bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of CCL1 antisense ODN on the M2bMφ elimination and subsequent generation of M1Mφ in bacterial translocation sites of mice irradiated with whole body 137Cs γ-rays (WBI-mice) (INC7P.411)
Authors: Kobayashi Makiko , Ito Ichiaki , Nishiguchi Tomoki , Cannon Krystal , Suzuki Sumihiro , Loucas Bradford , Suzuki Fujio 
Published: January 2023

Abstract
An essential host defense against gut-associated sepsis is a local antibacterial innate immunity mainly expressed by M1Mφ (IL-10-IL-12+ Mφ with marked bactericidal activity). Although Mφ are radioresistant, M1Mφ are not generated in bacterial translocation sites (lamina propria and mesenteric lymph nodes) of WBI-mice. M2bMφ (IL-10+IL-12-CCL1+ Mφ) appearing in response to radiation injuries inhibit Mφ conversion from resident Mφ to M1Mφ. CCL1 has been characterized as an essential chemokine for the maintenance of M2bMφ properties, and M2bMφ are shown to be major CCL1 producer cells. In this study, we tried to eliminate M2bMφ and induce M1Mφ in the bacterial translocation sites of WBI-mice by treatment with CCL1 antisense ODN. The ODN was administered s.c. to WBI-mice twice a day for 1 week starting 3 days after γ-irradiation (8-11 Gy). One day after the final treatment, these mice were stimulated orally with 107 heat-killed E. faecalis. Then, F4/80+ Mφ were isolated from the bacterial translocation sties of these mice and tested for their M1Mφ and M2bMφ properties. In the results, M1Mφ were not detected in the translocation sites of WBI-mice stimulated with the antigen. However, M1Mφ were isolated from WBI-mice treated with the ODN and stimulated with the antigen. M2Mφ were not isolated significantly from these mice. These results indicate that M1Mφ are inducible by the bacterial antigen in 8-11 Gy WBI-mice after treatment with CCL1 antisense ODN.
Effect of CCL1 antisense ODN on spontaneous bacterial translocation and subsequent sepsis in mice irradiated with whole body 137Cs γ-rays (WBI-mice) (INC7P.412)
Authors: Suzuki Fujio , Ito Ichiaki , Nishiguchi Tomoki , Cannon Krystal , Suzuki Sumihiro , Loucas Bradford , Kobayashi Makiko 
Published: January 2023

Abstract
Radiation victims who suffer from gastrointestinal injuries are extremely susceptible to gut-associated sepsis. In our previous studies, M1Mφ located in the lamina propria and mesenteric lymph nodes were shown to be major effector cells against bacterial translocation. However, M1Mφ were not generated in these organs of WBI-mice who are carriers of M2bMφ (inhibitor cells for Mφ conversion from resident Mφ to M1Mφ). CCL1 produced by M2bMφ is essential for the maintenance of M2bMφ properties. In this study, we examined the effect of CCL1 antisense ODN on gut-associated sepsis stemming from endogenous intestinal microflora in 9 Gy WBI-mice. CCL1 antisense ODN was administered s.c. to WBI-mice twice a day for 1 week starting 3 days post-exposure to γ-rays. This group of mice was designated as treated mice. The mortality rates and bacterial growth in the organs and blood of treated mice were compared to those of control mice (WBI-mice treated with scrambled ODN). In the results, control mice died within 14 days of γ-irradiation, while 30% of treated mice survived more than 30 days after the irradiation. Bacteria progressively grew in the mesenteric lymph nodes, liver and blood of control mice, while the pathogens were not isolated significantly from these organs of treated mice. These results indicate that sepsis stemming from spontaneous bacterial translocation is noticeably mitigated in WBI-mice treated with CCL1 antisense ODN.
Respiratory epithelial cell innate immunity against Mycobacterium tuberculosis is deregulated by air pollution particulate matter (PM) (INC7P.417)
Authors: Schwander Stephan , Cantarella Pasquale , Sarkar Srijata , Rockafellow Megan , Rivas-Santiago César 
Published: January 2023

