bluuebunny/crossref_metadata_2025_split huggingface dataset:
Dynamic decay of viral reservoirs in systemic and lymphoid compartments of SIV-infected rhesus macaques on long-term combined antiretroviral therapy
Authors: Wang Xiaolei , Ziani Widade , Shao Jiasheng , Veazey Ronald S. , XU HUANBIN 
Published: January 2023

Abstract

Material & Method
By quantitative real-time/digital PCR, we here longitudinally examined dynamic decay of viral parameters (cell-associated unspliced and multiply spliced viral RNA, episomal 2-LTR circular viral DNA and integrated proviral DNA) in systemic and lymphoid tissues in SIV-infected rhesus macaques on long-term combined antiretroviral therapy (cART, up to 20 months), and evaluated reservoir size and timing of viral rebound after treatment interruption. SIV RNA or DNA copy numbers were normalized and expressed as per 106 cell equivalents. Frequencies and absolute numbers of peripheral CD4+ T cells were analyzed before and after suppressive antiretroviral therapy.

Results
Our results showed that suppressive ART significantly reduced levels of cell-associated unspliced and multiply spliced SIV RNA/2-LTR DNA to undetectable, yet levels of integrated proviral DNA alone were stably maintained in systemic and lymphoid compartments throughout cART, which correlated with levels of viral replication prior to initiating cART. Following ATI, rapid clonal expansion of cells with integrated provirus were detected, as indicated by undetectable spliced tat/rev RNA and 2-LTR but increased levels of total proviral DNA, which may have contributed to subsequent viral rebound. Cell-associated SIV RNA and DNA levels completely recovered to pre-treatment levels in blood, spleen, lymph node, tonsil and gut-associated lymphoid tissues by 3 months after ATI.

Conclusions
These findings suggest that discriminating integrated HIV proviral DNA from non-integrated or defective proviruses in tissues are useful biomarkers for monitoring HIV persistence and latency for treatment and cure strategies.
Fibroblasts with trisomy 21 are more susceptible to damage from Herpes Simplex Virus-1 infection compared to typical fibroblasts
Authors: Singh Gurkirat , Bustillos-Saucedo Amalia , Royzman Kristina , Keefe Caitlin , Whitehead Tyler , Theard Curtis , James Stephanie 
Published: January 2023

Abstract
Down syndrome (trisomy 21; T21) is characterized as the presence of an extra copy of chromosome 21, resulting in cognitive and physical disorders. Individuals with T21 have a higher risk of mortality due to infection compared to typical individuals, despite having an extra interferon-alpha receptor (IFNAR), as this gene is located on q21. Herpes Simplex Virus 1 (HSV) causes the common cold sore, and can also result in serious disease such as encephalitis and periodontitis. There has been little research done with HSV in relationship to T21 although one study showed higher HSV burden in gum lesions in individuals with T21 compared to typical adults. The objective of this research was to measure differences in viral replication\protein expression and cellular responses to HSV infection in fibroblasts with and without T21. Viral replication was measured using qRT-PCR and protein expression by Western blot in cells infected with HSV, cellular gene expression was measured using mRNAseq. Viral replication and protein expression between the two groups was not significantly different although disomic cells had a slight increased rate of viral replication compared to cells with T21. However, it was observed that cells with T21 had increased cytopathic effects after 24 hours compared to typical cells. RNA sequencing data showed significantly different gene expression patterns between the two groups, notably in the EIF2 signaling pathway. Despite similar viral replication and protein expression, data presented here suggests that the fibroblasts with T21 are more susceptible to damage by viral infection. This may be due abnormalities in protein translation mechanisms yet to be delineated.
Demyelination and interferon signaling in axons cause retrograde upregulation of neuronal ISGylation pathway genes
Authors: Clarkson Ben D.S. , Johnson Renee , Howe Charles L 
Published: January 2023

