bluuebunny/crossref_metadata_2025_split huggingface dataset:
‘Plasma monocyte-derived extracellular vesicles are associated with neurocognitive impairment in HIV infected individuals.’
Authors: de Menezes Erika Grasiela Marques , Inglis Heather C. , D’Antoni Michelle , Shikuma Cecilia M. , Ndhlovu Lishomwa C. , Norris Philip 
Published: January 2023

Abstract
Monocytes contribute to the neuropathogenesis of HIV-related cognitive impairment (CI) as they enter the central nervous system and initiate a cascade of neuroinflammatory events. Extracellular vesicles (EVs) are potent vehicles of intercellular signaling and may reflect the state of cells in the brain under healthy and pathological conditions. We measured EVs in plasma from 79 HIV-infected adults enrolled in the Hawaii Aging with HIV Cohort, classified as having either normal cognition (n=16) or CI based on meeting criteria for HIV-Associated Dementia (n=25) or Mild Cognitive Motor Disorder (n=38). We tested whether EV phenotypes differed between those with and without CI. EVs expressing monocyte-associated markers (EVs/μl) were significantly higher in HIV+ donors with CI compared to those without CI: CD14 (819 vs. 118.5, p=0.017), CD4 (261 vs. 45, p=0.001), CD16 (898 vs. 215, p&lt;0.0001), CD163 (1660 vs. 55, p=0.010), and CCR5 (202 vs. 43.5, p=0.009). In addition, increased levels of EVs expressing neural markers were found in HIV+ adults with CI compared to those without CI: MAP2 (531 vs. 103.5, p=0.013), GFAP (1810 vs. 775, p=0.036), and CD200 (1216 vs. 36.5, p=0.005). There was no significant difference between the two groups in age, CD4 count, viral load, and, EVs expressing CD3, CD41a, CD62p, CD63, CCR2 and CD11b. Levels of CD16-expressing EVs negatively correlated with the degree of CI using a composite of 8 neuropsychological (NP) tests to compute a summary NPZ-8 score (r=−0.224, p=0.046). Our findings reveal that EVs expressing monocyte activation and neural damage markers correlate with CI in HIV+ adults. These techniques reveal novel circulating biomarkers of CI, and neuronal pathogenesis markers utilizing peripheral blood.
Enhanced immunoglobulin somatic hypermutation in human APOBEC3 transgenic mice
Authors: Jones Sean , Guo Kejun , Barrett Brad , Santiago Mario L. 
Published: January 2023

Abstract
Eliciting a broadly neutralizing antibody (bNAb) response remains a major hurdle in HIV-1 vaccine development, as a majority of these bNAbs are heavily mutated. Immunoglobulin (Ig) somatic hypermutation (SHM) is primarily driven by AID. However, our previous data in Friend retrovirus (FV) infected mice revealed that an evolutionarily-related deaminase, mouse APOBEC3 (mA3), may complement AID in mutating virus-specific Abs (Halemano et al, PNAS 2014). AID mutates primarily in an RC context (R=purine), whereas the A3s mutate in a YC context (Y=pyrimidine). Of note, mice encode only one A3 gene, but humans encode seven. To date, it remains unclear if the human APOBEC3 (hA3) deaminases can directly mutate Ig genes. To study the role of hA3s in Ig SHM, bacterial artificial chromosome (BAC)-transgenic mice expressing the 7 hA3s (hA3-Tg) were crossed onto an mA3 KO background. All hA3s were expressed in hA3-Tg B cells during FV infection. hA3-Tg (n=13) and mA3 KO (n=12) mice were vaccinated with live-attenuated FV and analyzed at 28 d. Surprisingly, the hA3-Tg mice have an impaired plasma NAb response (p=0.0003) despite similar virus-specific binding IgG titers (p&gt;0.1). Illumina sequencing of bone marrow IgG transcripts showed increased deamination mutations (C to T or G to A) in hA3-Tg versus mA3 KO mice (p=0.0043). Interestingly, this increase was accounted for by AID-type (RC), but not hA3-type (YC), mutations. Our findings reveal that hA3 can increase IgG SHM rates following vaccination, but the initial impact on the vaccine NAb response may be detrimental. Studies are ongoing to monitor the development of virus-specific antibodies in hA3-Tg mice, as harmful mutations may be more prominent during early time points post-vaccination.
Investigation of the immunological mechanisms underlying the attenuation of vaccinia virus lacking host-range factor C7
Authors: Deng Liang , Yang Ning , Dai Peihong , Wang Yi , Wang Weiyi , Merghoub Taha , Wolchok Jedd D. , Shuman Stewart 
Published: January 2023