Abstract
Epidemiological studies demonstrate a link between exposure to indoor air pollution and cigarette smoke and risk of development of Tuberculosis (TB). The role of ambient air pollution PM in human antimycobacterial immunity has not been studied. We examined the effects of PM (&lt;2.5 and &lt;10 μm diameter, from Mexico City) on Mycobacterium tuberculosis (M.tb)-induced expression of mRNA and protein of human β-defensin 2 and 3 (HBD-2,3, antimicrobial peptides), IL-8 and MCP-1 by qRT-PCR and ELISA, respectively. In addition, cell-mediated growth control of M.tb was measured by colony forming units (CFU) assay. Respiratory alveolar type II epithelial (A549) cells were exposed for 18 hours to PM at 0.1, 1, 10 and 50 μg/mL followed by M.tb exposure at a multiplicity of infection of 10 (MOI 10) for an additional 18 hours. We observed that PM exposure decreased the expression of HBD-2 and HBD-3 (p&lt;0.05), while IL-8 and MCP-1 expression remained unaltered in M.tb-infected A549 cells. Decreased antimicrobial peptide expression was not due to decreased cell viability upon PM exposure (shown by MTS and LDH assay). Interestingly, A549 cells exposed to PM exhibited a decreased capability to control M.tb growth (p&lt;0.05, CFU assays). We conclude that PM inhibits the gene expression and protein production of antimicrobial peptides in A549 cells, thus enhancing intracellular growth of M.tb that could potentially increase the susceptibility to M.tb infection.
Neutrophils prime a long-lived effector macrophage phenotype that mediates accelerated helminth expulsion (INC7P.423)
Authors: Chen Fei , Wu Wenhui , Millman Ariel , Craft Joshua , Chen Eunice , Urban Joseph , Kim Charlie , Gause William 
Published: January 2023

Abstract
The innate immune cells mediating macroparasite clearance remain largely undefined. We examined their role in acquired resistance to the parasitic nematode, Nippostrongylus brasiliensis (Nb) hypothesizing that they may mediate the markedly accelerated CD4+ cell-independent worm clearance occurring after secondary inoculation. After secondary inoculation, parasitic larvae in the lung are surrounded by macrophages and showed reduced ATP, indicating impaired metabolism. As late as one month after Nb inoculation, lung macrophages, transferred to naïve recipients, accelerated parasite clearance. Primed macrophages adhered to larvae in vitro and triggered increased mortality through CD11b-dependent mechanisms. Neutrophil depletion impaired the recall response and depletion of neutrophils in primed mice abrogated the protective effects of transferred macrophages in recipient mice and inhibited their binding to L3. Global transcriptome analyses of sort-purified lung neutrophils from Nb inoculated mice revealed a markedly different expression pattern from lung neutrophils isolated from LPS inculated mice, indicating that neutrophils, like macrophages, develop a distinct alternatively activated phenotype (N2) after helminth infection. These data thus indicate that differentially activated neutrophils in the context of a type 2 immune response prime a long-lived effector macrophage phenotype that binds metazoan parasites and directly mediates rapid worm damage and clearance.
Complementary immune roles for infected and uninfected cells during bacterial infection (INC7P.425)
Authors: Copenhaver Alan , Casson Cierra , Nguyen Hieu , Fung Thomas , Duda Matthew , Roy Craig , Shin Sunny 
Published: January 2023

Abstract
The innate immune system responds to virulent pathogens, yet many pathogens manipulate host signaling pathways which should limit immune activation. To understand how the immune system overcomes pathogenic manipulation we study the intracellular bacterium Legionella pneumophila, the cause of the severe pneumonia Legionnaire’s disease. Legionella utilizes a type IV secretion system to inject bacterial proteins into the cytosol of infected cells. Several of these bacterial effector proteins inactivate host cell factors involved in protein translation. Despite Legionella's ability to block host protein translation, inflammatory cytokines are still made against Legionella. It is unclear how infected cells can mount a cytokine response when host protein synthesis is blocked. Our studies demonstrate that infected cells do not produce many cytokines, such as IL-6 and TNF, critical for controlling Legionella infection. Instead, uninfected, bystander cells produce these cytokines. Infected host cells do produce IL-1 de novo. These data suggest that infected cells have mechanisms to overcome protein synthesis inhibition to produce IL-1 and that uninfected, bystander cells are important contributors to the immune response during infection with Legionella. This mechanism of immune activation has broad significance as many other bacterial pathogens manipulate immune cell signaling.
TLR7 and CD28 regulate plasmacytoid dendritic cell IFN-I induction and silencing during chronic viral infection (INC8P.439)
Authors: Macal Monica , Tam Hurtado Miguel , Chang Aaron , Hesser Charles , Zuniga Elina 
Published: January 2023

