bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mice deficient in Complement Receptor 1 produce normal to enhanced quantities of antigen specific immunoglobulin in response to naïve immune challenge (172.14)
Authors: Donius Luke , Handy Jennifer , Weis Janis , Weis John 
Published: January 2023

Abstract
Complement receptors (Cr)1 and 2 have been established to have important roles in control of complement activation in the immune microenvironment and B cell responses. Previous work investigating the effects of these receptors in vivo has relied on a double knockout mouse line in which each of these receptors was deleted. However, two drawbacks of this mouse must be acknowledged: 1) that both receptors are deleted and roles of each must be inferred, and 2) that the essential B cell signaling protein CD19 is expressed at a significantly higher level than WT cells. We have created a Cr1 deficient line of mice (expressing Cr2) by homologous gene targeting that expresses normal levels of Cr2 gene products on B cells. This animal has normal ratios of bone marrow, B2 cell, and B1 B cell populations. Cr2 negative marrow B cells express the same amount of surface CD19 as WT but mature Cr2-expressing B cells express less. These Cr2-only mice generate normal to slightly enhanced antibody titers to T-dependent and T-independent (both 1 and 2 antigens.) In conclusion Cr1 is not critical for development and maintenance of B cell populations nor for the generation of antibodies to naïve antigens. This mouse, along with the Cr1/2KO, will be a critical tool in delineating Cr1 specific immune response effects.
Lamprey variable lymphocyte receptors mediate complement-dependent cytotoxicity (172.20)
Authors: Yu Xiaoli , Wu Fenfang , Chen Liyong , Liu Xin , Wang Huaying , Su Peng , Han Yinglun , Li Mo , Feng Bo , Qiao Xu , Zhao Jing , Ma Ning , Liang Meng , Liu Huijie , Zheng Zhen , Li Qingwei 
Published: January 2023

Abstract
An alternative adaptive immune system is present in the most basal vertebrates, lampreys and hagfish, the only surviving jawless vertebrates. These eel-like fish use leucine-rich-repeat (LRR)-based receptors, called variable lymphocyte receptors (VLRs), for antigen recognition instead of the immunoglobulin (Ig)-based receptors used in jawed vertebrates. We showed that in Japanese lamprey (Lampetra japonica), antigen-specific VLRB antibodies interact with C1q-like and C3 proteins to mediate complement-dependent cytotoxicity for bacteria and tumor cells. The immune based lysis involved deposition of VLRB and C1q-like protein complex on the surface of target cells, activation of C3, and ultimate disruption of cell wall integrity. The demonstration of functional interaction between VLRB and complement components in lamprey provides evidence for the emergence of cooperative innate and adaptive immune responses at a pivotal point in vertebrate evolution, before or in parallel with the evolution of Ig-based antibodies and the classical complement activation pathway.
Complement mediates membrane rearrangements and lipogenesis (172.21)
Authors: Fattakhova Gulnar , Cascalho Marilia , Platt Jeffrey 
Published: January 2023

Abstract
The complement is a major effector part in innate immunity. Its main biological function is to recognize foreign particles and macromolecules, and to promote their elimination either by opsonization or lysis. In order to avoid potential risk of host cells lysis, an efficient system of regulatory proteins is involved. Additional protective mechanisms include removal of membrane attacking complex by both vesicle shedding and internalization followed by exocytosis. The aim of research is to investigate alterations in membrane biogenesis and lipid biosynthesis in cells targeted by complement. Treatment of mouse NIH 3T3 fibroblasts with complement induces early and late phase responses. The early phase response is characterized by increase in intracellular Ca2+, alterations in phospholipid packing order and plasma membrane asymmetry, as well as translocation of lysosomal vesicles to the cell surface followed by activation of acid sphingomyelinase and ceramide production. We demonstrated that prestimulation of cells with sublityc complement doses followed by treatment with lytic concentrations substantially reduces levels of both plasma membrane-associated acid sphingomyelinase and ceramide, and protects cells from lysis. During late phase response we observed alterations in mRNA levels of several SREBP-targeted genes and lipid biosynthesis. Taken together, our data link regulated lipid dynamics and metabolism to acquired cell resistance to complement attack (accommodation).
Chaperone like activity of HMGB1 reduces the formation of polyglutamine aggregates (172.24)
Authors: Min Hyun Jin , Shin Jeon-Soo 
Published: January 2023