Abstract
Cognitive impairment in MS is associated with diffuse gray matter injury, which remains poorly understood. Given that axons span both white and gray matter regions, it’s possible that signals within these axons contribute to the spread of diffuse injury in the brain. We show that retrograde IFNg signaling in axons (in vitro human and murine) and demyelination (murine in vivo) cause transcriptional / translational changes in neuronal cell bodies. Chief among the candidate signaling pathways identified is ISGylation, a process whereby a multitude of cellular proteins are modified by the attachment of one or more ISG15 molecules. Using custom adeno-associated viral vectors we tested how altering ISGylation in neurons affects neuronal synaptic function and how it affects neuronal responsiveness to specific inflammatory factors in cell cultures. We also measured ISGylation in MS brain tissues to see if neuronal ISGylation correlates with gray matter injury in MS. We report early evidence that increased neuron ISGylation alters the composition of neuron-derived extracellular vesicles. Microglia treated with extracellular vesicles from neurons overexpressing ISGylation pathway genes exhibited morphology consistent with increased phagocytic activity and increased mRNA expression of inflammatory cytokines including: CCL2, IL1b, TNFa, iNOS, CXCL10, CCL5, and IL6. At this point, it is still unclear how these changes are related to gray matter pathology. However, it is possible that microglial activation by these vesicles is involved with driving synapse loss as activated microglia are known to be capable of stripping neuronal synapses.
Chemokine CCL28 treats oropharyngeal candidiasis through rapid membrane disruption
Authors: He Jie , Thomas Monica A , Van Why Emma , Grayson Mitchell H , Volkman Brian F , Huppler Anna R 
Published: January 2023

Abstract
Candida albicans is a commensal organism that causes life-threatening or life-altering opportunistic infections. Treatment of these infections is limited by the paucity of anti-fungal drug classes. Antimicrobial peptides are a component of the immune response to fungi and promising agents for drug development. CCL28 is a CC chemokine that is abundant in saliva and has in vitro antimicrobial activity. In this study, we examine the spectrum of in vitro antimicrobial activity and the in vivo Candida killing capacity of CCL28 in oropharyngeal candidiasis (OPC), as well as the mechanism of anti-Candida activity. CCL28 is effective against multiple clinical strains of C. albicans, but less effective against C. glabrata. In the mouse model of OPC, application of wild type CCL28 reduces oral fungal burden in severely immunodeficient mice without causing excessive inflammation or altering tissue neutrophil recruitment. Both structured and unstructured CCL28 proteins show rapid and sustained fungicidal activity that is superior to clinical anti-fungal agents. Polyamine protein transporters are not required for CCL28 anti-Candida activity. Application of wild type CCL28 to C. albicans results in membrane disruption as measured by solute movement and enzyme leakage. Membrane disruption is reduced in CCL28 lacking the functional C-terminal tail. Our results indicate CCL28 functions in the immune response to disrupt fungal membranes and has potential for development as an anti-Candida therapeutic agent without inflammatory side effects.
Transcription factor Fli-1 impacts IL-17 expression and affects immune cell infiltration into the kidney in lupus-prone mice
Authors: Sato Shuzo , Zhang Xian , Matsumoto Haruki , Temmoku Jumpei , Fujita Yuya , Matsuoka Naoki , Furuya Makiko , Asano Tomoyuki , Kobayashi Hiroko , Watanabe Hiroshi , Migita Kiyoshi 
Published: January 2023

Abstract

Objectives
Transcription factor Fli-1 affects lupus nephritis development through regulating several cytokines and chemokines. Th17 immune response is important to maintain inflammation in lupus nephritis. The purpose of this study is to elucidate whether Fli-1 affects renal IL-17 expression and influence immune cell infiltration into the kidney.

Methods
Sera were collected from Fli-1-heterozygous (Fli-1+/−) MRL/lpr mice and wild-type (WT) littermates, and the concentration of IL-17 was measured by enzyme-linked immunosorbent assay. Expression of IL-17 and related molecules were measured by real-time polymerase chain reaction (rt-PCR) analysis. Kidneys from Fli-1+/− and WT MRL/lpr mice were stained and evaluated the grade of inflammation. IL-17 expression was detected using anti-mouse IL-17 antibodies. Immunofluorescence staining was performed using anti-CD3, CD4, and anti-IL-17 antibodies to detect T cells related to Th17 responses. In addition, to detect CCL20 expression as a chemoattractant of Th17 cells, anti-CCL20 antibodies and anti-CD11b antibodies were used to detect monocytes expressed CCL20.

Results
Fli-1+/− MRL/lpr mice had tendency of decreased serum IL-17 levels. Expression of IL-17 was significantly decreased in Fli-1+/− MR/lpr mice kidney interstitium, accompanied by decreased IL-6, STAT3, and IL-1β expression by rt-PCR. CD3 or CD4/IL-17 double-positive immune cells were significantly decreased in Fli-1+/− MLR/lpr mice; similarly, CCL20 positive monocytes were significantly decreased.