Abstract
Type I Interferons (IFN-I) are potent cytokines essential for immune surveillance against many pathogens including persistent viruses such as human immunodeficiency virus (HIV) in humans and Lymphocytic Choriomeningitis Virus (LCMV) in mice. Plasmacytoid dendritic cells (pDCs) are specialized to produce IFN-I but the molecular mechanisms underlying their initial IFN-I production and the subsequent exhaustion that occurs during chronic viral infection remain to be fully elucidated. Using chronic LCMV infection in mice, we demonstrated that pDCs initially sensed and produced abundant IFN-I in a TLR7-dependent manner. Furthermore, gene expression profile comparisons of pDCs and conventional (c) DCs identified CD28, a prototypic T cell stimulatory receptor, as highly expressed in pDCs. Strikingly, CD28 acted in a cell-intrinsic manner to negatively regulate the magnitude of pDC IFN-I production upon TLR stimulation in vitro and early after in vivo LCMV infection, but was dispensable for the subsequent IFN-I silencing. In contrast, cell-intrinsic TLR7 signaling was essential for the later pDC IFN-I exhaustion observed in chronic LCMV infection and compromised innate responses against secondary opportunistic pathogens. These results highlight a novel role for CD28 in limiting the magnitude of pDC IFN-I production and a dual role for TLR7 in mediating both induction and subsequent exhaustion of innate responses during in vivo chronic viral infection.
Critical role of Neutrophils in Respiratory Syncytial Virus (RSV) induced disease pathogenesis (INC8P.440)
Authors: Kolli Deepthi , Yueqing Zhang , Palkowetz Kimberly , Garofalo Roberto , Casola Antonella 
Published: January 2023

Abstract
Neutrophils comprise the largest number of immune cells in the human body, and being the first immune cells to the site of injury and infection, neutrophils act as the first line of defense against harmful pathogens. RSV has been identified as a leading cause of respiratory tract illness in children worldwide. RSV induced disease is characterized by epithelial desquamation, neutrophilic bronchiolitis and pneumonia. The role of neutrophils in innate immune responses to RSV infection has not been fully characterized. We have employed anti-Ly6G clone monoclonal antibody 1A8 to specifically deplete neutrophils in Balb/c mice to study the role of neutrophils in RSV induced disease pathogenesis. Neutrophil depletion in mice infected with RSV A2 or Long strain caused a significant increase in body weight loss, airway hyper responsiveness and lung inflammation. Even though no effect on peak viral replication was observed; a significant reduction in early viral load and a delayed viral clearance in mice lacking neutrophils were observed. While depletion of neutrophils has no effect on the production of BAL type I IFNs, a significant increase in Th17 group of cytokines (IL-17A, IL-17F, IL-22, IL-23p19 and CD40L), G-CSF, KC and MIP-1β was observed. Moreover, we observed a significant reduction in lung DCs, pDCs and virus specific CD4 and CD8 cells in neutrophil depleted mice. Our study demonstrates that neutrophils play a critical role in disease progression of RSV infections.
Antiviral regulation in porcine monocytic cells at different activation states (INC8P.430)
Authors: Sang Yongming , Rowland Raymond , Blecha Frank 
Published: January 2023

Abstract
Many pandemic viruses are monocytotropic, including porcine reproductive and respiratory syndrome virus (PRRSV), which directly infects subsets of macrophages and DCs and interferes in antiviral responses. To study antiviral responses in PRRSV-infected monocytic cells, we characterized inflammatory cytokine responses and genome-wide profiled signature genes to investigate response pathways in uninfected and PRRSV-infected monocytic cells at different activation states. Our findings showed suppressed interferon (IFN) production in macrophages at non-antiviral states and an arrest of lipid metabolic pathways in macrophages at antiviral states. Importantly, porcine monocytic cells at different activation states were susceptible to PRRSV and responded to viral infection differently. Based on gene ontology analysis, two approaches were used: 1) pharmaceutical modulation of cellular lipid metabolism and 2) in situ PRRSV replication-competent expression of IFN-α; both approaches significantly suppressed exogenous viral infection in monocytic cells. In particular, the engineered IFN-expressing PRRSV strain eliminated exogenous virus infection and sustained cell viability at 4 days post infection in macrophages. These findings suggest an intricate interaction of viral infection with activation status of porcine monocytic cells. An understanding and integration of antiviral infection with activation status of monocytic cells may provide a means of potentiating antiviral immunity.
The role of interleukin-27 in influenza A and Staphylococcus aureus co-infection (INC8P.435)
Authors: Robinson Keven , Lee Benjamin , Mandalapu Sivanarayana , Scheller Erich , Kolls Jay , Alcorn John 
Published: January 2023