Abstract
High mobility group box protein 1 (HMGB1), which functions as a DNA chaperone, has recently been shown to play intracytoplasmic roles as an autophagy regulator and a nucleic acid sensing molecule. In order to identify a new role for HMGB1 in cytoplasm, we studied the chaperone-like activity of HMGB1, since HMGB1 is a ubiquitous protein. Using light scattering after heat and chemical stress, we found that HMGB1 inhibited the chemically-induced aggregation of insulin and lysozyme as well as the heat-induced aggregation of citrate synthase, suggesting that HMGB1 has a chaperone-like activity in vitro. Also, HMGB1 restored the heat-induced suppression of luciferase activity as a reporter protein in O23 hamster fibroblast cells after co-transfection of firefly luciferase and HMGB1. Next, we demonstrated that HMGB1 inhibits the aggregation and therefore toxicity of elongated polyglutamine (polyQ), one of the main causes of Huntington’s disease. HMGB1 proved to be co-localized with polyQ in the co-transfection study, while over-expression of HMGB1 or exogenous treatment of recombinant HMGB1 protein remarkably reduced polyQ-induced aggregates in SHSY5Y neuroblastoma cells, upon filter trap and immunofluorescence assay. Taken together, we show that HMGB1 demonstrates chaperone-like activity and inhibits the cytotoxicity of polyQ-mediated disease.
Macrophage chitin binding proteins in phagocytosis and M1 activation in response to chitin microparticles (172.27)
Authors: Shibata Yoshimi , Kogiso Mari , Huang Chun-Jung , Nouri-Shirazi Mahyar , Mizoguchi Emiko , Dorey C Kathleen 
Published: January 2023

Abstract
Unlike soluble or non-phagocytosable 40 - 100 µm chitin, chitin microparticles (1 - 10 μm, CMP) induce MAPK activation within 20 min in a manner dependent on TLR2, phagocytosis and a cellular cholesterol-mediated mechanism, followed by a typical expression of M1 phenotypes. Based on the previous studies, we hypothesized that trans-membrane signal through surface TLR2 ligation is insufficient and additional chitin binding proteins (CBP) involved in the early CMP phagocytosis are obligatory for M1 activation; CBP are distinct between M1/M2 macrophages (MΦ). Peritoneal M1, M2 and control MΦ were isolated from mice 1 - 2 days after ip administration of CMP, 40 - 100 µm chitin, and saline, respectively. These MΦ were cultured with CMP; CMP with CBP were isolated shortly after phagocytosis, stained by specific antibodies and analyzed by FACS. Several CBP on CMP, including TLR2, CD206, CD62L, CHI3L1, and LAMP-1, exhibited distinct patterns between M1/M2 activations. For example, TLR2, fully engaged at the early phagocytosis, was increased in M1 and reduced in M2 relative to unstimulated MΦ. Our results suggest that, although exact mechanisms of CBP-mediated M1 activation still remain to be elucidated, the profiles of selected CBP that bind to CMP during phagocytosis are associated with the polarization of MΦ. These profiles offer a potential tool as diagnostic/prognostic biomarkers to monitor chronic inflammatory diseases and the efficacy of immunotherapy.
Direct regulation of Interleukin-6 expression by Notch signaling in macrophages (172.36)
Authors: Wongchana Wipawee , Osborne Barbara , Palaga Tanapat 
Published: January 2023

Abstract
Notch signaling is required for immune cell development. The intracellular domain of Notch (NIC) induces expression of target genes by binding to CSL, a DNA-binding protein. In macrophages, Notch signaling regulates inflammatory cytokines TNFα and IL-6. IL-6 is secreted by various cell types, including activated macrophages and functions in immune regulation and inflammation. The consensus CSL binding site in the promoter region of IL-6 was shown to negatively regulate IL-6 mRNA, but the role of Notch signaling in IL-6 expression in macrophages remained unclear. We investigated the role of Notch signaling in the regulation of the IL-6 transcript in macrophages. Our results showed that bone marrow derived macrophages (BMMΦ) and RAW264.7 cell lines activated with lipopolysaccharide (LPS) and interferon gamma (IFNγ) induced IL-6 mRNA expression and upregulation of Notch1. Inhibition of Notch signaling in BMMΦ using a gamma secretase inhibitor partially decreases the level of IL-6 mRNA and protein. In contrast, RAW264.7, transiently transfected with constitutively active Notch1, displayed a dramatic increase in IL-6 mRNA. Inhibition of NF-κB completely abrogated IL-6 mRNA expression induced by the over expression of NIC. Chromatin immunoprecipitation assay demonstrated that Notch1 binds to the promoter of IL-6 in activated RAW264.7 but not in unstimulated cells. These results strongly suggest that Notch1 positively regulates IL-6 expression via NF-κB in activated macrophages.
TGF-b/TGF-bRII-SOCS3 interactions signifies the tolerogenic pathway(s) for down-regulating CD11c+dendritic cells-mediated osteoclastogenesis & bone loss (172.38)
Authors: Teng Andy Yen-Tung , Liu Yen Chun 
Published: January 2023

