bluuebunny/crossref_metadata_2025_split huggingface dataset:
Equine bone marrow mesenchymal stromal cells inhibit reactive oxygen species production by neutrophils without affecting other important microbicidal functions
Authors: Henriquez Claudio , Espinosa Gabriel , Schenffeldt Andres , Alarcon Pablo , Gajardo Gonzalo , Plaza Anita , Caffi Valeria , Uberti Benjamin , Moran Gabriel 
Published: January 2023

Abstract
Neutrophils (PMN) are the largest leukocyte population in the blood of most mammals including horses, playing an important defensive role in many infectious diseases. However, the mechanisms that increase PMN as one of the main cellular subsets in the defense against pathogens, could also be involved in the pathophysiology of dysregulated inflammatory conditions. Mesenchymal stem/stromal cells (MSCs) are a heterogeneous population with a modulatory potential on the inflammatory response, and are known to interact with nearly all cells of the immune system including PMN. In this study, we investigate the in vitro modulation of equine bone marrow-derived MSCs on phagocytosis, ROS production, and NETs generation from equine PMN. We found that in co-culture with MSCs, unstimulated PMN produces less ROS (2.88% ± 1.43) than PMN in single culture (5.89% ± 2.63); interestingly, co-cultured PMN remains conditioned to produce fewer ROS after PMA stimulation in comparison to PMN not cultured with MSCs. We also found that inhibition of ROS production by co-cultured PMN occurred not only after prolonged PMN-MSC co-incubation (4 hours), but also when MSC were present in the culture at the moment of adding the stimuli (30% less than control), suggesting that MSCs don’t only modulate PMN functions prior to their arrival to the inflammatory site, but also at the inflammatory niche itself. Additionally, we found that incubation with MSC supernatant strongly inhibited ROS production (83% ± 6.35 less than control) without affecting phagocytosis or capacity for NETosis. These results support the potential use of MSCs in equine inflammatory conditions where neutrophils are the main effector inflammatory cells
Concomitant effector and LAG-3 regulatory responses of Th1 committed cells in acute influenza
Authors: Dutta Avijit , Huang Ching-Tai , Hung Chen-Yiu , Chen Tse-Ching , Lin Chun-Yen , Lin Yung-Chang , Chang Chia-Shiang , Chen Ting-An , Huang Yu-Lin 
Published: January 2023

Abstract
T-bet induced CD4+ T cells are committed to be IFN-γ competent Th1 cells. The Th1 response is essential for virus eradication, but is also a major cause of inflammation and disease aggravation in severe influenza. In our influenza hemagglutinin (HA) Ag-specific mouse experimental system, naïve CD4+ T cells respond to the infection with T-bet expression. However, they suppress cognate CD8+ T cell effector response-mediated inflammation and alleviate the disease. Here we report concomitant LAG-3 expression and IFN-γ competence in Th1 committed CD4+ T cells. LAG-3 expression increases with activation by virus as revealed by CFSE based cell division study. The Th1 committed cells with moderate LAG-3 expression produce IFN-γ and contribute to inflammation and aggravated disease. They can no longer produce IFN-γ with higher LAG-3 expression upon further activation. Th1 committed CD4+ T cells with high LAG-3 expression suppress cognate CD8+ T cells by decoupling their effector function from proliferative response, restrain lung inflammation and facilitate viral clearance. They are better than Foxp-3+HA-specific CD4+ T cells in disease amelioration. The Foxp-3+ cells suppress both effector function and proliferative response of CD8+ T cells. They moderate inflammation but impair viral clearance. Principal component analysis of genome-wide gene expression profiles further confirmed the stepwise evolution. Several regulatory function-associated genes such as Lag3, Pdcd1, Tigit, Ctla2a, Nr4a2, Klrc1, Rgs16, Sytl3, Il21, Il1r2 and Dusp4 are up-regulated graduallly as well from LAG-3Neg to LAG-3Med and LAG-3High cells.
Robust local immune response to vaccination with a live-attenuated HSV-1 strain (HSV-1 0ΔNLS) is delayed in mice vaccinated with the parental live virus
Authors: Carr Daniel J J , Gmyrek Grzegorz 
Published: January 2023

