bluuebunny/crossref_metadata_2025 huggingface dataset:
Cardiac fibroblast participates as an immune modulator in IL-17A-driven post-injury cardiac inflammation and remodeling
Authors: Chen Guobao , Wu Lei , Bracamonte-Baran William , Hou Xuezhou , Diny Nicola , Talor Monica , Cihakova Daniela
Published: January 2023

Heart diseases are the leading causes of mortality and morbidity worldwide. The post-injury cardiac inflammation and remodeling process determined the severity of cardiac damage and the likelihood of developing heart failure. We have adapted both experimental autoimmune myocarditis (EAM) and myocardial infarction (MI) mouse model to investigate the mechanism behind cardiac inflammation and remodeling. We showed that Interleukin 17 (IL-17A) signaling is required for adverse cardiac remodeling and fibrosis in both EAM and MI models. IL-17A induces cardiac fibroblast (CF) to produce granulocyte-macrophage colony stimulating factor (GM-CSF). GM-CSF in turn drives the development of adverse cardiac remodeling and fibrosis. Despite the heterogeneity of CF population, we successfully identified a specific subset of cardiac fibroblast to be the main recipient of IL-17A signal to produce GM-CSF, CCL2 and other cytokines/chemokines. The same CF subset was detected in heart failure patient biopsies from various etiologies including myocarditis, ischemia patients, etc. Further investigation also revealed that this subset is plastic in cytokine production. Lastly, both complete knockout of IL-17A receptor (R) and specific ablation of IL-17RA in this subset significantly reduced ventricular dilation and infarct scar size. Our findings provide an excellent opportunity to develop specific treatment for post-injury cardiac remodeling and fibrosis through specific targeting the IL-17A signaling to this CF subset or the downstream cytokine production by these cells.
Functional characterization of IRF5 exon 6 variants in SLE risk
Authors: Ozarkar Snehal , McFarland Adelle , Savan Ram
Published: January 2023

The goal of the proposed research is to determine how the interferon regulatory factor 5(IRF5) lupus risk variant translates into biological risk of systemic lupus erythematosus (SLE). Genome wide association studies (GWAS) have identified IRF5 as one of the most strongly associated genes with susceptibility to SLE. Although the genetic association of IRF5 with lupus is clear, the exact mechanism by which IRF5 risk variant promotes susceptibility for lupus is still unknown. We are investigating negative regulatory factors of IRF5 activity under normal conditions, which are unable to regulate IRF5 exon 6 risk variants thereby activating type I IFNs leading to aberrant immune activation. We have characterized IRF5 exon 6 variants in pDCs using molecular and biochemical approaches, and investigated the effect of negative regulators on IRF5 exon 6 variants. This will be a significant step to gain the mechanistic understanding of aberrant production type I IFNs and immune activation in lupus.
Increased NCR negative ILC 3 may be indicative for the early development of spontaneous colitis in the IL 10 knockout mouse
Authors: Glover Sarah C , Tang Ying , Li Jian , Doty Andria , Pope Jillian , Yang Ye , Jobin Christian
Published: January 2023

The IL10 knockout (KO) mouse is a model for environmentally and genetically triggered spontaneous colitis. The response of the adaptive immune system to these changes has been studied extensively in this model. However, the response of the innate immune system, specifically the response of innate lymphoid cell (ILCs) populations to the genetic background of the mouse and the environmental stimulus of introducing microbiota has not been well studied. Herein, we evaluated the response of ILCs and Lin marker positive lymphocytes to changes in genotype (wild type Mice B6 in SPF to IL10 KO B6 mice in SPF) and also compared IL10KO mice who were either germ free or SPF exposed. To accomplish this, IL10 knockout mice were exposed to SPF conditions for three weeks. LPMCs were isolated from the colon and small bowel. Innate lymphoid cells were identified as Lin− CD90.2+CD127+cells, and within total ILC, ILC2s were ST2+ cells, ILC1s were ST2−CD117− cells, and ILC3s were ST2−CD117+NKp46+/−cells. As expected, we saw a less developed adaptive immune response in GF IL10 KO mice, with up regulation in this compartment following SPF exposure (Lin+ population increase 1.87 fold). Upon comparison of WT SPF versus IL10 KO SPF, we observed a 5 fold increase in total ILCs and an 18% increase in NCR-ILC3 in the SPF. Comparison of IL10 KO GF versus SPF revealed a 1.8 fold increase in total ILC and a 3% increase in NCR-ILC3s. In conclusion, we show that both genetic and environmental factors play a role in the innate immune changes that occur prior to the development of spontaneous colitis in the IL10 KO mouse and that the shift towards NCR-ILC 3 maybe an early marker for the development of spontaneous colitis.
TLR7 enhances cross-presentation in Ly6C(+) monocytes by targeting endosomal properties.
Authors: Larson Sandy R , Atif Shaikh M , Jakubzick Claudia , Henson Peter M
Published: January 2023

Effector functions of monocytes beyond terminal differentiation has recently come to light. In the present study we have made the novel discovery that Ly6C(+) monocytes can acquire cell-associated Ag from apoptotic cells, termed as efferocytosis. This property was enhanced by the presence of pathogenic stimuli (TLR ligands) directly acting on monocytes. Furthermore, we have shown that monocytes can cross-present Ag acquired from those apoptotic cells, with differential ability of TLR4 and TLR7 ligands in enhancing that process. This has led us to investigate the role of various pathogenic stimuli beyond these two ligands. Our data has confirmed our previous findings and further established the fact that among known TLR ligands, TLR7 selectively enhanced the presentation of peptides on MHC-I molecules. To elucidate the mechanism, we performed RNA sequencing on monocytes treated with a TLR7 ligand showing the involvement of endosomal proteins in antigen presentation, specifically subunits of the NADPH oxidase. Our data clearly demonstrated the role of Ncf-1 in TLR7 mediated enhanced activation of antigen specific CD8+ T cells. Overall, this study defines a unique role for Ly6C(+) monocytes in promoting immunity against single-stranded virus by regulating endosomal pH, a characterized process required for Batf3-dependent DC cross-presentation.
MHC II binds to multiple sites on H1N1 hemagglutinin protein
Authors: Becker Tynan A , Ferrante Andrea
Published: January 2023