Abstract
Inflammation-induced bone-loss is linked to elevated osteoclasts(OC). Dendritic cells are antigen-presenting-cells involved in T-cell-immunity and can act as OC-precursors. We reported the development of functional-OC from CD11c+DC in-response-to RANKL plus microbes(Aa)-stimuli [DDOC]. Aims: to: i) identify molecules for regulating DDOC; ii) study the signals of modulating tolerogenic pathway that down-regulates DDOC-mediated osteoclastogenesis. Methods: DDOC are generated using co-cultures of CD11c+DC with 100ng/ml-RANKL (or-CD4+T/Foxp3+T-reg) & 10g/ml-Aa-sonicate. The results showed that: i) micro-arrays & functional analyses have identified key regulator(s) involved in DDOC-development, with a positive correlation to TGF-bRII. Neutralization of TGF-b abolishes DDOC development, based on TRAP & resorptive-pits (p&lt;0.02); ii) transfection of dominant-negative SOCS3-cDNA into CD11c+DDOC via lenti-viral-vector and adoptive-transfer yield a significant inhibition of osteoclatogenesis & bone-resorption (p&lt;0.05); iii) addition of Foxp3+CD4+T-reg into CD11c+DC plus Aa-Ag co-cultures results in reduced SOCS3-&-TRAP expressions & lowered-osteoclastogenesis in bone-resorption assays & NOD/SCID-mice (p&lt;0.05). Conclusion: i) TGF-b/TGF-bR-SOCS3 interaction is involved in modulating the tolerogenic pathway(s) of down-regulating DDOC activity, ii) Foxp3+T-reg may be an integral player underlying inflammation-induced osteoclastogenesis and bone-loss through DC-T-cell interactions.
Endothelial Cells Bind Immune Complexes in the Kidney (172.39)
Authors: Wenderfer Scott 
Published: January 2023

Abstract
Inflammatory kidney diseases often involve the deposition of antibodies (Ab) or antibody-antigen immune complexes (IC). The glomerulus is a highly evolved vascular system which filters blood to produce urine. Ab-mediated glomerular injury can lead to bleeding into the urine, protein wasting, hypertension, decreased urine output, edema, and kidney failure. However, the mechanisms by which IC deposit and the means by which they are cleared remain unclear. In order to model endothelium-Ab interactions, kidney endothelial cells (KEnCs) were isolated from C57BL/6 mice and cultured in vitro. Immuno-phenotyping was performed to assess expression of CD31, VEGF receptors, von Willebrand factor (vWF), and alpha smooth muscle actin. CD31+ vWF+ VEGF-R2+ KEnCs displayed a phenotype in culture similar to other microvascular cells. Monomeric and heat aggregated Ig (HA-IgG) and peroxidase-antiperoxidase IC (PAP-IC) were used for binding and stimulation assays. Effects of stimulation of KEnCs were identified using PCR arrays and multiplex cytokine analysis. KEnCs stained brightly with HA-IgG and PAP-IC, but only dimly with monomeric IgG. KEnCs responded to IC with robust chemokine and cytokine responses in a dose dependent manner. Competitive binding studies mapped the interaction to the Fc-portion of IgG, but suggest that the KEnCs bind independently of classical Fc-receptors. Results support an important role for KEnCs in the pathogenesis of IC kidney diseases.
MiR-106b family regulates autophagy via targeting ATG16L1 (172.41)
Authors: Lu Changming , Xu Zhi-xiang 
Published: January 2023

