bluuebunny/crossref_metadata_2025_split huggingface dataset:
Osteopontin receptor expression on immune effector cells in neonatal RSV infection (P3232)
Authors: Durre Kainath , Qureshi Nagib , Gevorkian Natalie , Qureshi Mahboob 
Published: January 2023

Abstract
Respiratory Syncitial Virus (RSV)-induced bronchiolitis-pneumonia during early infancy predisposes asthma in later life. Augmented Th2 response and skewed polarization of Dendritic cells (DC) towards plasmacytoid DC (pDC) are associated. In addition, Osteopontin (OPN) serves as an immunomodulator, which is relatively deficient in RSV- infected neonatal lungs. Exogenous OPN inhibits skewed pDC polarization in RSV- infected neonatal mice. In the present study, we examined the expression of OPN receptors (CD44 and CD51/61) by flowcytometry on pDC, myeloid DC (mDC), T and B cells in response to RSV infection in the presence and absence of NKT cells using wild type (WT) and NKT cell KO (NKT-/-) mouse models. Neonates and adults are shown to exhibit NKT and NK dominant responses, respectively, in RSV infections. Expression of CD44 and CD51/61 decreased on mDC, pDC, T and B cells in RSV infected WT pups compared to the uninfected pups particularly early on infection (day 2 and 4 post-infection). On the other hand, expression OPN receptors on DC, T and B cells were increased in the RSV-infected NKT-/- pups compared to the infected WT and uninfected NKT-/- pups. This augmented expression was abolished by day 10 post-infection. Increased expression of OPN receptors in absence of NKT cells signifies their inhibitory role. Further studies examining the roles of NKT cells in OPN treatment would help understand the underlying mechanisms for developing new therapeutic strategies.
IL-21 is required for maintaining mucosal Th1 and Th17 responses during Helicobacter pylori infection (P3184)
Authors: Carbo Adria , Hontecillas Raquel , Washington M. Kay , Chaturvedi Rupesh , Olivares-Villagómez Danyvid , Wilson Keith T. , Bassaganya-Riera Josep , Algood Holly M. 
Published: January 2023

Abstract
CD4+ T cells play a major role in the modulation of immune responses at the gastric mucosa during Helicobacter pylori infection. The relative contributions of CD4+ T cell subsets to gastritis and control of H. pylori colonization and clearance are not completely understood. Expression of IL-21, which is produced by a number of T cell subsets, increases during H. pylori infection. To elucidate the mechanisms of action of IL-21 in effector and regulatory CD4+ T cells during H. pylori infection, we combined computational modeling of CD4+ T cell differentiation and in vivo challenge studies in mice. Our computational modeling predicted activated production of IFNγ and IL-17 by IL-21, and the expression of T-bet, RORγt and phosphorylation of STAT3. To investigate the role of IL-21 in vivo, we infected IL-21-/- and wild-type mice with H. pylori SS1, and assessed colonization, gastric inflammation, cellular infiltration and cytokine profiles in the gastric mucosa. During chronic infection, IL-21-/- mice had higher H. pylori colonization, less gastritis and reduced expression of inflammatory cytokines and chemokines. Moreover, H. pylori infected IL-21-/- mice had reduced levels of CD4+ T cell specific RORγt and T-bet when compared to infected wild-type mice. Our results indicate that IL-21 contributes to the control of infection and severity of gastritis, suggesting a role for induction of gastritis in response to H. pylori through activation of both Th17 and Th1 effector responses.
Inflammasome activation in Histoplasma stimulated-dendritic cells (P3071)
Authors: Tzu-Hsuan Chang , Betty A Wu-Hsieh 
Published: January 2023

Abstract
Histoplasma capsulatum is an opportunistic dimorphic fungal pathogen. Dendritic cells take up Histoplasma yeasts and present antigens to activate both CD4 and CD8 T cells. Therefore, the interaction of dendritic cells with the fungus is critical to host defense against Histoplasma infection. Here, we investigated the mechanism of inflammasome activation in dendritic cells after infection with Histoplasma. Interaction of bone marrow-derived dendritic cells with Histoplasma activated caspase-1 and induced the secretion of IL-1β. Using blocking antibodies we also found that Dectin-2 regulated pro-IL-β gene transcription and caspase-1 activation after Histoplasma infection in dendritic cells. Histoplasma interaction with Dectin-2 triggered both the first (transcription of pro-IL-1β) and the second (activation of caspase-1) signals in response to Histoplasma stimulation. Cells with NLRP3-deficiency failed to activate caspase-1 and to process pro-IL-1β, showing that NLRP3 is an important pattern recognition receptor in the cytosol to control inflammasome activation by Histoplasma. Thus, our results demonstrated that both Dectin-2 and NLRP3 play important roles in inflammasome activation after Histoplasma infection.
IgM recognition of glycan antigens is impaired by IRAK-4/MyD88 deficiency (P3077)
Authors: Maglione Paul , Bossuyt Xavier , Meyts Isabelle , Picard Capucine , Cunningham-Rundles Charlotte 
Published: January 2023