The exact mechanisms of antigen processing within an antigen presenting cell (APC) are not well defined. The canonical pathway describes degradation of the antigenic proteins, competition between peptides for binding to MHC II, followed by presentation to T cells. Alternately, an entire protein might bind to MHC II first and be trimmed through the action of enzymes. Here we determined the binding affinity of HA to DR1, a human MHC II allele, in the presence and absence of the peptide-editing molecule DM and whether multiple DR1 can bind to a single HA. Binding affinities were determined by competitive binding assays with or without DM. Both native and denatured HA bind to DR1 with comparable affinities in the hundreds of nanomolar range. DM appears to decrease binding strength of HA irrespective of its state. Multiple binding events on individual HA were shown over time on blue native-PAGE gels. Bands of increasing molecular weight appear over a 24-hour time course in the blue native gels while amounts of HA and DR1 decrease over time. Taken together, these results support the hypothesis of an alternate pathway of antigen processing whereby the protein binds first to potentially multiple MHC II molecules and is then trimmed by enzymatic activity. Such MHC-mediated shielding mechanism may increase presentation of peptides by APCs that would be otherwise lost to cleavage.
Three unique interstitial macrophages in the murine lung at steady state
Authors: Gibbings Sophie L. , Thomas Stacey M. , Frasch Courtney S. , Atif Shaikh M. , McCubbrey Alexandra L. , Desch A. Nicole , Danhorn Thomas , Leach Sonia M , Bratton Donna L. , Henson Peter M. , Janssen William J. , Jakubzick Claudia V.
Published: January 2023

Rationale

The current paradigm in macrophage biology is that some tissues mainly contain macrophages from embryonic origin such as microglia in the brain, while other tissues contain postnatal-derived macrophages, such as the gut. However, in the lung and in other organs such as the skin, there are both embryonic and postnatal-derived macrophages.

Objectives

In this study, we demonstrate in the steady-state lung that the mononuclear phagocyte system is comprised of three newly identified interstitial macrophages (IMs), alveolar macrophages (AMs), dendritic cells and few extravascular monocytes.

Methods

We focused on similarities and differences between the three IM subtypes, specifically, their phenotype, location, transcriptional signature, phagocytic capacity, turnover and lack of survival dependency on CX3CR1.

Measurements and Main Results

Pulmonary IMs were located in the bronchial interstitium but not the alveolar interstitium. At the transcriptional level, all three IMs displayed a macrophage signature. All IMs expressed MerTK+CD64+ CD11b+ CX CR1+ and were furthermore distinguished by differences in cell surface protein expression of CD206, Lyve-1, CD11c, CCR2 and MHCII, along with the absence of Ly6C, Ly6G, and Siglec F. Ex vivo analysis revealed that all three IMs were highly phagocytic compared to AMs. Finally, similar phenotypic populations of IMs were present in other non-lymphoid organs, suggesting that these IMs may not be unique to the lung.

Conclusions

These findings promote future research to track four distinct pulmonary macrophages and decipher the division of labor that exists between them.
IgA You Assay? The Correlation between Sleep and Stress
Authors: Redmond Sarah , Biggs Erick , Blevins Kasey , Brimer Kelsey , Brown Cameron , Bryan Sandra , Burger Haley , George Anna , Hunt Mckenzie , Jannise Caitlin , Kesterson Melissa , Kwan Anthony , Marshall Jacob , Miller Kenzie , Mohamed Attia , Rajashekara Arpitha Mysore , Pagan Rachael , Peters Mary , Saenz Allison , Steeves Gregory , Stowers Samuel , Welch Nate , Williams Alexis , Williams Devonn , Wingfield Kyle , Zimmermann Krista
Published: January 2023

The most well known fact about sleep is that humans need it - while we are asleep our metabolic processes slow down and during long periods of sleep our bodies repair the damage that the stress our bodies have undergone throughout the day. Throughout the day our bodies are bombarded by signals from the environment, and undergoes biological changes in response to external and internal stimuli. Sleep is a period of time to restore the body to optimum condition to continue the process all over again the next day. The human body regulates stress by producing glucocorticoid steroid hormones, which also function as immune response suppressants. During sleep cortisol levels are decreased as they inhibit the body’s period of restoration. The production of immunoglobulin A, an antibody associated with mucosal membranes, is inhibited by glucocorticoids, and thus shows a relationship with sleep and other factors associated with stress. This experiment focused on correlations between sleep and stress found on the campus of Radford University. Our groups recorded the amount of sleep participants acquired the previous night and how long they had been awake, and correlated these with the concentration of salivary IgA. According to our initial data collection there was no significant correlation between the amount of sleep (p=0.305) or hours awake (0.670) to IgA levels. Additional sampling later in the semester will be used to supplement this data set and mapped using geospatial coordinates.
Regulation of Immunomodulatory Cytokine Secretion by IFNb-1a is Mediated through Up-regulation of IRF7-SOCS1 Signaling in Monocytes from Patients with Relapsing-Remitting Multiple Sclerosis
Authors: Zhang Xin , Tao Yazhong , Markovic-Plese Silva
Published: January 2023

Objective

To explore the mechanism by which IFNb-1a modulates cytokine production in monocytes from patients with relapsing-remitting multiple sclerosis (RRMS).