Abstract
Vaccinia virus C7 protein is an important host-range factor for the vaccinia virus life cycle in mammalian cells. Type I IFN plays an important role in host defense of viral infection. However, the role of C7 in immune modulation of the IFN pathway is unclear. Here, we find that C7 functions as a dual inhibitor of IFN production and IFN signaling by interfering with the activation of two transcription factors IRF3 and STAT2. Furthermore, vaccinia virus with deletion of C7 (VACVΔC7L) is non-pathogenic in wild-type C57BL/6J mice in an intranasal infection model. We elucidated the immunological mechanisms underlying the attenuation phenotype of VACVΔC7L, which can be summarized as follows: (i) intranasal infection of VACVΔC7L triggers the production of interferons (IFNs), proinflammatory cytokines and chemokines in bronchoalveolar lavage fluid (BAL), whereas WT VACV does not; (ii) intranasal infection of VACVΔC7L results in the recruitment of dendritic cells, inflammatory monocytes, neutrophils, CD8+, and CD4+ in the BAL, whereas WT VACV does not; and (iii) infection of primary type II lung alveolar epithelial cells (AECs) with VACVΔC7L induces IFN, proinflammatory cytokine and chemokine gene expression and protein secretion, whereas WT VACV does not. However, VACVΔC7L dramatically gained virulence in STAT2 or IFNAR1-deficient mice, with increased titers in the lungs and systemic dissemination of the viruses to the blood and distant organs. Taken together, our results provide evidence that vaccinia C7 is a key virulence factor that antagonize both IFN production and signaling and IFNAR1 and STAT2 are critical in restricting vaccinia viral replication in the lung alveolar epithelial cells.
Single cell analysis revealed distinct B-cell subpopulations that produce or respond to type I interferon in SLE
Authors: Hamilton Jennie A. , Wu Qi , Yang PingAr , Luo Bao , Liu Shanrun , Li Jun , Hong Huixian , Mattheyseys Alexa , Chatham Winn , Hsu Hui-Chen , Mountz John D. 
Published: January 2023

Abstract
We previously showed that B-cell endogenous expression of interferon beta (IFNβ) is associated with the development of autoreactive B cells in BXD2 autoimmune mice. As among all type I IFNs, IFNβ exhibits the highest affinity to IFNAR1 and IFNAR2 and IFNβ also can stimulate IFNαs, there is a pressing need for a better understanding of the source and responses of IFNβ in systemic lupus erythematosus (SLE). Flow cytometry and super-resolution imaging analysis showed increased expression of IFNβ in B cells obtained from PBMCs of SLE patients compared to healthy controls. The importance of SLE B-cell endogenous IFNβ in regulating TLR7-mediated responses was identified when CL264 (a TLR7 ligand) plus anti-Ig-induced CD69 and survival of purified B cells were significantly and equivalently suppressed by an anti-IFNβ or an anti-IFNAR1 blocking antibody. We next carried out single cell qRT-PCR analysis to determine if B-cell endogenous IFNβ acts in an autocrine or paracrine manner to modulate IFN stimulated genes (ISGs). Hierarchical analysis revealed three prominent and consistent clusters, consisting of an IFNB+ cluster, an ISG+ cluster, and an IFNA+ cluster in naïve B cells isolated from 3 SLE patients. Cells within the IFNB+ cluster expressed higher levels of IFNB, IFNA1, IFNA8, and TLR7. Cells within the ISG+ cluster expressed higher levels of IFNAR1, IFNAR2, BAFFR, MX1, RIG1 and TLR9. Cells within the IFNA+ cluster expressed higher levels of IFNA4, IFNA10, IFNA14, IFNA16, IFNA17, and TLR3. Together, the results reveal (1) B cells as an important source of IFNβ in modulating TLR7 responses in SLE; (2) a well-orchestrated program in modulating IFNβ donor versus responder B cells; and (3) some patients may benefit from specific targeting of IFNβ.
Loss of B cell Ezrin in lupus-prone Lyn-deficient mice downregulates B cell activation and autoimmune pathology
Authors: Gupta Neetu , Pore Debasis , Parameswaran Neetha , Huang Emily , Dejanovic Dina 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is an aggressive autoimmune disorder that afflicts 1.5 million Americans. Human GWAS studies have reported a strong association between the Src family tyrosine kinase Lyn and incidence of SLE, and B cells from SLE patients exhibit decreased Lyn expression. Genetic deletion of Lyn in mice results in loss of peripheral B cell tolerance and development of human SLE-like autoimmunity, characterized by hyperactive B cell antigen receptor (BCR) signaling, autoantibody production and glomerulonephritis. However, the molecular regulators of B cell pathogenesis in Lyn-deficient mice have not been fully characterized. We have previously shown that the membrane-cytoskeleton linker protein ezrin regulates multiple aspects of B cell function through its dynamic switching between phosphorylated and dephosphorylated states. We observed that ezrin is hyperphosphorylated in Lyn-deficient B cells, suggesting that it may impact B cell activation in this context. To test this hypothesis, we generated conditional deletion of ezrin in the B cells of Lyn-deficient mice (hereafter referred to as DKO mice). Compared to Lyn-deficient mice, the DKO mice exhibited a significant decrease in splenomegaly and reduction in autoantibodies against a variety of autoantigens. Glomerulonephritis and deposition of IgG and complement factor C3 in the kidneys of DKO mice were also markedly reduced. The DKO B cells were hyporesponsive to BCR stimulation and displayed a calcium mobilization pattern reminiscent of that observed in anergic B cells. Our data demonstrate that ezrin regulates B cell hyperactivation and associated pathology in the absence of Lyn, and suggest that ezrin may be a novel molecular target in SLE.
Characterization of MMP-28 in autoimmune neurodegenerative diseases
Authors: Tokmina-Roszyk Dorota , Onwuha-Ekpete Lillian C. , Refai Mohammed , Tokmina-Lukaszewska Monika , Bothner Brian , Fields Gregg 
Published: January 2023