Abstract
We have identified a highly efficacious vaccine 0ΔNLS (deficient in the nuclear location signal of ICP0) against herpes simplex virus type 1 (HSV-1) using an experimental ocular mouse model. In this study, a side-by-side analysis of the host immune response was conducted at the site of vaccination (footpad, FP) and the draining (popliteal) lymph node (PLN) of mice vaccinated with 0ΔNLS versus the parental, wild type HSV-1. Both viruses replicated locally with near equal efficiency through day 3 post infection (PI). By day 5 PI, 0ΔNLS titers recovered from the FP of mice were significantly reduced in comparison to the parental virus and this reduction carried through to clearance by day 9 PI. Even though viral titers were equivalent early post vaccination, 0ΔNLS vaccinated mice generated a more robust localized immune response in the FP in terms of a noticeable increase in the infiltration of myeloid cells and an increase in select cytokines/chemokines including G-CSF, CXCL1, CXCL9, CXCL10, CCL2, CCL5, IL-6, and TNF-α. However, at later time points (days 3–7 PI) the levels of select cytokines/chemokines were elevated in the wild type virus-infected FP with a significant drop in levels from the 0ΔNLS-infected FP. A similar profile was found in the PLN as that reported in the FP. In addition, there were significantly more germinal center B cells (CD19+GL-7+CD95+) and B cells that had undergone isotype switching in the PLN of 0ΔNLS vaccinated mice compared to mice vaccinated with wild type virus by day 7 PI. Collectively, there are unique differences in the host innate and adaptive immune response comparing mice vaccinated with the 0ΔNLS vaccine compared to those exposed to parental wild type virus that continues to be studied in this lab.
Serum IL-6 level may differentiate immune inactive phase from other phases in patients with chronic hepatitis B virus infection
Authors: LEE HYUN WOONG , Oh In Soo , CHANG HYE YOUNG , LEE JUNG IL , LEE KWAN SIK 
Published: January 2023

Abstract

Background and Aims
Previous studies have found that serum level of IL-6 was increased in HBV-infected patients. Here we evaluate the impact of IL-6 levels according to the natural history of chronic hepatitis B(CHB) infection.

Method
Patients(n=71) with hepatitis B virus(HBV) were enrolled between September 2016 and Feb. 2018. Treatment-naïve chronic HBV carries were recruited. IFN-γ, TNF-α, IL-2, IL-6, and IL-17A were measured by a cytometric bead assay according to manufacturer’s instructions. Soluble PD-1 and soluble CD14 were made by the enzyme linked immunosorbent assay (ELISA) technique. The assay procedures were followed according to the manufacturer’s instructions. We studied patients with immune inactive phase (HBV DNA&lt; 1,000 copies/ml, anti-HBe + and normal ALT, n=22), immune tolerant phase (HBV DNA &gt; 10,000 copies/ml, HBeAg + and normal ALT, n=36), and immune active phase (elevated ALT and HBV DNA&gt;10,000 copies/ml, n=13).

Results
Serum IL-6 levels in patients with immune inactive phase were significantly lower than patients with immune tolerant and immune active phases (267.3±245.6 pg/mL vs. 620.2±207.8 pg/mL and 1,815±803.2) (p&lt;0.001). Serum TNF-α levels in patients with immune inactive phase was slightly lower than patients with immune tolerant and immune active phases (3.36±4.40 pg/mL vs. 8.74±12.91 pg/mL and 111.9±377.4) (p=0.079). However, other cytokines (serum IL-2, IL-17A, IFN-γ), soluble CD 14, and soluble PD-1 levels might not be significantly related to the natural history of CHB phase.

Conclusion
These results demonstrate the potential to apply serum IL-6 as a biomarker for differentiating immune inactive phase from other phases in patients with chronic HBV infection.
Innate immune responses to Rift Valley fever virus in the brain
Authors: Weilhammer Dina R , Bourguet Feliza , Sanchez Kristina , Phillips Ashlee , Sebastian Aimy , Hum Nicholas 
Published: January 2023