Methods

91 RRMS patients and 77 control subjects were enrolled in the study. Magnetic resonance imaging (MRI) scans were used to detect the brain T2 and T1 lesion volume. Serum, cerebrospinal fluid (CSF) and peripheral blood CD14+ monocytes were collected for flow cytometry, RT-PCR, Western blotting and ELISA.

Results

As compared to the control subjects, IL-23 was increased in serum and CSF, while IL-27 was decreased in CSF of the RRMS patients. Serum IL-23 levels were positively correlated with brain MRI T2 and T1 lesion volumes in RRMS patients. CD14+ monocytes from RRMS patients showed decreased percentage of IL-27, IL-10 and IL-4, and increased percentage of IL-23; secreted lower levels of IL-27, IL-12p70 and IL-4, and higher levels of IL-23; and exhibited decreased gene expression of IFNb-1, IFNAR1, IFNAR2, SOCS1, TLR7 and MyD88. IFNb-1a in vitro treatment of monocytes from RRMS patients induced IL-27 and inhibited IL-23 production, increased gene expression of IRF7, MyD88 and SOCS1, and induced protein expression of IFNAR1, pSTAT1, IRF7, and SOCS1. STAT1 inhibitor diminished IFNb-1a-mediated STAT1 phosphorylation and up-regulation of IRF7 and SOCS1, while siRNA IRF7 abolished the IFNb-1a-mediated induction of SOCS1 in monocytes. Both STAT1 inhibitor and siRNA IRF7 impaired IFNb-1a-mediated regulation of IL-27 and IL-23.

Conclusion

Our study demonstrated that IRF7 is critical for the IFNb-mediated upregulation of SOCS1, which mediates IFNb-1a-induced up-regulation of IL-27 and down-regulation of IL-23 cytokine production in monocytes from RRMS patients.
Immune cross-reactivity between Dengue and Zika viruses in two pediatric studies in Nicaragua
Authors: Cruz Magelda Montoya , Puerta-Guardo Henry Nelson , Schildhauer Sam , Katzeltnick Leah , Balmaseda Angel Lazaro , Harris Eva
Published: January 2023

The 4 dengue virus (DENV) serotypes and Zika virus (ZIKV) are antigenically related mosquito-borne flaviviruses that cause human diseases of major medical and public health importance worldwide. The cross-reactivity between DENV and ZIKV has raised questions about cross-neutralization and concerns of cross-enhancement, yet few data exist characterizing the long-term antibody response. Here, we studied the extent of immunological cross-reactivity between DENV and ZIKV using samples from two long-term studies in Nicaragua: a hospital-based study with a longitudinal arm extending for 18 months and a 13-year ongoing community-based cohort study with annual healthy blood samples. We selected confirmed dengue cases, stratified by serotype (DENV1, 2, 3) and immune status (primary, secondary) (n=5/group), and analyzed plasma samples collected at acute, convalescent, 3-, 6-, 12- and 18-month time-points, as well as 1–3 years post-infection, using a Vero cell-based focus reduction neutralization assay. We found that primary DENV infections display lower levels of cross-neutralizing antibodies to ZIKV than secondary DENV infections, but that both maintain cross-reactivity over time. We noted differences by serotype, with less cross-neutralization in DENV1-immune plasma. However, when we compared these data with neutralization titers against DENV1-4, we found neutralizing antibody titers to ZIKV to be ~10-fold lower than to the homologous DENV. We are also testing Zika-positive samples with and without prior DENV exposure and samples negative for prior ZIKV or DENV infections. These findings improve our understanding of cross-neutralization over time between DENV and ZIKV, with implications for natural immunity and vaccines.
Effects of cell population rate of rat bone marrow derived mast cell on protein constituents analyzed by 2 dimensional electrophoresis
Authors: Cho Sung-Weon , Oh Haejin , Shin Kyung Sook
Published: January 2023

Mast cells are the immune cells for defense from infecting organism including helminthes. Rat bone marrow dervied mast cells (rBMMC) are the proliferating cells that are induced from rat hematopoietic stem cells (rHSC) cultured in the presence of both IL-3 and stem cell factor (SCF). rBMMC are suggested as the useful cells for the treatment of clonorchiaiss with the results of rBMMC effects on rat clonorchiasis in vivo. The effects of cell population rate on rHSC for rBMMC preparation were investigated by analysing protein constituents using two dimensional electrophoresis (2-DE) with silver staining visualization. The cells in 4 cell population rate conditions were analyzed with ordinary cell population rate (1.1 × 105/ml, O-CPR), high cell population rate (2.2 × 105/ml, H-CPR), very high cell population rate (5.5 × 105/ml, VH-CPR), and very high heavy cell population rate (11.1 × 105/ml, VHH-CPR). The spot ditribution were described as the patterns that have two main group regions with spot dozens, high molecular weight and low pH (HL), and low molecular weight and high pH (LH). The patterns were comparable among the four conditions of cell population rate. However, in comparison between individual spot, the density of each spot was observed in remarkable difference. The results were suggested that the treatment effects of rBMMC on rat clonorchiasis would be dependent on culture condition of cell population rate.
Lymphocyte aggregates accumulate in the skin of melanoma associated autoimmune vitiligo
Authors: Vella Jennifer L , Huang Yina H
Published: January 2023

