bluuebunny/crossref_metadata_2025_split huggingface dataset:
Activation of membrane TGFβ1 on alveolar macrophages by co-cultured alcohol primed alveolar epithelial cells (111.1)
Authors: Joshi Pratibha , Venado Aida , Curry-McCoy Tiana , Guidot David 
Published: January 2023

Abstract
Alcohol abuse impairs pulmonary innate immunity rendering individuals susceptible to pneumonia. We have previously shown that alcohol abuse causes oxidative stress in the lung and increases TGFβ1 expression. TGFβ1 downregulates GM-CSF receptors resulting in the alveolar macrophage and epithelial cell dysfunction. However, the exact mechanism of alcohol-mediated activation of TGFβ1 in vivo is still under investigation. We examined TGFβ1 expression on alveolar macrophages and epithelial cells from control- and alcohol-fed rats by real time PCR and flow cytometry. TGFβ1 gene expression was significantly higher in alcoholic macrophages and TGFβ1 protein was predominantly membrane bound in macrophages. In addition, as compared to control, alveolar epithelial cells from alcohol-fed rats had significantly higher expression of alpha5 integrin chain that is implicated in TGFβ1 activation. Therefore, we hypothesized that alcohol-induced increase in epithelial alpha5 would activate membrane TGFβ1 on macrophages. To examine this, macrophage (NR8383) and epithelial (L2) cell lines were co-cultured on transwell membranes to assess the changes in paracellular permeability. Only alcohol primed co-cultures showed a decrease in epithelial barrier function that was reversed by anti-TGFβ1 treatment. Taken together, these results suggest that cellular contact between alveolar macrophages and epithelial cells is essential for alcohol mediated activation of TGFβ1 in the lung.
SerpinB1 functions in generating neutrophil extracellular traps (111.11)
Authors: Farley Kalamo , Stolley James , Cooley Jessica , Remold-O'Donnell Eileen 
Published: January 2023

Abstract
SerpinB1 is a potent inhibitor of neutrophil serine proteases (NSPs), and mice deficient for serpinB1 showed reduced capacity to clear Pseudomonas aeruginosa lung infection, increase of proinflammatory cytokines and neutrophil death compared with wild type (WT) mice. Here we questioned whether this deficient anti-microbial protection might be caused by altered production of neutrophil extracellular traps (NETs). NETs, extruded chromatin associated with granule proteins including NSPs, capture and kill microbes, but excess NETs can cause tissue injury, e.g., impairment of lung function in patients with cystic fibrosis. On stimulation with phorbol myristate actetate (PMA) or platelet-activating factor (PAF), isolated bone marrow neutrophils of serpinB1-/- mice produced increased NETs compared with WT. Isolated lung neutrophils of LPS-treated serpinB1-/- mice also produced increased NETs compared with WT in response to PMA, and increased numbers of neutrophils with expanded nuclei. NET production was NADPH oxidase-dependent, and NADPH oxidase activity did not differ between the genotypes. Fluorescent microscopy showed that SerpinB1 translocated in PMA treated human neutrophils from the cytoplasm to the nucleus prior to nuclear membrane disintegration. SerpinB1 was also found as an extracellular component of NETs. These findings indicate that SerpinB1 contributes to regulation of NET formation and, by its presence in NETs, may protect against protease-induced tissue injury.
The role of type 1 IFN after oral Toxoplasma gondii infection (111.15)
Authors: Han Seong-Ji 
Published: January 2023

Abstract
The protective role of type I interferons (IFN) in host immunity to viral infections has been well characterized. However, the role of type I IFN following parasitic infection is less clear. To investigate this further, we used type I interferon receptor knock out (IFNAR -/-) mice and orally infected them with the protozoan parasite, Toxoplasma gondii (T. gondii). We observed a significant increase in the parasite load in the mesenteric lymph nodes (MLNs) of IFNAR-/- mice with a concomitant drop in survival relative to wild type mice. Furthermore, we observed decreased levels of serum CCL2 that correlates with reduced recruitment of CD11b+Ly6C+ inflammatory monocyte (IM) into the MLNs of IFNAR-/- mice. We observed that IFN-β in wild type mice is produced in two waves in the MLNs after oral infection and we are currently investigating which cells are responsible for type I IFN production at different time points. We present a model for the protective role of type I IFN during T. gondii infection and provide new insight into innate defense against protozoan parasites.
Effect of CCL1 antisense ODN on bacterial translocation in mice irradiated with whole body 137Cs γ-rays: elimination of M2b macrophages in translocation sites (111.19)
Authors: Suzuki Fujio , Patel Niket , Kogiso Mari , Cornforth Michael , Kobayashi Makiko 
Published: January 2023