Abstract
Rationale: Influenza A represents a significant cause of worldwide morbidity and mortality and results in increased susceptibility to secondary bacterial infections in the lung. We have previously shown that Influenza A impairs bacterial host defense through inhibition of Type 17 immunity. IL-27 is known to inhibit Type 17 immunity, suggesting a potential critical role for IL-27 in viral and bacterial co-infection. Methods: Wild-type and IL-27 receptor α knock-out (IL-27Ra-/-) mice were infected with Influenza A H1N1 (PR/8/34) or vehicle for 6 days followed by challenge with Staphylococcus aureus (ATCC 49775) or vehicle for 24 hours. Lung inflammation, bacterial burden, gene expression, and cytokine production were determined. Results: IL-27Ra-/- mice challenged with influenza A had increased weight loss compared to controls. In addition, IL-27Ra-/- mice displayed significantly decreased IL-10 production. IL-27Ra-/- mice co-infected with influenza and S. aureus had improved bacterial clearance compared to wild-type controls despite worsened lung pathology. Importantly, there were increased IL-17 and IL-22 expression and significantly decreased IL-10 production in the IL-27Ra-/- mice. Conclusions: These data suggest that IL-27 contributes to enhanced susceptibility to S. aureus pneumonia following influenza infection through the induction of IL-10 and regulation of Type 17 immunity.
NKT cells act as helper T cells to cytotoxic T cells by direct recognition of CD1d on T cells (IRC8P.487)
Authors: Oh Sejin , Park Se-ho 
Published: January 2023

Abstract
Cytotoxic T lymphocytes (CTLs), known as CD8+ T cells, recognize their targets by binding to antigens associated with MHC class I. NKT cells recognize foreign glycolipid antigens and endogenous glycolipid antigens presented by CD1d which is an MHC class I- like molecule. NKT cells have been shown to be essential for the optimal activation of CTLs. In this study, we investigated the mechanism how NKT cells provide help for the CTL responses. We observed significantly higher production of IFN-γ by antigen-stimulated OT-1 CD8+ T cells in the presence of NKT cells. This enhanced CTL activation by NKT cells was observed even in the antigen presenting cell (APC)-free conditions. We also found that CD8+ T cells start to up-regulate surface expression of CD1d reaching highest level at around 18 hours upon antigen recognition. These results suggest that not only APC, but also activated CTLs receive direct help from NKT cells by increasing CD1d expression on their cell surface when they recognize cognate antigens.
Role of Itk in Th17 mediated inflammation model hypersensitivity pneumonitis (IRC8P.491)
Authors: Carter Chavez 
Published: January 2023

Abstract
Hypersensitivity Pneumonitis (HP) is a lung disease caused by repeated inhalation of environmental antigens leading to inflammation, tissue scarring, and loss of lung function. This pathology is believed to be due to the increased IL-17A, a cytokine secreted primarily by Th17 cells that induces recruitment of inflammatory cells such as neutrophils. The thermophile Sacharopolyspora rectivigula (SR) causes HP, using a murine model of SR exposure; we study the molecular mechanism of this disease development. Using novel IL-17A-GFP reporter mice, our data suggests that the high levels of IL-17A induced in response to SR are produced in part by CD4+ T cells and not neutrophils. The Tec family tyrosine kinase Itk regulates T cell activation and cytokine production in conventional Th17 cells but Itk-/- mice still develop HP. Histology shows an increase in inflammatory cells in lung airways as well as decline of lung tissue architecture. Itk-/- lungs also have high levels of IL-17A mRNA expression, and an increase in the number of CD4+ IL-17A producing cells. However, Itk-/- mice lack IL-17A producing γδ T cells in the early stage of HP, which recovers in the later stage. This directly coincides with the emergence of IL-17A producing CD4+γδ T cells in the lungs of Itk-/- mice exposed to SR. We conclude that Itk regulated signals are not critical for the production of IL-17A in response to SR, and Itk may differentially regulate the production of IL-17A in different types of T cells.
Anti-CD3 antibody treatment induces hypoglycemia and super tolerance to glucose challenge in mice through enhancing glucose consumption by activated lymphocytes (IRC8P.474)
Authors: Xia ChangQing , Chernatynskaya Anna , Looney Benjamin , Clare-Salzler Michael 
Published: January 2023

