bluuebunny/crossref_metadata_2025_split huggingface dataset:
Andrographolide-triggered mitophagy-mediated NLRP3 inflammasome inhibition accounts for the prevention of colitis-associated cancer (TUM7P.919)
Authors: Sun Yang , Guo Wenjie , Xu Qiang 
Published: January 2023

Abstract
Nonresolving inflammation in the intestine predisposes individuals to the development of colitis-associated cancer (CAC). Inflammasomes are believed to mediate intestinial homeostasis, and their dysregulation contributes to inflammatory bowel diseases and CAC. However, few agents targeting inflammasomes to reduce CAC have been reported thus far. Here we show that small molecule andrographolide (Andro) protects mice against azoxymethane/dextran sulfate sodium-induced colon carcinogenesis through inhibiting NLRP3 inflammasome. Andro significantly attenuated colitis progression and tumor burden. Further study showed that Andro inhibited NLRP3 inflammasome activation in macrophages in vivo and in vitro. Furthermore, Andro triggered mitophagy in macrophage, which in turn inactivated the NLRP3 inflammasome. Moreover, downregulation of the PIK3CA-AKT1-MTOR-RPS6KB1 pathway accounted for Andro-induced autophagy. Finally, inhibition of IL-1β production and amelioration of murine experimental colitis induced by Andro were almost completely blocked by the knockdown of BECN1 expression, or by autophagy inhibitors 3-methyladenine, chloroquine, bafilomycin A1 and ammonium chloride in vitro and in vivo. Taken together, our findings demonstrate that Andro-driven mitophagy-mediated NLRP3 inflammasome inhibition is responsible for the prevention of CAC. Our data may help guide decisions regarding the use of Andro in patients with colitis, which ultimately reduces the risk of CAC.
Plasma-derived exosomes mediate immune suppression and serve as a biomarker of response to chemotherapy in Acute Myeloid Leukaemia (AML) (TUM7P.945)
Authors: Hong Chang-Sook , Muller Laurent , Boyiadzis Michael , Whiteside Theresa 
Published: January 2023

Abstract
Exosomes are virus-sized (30-100nm) vesicles formed within endocytic compartment and released into extracellular space. While all cells are capable of releasing exosomes under physiologic conditions, tumor cells are avid exosome producers. We have previously shown that plasma of AML patients at diagnosis contain high levels of exosomes relative to normal controls’ plasma (NC). AML exosomes carry various immunosuppressive molecules, including TGF-β1 related to NK-cell dysfunction in AML. Here we show that molecular content of exosomes isolated from patients’ plasma before, during and after chemotherapy (CT) reflects disease status. Thus, exosomes present in patients’ circulation carry biomarkers that can be useful in evaluating responses to therapy. AML exosomes carry different forms of TGF-β1 (pro-peptide, latency-associated peptide or active form) in different proportions. Only TGF-β1 pro-peptide was seen in exosomes obtained from NC or patients in remission. While TGF-β1 levels tracked protein levels in exosomes, elevated TGF-β1 levels discriminated better between patients receiving CT whose leukemia was likely to recur. In addition, co-incubation of normal NK cells with exosomes carrying active TGF-β1 resulted in down-regulation of NKG2D expression. In conclusion, elevations in exosomal protein and/or TGF-β1 levels during CT may reflect an incomplete response to CT and suggest the presence of residual disease in patients considered to have achieved complete remission.
An in vitro screen employing human neonatal dendritic cells identifies distinct synergy between Toll-like receptor and C-type lectin receptor agonists (VAC10P.966)
Authors: van Haren Simon , Foppen Willemina , Dowling David , Levy Ofer 
Published: January 2023

Abstract
Due to impairments in cell-mediated immunity, newborns are markedly susceptible to infection with intracellular pathogens. Impaired newborn immunity includes a reduced response to Toll-like Receptor agonists (TLRAs) by dendritic cells (DCs), placing them at risk for infection and limiting Th1-responses to many vaccines. Our hypothesis was that dual stimulation with TLRAs and C-type Lectin Receptor agonists (CLRAs) may overcome the reduced response of newborn DCs to common vaccine formulations. We screened TLRAs, CLRAs and their combinations for their ability to induce Th1-polarizing cytokine production from neonatal dendritic cells. Human adult- and cord blood monocyte-derived DCs (MoDCs) were generated in the presence of autologous plasma and stimulated for 18 hours. Cytokines and markers for cellular toxicity were measured to assess correlates of immunogenicity and reactogenicity. Dual activation of newborn DCs with a TLR8 agonist (R848) and a Mincle agonist (TDB) or with a TLR4 agonist (MPLA or GLA) and a Dectin-1 agonist (alkali-treated Zymosan) synergistically enhanced the production of the T-cell activating and -polarizing cytokines TNF-α, IL-12p70 and IL-1β. Our data suggest that targeting vaccinal antigens to the endocytic receptors Mincle or Dectin-1 is a powerful approach to induce Th1-mediated immunity in newborns. This study has the potential to inform development of novel adjuvanted neonatal vaccines and reduce infections in infants.
Anti-tumor response in T cells with CD8 independent high affinity TCR remains unaffected with aging (VAC11P.1009)
Authors: Kesarwani Pravin , Kristina Andrijauskaite , Rubinstein Mark , Mehrotra Shikhar 
Published: January 2023