Abstract
Mice irradiated with 5 Gy whole body 137Cs γ-rays (WBI-mice) are shown to be carriers of M2bMϕ (IL-10+CCL1+LIGHT+IL-12- Mϕ), which are inhibitory on the host antibacterial innate immunities in bacterial translocation. M2bMϕ maintain their suppressor cell properties through the production of CCL1. In this study, therefore, we tried to eliminate M2bMϕ from mesenteric lymph nodes (MLN) of WBI-mice by CCL1 antisense ODN gene therapy. WBI-mice were treated with CCL1 antisense ODN with various doses (2, 10, 50, or 250 μg/mouse), routes (i.p., i.m. or s.c.) and schedules (twice a day for 2 to 7 days, starting 1 to 4 weeks after γ-irradiation). F4/80+ MLN-Mϕ isolated from these mice were identified as M2bMϕ by their abilities to express intracellular IL-10 and LIGHT mRNA. As a control, MLN-Mϕ were isolated from WBI-mice treated with scrambled ODN and subjected to the same test. In the results, the numbers of MLN-M2bMϕ were greatly reduced in WBI-mice treated s.c. with 10 to 50 μg/mouse of CCL1 antisense ODN twice a day for 2 days, 5 days or 7 days, started 1 to 3 weeks after γ-irradiation. However, MLN-M2bMϕ were detected in WBI-mice treated with CCL1 antisense ODN twice a day for 2 days started 4 weeks after the irradiation. These results indicate that M2bMϕ are eliminated from MLN of WBI-mice s.c. treated with CCL1 antisense ODN twice a day for 2 days starting 1 to 3 weeks after γ-irradiation.
Fatal outcome of pandemic H1N1 2009 influenza infection is associated with neutrophil and macrophage lung infiltration but not enhanced viral burden in pregnant mice. (111.3)
Authors: Marcelin Glendie , Aldridge Jerry , Boon Adrianus , Duan Susu , Rehg Jerold , Webby Richard 
Published: January 2023

Abstract
Pandemic H1N1 2009 Influenza A virus (pH1N1) infection in pregnant or postpartum women is quite severe. The immunological mechanisms that affect infection outcome in these populations are not well understood. To address this, pregnant Balb/c and non-pregnant mice were inoculated with a wild-type pH1N1 virus. We determined that early cytokine responses were impaired in pregnant mouse lungs. Moreover, pulmonary infiltration of neutrophils and macrophages strongly correlated with an increase in mortality in pregnant mice. Increased mortality in pregnant animals was not associated with higher virus load because equivalent viral burden (virus titers and immunohistochemical staining) was observed in the nasal cavities and lower respiratory tracts of all mice. Pregnancy altered innate responses despite the presence of similar viral burden. Also, pregnant mice demonstrated equivalent influenza-specific activation of pulmonary CD8+ T lymphocytes however; they displayed elevated T-regulator lymphocyte (Tregs) responses in the lung and mediastinal lymph nodes. Despite these immunologic alterations, pregnant animals mounted equivalent hemagglutinin inhibition and IgG subclass titers in response to virus or immunization with monovalent inactivated pH1N1 vaccine. Therefore, the severity of pH1N1 in pregnant mice is associated with pulmonary neutrophil and macrophage invasion, dysregulated cytokine responses but not increased viral load.
IL-10 antisense oligodeoxynucleotides (ODN) improve the resistance of severely burned mice intradermally infected with MRSA (111.8)
Authors: Asai Akira , Kobayashi Makiko , Herndon David , Suzuki Fujio 
Published: January 2023