Conclusion
Fli-1 impacts IL-17 expression into the kidney of MRL/lpr mice by reducing CD3 and CD4 positive lymphocytes expressing IL-17 as well as CD11b positive monocytes expressing CCL20.
Understanding the role of STAT4 in CD4 T cell mediated neuroinflammation
Authors: Anderson Ashlyn E , Shin Boyoung , McWilliams Ian , Bryars Morgan , Harrington Laurie E 
Published: January 2023

Abstract
Multiple Sclerosis (MS) is an autoimmune disease that affects more than one million people in the United States and is characterized by the demyelination of axons in the central nervous system (CNS). While the etiology of the MS is unknown, CD4 T cells are central to disease pathogenesis. The transcription factor STAT4 is critical for Th1 cell development, is associated with risk for MS, and is required for experimental autoimmune encephalomyelitis (EAE), an animal model of MS. Interestingly, we and others find that many Th1 associated molecules such as IL-12, IFNγ and Tbet are dispensable for EAE. Hence, we hypothesize that STAT4 functions in effector CD4 T cells to promote neuroinflammation independent of the classical Th1 pathway. To test the potential role of STAT4 in Th17 cells, we employed the adoptive transfer model EAE. While we note that STAT4 expression does not impact the development of Th17 cells, it is required for the migration of Th17 cells into the CNS and for Th17 mediated demyelination. Moreover, RNA sequencing analysis shows that STAT4 regulates the expression of &gt;200 genes in the Th17 cells, many of which skew the balance between regulatory and inflammatory Th17 cell function such as Il10 and Tbx21, respectively. Together, these data reveal a novel role of STAT4 in controlling the functional properties of Th17 cells, and this may provide a promising therapeutic target for patients affected by Th17 driven autoimmune diseases.
Rapid and sensitive determination of cytokine release from cells without the need for sample transfer
Authors: Lazar Dan F , Kupcho Kevin R , Sondgeroth Casey A , Hsiao Kevin , Thompson David V , O’Brien Martha A , Gilden Julia K , Cali James J 
Published: January 2023

Abstract
We have utilized NanoLuc® Binary Technology (NanoBiT®) to develop a homogeneous and rapid assay method (≤ 70 min completion time) to measure cytokines released from cells in culture without the need for sample transfer and requiring only a standard, plate-reading luminometer for signal acquisition. Separate antibodies to a specific cytokine are labeled with the small, 11-amino acid subunit of NanoBiT luciferase (SmBiT) or its 17.6 kDa complementary subunit (LgBiT). When SmBiT- and LgBiT-labeled antibodies converge on the target cytokine, the resultant proximity of SmBiT and LgBiT reconstitutes a bright luciferase that produces light proportional to analyte levels. In this manner, immunoassays have been developed for several cytokines, including IL-1β, IL-2, IL-6 and IFN-γ. In general, these assays share LLODs &lt; 10 pg/ml and linearity over three logs of analyte concentration, mitigating the need for sample dilution. Following 24 h treatment of human PBMCs in 96-well plates with vehicle, LPS, R848, or a combination of PMA and ionomycin, cytokine detection reagents were added directly to the culture wells containing cells and medium. Depending on stimulus, maximal signal to background ratios (S/B) achieved for the cytokines were 347-, 450-, 580- and 655-fold for IL-1β, IL-2, IL-6 and IFN-γ, respectively. In a cell model comprised of activated T cells and target Raji B cells induced for 20 h with increasing bispecific T-cell engager Blincyto®, release of IL-2 and IFN-γ were observed with an EC50 of ~0.2 ng/ml and maximal S/B for IL-2 and IFN-γ of 82- and 168-fold, respectively. The implementation of this novel detection chemistry will enable rapid “add-and-read” assays for cytokine detection for both low- and high-throughput applications.
Serum Cytokines are regulated in African Americans with High HbA1c
Authors: Kimbro K Sean , Williams Ariel 
Published: January 2023

Abstract

Background
Low grade inflammation is associated with T2D resulting from a cadre of serum cytokines. IL3, IL4, IL7, IL-17A, GM-CSF, and GM-CSF play a coordinated pathogenic role in many inflammatory diseases. It has been previously reported high circulating HbA1c influences serum cytokine levels and cholesterol metabolism.

Methods
A total of 120 serum samples were collected from African Americans: 68 women and 52 men. This group consists of 45 normal HbA1c and 71 high HbA1c participants. Clinical metabolic parameters were measured. Cytokines were measured using Luminex X-MAP® technology. A series of unpaired t-test were computed to determine if there were significant differences between participants with and without elevated HbA1c. Pairwise Pearson Correlation (r) test was computed for each variable.