Abstract
Advancement in adoptive cell therapy has led to the use of T cells engineered with tumor antigen reactive high affinity T cell receptors (TCR) in clinical trials. However, T cell engineered with anti-tumor epitope reactive TCR from an aged individual could affect the clinical outcome. In order to study if aging affects the functional efficacy of a CD8 independent high affinity TCR, we compared anti-tumor T cell response from young/old TCR transgenic h3T(human TIL derived Tyrosinase TCR) mice developed in our lab using TIL1383I TCR obtained from TILs of a melanoma patient. In vitro comparison of h3T T cells from old/young mice shows that antigen specific stimulation of the T cells from old mice resulted in an increased expression of CD25 and increased cytokine(IL2,IFNy,TNFa) secretion as compared to T cells from young mice. Remarkably, an overall decreased immune response from old hgp100 reactive T cells from Pmel transgenic mice was observed. Further, adoptively transferred h3T old/young splenocytes in vivo showed comparable control of human melanoma tumor 624-MEL established in a xenograft model. Further, aged T cells engineered with TIL1383I TCR and transferred to treat established melanoma in aged HLA-A2 recipient also showed efficient tumor control. These results demonstrate that aging does not dampen the recognition and activation of CD8+T cells bearing CD8 independent high affinity TCR and thus its broad immunotherapeutic potential in treating patients across all ages.
Ad5 but not Ad26 induces an IFN-γ+IL-10+ CD4+ T cell population following immunization (VAC12P.1023)
Authors: Larocca Rafael , Provine Nicholas , Borducchi Erica , Barouch Dan 
Published: January 2023

Abstract
Adenovirus-based vaccines have arisen as an attractive method to elicit robust cellular and humoral immune responses. Multiple vector serotypes have been developed, but the CD4+ regulatory T cell responses generated by these vectors have not previously been compared. We therefore, compared the functionality of antigen-specific CD4+ regulatory T cells induced following Ad5 or Ad26 immunization. C57BL/6 mice were immunized intramuscularly with 1010vp of Ad5-SIVgag or Ad26-SIVgag. Spleen cells were harvest at day 10, and SIVgag-specific CD4+ regulatory T cell responses were evaluated. Immunization with 1010vp of Ad5-SIVgag induced a population of IL-10+ producing cells that comprised 30% of gag-specific IFN-γ+ CD4+ T cells. In contrast, the number of CD4+ T cells that co-produced IFN-γ and IL-10 was 8 times lower following Ad26 immunization. These cells were T-bet+ and FoxP3-. In addition, Ad5 immunization induced a substantially larger fraction of IL-10+ cells that co-express PD-1 than Ad26 immunization. Moreover, a larger fraction of IFN-γ+ CD4+ T cells were Tim-3+ following Ad5 versus Ad26 immunization. We also observed increasing frequency of IFN-γ+IL-10+ CD4+ T cells with increasing doses of Ad5. These data suggest that Ad5 vectors, but not Ad26 vectors, induce a large population of CD4+ T cells that co-produce IFN-γ and IL-10, an immunosuppressive cytokine that may potentially limit vaccine efficacy.
Development and characterization of mouse monoclonal antibodies reactive with chicken TL1A (VET2P.1029)
Authors: Lee Sung Hyen , Lillehoj Hyun , Jeong Misun , Sullivan Yvonne B. 
Published: January 2023