Abstract
Autophagy is a lysosomal degradation pathway that serves an adaptive role to protect organisms against diverse pathologies. Dysfunction of autophagy is implicated in the development of numerous human diseases. However, regulation of autophagy at the molecular level remains largely unknown. In this study, we demonstrated that microRNA (miRNA) 106b family members were a critical regulator of autophagosome formation as well as autophagy-mediated functions. We found that during the differentiation of human monocytes into dendritic cells, expression of autophagy-related genes (ATGs) and autophagosome were substantially induced, whereas a number of miRNAs were down-regulated. Silencing of dicer resulted in the increased expression of ATGs and autophagosome formation. Bioinformatic search uncovered that miR-106 family members were highly potential regulators for ATG proteins. Indeed, our experimental investigations demonstrated that miR-106b and miRNA-93 mimics remarkably decreased autophagosome induction under physiological and stress conditions. In contrast, administration of antagonists of miR-106b and miRNA-93 sensitized autophagy formation. Immunoblot and luciferase report assays demonstrated that ATG16L1 was a major component targeting by miRNA-106b family members. Over-expression of ATG16L1 partially rescued miRNA-106b transduction-induced autophagy suppression. Previous reports indicated that ATG16L1 suppression and autophagy deficit are potential factors that lead to inefficient elimination of bacteria in colon epithelial cells during the development of Crohn’s disease. Consistent with these reports and our aforementioned findings, miRNA-106b mimic dramatically reduced the clearance of LF82 bacteria in HeLa epithelial cells. Thus, we have identified a family of miRNA members (miR-106b) as a main inhibitor of autophagy, which may play a role in the eradication of intracellular bacteria and development of Crohn’s disease.
Potential contribution of the interaction between spinal cord-infiltrating Th1 cells and astrocytes to the development of nerve injury induced neuropathic pain (173.10)
Authors: Cao Ling , Draleau Kyle , Maddula Swathi 
Published: January 2023

Abstract
We previously demonstrated that CD4+ T cells contribute to the maintenance of L5 spinal nerve transection (L5Tx)-induced neuropathic pain in BALB/c mice, and T-Bet+ Th1 cells were found to be the dominant subtype of CD4+ T cells within the lumbar spinal cord following L5Tx. To investigate the specific downstream responses mediated by these infiltrating Th1 cells, we examined the cytokine expression of the spinal cord-infiltrating CD4+ T cells via flow cytometry. At day 7 post-L5Tx, the peak time for detecting the lumbar infiltrating CD4+ T cells, there were trends indicating increases in the numbers of IFNγ+ and TNFα+ lumbar spinal cord-infiltrating CD4+ T cells in the L5Tx group compared to the sham group. Further, we examined the expression of glial fibrillary acidic protein (GFAP), an indication of the level of astrocytic activation, in the L5 spinal cord via immunohistochemistry. As expected, in wild type BALB/c mice, L5Tx induced an increase in GFAP expression in the ipsilateral side of the spinal cord compared to sham-operated mice 7 days post-surgery. This increase appeared to be reduced in CD4 knockout mice post-L5Tx. Altogether our data suggest the involvement of the Th1 cytokines IFNγ and TNFα in the maintenance of L5Tx-induced neuropathic pain, which may have potential role in regulating spinal cord astrocyte activation.
Microfluidic Assay for Neutrophil Chemotaxis Towards Chemoattractant Gradients (173.13)
Authors: Jones Caroline 
Published: January 2023

Abstract
Chemoattractants represent an essential group of molecular guidance signals that regulate the transit of leukocytes out of the mainstream of blood and into tissues at sites of inflammation. In this project, we developed a microfluidic device that is simple to operate and allows for precise and robust measurements of neutrophil chemotaxis speed and directionality towards precisely defined chemoattractant gradients at single-cell resolution. These devices consist of non-planar, enclosed microenvironments of increasing complexity, from straight channels, to channels with posts and simple mazes. The cross-section of the channels (3 × 6 μm) is smaller than the human neutrophils, so that cells are mechanically confined and allowed to move only along the axis of the channels toward a gradient of chemoattractactant. This assay is currently being applied in a clinical setting to characterize the immune function in burn patients during treatment. Comparing samples from healthy donors to burn patients revealed impaired directional migration speed starting as early as 24 hours after burn injury, reaching a minimum at 72 hours, correlated to the size of the burn injury and potentially serving as an early indicator for concurrent infections. Further characterization of neutrophil chemotaxis using this new assay may have important diagnostic implications not only for burn patients, but also for patients afflicted by other diseases that compromise neutrophil functions.
Dynamin 2 regulates S1P1 signalling and T cell migration (173.15)
Authors: Willinger Tim , Ferguson Shawn , De Camilli Pietro , Flavell Richard 
Published: January 2023