While melanoma is not the most common form of skin cancer, it is responsible for the majority of skin cancer related deaths. A small percentage of people that develop melanoma simultaneously develop autoimmune vitiligo, resulting in a favorable prognosis for these individuals. Melanoma associated vitiligo is a CD8+ T cell mediated disease that results in the death of healthy melanocytes. The destruction of melanocytes generates the self antigen(s) required for memory CD8 T cells that persist long term and provide protection from melanoma. Although the link between autoimmune vitiligo and anti-melanoma immunity has long been established and well studied, the mechanisms driving vitiligo and tumor protection still remain unknown. Using a mouse model of melanoma-associated vitiligo we are investigating this intimate relationship. Using multiparameter immunohistochemistry, we have identified large aggregates of lymphocytes clustered throughout the initial site of depigmentation. These lymphoid aggregates are comprised of a number of different lymphocyte subsets including CD4 and CD8 T cells, dendritic cells and macrophages. However, they are distinct in composition, duration and location from tertiary lymphoid organs and other lymphocyte clusters identified in the setting of HSV infection. Only vitiligo-affected mice develop these aggregates suggesting they may be important in the vitiligo development or dissemination and/or tumor protection. Understanding the role these lymphocyte aggregates play in vitiligo will give us a better understanding on the mechanisms required for both autoimmune vitiligo and anti-melanoma immunity.
Factors controlling the tolerogenic phenotype of lymphatic endothelial cells.
Authors: McKenna Nicole , Berendam Stella , Rouhani Sherin , Turner Stephen , Koeppal Alex , Cummings Kara , Dekoter Rodney , Engelhard Victor H
Published: January 2023

The prevailing model of peripheral tolerance necessitates tissue-resident dendritic cells migrating to lymph nodes and cross presenting self-antigen to T cells, resulting in their deletion or anergy. Our lab has identified a novel mechanism of peripheral tolerance involving a subset of lymph node stromal cells called lymphatic endothelial cells (LEC). LEC in the lymph node induce peripheral tolerance by presenting epitopes derived from peripheral tissue antigens (PTAs) in conjunction with expression of PD-L1, leading to CD8 T cell deletion. However, PTAs and PD-L1 are not expressed on LEC in the colon or diaphragm. We performed RNAseq analysis on LEC from lymph node and diaphragm to identify differences in expression of transcription factors that might control expression of PD-L1 and PTA. We found that Spi-B is the most differentially expressed transcription factor in the lymph node compared to the diaphragm, with a fold change of over 500. Spi-B has been implicated in the development and function of medullary thymic epithelial cells, a key cell type in central tolerance. Lymph node LEC harvested from Spi-B deficient mice have a significant shift in markers associated with LEC tolerogenicity. Expression levels of PD-L1, ICAM-1, MAdCAM-1, and LTbR delineate different lymph node LEC subpopulations that vary in their ability to express PTAs and tolerize CD8 T cells. LEC from Spi-B deficient mice have decreased MAdCAM-1 expression and increased LTbR and ICAM-1 expression suggesting a population shift of non-tolerogenic lymph node LEC to tolerogenic lymph node LEC. This data suggests Spi-B may be involved in a negative feedback mechanism to limit the number of tolerogenic LEC during development.
TET2 deficiency promotes CD8+ T cell memory differentiation
Authors: Jordan Martha S , Carty Shannon A , Gohil Mercy , Cotton Renee , Johnson Matthew , Stelekati Erietta , Wells Andrew D , Wherry E. John , Koretzky Gary A
Published: January 2023

DNA methylation contributes to T cell differentiation. The ten-eleven translocation (TET) family of methylcytosine dioxygenases mediates active DNA demethylation. Here we demonstrate that TET2 regulates CD8+ T cell differentiation in vivo following acute viral infection. At steady-state, mice with a T-cell specific deletion of TET2 have intact thymic and peripheral T cell populations. However, following infection, TET2 loss promotes early acquisition of a memory CD8+ T cell fate in a cell-intrinsic manner without disrupting antigen-driven T cell expansion or effector function. Upon re-challenge, TET2-deficient memory CD8+ T cells exhibit superior pathogen control. Genome-wide methylation analysis of TET2-deficient versus WT CD8+ T cells identified multiple differentially methylated regions (DMRs). Analyses of DMRs suggest TET2 regulates T cell differentiation at regulatory genes, opposed to individual loci of differentially expressed memory markers. One hypermethyated region was the PRDM1 loci (encoding Blimp-1). This finding was associated with an increased Bcl6 to Blimp-1 ratio in vitro, a pattern predicted to favor memory cell fate. Taken together, our data indicate that TET2 is an important regulator of CD8+ T cell fate decisions.
Recombinantly engineered antibodies for reproducible and background free flow cytometry analysis
Authors: Fleischer Johannes , Singh Kalpana , Nölle Volker , Hellmer Jens , Schmitz Jürgen
Published: January 2023

Antibodies are key reagents for numerous cell analysis applications. Therefore, the contribution of antibodies in experimental irreproducibility is being critically looked at. The majority of antibodies used in basic research are still derived from hybridoma technology established in 1975. As highlighted during recent debates, hybridoma-based antibodies are prone to quality issues, i.e., lot-to-lot variability and lack of specificity. In addition, the technology itself doesn’t allow any improvements in the quality and the properties of the antibodies.