Abstract
The effect of IL-10 antisense ODN on the susceptibility of burned mice to intradermal (i.d.) infection of MRSA was studied. IL-10 antisense ODN (6 μg/mouse) was administered to mice twice a day for 3 days beginning 1 day after burn injury (3rd degree, 25% TBSA flame burn). Two days after burn injury, these mice were infected i.d. with 108 CFU/mouse of MRSA. Abscess formation in the infection site and mortality rates of these mice were tested. Also, F4/80+ Mϕ isolated from tissues surrounding the infection sites of these mice were biophysically characterized. In the results, sepsis caused by MRSA i.d. infection did not develop and abscess formation was demonstrated in burned mice treated with IL-10 antisense ODN. However, all burned mice treated with scrambled ODN (control group) died from sepsis, and abscess formation was not shown in these mice. Mϕ isolated from the infection site tissues of burned mice treated with IL-10 antisense ODN were identified as M1Mϕ, while those from control mice were shown to be M2Mϕ. Abscess formation was demonstrated in burned mice inoculated with M1Mϕ (but not M2Mϕ), and these mice resisted a lethal dose of MRSA i.d. infection. These results indicate that burned mice treated with IL-10 antisense ODN are resistant against i.d. infection of MRSA. M1Mϕ appearing in the infection site tissues of burned mice that are treated with IL-10 antisense ODN may play a role on abscess formation and inhibiting sepsis caused by MRSA i.d. infection.
Discrimination of membrane antigen affinity by B cells requires dominance of kinetic proofreading over serial triggering (112.10)
Authors: Tsourkas Philippos , Liu Wanli , Das Somkanya , Pierce Susan , Raychaudhuri Subhadip 
Published: January 2023

Abstract
B cells receptor (BCR) signaling in response to membrane-bound antigen increases with antigen affinity, a process known as affinity discrimination. However, if BCR molecules become signaling-capable immediately upon binding antigen, the decrease in serial engagement as affinity increases results in weaker signaling as affinity increases. We use computational modeling to show that a threshold time (kinetic proofreading) for antigen to stay bound to BCR before the latter becomes signaling-capable is needed to overcome the decrease in serial engagement with increasing antigen affinity, and replicate the monotonic increase in B cell signaling with affinity observed in experiments. This finding matches well with the experimentally observed timescale (~20 seconds) of antigen-mediated conformational changes to BCR that lead to Src-family kinase recruitment. The physical basis of the threshold time of antigen binding may lie in the formation timescale of BCR dimers. The latter decreases with increasing affinity, resulting in shorter threshold antigen binding times as affinity increases. Such an affinity-dependent kinetic proofreading requirement results in affinity discrimination very similar to that observed in experiments. We also develop a measure to characterize affinity discrimination in a quantitative manner. B cell affinity discrimination is critical to the process of affinity maturation, and thus our results have important implications in applications such as vaccine design.
ERK activation is required for PGE2-induced MMP-9 production in bone-morrow derived DCs (112.23)
Authors: Yen Jui-Hung , Kocieda Virginia , Ganea Doina 
Published: January 2023

Abstract
Dendritic Cells (DCs) play an important role in the initiation of the immune response. Immature DCs take up antigen, mature and migrate to the regional lymph nodes where they activate antigen-specific naïve T cells. Prostaglandins such as PGE2 are essential for the migration of DCs to lymph nodes. Matrix metalloproteinases (MMPs) play an essential role in DC migration, through the degradation of extracellular matrix (ECM) and basement membranes. MMP2, and particularly MMP9, are essential for dendritic cell migration in vitro, as well as in vivo to the draining lymph nodes. In this study, we show that PGE2-induced MMP9 expression is mediated through the EP2/EP4-cAMP-PKA/PI3K-ERK signaling pathway. EP2 and EP4 receptors as well as cAMP analog mimic the effect of PGE2 to upregulate MMP-9. In the presence of PKA, PI3K or ERK inhibitors, PGE2-induced MMP-9 production is abolished. Furthermore, PGE2-induced c-Fos expression mediated through ERK activation is required for the binding of AP-1 transcription factor to the MMP-9 promoter. In the presence of ERK inhibitors, PGE2-induced AP-1 binding is abrogated. Our results describe a new molecular mechanism for the effect of PGE on MMP-9 production in DCs, which could lead to a new therapeutic approach by using ERK inhibitors to regulate DC migration.
Mouse CD99 interact with adhesion molecule CD99L2 (112.27)
Authors: Lee Hye-Yeong , Ma Min-jung , Choi Eun Young 
Published: January 2023