Abstract
Two mouse monoclonal antibodies (mAbs) which specifically detect chicken TL1A (chTL1A) were developed and characterized. Both mAbs identified a 32 kDa E. coli-derived, poly-histidine-tagged fusion protein by Western blot analysis. Both mAbs identified TL1A-secreting cells in the chicken thymus, cecal tonsil, and bursa of Fabricius by immunocytochemistry, and were used to measure serum TL1A levels in normal and necrotic enteritis (NE)-afflicted chickens by antigen capture ELISA. These mAbs inhibited chTL1A-induced spleen lymphocyte proliferation, nitric oxide production by HD11 chicken macrophages, and blocked the cytotoxic effect of chTL1A against LSCC-RP9 chicken B tumor cells. These new mAbs detecting chTL1A will be important immune reagents for basic and applied research in poultry immunology
Characterizing immune cells of Atlantic Bottlenose Dolphins (VET2P.1047)
Authors: Nouri-Shirazi Mahyar , Bible Brittany , Zeng Menghua , Tamjidi Saba , Bossart Gregory 
Published: January 2023

Abstract
Marine mammals are ideal sentinel species for human health due to exposure to the same oceans and consumption of the same foods. There have been many studies which demonstrate that wild Atlantic Bottlenose Dolphins are exposed to high levels of contaminants which lead to a suppressed immune system and are therefore more susceptible to opportunistic infections, many of which are zoonotic diseases. However, nearly no research has been done on determining defects in the immune cell population of dolphins, especially DCs which are essential for initiating an immune response. We hypothesize phenotypic and functional differences in the PBMC, including DC precursors, of wild dolphins as compared to managed dolphins. Specifically in this study, we have used terrestrial-specific antibodies and growth factors to characterize immune cells in PBMC and to generate monocyte-derived DCs. We have identified cross-reactive terrestrial antibodies that could detect immune cell subsets within PBMC, including B cells, T cells, NK cells, monocytes and APCs. Interestingly, using these antibodies we found significant changes in immune cell subsets within PBMC of wild and managed dolphins. Finally among the terrestrial DC growth factors tested we found rat GM-CSF and IL-4 generated DCs expressing higher levels of CD11c, CD14, CD40, CD80, CD86, MHC I and MHC II. Our findings allow us to further study defects in the immune cells, especially DCs, in response to environmental contaminants.
Evaluation of Montanide™ ISA 71 VG adjuvant against coccidiosis (VET2P.1030)
Authors: Lillehoj Hyun , Lee Sung Hyen , Dupuis Laurent , Ben Arous Juliette 
Published: January 2023

Abstract
Chickens were immunized subcutaneously with an Eimeria recombinant profilin protein plus MontanideTM ISA 70 VG (ISA70) or MontanideTM ISA 71 VG (ISA 71) water-in-oil adjuvants, or with profilin alone. While immunization with profilin/ISA 70 vs. profilin alone altered the levels of more total transcripts compared with profilin/ISA 71 vs. profilin alone (509 vs. 296), the latter was associated with a greater number of unique biological functions, and a larger number of genes within these functions, compared with the former. Canonical pathway analysis identified 10 pathways related with genes encoding the altered transcripts in animals immunized with profilin/ISA 71 vs. profilin alone, compared with only 2 pathways in profilin/ISA 70 vs. profilin alone. Vaccination with profilin/ISA 71 was associated with greater body weight gain following E. acervulina infection, and decreased parasite fecal shedding after E. maxima infection, compared with profilin alone. Anti-profilin antibody levels were higher in sera of E. maxima- and E. tenella-infected chickens vaccinated with profilin/ISA 71 compared with profilin alone. The levels of transcripts encoding interferon-γ, interleukin (IL)-2, IL-10, and IL-17A were increased in intestinal lymphocytes from E. acervulina-, E. maxima-, and/or E. tenella-infected chickens vaccinated with profilin/ISA 71 compared with profilin alone.
Semi-quantitative analysis of T cell receptor (TCR) β and TCR γ gene expression in xenopus laevis: a novel animal model to study γδ T cells (VET2P.1036)
Authors: Kongprachaya Yuthana , Wilson Darayu , Appasamy Pierette 
Published: January 2023

Abstract
The polypeptide chains of heterodimeric TCRs divide T cells into two classes: αβ or γδ T cells. The functions of γδ cells overlap with cells of the innate and adaptive immune system and include cytokine production, maintaining tissue homeostasis and antigen presentation. γδ TCR ligand recognition is MHC-independent and TCRs bind not only peptides, but also other soluble, non-soluble and membrane-bound antigens. In humans and mice, γδ T cells represent only five percent of the total T cell population, although the percentage is higher in certain epithelial tissues such as the intestines and urogenital tract in adult animals. Phylogenetic analysis of the amino acid sequence of γδ TCR suggests that γδ T cells evolved prior their αβ counterparts and it is likely that γδ T cells may be more abundant in relatively primitive vertebrates like frogs and fish. We developed a semi-quantitative RT-PCR assay to compare levels of γ-TCR and β-TCR RNA in adult Xenopus laevis tissues, as markers for the presence of αβ and γδ T cells, respectively. Preliminary data indicate that γδ cells are as abundant as αβ cells in adult X. laevis spleen; ongoing experiments are evaluating γ-TCRC and β-TCRC RNA in X. laevis intestines, lungs, thymus and skin and in these tissues during metamorphosis. These studies will provide valuable information to establish Xenopus laevis as a novel animal model to study the development and function of γδ TCR-expressing cells.
A few changes in wild-type picornavirus genome lead to immunosuppression (VIR8P.1068)
Authors: Cusick Matthew , Libbey Jane , Fujinami Robert 
Published: January 2023