Abstract
About 75% of patients with IRAK-4 or MyD88 deficiency develop overwhelming infections caused by S. pneumonia or S. aureus in early childhood. While IgG production is not compromised, TLR signaling mediated by IRAK-4 and MyD88 has broad effects upon the immune system, including stimulation of T-independent IgM antibody. In fact, it has recently been shown that IRAK-4/MyD88 deficiency reduces a putative reservoir of IgM production, circulating CD27+, IgD+, IgM+ B cells. Using an array of 610 glycans covering diverse structures (functionalglycomics.org), we measured serum IgM binding glycans from 8 age-matched control, 8 IRAK-4-deficient, and 3 MyD88-deficient patients. Serum IgM from these patients bound a significantly reduced number of glycans relative to controls (p &lt; 0.05). Using a microbial glycan database (csdb.glycoscience.ru) we found that control IgM bound 18 glycans expressed by S. pneumonia or S. aureus, while IgM from IRAK-4/MyD88-deficient subjects bound only 1. The reduced recognition of glycans by IgM in IRAK-4/MyD88-deficient patients was supported by ELISA, with significant reduction in IgM binding S. pneumonia capsular glycans (p = 0.004), cell wall glycan (p &lt;0.05), and phosphorylcholine (p &lt;0.05), as well as S. aureus teichoic acid (p &lt; 0.0001). These results indicate that IRAK-4/MyD88 deficiency impairs IgM recognition of glycans expressed by S. pneumonia and S. aureus, providing an explanation for heightened susceptibility to these encapsulated pathogens.
Whole Blood Assay detects endotoxin and non-endotoxin pyrogens (P3098)
Authors: Vysyaraju Kranthi , Rivera-Mariani Felix , Negherbon Jesse , Hogberg Helena , Hartung Thomas 
Published: January 2023

Abstract
Pyrogenic contaminations may precipitate adverse effects that range from fever to death. There is limited surveillance on non-endotoxin pyrogens and a sensitive technique to detect them is essential. Cells of the immune system come in contact with pyrogens; this causes the release of mediators that induce fever. The Whole Blood Assay (WBA) exploits this human physiological response; human whole blood is incubated with test samples, and interleukin-1 beta (IL-1β) induced in the presence of pyrogenic contamination is measured by ELISA. In this study, we demonstrate the ability of WBA to detect non-endotoxin pyrogens. Human whole blood was incubated with different doses of the following non-endotoxin pyrogenic stimuli: lipoteichoic acid (gram positive bacteria cell wall), Pam2CSK4, FSL-1 (bacterial diacylated lipoproteins), zymosan (fungal [1, 3] - beta glucan) and fungal spores. Levels of induced IL-1β were compared with endotoxin stimulated blood and unstimulated blood as positive and negative controls respectively. All tested stimuli showed pro-inflammatory response compared to the negative control (p &lt; 0.05) confirming that WBA effectively detects non-endotoxin pyrogens. In conclusion, WBA is a sensitive and valuable assay relevant to the human response to a broad range of pyrogenic stimuli. It can also be easily adapted for application in quality assessment of biotherapeutics and immunotoxicology.
Bioinformatic meta-analysis identifies a new role for macrophage E2F transcription factors during interaction with intracellular pathogens. (P3102)
Authors: Gregory David , Kramnik Igor , Kobzik Lester 
Published: January 2023

Abstract
We sought to identify the portion of the macrophage genome that is involved - for good or ill - in the macrophage’s interactions with intracellular pathogens. We therefore analysed publically available gene expression datasets for macrophages infected with Mycobacterium tuberculosis, Listeria monocytogenes, Francisella tularensis, Bacillus anthracis, Yersinia spp., Brucella melitensis, or Leishmania ssp.. To reflect the diversity of interactions and contexts that may influence the importance of different genes, we included data from a variety of human and mouse primary cells and lines, along with experiments comparing resistant to susceptible genetic backgrounds or the effect of IFNγ. We combined these diverse datasets to identify genes that were significantly regulated in multiple contexts. We define this group of ~3500 genes as the MacPathome: the subset of the genome particularly relevant to macrophages and their intracellular pathogens. A search for enriched regulatory motifs in the promoters of these genes found, in addition to known immune regulators, an unexpected over-representation of binding sites for the E2F transcription factor family. Initial validation by qPCR showed inhibition of E2F expression following infection of macrophages with various intracellular bacteria. An inhibitor of E2F activity (HLM006474) decreased bacterial load. These data indicate a novel role for E2F family members in the macrophage response to intracellular pathogens.
Acute cytokine responses of human monocytes to Toxoplasma gondii (P3104)
Authors: Bagriacik Emin , Hasgur Suheyla , Yavuz Emine , Atak Aysegul , Delibasi Tuncay 
Published: January 2023

