bluuebunny/crossref_metadata_2025_split huggingface dataset:
Extracellular bactericidal functions of porcine neutrophils (133.20)
Authors: Scapinello Sarah E. , Brooks Andrew S. , Caswell Jeff L. , Macinnes Janet I. , Hammermueller Jutta , Smith Alexandra K. 
Published: January 2023

Abstract
Neutrophils are one of the main effector cells of innate immunity and were shown to kill bacteria via phagocytosis more than 100 years ago. New developments, however, also show that neutrophils are capable of antimicrobial activity by producing extracellular structures named neutrophil extracellular traps (NETs). The antimicrobial effects of NETs can be attributed to cathelicidins, a category of antimicrobial peptides stored as inactive proforms. These proforms are cleaved by neutrophils elastase during cell activation in response to pathogens. This project focuses on the cathelicidin peptides contained within NETs and their effectiveness in killing several common bacterial pathogens of swine.
Subsequent investigations address the contribution of cathelicidins to extracellular bactericidal activity of porcine neutrophils. Activated secretions of neutrophils are effective in killing 100% of Escherichia coli K12, 21% of Streptococcus suis, and 74% of Actinobacillus suis. Pasteurella multocida appears to survive in the presence of activated neutrophil secretions demonstrating a 38% increase in growth when compared to P. multocida exposed to non activated neutrophils secretions. Identification of the contribution of cathelicidins to this bactericidal activity is pending. These data suggest that activated neutrophil secretions have bactericidal activity against common pathogens in swine.
Differences between pathogenic and non-pathogenic microsporidial-induced signaling in macrophages (133.30)
Authors: Mathews Amber , Fischer Jeff L , Hotard Annie , Donze Hollie Hale 
Published: January 2023

Abstract
Microsporidia are ubiquitous, obligate intracellular eukaryotes that cause chronic diarrhea in humans, especially in immunocompromised individuals. Little is known about the innate immune response to these parasites. Macrophages are a source of interleukin-12 (IL-12) and play an essential role in the link between innate and adaptive immunity. Previous studies from our lab have shown that pathogenic species, E. cuniculi and E. intestinalis, are preferentially recognized by TLR2 in primary human macrophages and produce IL-10 (Th2) responses. In this study, we are investigating the signaling cascades regulated by TLR2 and TLR4 engagement with pathogenic and non-pathogenic species of microsporidia. Because TLR4 engagement triggers MAPK activation leading to IL-12 production, we hypothesize that differences in TLR activation by non-pathogenic species will lead to activation of p38 MAPK and IL-12 production. Using small interfering RNA, p38 MAPK was knocked down in human macrophages and resulted in a decrease in IL-12p40 and tumor necrosis factor alpha (TNF-α) production when infected with Vairimorpha necatrix, a non-pathogenic species, and a decrease in TNF-α production when infected with E. cuniculi. Using western blot, we found that activated p38 MAPK is continuously expressed from 3h to 24h post infection of human macrophages with V. necatrix. Expression of activated p38 MAPK is transient when infected with E. cuniculi as we only detected activation at 3h and 6h post infection. These data suggests that activation of p38 MAPK is necessary for the appropriate innate immune response to microsporidia and for control of infection. This work is supported by NIH grant AIO71778.
Ly49H signaling through both DAP10 and DAP12 is required for optimal NK cell responses to mouse cytomegalovirus infection (133.39)
Authors: Orr Mark T , Lanier Lewis L 
Published: January 2023

Abstract
The activating natural killer (NK) cell receptor Ly49H recognizes the mouse cytomegalovirus (MCMV) m157 glycoprotein expressed on the surface of infected cells and is required for protection against MCMV. Although Ly49H has previously been shown to signal via DAP12, we now show that Ly49H must also associate with and signal via DAP10 for optimal function. In the absence of DAP12, DAP10 enables Ly49H-mediated killing of m157-bearing target cells, proliferation in response to MCMV infection, and partial protection against MCMV. DAP10-deficient Ly49H+ NK cells, expressing only Ly49H - DAP12 receptor complexes, are partially impaired in their ability to proliferate during MCMV infection, display an aberrant signaling profile, produce less IFN-γ upon Ly49H engagement, and demonstrate reduced control of MCMV infection. Deletion of both DAP10 and DAP12 completely abrogates Ly49H surface expression and control of MCMV infection. Thus, optimal NK cell-mediated immunity to MCMV depends on Ly49H signaling through both DAP10 and DAP12.
Peripheral blood monocytes (PBMCs) display innate antiviral response to rhinovirus (RV) infected bronchial epithelial cells (133.44)
Authors: Wark Peter AB , See Hayley V , Simpson Jodie L , Forbes Rebecca L , Hansbro Phil 
Published: January 2023