Results
In women, IL-3 expression was upregulated 1.7-fold in the presence of high circulating glucose. In men, IL-3 expression was downregulated 1.5-fold. Individuals with high HbA1c exhibit a negative correlation between IL-3 and BMI in men and women. Male serum IL-7 levels decrease in presence of circulating high glucose while female serum IL-7 levels increase. Normal HbA1c men have a 2.7-fold higher serum IL-7 level than women with normal HbA1c. Women participants have a positive correlation with serum IL-7 and HbA1c, as well as BMI. These correlations are lost in men. In the presence of high HbA1c, there is an increase in serum levels of IL-17A, G-CSF, and GM-CSF in women with high HbA1c.

Conclusion
An increase in HbA1c increases the expression of inflammatory cytokines and the risk of metabolic disease. Our data suggests that, in a sex specific manner, several cytokines may regulate cholesterol and lipid management in T2D.
Roles of the RNA binding protein Arid5a in IL-17-mediated diseases
Authors: Taylor Tiffany , Amatya Nilesh , Li Yang , Coleman Bianca , Zhou Chunsheng , Gingras Sebastien , Gaffen Sarah L 
Published: January 2023

Abstract
Interleukin 17 is a proinflammatory cytokine required for host defense against extracellular microorganisms but also contributes to the pathogenesis of certain autoimmune diseases. Candida albicans is a commensal fungus that colonizes mucosal tissues. During immunosuppression, C. albicans can cause superficial infections, such as oropharyngeal candidiasis (OPC). Defects in IL-17 signaling increase susceptibility to OPC in both humans and mice. IL-17 receptor signaling involves RNA binding proteins (RBP) that regulate translation and stability of target mRNA transcripts. During OPC, IL-17 induces several RBPs including Arid5a. This RBP interacts with AU-rich elements located in the 3′ UTR of many inflammatory mRNAs. In T cells, loss of Arid5a reduces expression of IL-6 and STAT3 and leads to decreased frequency of Th17 cells. Consequently, Arid5a−/− mice are resistant to experimental autoimmune encephalomyelitis (EAE), an IL-17-driven model of multiple sclerosis. Although made by T cells, IL-17 signals on non-hematopoietic cell types. We reported an in vitro role for Arid5a in the downstream IL-17 receptor pathway. Specifically, Arid5a regulates stability and/or translation of several IL-17-dependent mRNAs including Il6. Accordingly, we postulated that Arid5a plays a critical role in protection against OPC by stabilizing and increasing expression of IL-17-responsive genes. To test this hypothesis, CRISPR/Cas9 was used to generate an Arid5a knockout mouse. We confirmed that Arid5a−/− mice are resistant to EAE. To ascertain the effects of Arid5a in OPC, we infected mice orally with C. albicans and assessed fungal loads and weight loss. Surprisingly, Arid5a−/− mice were resistant to OPC, in contrast to Il17ra−/− mice.
Neutrophilic granule protein is a novel murine LPS antagonist
Authors: Hong Jaewoo , Qu Peng , Wuest Todd , Lin P Charles 
Published: January 2023

Abstract
Neutrophilic Granule Protein (NGP) was previously reported as a granular protein of neutrophils in mouse, but the function has not been known clearly. We found the presence of the possible signal peptide in NGP and validated this protein is circulating in the bloodstream. In our findings, NGP is being modified post-translationally in Golgi apparatus and endoplasmic reticulum, which is a universal character of secretory molecules with a signal peptide. The secreted NGP protein could be detected both in vitro and in vivo. NGP has sequence similarity with an antimicrobial protein cathelicidin, and we observed the aspect of inflammation of NGP. Interestingly, NGP interacts with the complex of LPS and LBP. This interaction blocks the binding of the complex of LPS and LBP to Toll-like receptor 4 (TLR4) and the downstream inflammatory signals. Furthermore, the inhibitory function of NGP against the inflammatory effect of LPS could be observed in both in vitro and in vivo. With these findings, we report NGP is a novel secretory protein to mask LPS and inhibit its function.
Type 1 interferon expression and signaling occur in distinct regions in granulomas during Mycobacterium tuberculosis infection
Authors: Jespersen Alyssa M , Junecko Beth A , Talukdar Priyanka , Mattila Joshua T 
Published: January 2023

