs) infiltrating the CNS can potentially encounter HIV from infected perivascular macrophages lining the BBB. They can thereafter carry HIV to cervical lymph nodes (CxLNs), where HIV particles will be transmitted to CD4 T cells and eventually get trapped in CD35+ follicular DCs (fDCs) to create a viral reservoir within the germinal centers. HIV patients using drugs of abuse such as morphine can further compromise the BBB and enhance infectivity of the CNS. Research demonstrating the presence and role of DCs in the CNS and lymphoid organs during HIV infection has not well established. In this respect, we explored the presence of DCs in the brain parenchyma and CxLNs of rhesus macaques infected with SIV and administered with morphine. Cells positive for cDC markers were consistently found in the brain parenchyma of SIV-infected macaques and enhanced within infected macaques given morphine. Examination of CxLNs in SIV-infected macaques confirmed presence of SIV p27+ cDCs, and CD4+ T-cells entering CD20+ germinal centers further trapping of virus within CD35+ fDCs. Ongoing studies are assessing the effect of SIV chronic infection and Morphine on fDC viral entrapment. These results provide first evidence of DC trafficking and containment of SIV particles within fDCs of cervical lymph nodes in the context of NeuroAIDS vis-à-vis drugs of abuse.
Distinct impact of SIV infection in interstitial and alveolar lung macrophages on the pathogenesis of lung disease in rhesus macaques (VIR9P.1147)
Authors: Kuroda Marcelo , Cai Y , Sugimoto C. , Arainga M , Midkiff C. , Walker E , Haupt E , Alvarez X , Lackner A , Kim W. , Didier Elizabeth 
Published: January 2023

Abstract
Declining CD4+ T cells during HIV infection contributes to immunodeficiency, but we recently reported that increasing monocyte turnover better predicted onset of terminal disease progression to AIDS in SIV-infected rhesus macaques. Here we describe the kinetics of SIV infection on distinct lung macrophage subsets and on monocyte/macrophage turnover rates in relation to pulmonary disease progression in rhesus macaques. Monocyte and tissue macrophage turnover were monitored by in vivo BrdU injection and cell uptake. Cell-associated SIV DNA in lung tissue was quantified by qPCR and in situ hybridization. Massive SIV infection was associated with increasing death rate of shorter-lived lung interstitial macrophages (IM) with elevated monocyte turnover and progression to AIDS. Conversely, the turnover of alveolar macrophages (AM) and decreasing numbers of CD4+ T cells in lung tissue did not directly correlate with disease progression. SIV DNA levels within IM and AM of the lung increased as monocyte turnover increased, but did not change within lung CD4+ T cells regardless of the stage of disease. These data suggest that SIV infection and concurrent destruction of lung IM contribute to pulmonary pathogenesis during AIDS progression while the longer-lived AM that become infected may contribute to establishing a virus reservoir. Also, bronchoalveolar lavage (BAL) specimens contain AM but not IM, so may be insufficient for fully evaluating macrophage-mediated responses in the lung.
Lactobacilus johnsonii supplemented mice modulate RSV-induced disease and is associated with increased DHA metabolite that alters DC activation through PPAR- γ. (VIR9P.1148)
Authors: Fonseca Wendy , Jang Sihyug , Rasky Andrew , Morris Susan , Ptaschinski Catherine , Fuimura Kei , Lynch Susan , Lukacs Nicholas 
Published: January 2023

Abstract
The gut microbiome contributes to a variety of mammalian processes, including modulation of host immune responses. We have found that L. johnsonii-supplemented mice displayed altered gut microbiome profiles and reduced RSV-induced pulmonary immunopathology. Metabolomics analysis of plasma from L. johnsonii-supplemented mice identified docosahexaenoic acid (DHA) and 2-hydroxyisobutyrate (butyrate precursor) as increased metabolites. We hypothesized that DHA and butyrate played a role in the modulation of the immune response in lung. We found that bone marrow-derived dendritic cells (BMDC) treated with DHA or butyrate expressed and produced reduced levels of IL-12, IL-6, IL-1β, IFN-β and TNF-α when stimulated with LPS, CpG or RSV compared to untreated cells. DHA treatment of RSV-infected BMDC prior to their co-culture with T cells isolated from RSV-infected mice significantly reduced their production of Th2 cytokines and IFNΥ. Then, we determined whether DHA was activating the nuclear receptor peroxisome proliferator-activated receptor gamma (PPAR-γ) in BMDC, which has been described as an inflammatory response inhibitor. We observed that BMDC treated with DHA and PPAR- γ antagonist (GW9662), had altered expression of mRNA and production of cytokines compared to DHA treated cells. These results suggest that DHA is modulating the immune response through PPAR-γ in dendritic cells and may be one mechanism of attenuating RSV immunopathology in mice supplemented with L. johnsonii.