Recombinantly generated antibodies possess highly defined characteristics and are produced under standardized culture conditions leading to products that are reproducible in nature. This saves users the time associated with standardizing analysis and decreases the financial burden associated with working with inconsistent reagents. Recombinant technology also opens up myriads of possibilities to improve antibody properties. REAfinity™ Antibodies provided by Miltenyi Biotec are recombinantly engineered to lack background binding and share a single isotype. This solves the two common problems associated with flow cytometry: non-specific signals and complicated experiment planning with multiple antibody isotypes.
Fast isolation of mouse XCR-1+ dendritic cells
Authors: Huang Yi-Ju , Morrissey-Wettey Frank , Andreoni Cristina Conforti , Dzionek Andrzej
Published: January 2023

Cross-presenting CD8-+ conventional dendritic cells (cDCs) are important for the eradication of cancers and viral infections due to their ability to induce cytotoxic T lymphocyte (CTL) responses. Cross presenting DCs are a very rare subset and in the past, studies of these cells have been limited by their scarcity of specific cell surface markers. Therefore, methods for the detection and isolation of these cells were commonly based on a multitude of immunophenotypic criteria, such as the expression of CD11c and CD8- and the absence of CD3, CD4, SIRP-- and CD11b. Recently, it was demonstrated that crosspresenting cDCs in lymphoid and non-lymphoid tissues specifically express XCR1, which correlates with the ability to take up and cross-present exogenous antigens. Combining our recombinant REAfinity™ Anti-XCR1 mouse mAb with MACS® Technology based on UltraPure MicroBeads, we developed a new method for the fast and easy isolation of cross-presenting DCs. Using this method XCR1+ DCs can be routinely enriched with high recovery and purity without the need for time-consuming laborious flow sorting. This will facilitate the study of cross-presenting DC subset, which will ultimately help to develop new therapeutic strategies employing DCs in the future.
Regulatory T cells exert differential functions during the early and late stages of the immune response to respiratory virus infection
Authors: Rogers Meredith C , Lamens Kristina D , Shafagati Nazly , Williams John V
Published: January 2023

Human metapneumovirus (HMPV) is the second leading cause of viral lower respiratory illness in children, and can be severe in premature infants, immunocompromised persons, and the elderly. Clearance of HMPV is predominantly mediated by cytotoxic CD8+ T lymphocytes (CTLs). HMPV and other respiratory viruses induce CTL impairment, where virus-specific CTLs are unable to function for an optimal antiviral immune response. We recently demonstrated that the inhibitory receptor Programmed Cell Death-1 (PD-1) mediates early functional impairment of the CTL response to HMPV, influenza, and other respiratory viruses early in infection. However, CTLs became impaired by day 10 post-infection in PD-1-deficient mice, indicating that other immunoregulatory cells or molecules may be acting during HMPV infection. CD4+ regulatory T cells (Tregs) are suppressive in cancer and some infections. We found that Tregs were increased in both number and percentage and activated during HMPV infection. Treg depletion throughout infection significantly increased CTL functionality and reduced peak virus titer. Early depletion of Tregs led to reduced recruitment of HMPV-specific CTLs to the lung, while depletion late after infection maintained the proportion of HMPV-specific CTLs and accelerated virus clearance. These data suggest that Tregs may perform distinct functions at different stages of lung infection. Tregs may be beneficial during the priming stage of the immune response, but are overall detrimental in the setting of HMPV infection and could be a target for therapeutics.
Noncytopathic bovine viral diarrhea virus 2 impairs virus control in a mouse model
Authors: Kyoung-Seong Choi , Seong Giyong , Shin Seung-Uk
Published: January 2023

Objective

Bovine viral diarrhea virus (BVDV) is an economically important viral pathogen. BVDV is classified into two distinct species: BVDV1 and BVDV2. Both these species consist of two biotypes: cytopathic (cp) and non-cytopathic (ncp), differing in their ability to induce cytopathogenic effects in cultured cells. In this study, we investigated the immunological differences between ncp BVDV1 and ncp BVDV2 infections.

Materials and methods

Specific pathogen-free BALB/c mice (6–8 wk old) were used. The challenge was performed by IP injection of 5 ′ 105 TCID50 of ncp BVDV1 or ncp BVDV2. Each BVDV strains- and mock-inoculated mice were sacrificed on days 4, 7, 10, and 14 post-inoculation (pi). Each tissues and blood were harvested. BVDV antigen detection, immunohistochemical staining was performed on mouse tissues. Spleens were used for flow cytometry. Cytokines and chemokines were detected separately by using a mouse CBA inflammation kit.

Results

Viral antigens were detected in the spleens of all infected mice from days 4 through 14 and also found in the mesenteric lymph nodes, GALT, heart, kidney, intestine, and BALT of some infected mice. In ncp BVDV2-infected mice, flow cytometric analysis revealed markedly fewer CD4+ and CD8+ T lymphocytes and lower expression of CD80/CD86 and I-A/I-E than those in ncp BVDV1-infected mice. IL-6 and MCP-1 was higher in the plasma of ncp BVDV2- than that in the plasma of ncp BVDV1-infected mice.