Abstract
Toxoplasma gondii is an intracellular protozoan parasite. The parasite is capable of penetrating to human monocytes. Some studies showed immune responses to Toxoplasma gondii infection. However, acute cytokine responses of monocytes from sero negative individuls to the parasite are still elusive. Therefore, we studied cytokine responses of enriched human monocytes to tachyzoites in culture medium as an in vitro model of acute Toxoplasma gondii infection. Human monocytes were stimulated by tachyzoites in culture medium. Cytokine productions were determined by ELISA. Viability of cells was determined to find whether the monocytes were alive and functioning following the parasite infection. Infected monocyte viability was more than 98% after 18 hours indicating that they were functional cytokine secreting cells. Infection by the parasite increased proinflammatory cytokines such as IL-6 and TNFa significantly. Similarly IL-17 production increased after the infection. Anti-inflammatory cytokines such as IL-10 and TGFb productions were altered significantly as well. These results indicated that cytokines released by infected monocytes might contribute regulatory mechanisms of T cell activations during acute infections of Toxoplasma gondii.
CD40 induces anti-T. gondii activity in non-hematopoietic cells via the autophagy pathway (P3050)
Authors: Muniz-Feliciano Luis , Van Grol Jennifer , Subauste Carlos 
Published: January 2023

Abstract
Toxoplasma gondii infects both hematopoietic and non-hematopoietic cells and can cause cerebral and ocular toxoplasmosis. Host protection likely acts at both cellular levels to control the parasite. CD40 is key for protection against cerebral and ocular toxoplasmosis. We determined if CD40 induces anti- T. gondii activity at the level of non-hematopoietic cells. Engagement of CD40 on various endothelial cells including human brain microvascular endothelial cells, human umbilical vein endothelial cells, a mouse endothelial cell line, as well as human and mouse retinal pigmented epithelial cells resulted in killing of T. gondii. CD40 stimulation increased expression of the autophagy proteins Beclin 1 and LC3 II, enhanced autophagy flux and led to recruitment of LC3 around the parasites. The late endosomal/lysosomal marker LAMP-1 accumulated around the parasite in CD40 stimulated cells. This was accompanied by killing of T. gondii dependent on lysosomal enzymes. Accumulation of LAMP-1 and killing of T. gondii were dependent on the autophagy proteins Beclin 1 and Atg7. Together, these studies revealed that CD40 induces toxoplasmacidal activity in various non-hematopoietic cells dependent on proteins of the autophagy machinery.
Dual role of type I IFN during plasmodium infection (P3056)
Authors: Chan Jennie , Liehl Peter , DeOliveira Rosane , Sharma Shruti , Golenbock Douglas , Mota Maria , Fitzgerald Katherine 
Published: January 2023

Abstract
The molecular mechanisms regulating the inflammatory response during malaria are still poorly defined. Inflammatory cytokines and type I IFNs induced when innate immune sensors recognize components of the malaria parasite can contribute to clearance of the parasite and in some circumstances these same effectors lead to experimental cerebral malaria (ECM). Infection of C57BL/6 mice with Plasmodium berghei ANKA (PbA) leads to ECM where animals succumb to infection and die. Upon infection with PbA infected red blood cells (iRBCs), C57/Bl6 mice succumb to death 6-12 days post-infection. Recently, we have found that mice lacking the type I IFN receptor, IFNAR-/-, are protected from ECM-mediated death, implicating an important role of type I IFNs in exacerbating the ECM phenotype. Alternatively, when mice are infected with liver-tropic PbA sporozoites, a type I IFN response is induced while the parasites develop inside hepatocytes. This host response is responsible for upregulating interferon stimulated genes (ISGs) and limit the parasite load in the liver. The expression of ISGs are abrogated in IFNAR-/- mice. This protective phenotype is dependent on IRF3/7 and the adaptor MAVS suggesting that parasite RNA is recognized by host cells. Collectively, these findings reveal a dual role of type I IFNs that contribute to ECM-mediated death during PbA blood stage infections, but are also responsible for reducing parasite load during a liver stage infection.
Medullublastoma expresses CD1d and can be targeted for immunotherapy with NKT cells (P4453)
Authors: Liu Daofeng , Song Liping , Brawley Vita , Robison Nathan , Wei Jie , Gao Xiuhua , Ahmed Nabil , Asgharzadeh Shahab , Metelitsa Leonid 
Published: January 2023