Abstract
Rift Valley Fever Virus (RVFV) is a category A biodefense pathogen due to the potential for intentional spread by aerosol and the lack of any licensed vaccine or antiviral therapeutics. Despite the brain being a major target of viral replication and tissue damage following aerosol infection, little is known about innate immune responses to RVFV in the brain. Thus, we are investigating the responses of resident innate immune cells of the brain to RVFV infection. Microglia and astrocytes mount a robust response to RVFV infection that is dependent upon MAVS and independent of TLR3 and TLR7. MAVS KO mice have significantly higher viral titers in the brain following intranasal infection, but surprisingly many markers of immune activity were similarly upregulated in the brains of WT and KO mice, including infiltration of CD45+ cells, upregulation of inflammatory genes, and cytokine expression. Indeed, many inflammatory and cytokine genes were upregulated more significantly in infected KO brains than in WT. This is in contrast to the in vitro response of microglia and astrocytes, where MAVS KO cells were virtually unresponsive to RVFV infection. RNA-seq analysis of infected brain tissue revealed that key signal transduction pathways are not activated in MAVS KO animals. These results are an important step towards understanding the precise molecular pathways responsible for controlling RVFV infection in the brain, and will be critical to informing the development of vaccines and antiviral therapeutics that are successful in preventing encephalitis caused by RVFV.
IL-36γ regulates neutrophil infiltration and limits neuroinvasion in genital HSV-2 pathogenesis
Authors: Gardner Jameson K , Herbst-Kralovetz Melissa M. 
Published: January 2023

Abstract
Herpes simplex virus 2 is a neurotropic virus that causes a persistent, life-long infection that increases risk for other sexually transmitted infections. The vaginal epithelium is the first line of defense against HSV-2 and secretes immune mediators, including the pro-inflammatory cytokine IL-36γ, to coordinate the immune response. Previously, we showed that IL-36γ treatment promoted polymorphonuclear cell infiltration to the vaginal cavity and protected against lethal HSV-2 challenge. Herein, we extended these findings using flow cytometry to profile the cellular infiltrate. IL-36γ significantly (p &lt; 0.0001) induced transient neutrophil infiltration, but did not impact monocyte or macrophage recruitment. Using IL-36γ−/− mice in a lethal HSV-2 challenge model, we show that neutrophils were significantly (p &lt; 0.0001) depleted at 1- and 2-days post infection, and that mature neutrophil recruitment was impaired. In addition, IL-36γ−/− mice develop genital disease more rapidly (p &lt; 0.0001), have significantly (p &lt; 0.0001) reduced survival time, and display an increased incidence (p &lt; 0.0001) of neurologic disease compared to wild type mice. IL-36γ−/− mice exhibited a significant (p &lt; 0.05) delay in clearance of the virus from the vaginal epithelium, and analysis of virus spread to the spinal cord, bladder, and colon demonstrated that HSV-2 spread significantly (p &lt; 0.05) more quickly in IL-36γ−/− mice. Further, we detected productive HSV-2 infection in the brainstem of IL-36γ−/− mice significantly (p &lt; 0.0001) more frequently than in wild type mice. Ultimately, we demonstrate that IL-36γ is a key regulator of neutrophil recruitment in the vaginal microenvironment and may function to limit HSV-2 neuroinvasion.
Cytomegalovirus infection drives avidity selection of natural killer cells
Authors: Adams Nicholas M , Lumaquin Dianne , Santosa Endi K , Whitlock Benjamin M , Jackson Benjamin T , Le Luduec Jean-Benoît , Sottile Rosa , Weizman Orr-El , Huse Morgan , Hsu Katharine C , Sun Joseph C 
Published: January 2023

Abstract
Adaptive lymphocyte clones with the greatest receptor affinity tend to dominate primary and secondary immune responses. Strength of receptor signal and receptor abundance, along with competition for antigen and cytokines, drive preferential outgrowth of certain clones, shaping the effector and memory pool in selection processes known as affinity and avidity maturation. Natural killer (NK) cells are innate lymphocytes capable of “adaptive” responses following infectious challenges. However, whether NK cells undergo a similar selection process on the basis of their avidity for cognate ligand is not known. Here, we show that NK cells with a broad range of avidities for the mouse cytomegalovirus (MCMV) glycoprotein m157 are initially recruited after virus infection, but those with highest avidity are selected to undergo the greatest clonal expansion to comprise the memory NK cell population. Furthermore, pre-established levels of Ly49H receptor expression within the Ly49H+ NK cell pool dictated the functional contribution of a given NK cell during MCMV infection, with lower avidity NK cells possessing greater capacity for IFN-γ production, and higher avidity NK cells possessing greater capacity for cytotoxicity and adaptive responses. Moreover, we provide evidence for avidity selection also occurring in human NK cells during human CMV (HCMV) infection. These results provide a mechanistic understanding of how heterogeneity in NK cell avidity underlies the diversification of NK cell effector function during a primary antiviral immune response, and how the process of avidity selection may serve to produce the most potent memory NK cell pool.
IL-18 defines exclusive ‘memory-like’ NK cell populations
Authors: Anderko Renee R. , Rinaldo Charles R. , Mailliard Robbie B. 
Published: January 2023