Abstract
Rhinovirus (RV) infection is an important trigger of acute asthma and COPD.
Study objectives; were to characterise the response of PBMCs and CD8 lymphocytes when exposed to RV infected bronchial epithelial cells.
Methods; Human Calu3 cells were used as a model for bronchial epithelial cells and infected with RV-43 and RV-1B. PBMCs were obtained from healthy individuals and separated by Ficoll-Paque centrifugation. CD8 lymphocytes were identified as CD3+, CD8+ by flow cytometry. PBMCs were exposed to infected Calu3 cells, cells exposed to UV inactive RV, PHA and TLR3 agonist PolyI:C. Response was measured by ELISA for release of interferon(IFN)-γ and TNF-α. CD8 activation with expression of perforin or proliferation measured by carboxyfluorescein succinimidyl ester assay.
Results: Exposure of PBMCs to Calu3 cells infected with both RV led to significant release of IFN-γ and TNF-α, smaller release was seen in response to UV inactive RV. Exposure to PolyI:C also led to a similar release. CD8+ cells were found to be the primary source of IFN-γ release. CD8+ cells did not show an increase in perforin expression or evidence of proliferation.
Conclusions; exposure of PBMCs and CD8+ cells to an RV infected bronchial epithelium alone results in an innate immune response mediated by TLR3 without evidence of a CD8 adaptive immune response.
TLR-7 Mediated Responses to Respiratory Syncytial Virus (RSV) (133.45)
Authors: Lindell Dennis M , Morris Susan , Mukherjee Sumanta , Lukacs Nicholas W 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) infection results in lung disease characterized by mucus hypersecretion and inflammation. RSV has also been implicated in both the initial development of asthma, as well as the exacerbation of existing asthma. Previous studies in our laboratory have demonstrated roles for TLR3 and the adapter molecule MyD88 in the host response to RSV. In these studies, we investigated the role of TLR 7 in the recognition of RSV and RSV-induced pulmonary disease. TLR7 -/- and WT C57 Bl/6 mice were infected intratracheally with 10^5 PFU RSV Line 19. Compared to control mice, TLR 7-/- mice had significantly increased mucus hypersecretion, as assessed via PAS staining of lung sections and the expression of Muc5ac and Gob5. Mucus hyperresponsiveness was associated with increased IL-17 and IL-13 (as well as other cytokines). Cell type specific studies revealed altered responses to RSV in TLR7 -/- B cells, plasmacytoid dendritic cells (pDC), and surprisingly, classical myeloid dendritic cells (mDC). Studies in bone marrow-derived dendritic cells revealed TLR-7 dependent effects on IL-12 signaling. In response to the TLR7/8 specific agonist 3M-011, BMDC dramatically up-regulated IL-12 p35. In response to RSV, TLR7 -/- BMDC were significantly impaired in the induction of IL-12 p35 in response to RSV, but exhibited significantly higher IL-23 p19. Thus, TLR-7 regulates IL-12/IL-23 responsiveness to RSV.
Supported by P01HL031963 (NWL), R01AI073876 (NWL)
Evidence of Murine Alternatively Activated Lung Macrophages are Associated with Failure to Clear Pulmonary Cryptococcus neoformans (133.54)
Authors: Quintero Eric A , Gott Katherine A , Wilder Julie A 
Published: January 2023

Abstract
We have previously shown that murine pulmonary immunity to the fungal pathogen Cryptococcus neoformans (Cne) is genetically regulated. C.B-17 mice efficiently clear the organism from the lung following intranasal inoculation whereas C57BL/6 mice do not. The ability to clear Cne from the lungs of C.B-17 mice is dependent upon the development of a Type 1 immune response. In contrast, infected C57BL/6 mice mount a Type 2 immune response. As alternatively activated macrophages (AAMs) have been associated with the development of a Type 2 immune response, we reasoned that Cne-infected C57BL/6 mice may have more AAMs in their lungs as compared to infected C.B-17 mice. We tested our hypothesis by analyzing the expression of four transcripts generally over-expressed in AAMs (YM1, FIZZ1, ARG1, and ARG2) in whole lung homogenate samples via PCR. Our data show increased expression of these markers over time post-Cne infection in both strains (increasing from 6 hours - 14 days) with greater expression in C57BL/6 mice at all time points. Lungs from naïve C57BL/6 mice also showed higher expression of ARG1, FIZZ1 and YM1 compared to naïve C.B-17 lungs. These data support our hypothesis that there are more AAMs in the lungs of Cne-infected C57BL/6 mice as compared to similarly infected C.B-17 mice. This research was supported by Lovelace Respiratory Research Institute internal funds.
Bacillus anthracis lethal toxin abrogates TCR-induced cytokine production by CD1d-restricted Natural killer like - T cells (134.15)
Authors: Joshi Sunil K , Lang Gillian , Devera T. Scott , Aye Lindsay M , Shah Hemangi B , Ballard Jimmy , Lang Mark 
Published: January 2023