Abstract
Medulloblastoma (MB) is the most common brain tumor of childhood. Current therapies are toxic and often not curative that necessitates development of targeted immunotherapy. However, little is known about immunobiology of this tumor. In this study, we show that MB cells in 9 of 20 primary tumors express CD1d, an antigen-presenting molecule for Natural Killer T cells (NKTs). Quantitative RT-PCR analysis of 61 primary tumors revealed a high level of CD1d mRNA expression in a molecular subgroup characterized by overactivation of Sonic Hedgehog (SHH) oncogene. CD1d-positive MB cells cross-presented glycolipid antigens to activate NKT-cell cytotoxicity. Intracranial injection of human NKTs resulted in regression of orthotopic MB xenografts in NOD/SCID mice. Importantly, the numbers and functional properties of peripheral blood type-I NKTs were preserved in MB patients. Therefore, CD1d is expressed on tumor cells in a subset of MB patients and represents a novel target for immunotherapy of this brain tumor.
A novel CEA-based cancer vaccine that blocks tumor implantation and tumor foci formation in vivo (P4459)
Authors: Gariepy Jean , Mallick Amirul , Huang Eric , Abdul-Wahid Aws 
Published: January 2023

Abstract
Metastatic forms of cancer account for most cancer-related deaths. Essentially all patients with advanced metastatic cancer are given palliative care in the absence of any useful curative strategies. Since tumour foci formation and expansion are directly linked to cell-cell adhesion events, we have developed a human carcinoembryonic antigen (CEA)-based subunit vaccine using only an altered-self form of its IgV like N domain responsible for the homotypic adhesion of CEA-expressing cancer cells. We now report that simply vaccinating CEA-expressing transgenic mice intramuscularly with this subunit and MF59 (squalene) engenders anti-adhesive as well as cytocidal (CDC and ADCC) antibodies in these transgenic animals that effectively block the subsequent lodging and formation of tumour foci at sites such as the peritoneal cavity and kill CEA-expressing murine tumour cells. This vaccine formulation may represent a simple, safe, preventive strategy against the long-term establishment of metastases in patients diagnosed with CEA-expressing colon, lungs, ovarian and breast cancer.
CD4 T cells control the induction, magnitude, and phenotype of the primary CD8 T cell response following adenovirus vector immunization (P4465)
Authors: Provine Nicholas , Penaloza-MacMaster Pablo , Larocca Rafael , Borducchi Erica , McNally Anna , Parenteau Lily , Barouch Dan 
Published: January 2023

Abstract
Despite the widespread use of replication-incompetent recombinant Adenovirus (Ad) vectors as candidate vaccine platforms for the elicitation of CD8 T cell responses, the mechanism by which these vectors elicit a CD8 T cell response remains poorly understood. Our data demonstrate that the induction of a primary CD8 T cell response by Ad immunization is dependent on CD4 T cell help. CD4 and MHC class II knockout mice had substantial defects in the primary CD8 T cell response following Ad vaccination. Furthermore, The CD8 T cell response generated in the absence of CD4 T cells exhibited a less effector-like phenotype, as assessed by surface staining, cytokine production, and expression of transcription factors. The unhelped CD8 T cell population also exhibited increased apoptosis and increased PD-1 expression. Intriguingly, longitudinal removal of the CD4 T cell population by monoclonal antibody depletion demonstrated that CD4 T cell help is required for at least eight days post-immunization to facilitate normal expansion and effector differentiation of the responding CD8 T cell population. Collectively, these data suggest that CD4 T cell help is needed not only at the time of initial antigen exposure but also during the first week of induction of CD8 T cell responses. Future experiments will aim to identify the mechanism(s) by which CD4 T cell help is provided in this system.
Neutrophil Gelatinase Associated Lipocalin (NGAL) regulates the onset of serum autoantibodies in pristane induced lupus (P4059)
Authors: Pawar Rahul , Goilav Beatrice , Xia Yumin , Zuang Haoyang , Reeves Westley , Putterman Chaim 
Published: January 2023