Discussion

Our results demonstrate that ncp BVDV1 and ncp BVDV2 interact differently with the host innate immune response in vivo. These findings highlight an important distinction between ncp BVDV1 and ncp BVDV2, and suggest that ncp BVDV2 impairs host’s virus control and enhances virus dissemination.
MMPs/TIMPs imbalances in the peripheral blood and cerebrospinal fluid are associated with the pathogenesis of HIV-1-associated neurocognitive disorders
Authors: Xing Yanyan , Shepherd Nicole , Lan Jie , Li Wei , Gupta Samir , Zhang Shanxiang , Dong Jun , Yu Qigui
Published: January 2023

HIV-1-associated neurocognitive disorders (HAND) continue to be a major concern in the infected population, despite the widespread use of antiretroviral therapy. Growing evidence suggests that an imbalance between matrix metalloproteinases (MMPs) and endogenous tissue inhibitors of MMPs (TIMPs) contributes to the pathogenesis of HAND. In our present study, we examined MMPs/TIMPs protein levels and MMPs enzymatic activities in both plasma and cerebrospinal fluid (CSF) samples from HIV-1 patients and HIV-1-negative controls. We found that imbalances between MMPs and TIMPs occurred in both the plasma and CSF of HIV-1 patients, especially those with neurocognitive disorders. In the plasma, the protein levels of MMP-2, TIMP-1, and TIMP-2, but not MMP-9, were increased in all HIV-1 patients when compared with HIV-1-negative controls. The enzymatic activity of both MMP-2 and MMP-9 was increased in all HIV-1 patients. Particularly, the ratio of MMP-2/TIMP-2 was significantly increased in HAND patients, not in patients without neurocognitive disorders. In the CSF, MMP-2 activity was increased, but the ratio of MMP-2/TIMP-2 was not altered. De novo induction and activation of MMP-9 in the CSF of HAND patients was particularly prominent. The imbalances between MMPs and TIMPs in the blood and CSF were related to the altered profiles of inflammatory cytokines/chemokines and monocyte activation. In addition, HIV-1 plasma directly damaged a blood brain barrier (BBB) model, leading to increased BBB permeability and vigorous transmigration of monocytes. Thus, the increased ratio of MMP-2/TIMP-2 in the peripheral blood and de novo induction and activation of MMP-9 in the CSF are significant in HAND patients and may be biomarkers for HAND.
The role of meningeal lymphatics in viral meningitis
Authors: Herz Jasmin , Lupi Alexander , McGavern Dorian B , Kipnis Jonathan
Published: January 2023

Discovery of the meningeal lymphatic vessels in the brain meninges (coverings of the brain) led to the hypothesis that they carry macromolecules and immune cells from cerebrospinal fluid (CSF) to cervical lymph nodes (cLN). Our new findings suggest that the lymphatic system controls immune responses through guiding antigen-presenting cells, such as dendritic cells (DC) from the meninges to draining lymph nodes. By using surgical and pharmacological methods to interfere with meningeal lymphatic vessels, we found lymphatics to serve as a major conduit by which DC leave the meninges, traffic to cLN and promote the generation of antigen-specific T cells. Inflammation-induced migration of DC is primarily regulated through a CC-chemokine receptor 7 (CCR7) dependent pathway. Co-injection of equal parts of differentially labeled CCR7-deficient and wild-type (WT) DCs into the CSF of the brain did allow translocation of CCR7-competent DC into the T cell zone of the draining lymph nodes but not CCR7−/− DC. Subsequently, antigenic stimulation by CCR7−/− DC did not induce a T cells response in the cLN, whereas WT DC enforced the majority of antigen-specific T cells to proliferate and increase their effective trafficking through integrin expression. To gain insights into how T cell responses are elicited when the meninges are infected with a virus, we currently set out to understand the lymphatics function in a mouse model of virus-induced meningitis. Injection of Lymphocytic choriomeningitis virus into the CSF induces rapid, uniform and fatal inflammation that is dependent on the generation of viral specific CD8+ T cells and potentially the exit of migratory DC through meningeal lymphatic vessels.
TET2 and 3 proteins control iNKT cell lineage specification and TCR mediated expansion
Authors: Tsagaratou Ageliki , Avalos Edahi Gonzalez , Rautio Sini , Browne James Scott , Togher Susan , Pastor William , Rothenberg Ellen V. , Lahdesmaki Harri , Rao Anjana
Published: January 2023

TET proteins are 2-oxoglutarate- and Fe(II) dependent dioxygenases that catalyze the hydroxylation of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) and further oxidation products (5-formylcytosine and 5-carboxylcytosine) in DNA. They can act as mediators of “active” (replication-independent) DNA demethylation, achieved through excision of 5fC and 5caC by thymine DNA glycosylase (TDG) followed by replacement with an unmethylated cytosine by base excision repair. They are also novel epigenetic marks that are recognized specifically by readers.

The role of TET proteins in developing thymocytes remains elusive. In the present study, we show that simultaneous deletion of TET2 and TET3 in mice (DKO) results in aberrant development of invariant NKT cells that are skewed towards the NKT17 lineage. We also found upregulation of proliferation controlling genes as well as genes that are normally expressed in precursor cells. This results in an unprecedented expansion of iNKT cells, leading to lethality. The expansion is driven by antigen recognition, since the disease is effectively transmitted to fully immunocompetent recipient mice only if they express CD1d, a non-classical MHC protein that presents lipid antigens to iNKT cells. Molecular analysis revealed that TET2 and TET3 drive iNKT lineage specific DNA demethylation and control gene expression via DNA demethylation and/or chromatin accessibility of key lineage specifying factors such as RORgt, Tbet and ThPOK.