Abstract
Matrix metalloproteinases (MMPs) are a family of proteolytic enzymes that mediate the degradation of extracellular matrix components and are crucial for normal physiological processes; however, their dysregulation has been implicated in numerous pathologies associated with autoimmunity as well as neurological disorders. MMP-28 is the least characterized member of this family and it has been implicated as a regulator of myelination during neural development and repair. Moreover, recent studies have suggested a role for MMP-28 in determining the strength of the immune response. To unravel the complex role of MMP-28 in an autoimmune neurodegenerative disease such as Multiple Sclerosis (MS) we utilized the Experimental Autoimmune Encephalomyelitis (EAE) murine model. We compared disease progression in both WT and MMP-28 null mice and did not observe significant difference between both groups. However, weight loss and therefore disease severity was overall higher in MMP-28 null mice. Subsequently, we subjected brain samples from both WT and MMP-28 null to global protein profiling using a shotgun proteomics approach and corresponding protein profiles were compared. Our findings, provide further evidence that MMP-28 is an important factor in the autoimmune response. Moreover, in addition to the expected impact of MMP-28 on various proteins associated with myelin, we identified mitochondrial protein complexes that were upregulated in samples derived from MMP-28 null mice. These data provide evidence of a previously unknown intracellular role of MMP-28 and suggest a possible novel mechanism of immune regulation. Our data suggest that MMP-28 might be an important antitarget in autoimmune neurodegenerative diseases such as MS.
A detrimental effect of interleukin-22 on salivary gland tissue integrity and function
Authors: Yoo Bora , Zhou Jing , Yu Qing 
Published: January 2023

Abstract
Interleukin-22 (IL-22) has well-documented effects in promoting epithelial wound healing and protecting tissue damage in the intestine and lung. However, in certain inflammatory diseases such as psoriasis, IL-22 can also exert tissue-detrimental effect. Although the up-regulation of IL-22 has been reported in patients with Sjögren’s syndrome (SS), the role of IL-22 in this disease is not well-characterized. Herein, we investigated the effects of IL-22 on SS-like salivary gland disorder using both in vivo and in vitro approaches. To assess the effects of exogenous IL-22 on salivary gland tissues in vivo, IL-22 was directly injected into submandibular glands (SMGs) of 7–12 weeks old female C57BL/6 and the non-obese diabetic (NOD) mice, a model of SS disease. The salivary flow rate was decreased and the caspase-3 level was increased in the SMGs of the IL-22-treated C57BL/6 mice. IL-22 treatment also decreased the salivary flow rate and increased the level of capase-3 in the SMGs in the NOD mice. We determined the effects of endogenous IL-22 using an anti-CD3 antibody-induced acute exocrinopathy model. Anti-CD3 antibody was intraperitoneally injected into 8-week-old female C57BL/6 mice every other day for 3 days, together with a neutralizing anti-IL-22 antibody or its isotype control IgG. Neutralization of endogenous IL-22 improved saliva production and reduced caspase-3 activation in the SMGs in anti-CD3-treated C57BL/6 mice. Moreover, in vitro IL-22 treatment of a human salivary gland epithelial cells induced STAT3 activation and inhibited cell expansion. Taken together, these results provided strong evidence that IL-22 can directly act on salivary gland epithelial cells and promote damage and dysfunction of salivary gland tissues.
Hemopexin deficiency prevents joint injury following collagen antibody-induced arthritis
Authors: Baig Sahar , Du Yong , Ling Qin , Paglicawan Laura , Vanarsa Kamala , Mohan Chandra 
Published: January 2023