Abstract
NGAL (lipocalin-2), a member of the lipocalin superfamily, is expressed by neutrophils, hepatocytes, and renal resident cells following inflammatory conditions, and is involved in innate immunity during infection. Previously, we found that NGAL worsens renal disease in nephrotoxic nephritis. However, its relevance to adaptive immunity has not been fully explored. To investigate a possible role for NGAL in autoreactive antibody responses in experimental lupus, we injected pristane into B6 (n=10) and NGAL-deficient B6 (NGAL-/-) (n=10) mice. Analyzing the levels of serum autoantibodies at 3 months post injection, we found that B6 NGAL-/- mice had significant increases in IgG2a and IgG2b anti-double stranded DNA antibodies. Similarly, the titers of IgG2a and IgG2b antibodies to single stranded DNA and histone were significantly increased in pristane challenged NGAL-/- mice as compared to NGAL sufficient mice. Furthermore, elevated serum anti-ribonucleoprotein antibodies were found in pristane treated NGAL-/- mice by immunoprecipitation. Although autoantibody titers in NGAL-/- mice were not increased at baseline, our results are consistent with the apoptotic defects and accumulation of lymphoid cells reported previously in these mice. We also found significant upregulation of IRF5, IP-10, CXCL13, CXCR5, CxCR3, PD-1, PD-L1, and activation induced cytidine deaminase (AID) in spleens of NGAL-/- mice compared to WT mice. In vitro, we found that pristane and other inflammatory mediators promoted NGAL secretion by splenocytes. In conclusion, NGAL deficiency accelerates the onset of anti-nuclear antibodies in pristane induced lupus, suggesting that NGAL may regulate the spontaneous or induced humoral autoimmunity.
Early events in immunologic tolerance to apoptotic cells are dependent on marginal zone macrophage CCL22 production. (P4081)
Authors: Ravishankar Buvana , Chandler Philip , McGaha Tracy 
Published: January 2023

Abstract
Apoptotic cells (ACs) promote immunologic tolerance but the early innate mechanism(s) involved in the process are not known. Here we report that administration of ACs i/v induced rapid splenic expression of the regulatory T cell chemokine CCL22 by CD169+ marginal zone macrophages (MZMs). Similarly, in-vitro culture with ACs lead to expression of CCL22 in purified MZMs, but not CD11c+ dendritic cells (DCs) and conditioned media from MZM cocultures induced migration of Tregs in a CCL22 dependent manner. Administration of a soluble antagonist for the CCL22 receptor (i.e. CCR4) skewed the early innate immune response to ACs in-vivo shifting the balance from regulatory (IL-10, TGF-β) to inflammatory (TNF-α, IL-6, IL-12) cytokine production in splenic DCs and macrophages. Similarly, CCR4 blockade inhibited splenic accumulation of Tregs after apoptotic cell injection i/v and enhanced effector T cell responses to AC-associated antigens. Short-term CCR4 inhibition at the time of i/v apoptotic cell challenge (i.e. blockade only at the time of apoptotic cell administration) reversed AC-mediated tolerance to skin allografts in both primary recipients and in Treg adoptive transfers to secondary recipients instead promoting more rapid rejection. Suggesting a primary role for CCL22 in AC tolerance. Thus, we show for the first time that selective induction of CCL22 by MZMs is an essential early innate step in the generation of infectious immune tolerance to apoptotic self.
Interferon alpha deregulates endothelial nitric oxide synthase (P4085)
Authors: Buie Joy , Oates James 
Published: January 2023

Abstract
Background: Recent in vitro studies suggest that Type I interferons promote endothelial progenitor cell dysfunction and apoptosis while others have shown that Type I interferon gene signature correlates with increased endothelial dysfunction in systemic lupus erythematosus patients. Although causes of ED are likely multifactorial, all pathways converge on the diminished activity of endothelial nitric oxide synthase. We examined the effects of IFN-alpha on eNOS gene expression, phosphorylation, and NO production. Methods: Changes in eNOS expression in response to IFN-a (100, 1000 IU) were quantified by real-time PCR. Functional changes in response to vascular endothelial growth factor were assessed by immunoblot. We also evaluated the effect of serum from patients that induce a type I IFN signature from 5 patients and 2 controls on eNOS expression and VEGF mediated function. Gene expression with PCR and phosphorylated serine 1177 eNOS expression with immunoblot were assessed . Type I IFN neutralization studies (IFN-a Ab, IFNAR Ab) were also performed and eNOS expression was assessed. Results: HUVECs treated with IFN-a at 100IU and 1000IU exhibited a significant reduction in eNOS gene expression, VEGF induced phosphorylation at the serine 1177 site after 24 hours, and NO production. SLE serum also caused a reduction in eNOS gene and phosphorylated eNOS protein expression. Moreover, addition of neutralizing antibodies partially reversed observed affects on eNOS gene changes.