Abstract
CD99 and CD99-like-2 protein (CD99L2) are membrane glycoprotein leukocyte antigens that do not belong to any known protein family. They are expressed on leukocytes and at the endothelial cell junction, and participate in leukocyte diapedesis process. Based upon the similarity in expression pattern, we tested whether they would interact each other, and show here that the interaction of the two molecules is mediated via cytoplasmic domain of both molecules. In bimolecular fluorescence complementation (BiFC) assay, co-expression of CD99 and CD99L2 emitted fluorescence suggesting the interaction between the two molecules and heterodimer formation. The interaction was further confirmed by fluorescence resonance energy transfer (FRET) assay. Previously, we have reported that CD99 forms CD99-CD99 homodimer. Unlike CD99, CD99L2 does not form homodimer, rather it binds to CD99 and forms CD99-CD99L2 heterodimer. CD99-CD99 homodimer and CD99-CD99L2 heterodimer exist in a cell without competition. With cytoplasmic domain mutant of CD99, CD99L2 could not form heterodimer, indicating two molecules join through cytoplasmic domain. In reverse case when cytoplasmic domain mutant of CD99L2 was used for BiFC, CD99 could not form heterodimer. The interaction between CD99 and CD99L2 was verified by co-immunoprecipitation using mouse spleen cells. Our results reveal that mouse CD99 interact with CD99L2 and form heterodimer through its cytoplasmic domain.
Autocrine function of CD8+ T cells by modulation of CCR5 expression (112.3)
Authors: Carreno Roberto 
Published: January 2023

Abstract
Although we have shown CCR5+ T cells are known to play a role in the pathogenesis of graft-versus-host disease (GVHD), the detailed biofunction of CCR5 remains uncertain. Recently we have shown that CCR5+CD8+ T cells exhibit an effector function due to the secretion of cytolytic cytokines TNF-α and INF-γ. In addition, CCR5+CD8+ T cells have been shown to proliferate upon DC stimulation and also secrete the proliferative cytokine IL-2. Thus CCR5+CD8+ T cells maintain a polyfunctional state including both effector and memory. Moreover, in effector CD8+ T cells (CD45RA+CD27- [M3]) there is an upregulation of MIP1-β, the ligand for CCR5. These works indicate that CCR5 plays a role in memory/effector phenotype and function. Here we use CCR5 to further define memory and effector CD8+ T cells, and correlate this to their function. Using FACs we characterized central (CD45RA-CD27+ [M1]) and effector (CD45RA-CD27- [M2]) memory CD8+ T cells have high CCR5 expression. We also observed a decline in CCR5 expression in M3 CD8+ T cells. As a result, we propose that CD8+ T cells utilize autocrine signaling by promoting lymphocyte migration and infiltration to sites of inflammation by modulating the expression of CCR5 and its ligands. Further understanding of this autocrine type signal may help understand the role of CCR5 in GVHD pathogenesis.
Association of elevated expression of Toll-like receptor-2 in carotid endarterectomy specimens with the severity of atherosclerotic disease (112.9)
Authors: Tan Xiaoyu , Cherian Greoge , Cherian Rachel , Stechschulte Daniel , Dileepan Kottarappat 
Published: January 2023

Abstract
Significant interest is now focused on the role of Toll-like receptors (TLR) and cyclooxygenases (COX) in vascular disease. In this study we determined whether TLRs and COX gene expression in carotid atherosclerotic lesions is associated with clinical manifestations of the disease. Carotid endarterectomy specimens collected during surgery in nine patients with clinically significant carotid artery occlusive disease were analyzed for the expression of TLR2, TLR4, COX1 and COX2 mRNA by real-time PCR. Immunohistochemical evaluations of the specimens were carried out to quantify the presence of mast cells, lymphocytes, and macrophages using CD117, CD3, and CD68 antibodies. The level of expression of specific genes and density of the inflammatory cell population in the plaques were compared with the characteristics and location of plaques, and the severity and distribution of the clinical disease. The analysis of the expression of TLR2 with the angiographic location of the plaque revealed that the plaques expressing the higher levels of TLR2 were located at or near the carotid bifurcation whereas the plaques with the lowest expression of TLR2 were located distal to the carotid bifurcation. The plaques with the higher levels of TLR2 mRNA also showed relatively increased numbers of mast cells, T lymphocytes, and macrophages. The magnitude of TLR2 expression in the plaque appears to be associated with the severity of the atherosclerotic disease.
CD180 (RP105) intrinsically activates B cells to produce polyclonal Ig and integrates diverse TLR signals to modify cell fate (113.15)
Authors: Chaplin Jay , Kasahara Shinji , Clark Edward , Ledbetter Jeffrey 
Published: January 2023