Abstract

Background
Rheumatoid arthritis is an autoimmune disorder in which the immune system attacks its own tissues, resulting in swelling and inflammation of the joints. Hemopexin (Hx), an acute-phase protein synthesized by hepatocytes in response to proinflammatory cytokines, binds to heme, and controls heme-iron availability in tissues and T lymphocytes. Its absence has been associated with increased inflammatory properties of high-density lipoproteins and the development of autoimmune disease, such as experimental autoimmune encephalomyelitis (EAE). The aim of this study is to investigate the role of Hx in the development of arthritis using the collagen antibody-induced arthritis (CAIA) mouse model.

Methods
CAIA was induced in Hx-deficient (Hx−/−) mice and wild-type (WT) littermates, via i.p injection of antibodies directed against type II collagen, followed by the administration of lipopolysaccharide (LPS). Paw thickness and disease severity was evaluated daily. Blood samples were collected to measure the serum Hx levels using ELISA.

Results
During CAIA, serum Hx level became elevated and remained high. When CAIA was induced in Hx−/− mice, they exhibited reduced arthritis compared to WT mice, as arthritic Hx−/− mice had lower average paw thickness (1.32 ± 0.02 vs 2.67 ± 0.05, 2-tail t test, P &lt; 0.05 ) and reduced arthritis scores ( 2.0 ± 0.5 vs 7.3 ±1.25, 2-tail t test, P &lt; 0.01) at the end of the experiment compared to WT mice. Pathology studies are in progress.

Conclusion
Hx−/− mice are protected from CAIA, with reduced arthritis, indicating a pro-inflammatory role for Hx in the development of arthritis. Further studies are warranted to unravel the pathogenic mechanisms involved and clinical relevance of this molecule.
Multi-parametric Human Cell-Based Inflammation Assay for Cytokines and Cell Surface Antigens
Authors: Cromwell Evan , Hoxha Ori , Hammer Matthew , Sirenko Oksana 
Published: January 2023

Abstract
Inflammation is a complex event in which cells respond to various endogenous and exogenous stimuli. Factors such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and interferon (IFN)-γ activate signaling pathways leading to the expression of cytokines and cell-surface antigens. The ability to monitor up-regulation of these molecules provides an important physiological read-out for cell-based models of inflammation. We present results from a multiparametric cell-based assay that uses immunoassays for secreted cytokines and fluorescence read-outs of cell surface markers to evaluate the effect of different mediators on inflammatory response. Primary human umbilical vein endothelial cells (HUVEC) were stimulated with inflammation cytokines (TNF-α, IFN-γ, and IL-1β) for 20 hours. Then, a newly developed microfluidic-based assay system, PuMA, was used to quantify amount of MCP-1, IL-8, and IL-6 in cell supernatants. Expression of adhesion molecules VCAM and HLADR was quantified by measurement of total fluorescence intensity after staining cells with directly conjugated antibodies using an ImageXpress Nano Automated Imaging System. Concentration dependent inhibition of the inflammation responses by the compounds AG126 and MG132 were measured. Clear differences in cytokine expression were seen between the compounds consistent with their reported mechanisms of action. The combination of imaging and microfluidic-based assays provides an efficient multiparametric assay system that can be used to test the efficacy of anti-inflammatory compounds versus toxicity and also provide significant insight into the mechanism of action by selective inhibition of markers triggered by different signaling pathways.
Aortic intima microenvironment instructs macrophage gene expression signature in steady state and progressing atherosclerosis
Authors: Williams Jesse W. , Zaitsev Konstantin , Kim Kiwook , Kim Kyeongdae , Zinselmeyer Bernd , Choi Jae-Hoon , Artyomov Maxim N. , Randolph Gwendalyn J. 
Published: January 2023