Abstract
Viruses, such as human immunodeficiency virus and poliovirus, use a variety of mechanisms to suppress the immune response and evade clearance by the host. Therefore, investigating how a few changes in the viral genome of a non-lethal virus can lead to an alteration in disease, from survivable to immunosuppression and death, would provide valuable information into viral pathogenesis. Here we investigate a picornavirus, the Theiler’s murine encephalomyelitis virus (TMEV), in its natural host, the mouse. C57BL/6 mice infected intracerebrally (i.c.) with the DA strain of TMEV develop acute encephalitis; mice survive the acute disease and clear the virus. A mutant of the DA strain of TMEV was inadvertently created, called H101, which encodes a point mutation (T101I) in VP1. Additional sequencing showed several nucleotide substitutions in the 5’ untranslated region as well as additional amino acid substitutions in the capsid protein coding region, suggesting that there are a number of perturbations in the viral genome. C57BL/6 mice infected with the H101 mutant via a peripheral route [intraperitoneal (i.p.)] become immunosuppressed through specific depletion of T cells. For C57BL/6 mice infected with the H101 mutant virus by either the i.c. or i.p. route, the immunosuppression is profound. This study provides experimental evidence that infection by a virus having just a few changes in the viral genome can now result in profound immunosuppression due to T cell depletion.
Defining the targets of Dengue virus infection in human skin (VIR8P.1069)
Authors: Duangkhae Parichat , Smith Amanda , Marques Ernesto , Ryman Kate , Barratt-Boyes Simon 
Published: January 2023

Abstract
Dengue is the most important arboviral infection of humans. Dengue virus (DENV) is introduced into human skin by the bite of infected mosquitoes. While Langerhans cells (LCs) have been implicated as the main targets of DENV infection the dynamics of infection of human skin remain ill-defined. Here, we exposed skin explants from healthy human donors to DENV to determine the targets of infection. Skin explants were inoculated with DENV and analyzed by immunofluorescence after intervals of incubation using antibodies to cell-specific markers and viral protein NS3. Time course experiments showed that the first targets of DENV infection were basal keratinocytes, with infection detected within 8 h. From 12-48 h of infection abundant virus replication was detected in epidermal LCs and dermal macrophages (MØs) as well as tubular structures consistent with lymphatic endothelium. Quantitative analysis indicated that exposure to DENV resulted in significant infiltration of MØs into dermis. These preliminary data suggest that DENV initially infects basal keratinocytes which may release factors promoting the mobilization of infected LCs out of the epidermis, and the influx of MØs into the dermis, which subsequently replicate DENV to high levels. Ongoing experiments are designed to determine the mechanism for MØ recruitment. These studies are revealing that DENV infection of human skin is a dynamic process involving sequential infection and recruitment of distinct cellular targets.
Hepatocytes intrinsic STAT4 regulates auto-immune liver inflammation (BA3P.209)
Authors: Kumar Pawan , Chen Kong , Pociask Derek , Kolls Jay 
Published: January 2023