Abstract
CD180 is homologous to TLR4 and regulates TLR4 signaling, yet its function is disputed. In this work, injection of anti-CD180 mAb into mice induced rapid Ig production of all classes and subclasses except IgA and IgG2b, with up to 50-fold serum increases of IgG1 and IgG3. IgG production after anti-CD180 injection was not due to reactivation of memory B cells and was retained in T cell-deficient (TCR KO) and MyD88 KO mice. Expansion of splenic B cells occurred in WT and TCR KO mice whereas T cell expansion occurred in WT but not in B cell-deficient (μMT) mice, indicating a direct role for B cells but not T cells in CD180 stimulation in vivo. Combinations of anti-CD180 with various TLR ligands biased B cell fate as co-injection diminished Ig production but purified B cells exhibited synergistic proliferation. Anti-CD180 had no effect on cytokine production from B cells but increased IL-6, IL-10, and TNF-α production in combination with LPS or CpG. Thus CD180 stimulation induces intrinsic B cell proliferation and differentiation, causing rapid increases in IgG, and integrates MyD88-dependent TLR signals to regulate proliferation, cytokine production, and differentiation.
Deletion of factors in the Id3 pathway results in a dramatic phenotype where Vγ1Vδ6.3 (γδ NKT) T cells become the dominant T lymphocyte (113.2)
Authors: Alonzo Eric , Sant'Angelo Derek 
Published: January 2023

Abstract
Recent work from our lab and others has shown a significant expansion of γδ NKT cells in mice that lack the HLH-protein Id3. In contrast to WT mice, nearly all Id3-deficient γδ NKT cells express high levels of the innate T cell determinant, PLZF and produce copious amounts of IL-4 upon activation. We have been exploring the transcriptional network that controls the development of γδ NKT cells. In an effort to identify additional factors that play a key role in expansion of γδ NKT cells, we have undertaken a candidate approach to evaluate genes that function in the Id3 pathway. To do this we are studying single and double gene deficient mice to determine the interrelation of different transcription factors. These studies have led to the discovery of a gene combination that results in a dramatic alteration of thymopoiesis. In these mice, while thymic cellularity was not affected, the frequency of γδ T cells versus αβ T cells was reversed. γδ T cells were found to be the dominant lymphocyte, representing ~65% of the cells. Furthermore, nearly all of the γδ T cells were now found to express PLZF. This dramatic phenotype appears to be T cell-specific, as both myeloid and lymphoid lineages are present. Analysis of thymic progenitors revealed major alterations in DN2-DN4 stages of development. Our findings uncover a fundamental regulatory pathway that favors the development of γδ NKT cells, while restricting the maturation of pro-T cells into the conventional T cell lineage.
Acute stress-induced thymic atrophy is mediated by intrathymic inflammatory and wound healing responses (113.25)
Authors: Billard Matthew , Gruver Amanda , Sempowski Gregory 
Published: January 2023

Abstract
Productive thymopoiesis is essential for a robust and healthy immune system. Thymus unfortunately is acutely sensitive to stress resulting in involution and decreased T cell production. Thymic involution is a complication of many clinical settings, including infection, malnutrition, and irradiation or immunosuppressive therapies. Little is known, however, about intrathymic mechanisms that may actively contribute to thymus atrophy or initiate thymic recovery following stress events. In this work, phenotypic, histologic and transcriptome/pathway analysis was performed to identify putative intrathymic mechanisms that drive endotoxemia-induced involution of murine thymic tissue. Thymus atrophy in this murine model was confirmed by down-regulation of genes involved in T cell development, cell activation, and cell cycle progression, correlating with observed phenotypic and histologic thymus involution. Significant gene changes support the hypothesis that multiple key intrathymic pathways are differentially activated during stress-induced thymic involution. These included direct activation of thymus tissue by LPS through TLR signaling, local expression of inflammatory cytokines, inhibition of T cell signaling, and induction of wound healing/tissue remodeling. Taken together, these observations demonstrated that both systemic and direct intrathymic responses to endotoxin challenge concurrently contribute to thymic involution during endotoxemia.