Abstract
While NK cells are well known for their killing effector function, they also play a critical role as immune helper cells, providing innate alarm signals that shape and regulate the adaptive immune response. In chronic HIV-1 infection, an expanded population of CD56dim FcRγ deficient NK cells persists, similar to the ‘memory-like’ FcRγ− NK cell type identified in cytomegalovirus infection. The purpose of this study was to explore the phenotypic and functional relationship between these FcRγ− NK cells and the previously described IL-18-induced ‘memory-like’ NK helper cells in the setting of HIV-1 infection. We utilized HIV-1 seropositive participants of the Multicenter AIDS Cohort Study to measure baseline frequencies of peripheral blood FcRγ− NK cells by flow cytometry. We also treated NK cells for 24h with IL-18 alone, or in combination with IL-12, and assessed the phenotypic and functional impact on the FcRγ− and FcRγ+ subsets by flow cytometry. The frequencies of FcRγ− NK cells varied widely in HIV-1 infected men. IL-18 drove the differentiation of CD25+/CD83+ NK helper cells, which produced IFNγ and strongly diminished CD16 expression in response to IL-12. In contrast, FcRγ− NK cells did not respond to IL-18, illustrated by failure to express CD25 or CD83, to downregulate CD16 expression, and to produce IFNγ. FcRγ deficiency was also characterized by an attenuated capacity to express IL18Rα upon IL-18 and IL-12 co-activation. Our results indicate that ‘memory-like’ NK helper cells and FcRγ− NK cells are exclusive populations and suggest that HIV-1 drives the development of cytokine-exhausted FcRγ− NK cells that are unable to provide proper immune ‘help’ in response to innate activation signals.
Understanding the role of hnRNPA1 on T cell differentiation and function in the Gut
Authors: White Theresa L , Gable Matthew , Jin Ye , Morel Penelope A 
Published: January 2023

Abstract
Regulatory T cells (Treg) play a critical role in preventing autoimmunity diseases and maintaining tolerance in various tissues, particularly the gut. Peripheral derived Tregs (pTregs) are especially crucial in gut homeostasis and tolerance to commensal and food antigens. We have previously shown that knockdown of the RNA-binding protein, heterogeneous nuclear ribonucleoprotein (hnRNP) A1, reduces in vitro pTreg induction upon low TCR stimulation. In addition, we have shown that, under conditions of low TCR stimulation, in which pTreg are induced, hnRNP A1 is phosphorylated by Akt. In order to explore the impact of hnRNP A1 phosphorylation by Akt on pTreg induction in vivo we have generated a mutant mouse model, hnRNPA1-S199 in which the known Akt phosphorylation site on hnRNP A1 is mutated to Alanine. Extensive characterization of immune cells in this mutant mouse revealed no abnormalities in the number or frequency of major immune cell types at steady state. However, we identified abnormalities in the number of Treg in the mesenteric lymph nodes (mLN) in mutant mice, prompting further investigation. Using the Cytex Aurora flow cytometer we developed a 19 color T cell panel to characterize T cells in the mLN, Peyer’s patches and lamina propria. Detailed characterization of the pTreg and tTreg populations in the gut of WT and mutant mice will determine whether the abnormalities observed in the mLN are reflected in different intestinal areas. In addition, we are examining the induction of pTreg using an adoptive transfer model of OT-II TCR transgenic mutant and WT CD4 T cells followed by oral OVA administration. Further studies will focus on the mechanisms by which hnRNP A1 impacts pTreg induction.
Genetics but not intestinal microbiota determines the onset of systemic autoimmune disease in BXD2 mice
Authors: Hong Huixian , Wu Qi , Yang PingAr , Duck Wayne , Morrow Casey D. , Foote Jeremy B. , Schoeb Trenton R. , Elson Charles O. , Hsu Hui-Chen , Mountz John D. 
Published: January 2023