Abstract
Macrophages reside in the vascular intima of the aorta in plaque prone regions. Intima macrophages were found to be ‘tissue resident’, possessing the ability to self-renew independent of circulating monocytes and having a unique dependence on Csf1 and Flt3 pathways for development. Lineage tracing and depletion experiments showed these cells were regulators of early plaque development in models of atherosclerosis. Using bulk RNAseq on intima resident macrophages and single cell sequencing data from total leukocytes from C57Bl/6 mouse aorta, we identified a core gene expression profile associated with intima resident macrophages. Comparing this gene signatures to single-cell gene expression data obtained from total leukocytes from atherosclerotic aortas identified overlapping gene expression with foamy macrophages, despite foamy macrophages developing exclusively from recruited monocytes at this stage of disease. The intima resident gene signature was expressed in foamy cells in addition to cholesterol handling genes, but not in newly recruited monocytes. Importantly, the intima gene signature was only associated with cells lacking expression of inflammatory genes. Together this suggests there is a niche-driven gene expression program within the aortic intima influencing macrophage differentiation and function. Targeting this newly identified gene set may be important for regulation of the inflammatory pathways and foamy macrophage differentiation in atherosclerosis.
Dendritic cell derived IL-12p40 binds extracellular proteins to make heterodimeric cytokines
Authors: Gerber Allison N. , Abdi Kaveh , Singh Nevil J. 
Published: January 2023

Abstract
IL-12 and IL-23 are heterodimeric cytokines which share a common subunit, p40. Binding of p40 to p35 results in IL-12, while binding to p19 generates IL-23. Assembly of these cytokines is thought to occur only when both relevant subunits are expressed inside a dendritic cell (DC) allowing for covalent linkage and secretion. Intriguingly, the p40 subunit is also released from DC as a monomer with unknown function. We previously hypothesized that monomeric p40 can bind p35 (or p19) released by another cell in the extracellular space. Consistent with this, recombinant p40 and p35 were shown to assemble in cell free extracts. To test this hypothesis in a physiological setting, in vitro and in vivo assays were performed by mixing p40−/− and p35−/− cells. In these assays, since no one cell can make both subunits, the only way IL-12 can be present is through extracellular assembly. In vitro bioassays for IL-12 function showed proliferation and IFNγ production in response to mixing. In vivo, mixed bone marrow chimeras infected with Leishmania major were able to induce IFNγ production by T cells. These results suggest that functional IL-12 assembly can occur in the extracellular milieu from component peptides. Importantly, this mechanism may allow even non-hematopoietic tissues to direct T cell differentiation by releasing proteins capable of binding to DC-derived p40. In order to identify novel heterodimeric cytokines capable of fulfilling that paradigm, we have used a panel of proteins that can bind p40, identified from an unbiased mass spectrometry based analysis. Recombinant versions of these proteins expressed as dimers with p40 are being evaluated for functional activity.
Response gene to complement 32 interacts with SMAD3 to promote TGF-β-induced extracellular matrix production in multiple sclerosis
Authors: Tatomir Alexandru , Boodhoo Dallas , Tegla Cosmin , Rus Violeta , Rus Horea 
Published: January 2023

Abstract
We have previously shown that response gene to complement 32 (RGC-32) mediates transforming growth factor-β (TGF-β)-induced astrocyte reactivity and extracellular matrix production. However, the molecular mechanisms underlying RGC-32 expression and function remain to be elucidated. In the present study, we found that TGF-β induced expression and nuclear translocation of both RGC-32 and SMAD3 in astrocytes. Using specific pathways inhibitors, we have found that RGC-32 induction is mediated by SMAD-dependent and independent mechanisms. We investigated whether RGC-32 interacts with SMAD2 or SMAD3 in astrocytes and if this association plays a role in extracellular matrix synthesis. Co-immunoprecipitation showed that SMAD3, but not SMAD2 physically interacts with RGC-32 in astrocytes. SIS3, an inhibitor of SMAD3 phosphorylation, significantly reduced RGC-32 translocation to the nucleus, indicating that SMAD3 phosphorylation is required for RGC-32 nuclear translocation. On the other hand, silencing RGC-32 with siRNA had no effect on SMAD3 nuclear translocation. Y27632, a selective ROCK inhibitor was also able to significantly inhibit RGC-32 nuclear translocation, while the SMAD3 nuclear translocation was unaffected. The inhibition of SMAD3 phosphorylation also blocked the RGC-32-induced collagen type I expression, indicating a synergistic action between RGC-32 and SMAD3 to induce the expression of collagen type I. Taken together, our data demonstrate for the first time that RGC-32 interacts with SMAD3 to mediate the extracellular matrix production in astrocytes. These data suggest that a similar mechanism might be involved in mediation of gliosis seen in chronic multiple sclerosis lesions.