Abstract
Anti-CD3 antibody has been employed in several clinical settings. However, its effect on glucose metablolism has not been well investigated. In the current study, we investigated how anti-CD3 treatment affected blood glucose levels in mice. We found that anti-CD3 treatment induced immediate reduction of blood glucose as early as 3-4 hours post-treatment. Furthermore, we observed that a single dose of anti-CD3 treatment was able to correct hyperglycemia in all diabetic NOD mice. This glucose-lowering effect was not attributable to major T cell cytokines secreted by anti-CD3 activated T cells. Of interest, when tested in a normal strain of mice (C57BL/6), the C-peptide levels in anti-CD3-treated animals were significantly lower than those of control mice. Paradoxically, the anti-CD3-treated animals were highly tolerant to the exogenous glucose challenge. In addition, we found anti-CD3 treatment significantly induced activation of T and B cells in vitro and in vivo. Our further studies showed that anti-CD3 treatment lowered the glucose levels in T cell culture media, and increased the intracellular transportation of 2-NBDG particularly in activated T and B cells. In addition, injection of anti-CD3 antibodies upregulated Glut1 expression in spleen cells. The present study suggests that anti-CD3 therapy-induced hypoglycemia likely results from increased glucose transportation and consumption by the activated lymphocytes.
Late antigen recognition at the memory checkpoint is necessary for CD4+ T cell memory formation. (IRC9P.708)
Authors: Bautista Bianca , McKinstry Kai , Strutt Tara , Swain Susan 
Published: January 2023

Abstract
During the contraction phase of an influenza immune response, the majority of responding T cells undergo Bim-driven apoptosis leaving a small, heterogeneous population of T cells that will survive to form memory. The regulation of this process is poorly understood. Recently, our lab defined a “memory checkpoint” during which highly differentiated effector CD4+ T cells require autocrine IL-2 immediately preceding contraction to become memory. To determine whether antigen recognition is required for IL-2 production during the memory checkpoint, we developed an adoptive transfer model to modulate antigen. Donor CD4+ effectors transferred to hosts without antigen underwent immediate cell death and failed to form memory, suggesting that antigen recognition is indeed the key event at this checkpoint. The ex vivo evaluation of CD4+ effector cells at the memory checkpoint revealed that both co-stimulation and IL-2 signaling contribute to survival. TCR stimulation during the memory checkpoint resulted in higher sustained expression of memory markers CD127 and CXCR3. The survival to memory is mediated in part by a sustained reduction in Bim expression, as the requirement for late antigen interactions was eliminated in Bim heterozygous CD4+ T cells. These findings demonstrate that effectors generated by influenza must recognize antigen at the memory checkpoint to become memory.
Tapeworm antigens block LPS-induced inflammatory mediator production by macrophages via IL-10 and recruit myeloid derived suppressor cells (IRC10P.463)
Authors: Reyes Jose , Fernado Maria , Matisz Chelsea , McKay Derek 
Published: January 2023

Abstract
Parasites have been shown to suppress concomitant inflammatory disease in mouse models: mice infected with the rat tapeworm, Hymenolepis diminuta, are protected from colitis. Assessing the mechanism by which this worm may inhibit inflammation the objective was to determine if a PBS-soluble extract of H. diminuta antigens (HdAg) directly affected innate immune cell activity. Bone marrow macrophages (BMMs) exposed to LPS ± HdAg for 48 hr and IL-1β, IL-6, TNFα, NO and IL-10 measured in culture supernatants 1, 6, 24 and 48 hr later. Additional studies included the addition of an anti-IL-10 antibody to the cell culture. In other experiments, mice received HdAg (500 μg, ip) and 24 hr later peritoneal exudate cells were analyzed by flow cytometry. LPS-induced IL-1β and TNFα, but did IL-6 or NO, were significantly reduced by ~50% by HdAg co-treatment. BMM treated with HdAg showed a dose- and time-dependent increase in IL-10, paralleled by increased STAT3 phosphorylation suggesting IL-10 feedback on the BMM: anti-IL-10 prevented the HdAg inhibition of LPS-evoked TNFα by BMM. Mice given HdAg recruited a F480+Gr1+ cell population that boosted IL-10 production by anti-CD3/CD28-activated T cells. In conclusion, HdAg enhanced IL-10 production by LPS-stimulated BMMs which in turn down-regulated TNFα and IL-1β secretion. This, coupled to the recruitment/generation of MDSC, likely contributes to the suppression of pro-inflammatory events in H. diminuta infected mice.