Abstract
Type 1 interferon (IFN1s) expression is associated with progression of tuberculosis (TB), the disease caused by Mycobacterium tuberculosis (Mtb). This relationship has been assessed in murine models of TB and in human peripheral blood and airway cells, but IFN1 expression in human granulomas, the lesions associated with TB, and disease has not been addressed. To determine if IFN1s are expressed in granulomas, which cells express them, and which cells undergo IFN1-regulated signaling, we investigated these factors in granulomas from macaques, an animal model that recapitulates human TB. We found that IFN1s and interferon stimulated genes (ISGs) were expressed at higher levels in granulomas than uninfected lung. In granulomas, the strongest expression was observed within epithelioid macrophages, a subset of cells that are in close proximity to Mtb, but other myeloid cells including some neutrophils and alveolar macrophages also expressed IFN1s. The lymphocyte cuff showed greater expression of the IFN1 receptor, IFNAR1, and also expressed more of the ISG MX1 than the epithelioid macrophage region. These data indicate that IFN1 is expressed by epithelioid macrophages and IFN1-related signaling occurs within the lymphocyte cuff. These results suggest that IFN1 expression may be due to the presence of Mtb or Mtb antigens and that IFN1-regulated signaling and responses are spatially distinct from the regions and cells where IFN1 expression occurs.
Interleukin-22 receptor signaling in Paneth cells is critical for their maturation and antimicrobial function
Authors: Gaudino Stephen , Beaupre Michael , Lin Xun , Rathi Sonika , McLaughlin Patrick , Mehta Neil , Eskiocak Onur , Yueh Brian , Van der Velden Adrianus , Beyaz Semir , Kumar Pawan 
Published: January 2023

Abstract
Interleukin-22 (IL-22) acts in the intestine to promote critical tissue protective functions. However, since a diverse array of intestinal cell types (absorptive, secretory, and stem cells) express IL-22Ra1, a receptor for IL-22, it has been difficult to determine what cell type(s) specifically respond to IL-22 to mediate mucosal host defense. To address this question, we used entire gut epithelium, intestinal stem cell (ISC)-specific, and Paneth cell-specific IL-22Ra1 knockout mice. Entire epithelium-specific IL-22Ra1 knockout (Il22Ra1fl/fl;Villin-cre) mice displayed defects in Paneth cell function. Using ISC-specific IL-22Ra1 knockout mice (Il22Ra1fl/fl;Lgr5-EGFP-creERT2) and lineage tracing mice, we ruled out the involvement of Lgr5+ ISC-dependent IL-22Ra1 signaling in regulating the lineage commitment of epithelial cells, including Paneth cells. Using novel Paneth cell-specific IL-22Ra1 knockout mice (Il22Ra1fl/fl;Defa6-cre), we show that IL-22Ra1 signaling in Paneth cells is required for small intestinal host defense. We show that Paneth cell maturation, antimicrobial effector functions, gene expression of specific WNTs (Wnt6 and Wnt9b), and enteroid morphogenesis are dependent on cell-intrinsic IL-22Ra1 signaling. Furthermore, IL-22 signaling in Paneth cells regulates the intestinal commensal bacteria and microbiota-dependent IL-17A immune responses. Finally, we show Paneth cell-specific IL-22Ra1 signaling helps provide immunity against Salmonella typhimurium. Collectively, our findings provide a unique and novel role of IL-22 in Paneth cell-mediated small intestinal host defense.
Differential regulatory mechanisms underlie the control of Vγ4 and Vγ6 γδT17 development by HEB transcription factors
Authors: Anderson Michele K , Selvaratnam Johanna , Wong Alison 
Published: January 2023

Abstract
IL-17-producing γδ T-cells (γδT17) are critical components of the innate immune system, but the pathways that control their development are not well understood. We have previously shown that HEB transcription factors operate upstream of Sox4 and Sox13 to initiate the γδT17 program. However, the defects in fetal Vγ4+ γδT17 cells appear to be distinct from those in the Vγ6+ γδT17 cells. In the absence of HEB, fetal Vγ4+ γδT17 cells are unable to develop, whereas Vγ6+ γδT17 cells develop but are not able to produce IL-17. To further understand the molecular mechanisms that underlie these defects, we performed single cell RNA-seq on E18.5 γδTCR+ cells from WT and HEB-deficient mice, and did differential gene expression analysis on clusters expressing Vγ5, Vγ6, or Vγ4 mRNA. Our analysis revealed that in addition to Sox4 and Sox13, an entire suite of genes associated with the development and function of γδT17 cells was downregulated in HEB-deficient cells. Furthermore, Vδ4, which is normally restricted to cells expressing Vγ6 or Vγ5, was expressed at high levels in nearly all clusters, whereas the levels of Vγ4 and Vδ5 mRNA were severely decreased. These results suggest that direct control of Vγ and Vδ chain mRNA expression underlie the defect in Vγ4+ cell development, whereas Vγ6+ cells require a direct input from HEB into the gene program controlling γδT17 differentiation even in the presence of proper γδTCR pairing.