Collectively, TET2 and TET3 are fundamental regulators that seal iNKT lineage fate, ensure proper development and maturation and safeguard aberrant TCR mediated expansion.
Defining group 2 innate lymphoid cell tissue niches
Authors: Molofsky Ari B
Published: January 2023

Group 2 innate lymphoid cells (ILC2) are tissue resident cells that are key instigators of type 2 allergic immune responses. ILC2 can promote beneficial type 2 immunity in the context of helminth infection, but are also key contributors to the allergic pathology associated with asthma and atopic dermatitis. ILC2 are developmentally deposited in tissues where they locally proliferate, and recent work suggests their contribution to physiologic tissue responses during tissue development and tissue remodeling. Our group’s prior work indicates adipose tissue ILC2s are key coordinators of a mixed regulatory/type 2 immune response which promotes adipose tissue function and is protective in models of obesity and type 2 diabetes. However, the cells and signals that regulate ILC2 in tissues are only beginning to be explored. In adipose tissue and elsewhere, the IL-1 family cytokine IL-33 is a dominant positive regulator of ILC2 function. As such, we have pursued the regulation and cellular sources of IL-33 that are proximal to tissue-resident ILC2. Using high-resolution imaging and genetic approaches, our studies suggest ILC2 are maintained in specific micro-anatomic tissue niches that are defined by specific non-hematopoietic IL-33 expressing cells. By defining these ILC2 niches, we hope to better understand the cells and signals that regulate ILC2 so that we may ultimately exert control over both beneficial and pathologic type 2 immune responses.
Beta-2 adrenergic receptor signaling prevents hyperinflammation through early release of IL-10
Authors: Agac Didem , Estrada Leonardo , Farrar David
Published: January 2023

Innervation of the lymphoid tissues by catecholaminergic nerve endings allow the sympathetic nervous system to communicate with immune cells through the release of norepinephrine (NE). NE signals through adrenergic receptors, which are widely expressed by immune cells. We have identified a novel pathway where NE can suppress pro-inflammatory cytokine secretion from bone marrow-derived macrophages in response to multiple TLR agonists. This pathway operates through the beta2-adrenergic receptor (ADRB2), and the suppressive effect of NE depends on the early induction of the anti-inflammatory cytokine IL-10. We assessed the in vivo role of this pathway using a LPS endotoxemia model and found that ADRB2 knockout (KO) animals rapidly succumbed to a sub-lethal LPS challenge. This sensitivity is correlated with elevated serum levels of TNFα and suppressed IL-10 in ADRB2 KO animals. The lethality in the ADRB2 KO animals was rescued by administering exogenous IL-10. Additionally, the ADRB2-specific agonist salmeterol rescued wild-type animals from a lethal LPS challenge, which was reversed by neutralizing anti-IL-10 antibody. These observations suggest that ADRB2 signaling is critical for controlling inflammation through the rapid induction of IL-10. Through transcriptome analysis, the NR4A nuclear orphan family members were induced by NE, and the presence of several putative NR4A binding sites within the IL-10 promoter suggests that these factors may directly regulate IL-10 expression in response to ADRB2 signaling. Further studies will directly test the control of IL-10 expression by NR4A factors. Understanding this pathway will provide new insights into how the nervous and immune systems communicate through ADRB2 signaling.
Primary cilia regulate accumulation of innate and adaptive immune cells following injury
Authors: Zimmerman Kurt A , Song Cheng , Yoder Bradley K
Published: January 2023

Primary cilia are small microtubule based appendages found on the cell surface of almost every cell type. Mutations in proteins localized to the primary cilia or proteins required for ciliary assembly lead to polycystic kidney and liver disease with fluid filled cysts and fibrosis in both organs. Recent studies have found that tissues in which cilia function on epithelial cells is disrupted have an altered response to injury; however, the signaling pathways, cytokines, and cell types involved remain poorly defined although some studies suggest that macrophages are important. We hypothesize that cells containing a dysfunctional primary cilium are unable to elicit or resolve a normal injury response and produce cytokines that recruit immune cells thereby promoting cystic disease. Our data show that cilia mutant mice have an abnormal accumulation of neutrophils, bone marrow derived monocytes, tissue resident macrophages, and T cells in the kidney and liver following injury. In addition, qRT-PCR analysis of epithelial cells containing dysfunctional primary cilia shows increased mRNA levels of cytokines important for immune cell recruitment and activation including MCP-1, CSF-1, CXCL-1, and CX3CL- 1. Genetic reduction of monocytes (CCR2−/− mice) or a specific subset of tissue resident macrophages (CD11c+) in the kidney of cilia mutant mice did not affect cyst formation. However, our preliminary studies indicate that pharmacological inhibition of tissue resident macrophages (CSF1R inhibitor) or genetic ablation of adaptive immune cells (Rag1−/− mice) reduces the severity of cyst formation in cilia mutant mice. Targeting certain immune populations may prove of therapeutic benefit for polycystic kidney and liver disease patients.
Alpha-7 nicotinic acetylcholine receptor agonist GTS-21 influences the inflammatory response in an animal model of heart failure
Authors: Kha Mindy , Chow Sheryl , Cameron D. Joshua , O’Barr Stephen A
Published: January 2023

Cardiovascular disease, specifically heart failure affects nearly 6 million Americans, and remains to be the number one cause of hospitalization and death globally. Multiple studies have suggested that inflammation plays a pivotal role in the pathogenesis of this disease. There is a novel link between the vagus nerve and the inflammatory responses, where the vagus nerve can limit inflammation via the alpha-7 nicotinic acetylcholine receptor (α7 nAChR) in relation to the “cholinergic anti-inflammatory pathway (CAP)”. Selective pharmacological stimulation of the α7 nAChR may have therapeutic potential for the treatment of inflammatory conditions. We determined the anti-inflammatory potential of 3-(2,4-dimethoxybenzylident)-anabaseine (GTS-21), an α7-selective partial agonist, in an in vivo zebrafish heart failure model. To investigate the possible therapeutic benefit of GTS- 21, we used a chronic exposure system in which animals are pre-treated with GTS-21 for 20 minutes per day for four consecutive days. On day four after a short washout period, heart failure is chemically induced by Oligo-[2-(2- ethoxy)-ethoxyethyl)-guanidinium-chloride] (PGH) and survival is recorded. We show that GTS-21 has a profound anti-inflammatory effect in improving survivability in a concentration dependent manner. Moreover, rtPCR analysis on heart tissue shows that pre-treatment with GTS-21 induces modifications in levels of gene regulatory proteins leading to altered cytokine expression within the myocardium. In all, these data indicate that GTS-21 may be a promising therapeutic as it influences the inflammatory response in heart failure.
PLAG reduces the infection risk via acceleration of bacteria clearance through NF-κB activation in Pseudomonas aeruginosa induced pneumonia model with chemotherapy-induced neutropenia
Authors: Kim Yong-jae , Jeong Jinseon , Yoon Sun Young , Sohn Ki-Young , Kim Heung-Jae , Kim Jae Wha
Published: January 2023

Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen and can cause an acute and chronic infection disease in immunocompromised condition. P. aeruginosa has a potent pathogen; causes invasion diseases in hospitalized patients and complicates 90% of cystic fibrosis deaths. NF-κB and STAT3 are important signal molecules in inflammatory responses. Recently, research on the interaction between NF-κB and STAT3 in inflammation has been actively conducted. Previously, we reported that PLAG (1-palmityoyl-2-linoleoyl-3-acetyl-rac- glycerol) has an effect on regulation of STAT3 activation in diverse disease model. Therefore, we investigated whether PLAG has an effect on bacteria-infected mice with chemotherapy-induced neutropenia. To establish neutropenic mouse model, we used Cyclophosphamide and doxorubicin (AC regimen). And P. aeruginosa was administrated by intranasal injection in AC regimen induced neutropenic mice and followed by oral administration of PLAG during the experiment. We observed that survival rate of P. aeruginosa infected neutropenic mice is increased and bacteria in BALF is decreased by PLAG at early time. To reveal the function of PLAG on increase of infected bacteria elimination, we checked out neutrophil recruitment and NETosis. We observed that neutrophil recruitment in BALF was not increased significantly, but NETosis occured by P. aeruginosa at early time. And PLAG enhanced NETosis through NF-κB activation via down-regulation of STAT3. Based on these data, we suggest that role of PLAG on bacteria clearance, regulates interaction between NF-κB and STAT3 activation through P. aeruginosa induced pneumonia model with CIN and PLAG could be help immunocompromised patients.
The mitochodnrial E3 ligase MARCH5 resolves RIG-I and MAVS aggregates in innate immunity
Authors: You Young Suk , Kim Jae-Hoon , Lee Jong-Soo , Cho Hyeseong
Published: January 2023

Ubiquitin proteasome system (UPS) on the mitochondrial is one of quality control systems that works as a first line of defense barrier against aggregated or misfolded proteins. In innate immunity formation of the MAVS(Mitochondrial anti-viral signaling protein) oligomers elicits robust type-I interferon induction upon viral infection and however, persistent RIG-I and MAVS complex rather leads to host immunopathology. We recently reported that mitochondria-resident E3 ligase, MARCH5, recognizes the oligomeric form of RIG-I and MAVS complex. MARCH5+/− mice and MARCH5 deficient immune cells exhibited low viral replication and elevated type-I interferon response to viral infection. MARCH5 bound RIG-I and MAVS complex only during viral infection when MAVS forms oligomer. MARCH5 mediates Lys-48-linked poly-ubiquitination chain addition to RIG-I and MAVS oligomer and induces the RIG-I/MAVS monomer. Next, we studies have suggested that a novel function of MARCH5 in inflammation signaling. Together, these data demonstrates that MARCH5 regulates oligomeric RIG-I and MAVS complex.
PTGER4 inhibition in non-obese diabetic mice restores impaired IFNAR signaling, decreases chronic inflammatory signals and inhibits autoimmunity
Authors: Rahman M. Jubayer , Rodrigues Kameron , Liu Yi , Zhao Yongge , Quiel Juan A. , Tarbell Kristin V
Published: January 2023

We recently demonstrated that dendritic cells (DCs) from autoimmune-prone non-obese diabetic (NOD) mice at prediabetic age (8–10 week) have a diminished IFN-alpha Receptor (IFNAR) response despite increased production of type 1 IFN. This impaired IFN response may result from chronic activation of inflammatory pathways that contribute to dysregulated innate activation. qPCR of innate targets showed that younger NOD mice (3 week old) did not yet display impaired IFNAR signaling, but already had higher IL-1. To understand the interplay between different innate signals, we compared gene expression of 5 key DC populations from prediabetic NOD mice and diabetes-resistant B6.g7 mice. Several inflammatory pathways including IL-1, Eicosanoids, and NFκB were higher in NOD DCs whereas type 1 IFN-response genes, especially those associated with tonic signal, were lower in prediabetic NOD mice. Analysis of potential upstream regulators of these changes identified increased PTGER4 signal and decreased IRF7. PTGER4 binds prostaglandin E2 (PGE2) and mediates specific aspects of both inflammatory and regulatory eicosanoid signaling. Interestingly, NOD mice treated with an inhibitor of PTGER4 displayed increased IFNAR signaling in DCs, but decreased IL-1, Nlrp3 and IFN-gamma production. These changes suggest restoration of cross-talk between key inflammatory mediators, namely that IFN-gamma and IL-1 can inhibit type 1 IFN and vice versa. Furthermore, mice treated with the PTGER4 antagonist display reduced autoimmune pathology, namely an increase in the number and percentage of healthy islets. These findings highlight PTGER4 as a potential target in type 1 diabetes, affecting several inflammatory pathways including IFNAR signaling.