Abstract
Sphingosine 1-phosphate (S1P) is essential for T cell egress from lymphoid tissues by acting on its receptor S1P1. S1P agonism on S1P receptors is now used to treat autoimmune diseases by sequestering lymphocytes in lymphoid organs. However, not much is known about molecules that mediate T cell egress downstream of S1P1. Dynamin 2 is a large GTPase that is required for vesicle scission during endocytosis. Mice with a T cell-specific deletion of dynamin 2 had profound lymphopenia and an accumulation of mature thymocytes. Intrathymic dye injection revealed that thymic egress was inhibited in the absence of dynamin 2. Dynamin 2 regulated T cell migration downstream of S1P-S1P1 since dynamin 2-deficient thymocytes did not migrate in response to S1P in vitro. This was not due to a general defect in cell motility because T cells lacking dynamin 2 had a normal migratory response to CCL21. Dynamin 2 was required for ligand-induced S1P1 downmodulation, consistent with its known function in receptor-mediated endocytosis. In addition, dynamin 2 knockout thymocytes co-expressed S1P1 and CD69 on the cell surface demonstrating impaired S1P1 signalling in vivo. Surprisingly, the critical role of dynamin 2 in S1P1 signalling was independent of receptor internalization. Instead, dynamin 2-deficient T cells showed faster desensitization of S1P1 after agonist stimulation in vitro. Our data reveal an unexpected role for dynamin 2 in promoting T cell egress by regulating S1P1 signalling.
KLF2 and S1Pr1: Aiding in memory T cell trafficking and retention in non-lymphoid tissue (173.16)
Authors: Skon Cara , Lee June-Yong , Jameson Stephen 
Published: January 2023

Abstract
Memory T cells, unlike naïve T cells, have the capacity to traffic and be retained in non-lymphoid tissue (NLT). Because most infections originate in NLT and early detection of pathogens can lead to a more effective immune response, identifying factors contributing to memory T cell residence in NLT is an important aspect of memory T cell potential. The transcription factor kruppel-like factor 2 (KLF2) controls naive T cell trafficking (primarily through inducing sphingosine 1 phosphate receptor 1 (S1Pr1)), yet KLF2 expression in memory T cells has not been extensively addressed. Using eGFP-KLF2 reporter mice we report that almost all CD8+ memory T cells in lymphoid tissue express KLF2, yet the vast majority of CD8+ memory T cells in NLT (including small intestine, salivary glands, kidney and brain) are KLF2lo. Forced-expression of the KLF2 target gene S1Pr1, using retroviral transduction, caused a decrease in memory CD8+ T cells in NLT versus spleen, confirming a functional consequence of S1Pr1 down-regulation. We also showed that inflammatory cytokine milieu causes a decrease in KLF2 protein in the absence of antigen. Our data suggest that local cues from the non-lymphoid environment, including cytokines, cause a decrease in KLF2 and S1Pr1 leading to a cessation in CD8+ memory T cell trafficking that aids in memory T cell retention in NLT.
Tissue exit - a novel control point in the accumulation of antigen-specific CD8 T cells in the influenza A virus-infected lung (173.20)
Authors: Jennrich Silke , Lee Michael , Lynn Rachel , Dewberry Kristofer , Debes Gudrun 
Published: January 2023

Abstract
Memory/effector T cells efficiently migrate into extralymphoid tissues and sites of infection, providing immunosurveillance and a first line of defense against invading pathogens. Even though it is a potential means to regulate the size, quality, and duration of a tissue infiltrate, T cell egress from infected tissues is poorly understood. Using a mouse model of influenza A virus infection, we found that CD8 effector T cells egressed from the infected lung in a CCR7-dependent manner. In contrast, following antigen-recognition, effector CD8 T cell egress decreased and CCR7 function was reduced in vivo and in vitro, indicating that the exit of CD8 T cells from infected tissues is tightly regulated. Our data suggest that the regulation of T cell egress is a mechanism to retain antigen-specific effectors at the site of infection to promote viral clearance, while decreasing the numbers of bystander T cells and preventing overt inflammation.
A cyclic AMP signal participates in T cell tissue retention by selectively blocking tissue egress. (173.21)
Authors: Lees Jason , Li Xin 
Published: January 2023

Abstract
Accumulation of effector T cells within target tissue(s) is a critical step in the induction of tissue directed immunity. Cyclic AMP (cAMP) is a known regulator of T cell activity and function. In this study we examined the effects of cAMP on T cell trafficking and retention. We found that increasing levels of cellular cAMP, either through direct uptake of a cAMP analogue or induced by treatment with nonanedioic acid, decreased the ability of T cells in vitro to traffic to CXCL12 and to suboptimal, but not optimal, doses of CCL21. Levels of cAMP that decreased chemotaxis had no effect on T cell viability, activation, proliferation, or cytokine production. To determine the role of cAMP on T cell retention, we examined the capacity of transferred T cells to traffic from the foot pad to the draining lymph node. Increasing T cell cAMP levels significantly decreased T cell movement to the lymph node but not the capacity of cells to enter lymph nodes via high endothelial venules. We suggest that cAMP can act to increase T cell tissue retention by blocking responses to suboptimal levels of chemokines.