Abstract
Autoimmune hepatitis is a condition where immune cells target and destroy liver cells. The role of STAT4 in autoimmune hepatitis is ill-defined. Therefore, we investigated the role of STAT4 signaling in Concanavalin A (ConA)-induced auto-immune hepatitis model. Our data indicate Stat4-/- mice but not WT, Il12p35-/- or Il12p40-/- mice were resistant to ConA-induced hepatitis as measured by liver pathology and the serum alanine aminotransferase (ALT) level. Furthermore, no significant difference in serum ALT and necrotic lesions were observed when WT or Stat4-/- spleenocytes were adoptive transfer into lymphocyte deficient Rag2gc-/- mice, suggesting that protection in Stat4-/- is independent of lymphocyte STAT4. Interestingly, serum IFNg level were significantly reduced in both Il12p35-/- and Stat4-/- mice, and injection of recombinant IFNg into Stat4-/- mice increase the serum ALT, but it does not reach the level observed in WT mice, suggesting a non-IFNg mediated protection in Stat4-/- mice. Surprisingly, there were no significant difference in FAS and FasL transcript in ConA injected liver tissue in WT and Stat4-/- mice. In addition, anti-FAS antibody injections lead to significant necrotic changes in WT, but not in Stat4-/- mice. Our data strongly indicate absence of STAT4 signaling potentially in hepatocytes regulates cell survival, which is further confirmed by significantly more pSTAT3 staining in liver tissue of Stat4-/-.
The nuclear receptor Nur77 suppresses CNS autoimmunity by limiting myeloid catecholamine production (BA3P.134)
Authors: Shaked Iftach , Hanna Richard , Ley Klaus 
Published: January 2023

Abstract
In multiple sclerosis (MS) and its mouse model, experimental autoimmune encephalitis (EAE), recruitment of peripheral autoreactive leukocytes to the central nervous system (CNS) is central for pathogenesis. Although sympathetic norepinephrine (NE) has been described as a key driving force of leukocyte recruitment, molecular mechanisms that control this process have not been fully elucidated. Macrophages can also produce NE, but the importance of this activity has not been addressed in the MS context. Nur77, an orphan nuclear receptor involved in differentiation of monocytes, can be activated by both inflammatory and sympathetic stimuli. Here we show that in macrophages, Nur77 is essential to regulate expression of tyrosine hydroxylase (TH), a rate-limiting enzyme in catecholamine biosynthesis. Mice with global or myeloid-specific Nur77 deletion exhibited enhanced TH expression and elevated blood NE levels, which triggered excessive production of IL-6, CXCL1, CCL2 and CCL5, ultimately leading to accelerated and exacerbated EAE. Live cell imaging of the spinal cord revealed enhanced infiltration of encephalitogenic 2D2 T cells and monocytes, and hastened activation of microglia in Nur77-deficient mice. Consistently, exacerbated EAE in Nur77-deficient mice was ameliorated by TH inhibition or by α1 adrenergic receptor blockade. Our data identify Nur77 as a key nuclear receptor regulating myeloid catecholamine production and CNS autoimmunity.
X chromosome dose and sex bias in autoimmune diseases: increased 47,XXX in systemic lupus erythematosus and Sjögren’s syndrome (BA4P.211)
Authors: Liu Ke , Kurien Biji , Zimmerman Sarah , Kaufman Kenneth , Mariette Xavier , Kottyan Leah , Jonsson Roland , Ng Wan-Fai , Omdal Roald , Rischmueller Maureen , Wahren-herlenius Marie , James Judith , Lessard Christopher , Thompson Susan , Gaffney Patrick , Montgomery Courtney , Siminovitch Katherine , Edberg Jeffrey , Kimberly Robert , Tsao Betty , McCune W. , Salmon Jane , Sivils Kathy , Harley John , Scofield R. 
Published: January 2023

Abstract
Mechanism for female predominance in autoimmunity is unknown. We suspected an X chromosome dose effect and predicted if so, triple X (47,XXX, 1 in ~1,000 live female births) would be increased in female predominant diseases (systemic lupus erythematosus [SLE], primary Sjögren’s syndrome [SS], primary biliary cirrhosis [PBC] and rheumatoid arthritis [RA])compared to diseases without female predominance (sarcoidosis, granulomatosis with polyangiitis [GPA]) and healthy controls. We used single nucleotide polymorphism (SNP) arrays to identify 47,XXX and fluorescent in situ hybridization, or q-PCR to confirm when possible. 47,XXX was found in 7 of 2,948 SLE and 3 of 1,053 SS female patients, but in none of the 4,822 female controls (OR≥21.31, 95% CI: 2.36-∞, p=0.001 and OR≥22.95, 95% CI: 1.89-∞, p=0.006, respectively). One 47,XXX was present for every ~421 SLE women and ~351 SS women. In addition, we identified one 47,XXX from each of 1,159 women with PBC and 943 with sarcoidosis. No 47,XXX was identified among 453 women with RA or 247 with GPA. In conclusion, 47,XXX was present in excess among SLE and SS subjects as predicted by X chromosome dose effect. These estimated prevalence of SLE and SS with 47,XXX being respectively ~2.4 and ~2.8 times higher than in women with 46,XX and ~24 and ~39 times higher than in men with 46,XY. There was no increase of 47,XXX is other female-biased diseases, suggesting multiple pathways to such a bias in autoimmunity.