Abstract
Exogenous CD1d-binding glycolipid (α-Galactosylceramide, α-GC) stimulates TCR signaling and activation and proliferation of type-1 natural killer like - T (NKT) cells. These activated NKT cells play a central role in regulation of adaptive and protective immune responses against pathogens and tumors. In the present study, we tested the direct in vivo and in vitro effect of Bacillus anthracis lethal toxin (LT) on NKT cells. LT is a complex protein formed by pore-forming protective antigen (PA) upon binary combination with lethal factor (LF) subunit. A sub-lethal dose of LT administered in vivo in C57BL/6 mice altered expression of the activation receptor NKG2D by NKT cells but not by NK cells. We observed that NKT cells expressed anthrax toxin receptors (CMG-2 and TEM-8) and bound PA more than other immune cell types. The in vivo administration of LT led to decreased TCR-induced cytokine secretion but did not affect TCR expression. We propose that Bacillus anthracis-derived LT contributes to immune evasion by altering NKT function.
This work was supported by Defense Threat Reduction Agency (DTRA) grant CBDIF-07-PRET-01 to M.L.L and by a pilot award to M.L.L. as part of NIH grant 5 U19 AI062629-05.
Post-translational regulation of the NKG2D ligand Mult1 in response to cell stess (134.2)
Authors: Nice Timothy J , Coscoy Laurent , Raulet David H 
Published: January 2023

Abstract
NKG2D is a major stimulatory receptor expressed by natural killer cells and some T cells. The receptor recognizes MHC class I-like cell surface ligands that are poorly expressed by normal tissues but often induced in transformed and infected cells. Induction of NKG2D ligands during transformation or infection leads to activation of NKG2D-expressing immune cells and elimination of the diseased target cells. The existence of several NKG2D ligands in each individual, some with strikingly divergent protein sequences, raises the possibility that different ligands are regulated by distinct disease-associated stresses. The transcripts for some ligands, including murine UL16-binding protein-like transcript 1 (Mult1), are abundant in certain normal tissues where cell surface expression is absent suggesting the existence of translational or post-translational regulation. We report that under normal conditions, Mult1 protein undergoes ubiquitination dependent on lysines in its cytoplasmic tail and is degraded by the lysosome. Mult1 degradation and ubiquitination is reduced in response to stress imparted by heat shock or ultraviolet (UV) irradiation, but not by other forms of genotoxicity, providing a novel mechanism for stress-mediated cellular control of NKG2D ligand expression.
TLR3 Signaling in iNKT cell Promotes Apoptosis of γδ T cells (134.27)
Authors: Ajuebor Maureen N , Jin Yijun , Chen Qingling , Gardner Tommy 
Published: January 2023

Abstract
Polyinosinic-polycytidylic acid (poly I:C, a specific ligand for TLR3) activate immune cells (such as DC and NK cell) in a TLR3-dependent manner to induce cytokine production. TLR3 expression by iNKT cells has not been determined. The main objective of this study was to determine TLR3 expression by iNKT cell and their functional responsiveness after stimulation with poly I:C. We hypothesize that TLR3 signaling in iNKT cells suppress γδT cell functions by induction of apoptosis. Our hypothesis is based on the following observations: First, poly I:C administration into C57BL6 mice was associated with extracellular TLR3 and intracellular cytokine (IFN-γ and TNF-α) expression by hepatic iNKT cells. Second, activated hepatic iNKT cells promote apoptosis of γδT cells via induction of IFN-γ and TNF-α since pretreatment of C57BL6 mice with anti-TNF-α or anti-IFN-γ Abs prior to poly I:C administration caused a significant decrease in the frequency of active caspase 3 expression by hepatic γδT cells. Finally, the frequency of γδT cells accumulation in the liver was significantly increased by iNKT cell deficiency (vs. WT mice) after poly I:C administration. Moreover, hepatic γδT cells derived from iNKT cell KO mice were resistant to apoptotic death (and thus associated with increased survival) relative to hepatic γδT cells derived from WT mice post-poly I:C administration. It is noteworthy that serum levels of TNF-α and IFN-γ were also ablated by iNKT cell deficiency. In summary, our results support the notion that (i) hepatic iNKT cells directly respond to TLR3 signaling to activate γδT cells; (ii) the activation depends on cell-cell contact whereby iNKT cell-derived cytokines induce the programmed cell death of γδT cells; an effect that may attenuate potential effector functions of γδT cells.
Autocrine signaling produced by M2bMφ is required for the prolongation of properties (134.29)
Authors: Asai Akira , Shigematsu Kenji , Kobayashi Makiko , Herndon David N , Suzuki Fujio 
Published: January 2023

Abstract
Three different subtypes of M2Mφ (M2aMφ, M2bMφ and M2cMφ) have recently been demonstrated in mice with severe burn injuries. M2aMφ and M2cMφ appeared 1 to 4 days after burn injury, while M2bMφ appeared in mice 1 to 3 weeks after burn injury. In this study, how M2bMφ with suppressor cell activities are prolonged is examined. M2bMφ were peritoneal Mφ isolated from burned BALB/c mice (25% TBSA, 3rd degree flame burn) using biotin-conjugated anti-mouse F4/80 mAb and streptavidin magnetic beads. M2bMφ (1 x 106 cells/ml) isolated from mice 16 days after burn injury were cultured under frequent medium changes (every 12 hrs for 3 days). As a control, the same M2bMφ preparation was cultured for 3 days without any medium changes. Mφ harvested were assayed for LIGHT mRNA by RT-PCR. Also, obtained Mφ were cultured an additional 120 hrs and culture fluids harvested were assayed for CCL1 using ELISA. LIGHT mRNA and CCL1 were typical biomarkers for M2bMφ. In the results, M2bMφ cultured for 3 days without any medium changes expressed LIGHT mRNA. However, this mRNA was not expressed by M2bMφ cultured with frequent medium changes. CCL1 was detected in the culture fluids of M2bMφ that had been previously cultured without any medium changes. When M2bMφ were cultured for 3 days under the frequent medium changes, the ability of these cells to produce CCL1 was reduced by 70%. These results indicate that M2bMφ cultured under the frequent medium changes lose their M2bMφ properties. Some soluble factors in the culture fluids of M2bMφ were shown to be involved in the persistence of M2bMφ properties. This study was supported by SHC NA #8620.
Heterogeneity of COX-2 macrophages in acute and chronic mycobacterial inflammation (134.30)
Authors: Shibata Yoshimi , Kogiso Mari , Pantuso Traci , Shinohara Tsutomu , Dorey C. Kathleen , Henriksen Ruth Ann 
Published: January 2023

Abstract
Persistent up-regulation of cyclooxygenase-2 (COX-2) and over-production of PGE2 by activated macrophages (MØ) compromise host immunity in cancer immunotherapy, autoimmune diseases, and chronic mycobacterial inflammation such as TB. Previous studies showed that a population of COX-2+ MØ is induced in the spleen of mice given i.p. HK-Mycobacterium bovis BCG. These MØ are derived from radiosensitive immature bone marrow cells that migrate and localize in the spleen. Mature spleen MØ express active COX-2 (subcellularly associated with the nuclear envelope (NE)), and release increased PGE2 from day 5 to day 21 after BCG administration. To determine if local sites of inflammation caused by BCG accumulate similar COX-2+ MØ, C57Bl/6 mice were given i.p. or i.n. BCG (1 and 0.5 mg doses, respectively). We found that 1 - 2 days after i.p. administration, peritoneal and splenic MØ transiently exhibited COX-2 which was dissociated from the NE, inactive, and released no PGE2. However, unlike splenic MØ, neither active nor inactive COX-2 was found in peritoneal MØ over days 3 - 28. After i.n. administration, alveolar MØ persistently expressed inactive COX-2 over days 1 - 28. No alveolar MØ with active COX-2+ were detected. Normal splenic, peritoneal, and alveolar MØ treated in vitro with BCG expressed active COX-2 and increased PGE2 release within 24 h, indicating that in vitro MØ activation does not always mimic in vivo responses. Our results indicate that in vivo BCG preferentially induces active COX-2 in the splenic MØ. In addition, without suppression by PGE2, MØ with inactive COX-2 may be "hyper-active" and regulate tissue-dependent acute and chronic inflammation. (Supported by NIH HL71711, DOD DAMD 17-03-1-0004, BCCRP 06BS-04-9615)
Serum Amyloid P inhibits Renal Fibrosis through an Fc-gamma receptor dependent mechanism. (134.33)
Authors: CASTANO ANA P , Lupher Jr Mark , Duffield Jeremy S 
Published: January 2023

Abstract
Serum amyloid P (SAP) inhibits fibrocyte outgrowth from PBMCs and may therefore play a role in fibrosis. We studied the role of SAP in regulating fibrosis in the kidney where there are very few fibrocytes. In the UUO model of kidney fibrosis, IP or IV SAP (20µg/g) significantly inhibits fibrosis. (11.3% at 14d vs. 5.4%). Similar findings were seen in other models of kidney fibrosis. In transgenic Col1-GFP mice, SAP reduced myofibroblast GFP by 50%, but myofibroblast numbers were not affected, indicating SAP switches off collagen production. SAP does not bind to fibroblasts or reduce collagen production by fibroblasts in vitro. SAP binds to monocytes, Møs and apoptotic cells. Møs are fibrogenic through paracrine signaling to fibroblasts. SAP binds FcγRI, II, III, IV, in mice and inhibits Monocyte/Mø activation, assessed by inflammatory gene transcript levels, and cytokine production in vitro. Primed Møs are susceptible to soluble IgG-mediated activation suggesting SAP inhibits kidney Mø activation by an Fcγ-dependent mechanism. Purified UUO kidney Møs were also less activated and produced more IL10 in mice treated with SAP. To test the role of FcγRs in kidney Mø activation we induced UUO fibrosis in FcRγ-chain KO mice. Compared with controls, fibrosis was attenuated, confirming a role for FcγR-mediated activation of kidney Møs in the development of fibrosis. Our studies point to SAP as a potential therapeutic in the management of progressive fibrosis.
Defective Dendritic Cell Phagocytic Function in Aging (134.36)
Authors: Julius Annabelle , Abernathy Lisa , Yung Raymond 
Published: January 2023

Abstract
Dendritic cells (DCs) play an important role in many immune-mediated processes in aging. One of their major functions is the phagocytic clearance of pathogens and tumor cells. Currently, very little is known about the role of aging in determining DC phagocytic function. Using murine CD11c+ bone marrow-derived DCs from young (3-6 months) and old (21-23 months) C57BL/6 mice, we compared the phagocytic function of DCs. We found that old DCs have decrease capacity to internalize exogenous FITC-dextran compared with young DCs. Similarly, old DCs displayed a decrease in the global uptake of E. coli bacteria per cells (398 ± 32.6 fluorescence units) when compared to young DCs (598.7 ± 13.7). This impaired phagocytic activity in old DCs was further confirmed by quantifying the number of bacteria ingested per cell by confocal microscopy. Intriguingly, aging does not affect the ability of DCs to ingest zymosan or Gram positive bacteria such as S. Aureus, suggesting a decrease in the expression and/or function of selective receptors associated with gram negative bacteria phagocytosis in aging. Cell surface expression analysis of several receptors including CD206, CD205, TLR2, TLR4 and TLR5 were similar in both aged group. However, aging impacts the expression of several mediators of TLR4 Myd88 independent but not dependent signaling, including Ticam2 and IRF3, which may contribute to the observed impaired gram negative DC-mediated phagocytosis in aging. A better understanding of the effect of age on DC phagocytosis may improve our understanding on why selected infectious diseases are more prevalent in the elderly and provide us with new treatment options.
Thapsigargin and flavin adenine dinucleotide ex vivo treatment improves leukocyte microbiocidal activity through rescuing trafficking-defective gp91-phox in chronic granulomatous disease (134.37)
Authors: Huang Ya-Fang , Liu Si-Yen , Yang Pei-Wen , Yen Chia-Liang , Shieh Chi-Chang 
Published: January 2023

Abstract
Mutations in leukocyte NADPH oxidase genes lead to defective respiratory burst in white blood cells and cause chronic granulomatous diseases (CGD) in affected patients. The most common form of CGD is caused by mutations in the membrane-bound component of gp91-phox, which is encoded by the CYBB gene located on the X chromosome. We previously reported a patient with CYBB mutation (H338 to Y) which prevents the intracellular trafficking and expression of gp91-phox on the leukocyte surface. By treating the leukocytes with thapsigargin and flavin adenine dinucleotide (FAD), we rescued the capacity of the leukocytes to produce reactive oxygen species (ROS). We visualized the enhanced cell surface expression of the mutant protein after thapsigargin and FAD treatment with GFP-tagged gp91-phox expression with the CGD mutation. Protein and cellular analyses showed that treated cells expressed more endo-H resistant gp91-phox protein and are more effective in killing bacteria. Thapsigargin and FAD-treated CGD leukocytes had enhanced activity in protecting mice from S. aureus-induced peritoneal abscess formation in a mouse model of CGD. These results indicate that thapsigargin and FAD ex vivo treatment is effective in rescuing the ROS-producing activity of leukocytes in selected CGD patients.