Abstract
The onset and severity of autoimmunity can be attributed to both genetic predisposition and environmental triggers including intestinal microbiota dysbiosis. To determine if development of spontaneous autoimmunity is dependent on gut microbiota, we generated germ-free (GF) BXD2 lupus mice which otherwise develop spontaneous germinal centers (GCs) and high titers of serum autoantibodies. The microbiome and immune phenotypes in specific pathogen free (SPF) and GF BXD2 and normal B6 strains of mice were analyzed. The GF status was confirmed by gut bacterial culture. Surprisingly, the GF status did not affect disease onset and severity in 6-mo-old mice. There were comparable serum levels of IgG or IgM autoantibodies against lupus autoantigens (DNA, histone, and RNP) in SPF versus GF BXD2 mice. Flow cytometry analysis showed that the frequency and absolute numbers of GC B cells, CD4+ T cells, follicular T-helper (Tfh) cells in the spleen were also indistinguishable between SPF and GF BXD2 mice although GF BXD2 mice exhibited a higher number of CD19+ B cells and Tregs, compared to the SPF counterpart. Histologic examination of kidney and lung also showed similar disease scores between SPF and GF BXD2 mice. At older age (12-mo-old), however, there were diminished GC B cells and serum levels of autoantibodies in GF BXD2 mice compared to SPF mice. The results suggest a model in which genetic factors play a dominant role in determining the onset and susceptibility of autoimmunity and that endogenous self-antigens are the primary driver of autoreactive T and B cells leading to spontaneous GCs in BXD2 mice. In contrast, environmental factors such as gut microbiomes may play a modifying role in regulating the persistence of systemic autoimmunity.
Single cell RNA sequence analysis reveals novel clusters unique to post-infectious myocarditis in the mouse model of Coxsackievirus infection
Authors: Reddy Jay , Lasrado Ninaad , Arumugam Rajkumar , Jayaraja Sabarirajan , Thibivilliers Sandra , Libault Marc , Starr Timothy K 
Published: January 2023

Abstract
Myocarditis is a common occurrence in children and adolescents that can lead to dilated cardiomyopathy. Since it is difficult to identify the triggers of this complex disease, mechanistic understanding of DCM pathogenesis may create avenues to develop new treatment strategies. We have been engaged in delineating the autoimmune pathways underlying the development of DCM using various mouse models of myocarditis that resemble human disease. One such model is Coxsackievirus B3 (CVB)-induced myocarditis in A/J mice that show two distinct phases of disease, but occur in continuum. While, the acute, infectious phase (up to ~10 days post-infection) is marked by viremia and infiltrations in the hearts, infectious virions become undetected during the chronic phase (~beyond 18 days post-infection), and animals develop DCM. The latter phenotype can occur over a period of ~60 to 90 days, and hitherto, it has been a challenge to determine the underlying mechanisms. To dissect this complex nature of disease, we recently performed single cell RNA sequence analysis on heart infiltrates obtained from CVB-infected mice. Using panels of 15 to 30 marker genes for various immune and non-immune cells, we identified 18 and 26 clusters respectively in healthy and infected hearts that also include few unknown clusters unique for each group. Likewise, transcriptome analyses led us to identify a total of 14,051 genes to be differentially expressed, and of these, 206 and 345 transcripts were unique to healthy and CVB-infected groups, respectively. The data suggest that uniquely expressed genes in the CVB-infected hearts may have a role in the development of post-infectious myocarditis leading to DCM.
Cardiac myosin-specific TCR transgenic mouse models for studying dilated cardiomyopathy
Authors: Reddy Jay , Massilamany Chandirasegaran , Arumugam Rajkumar , Lasrado Ninaad , Gangaplara Arunkumar , Jayaraja Sabarirajan , Steffen David , Gurumurthy Channabasavaiah 
Published: January 2023

Abstract
We previously reported that the cardiac myosin heavy chain-alpha (Myhc) 334–352 encompasses epitopes for both CD4 and CD8 T cells that can induce autoimmune myocarditis/dilated cardiomyopathy (DCM) in A/J mice expressing the IAk and IEk molecules. More recently, we made an unexpected observation that, Myhc 334–352 although by being a poor binder of IAb molecule in C57Bl/6 mice, could induce strong T cell responses, but myocarditogenicity was severely dented. In our efforts to delineate the roles of Myhc-specific, CD4 and CD8 T cells in the DCM pathogenesis, we have now created the T cell receptor (TCR) transgenic (tg) mice on C57Bl/6 background for Myhc 334–352, and demonstrate that the Myhc 334–352-specific TCR was found common to both CD4 and CD8 T cells. While, the naïve tg T cells do not respond, antigen-primed CD4 and CD8 T cells from immunized mice respond to Myhc 334–352 that produce predominantly, interferon-γ. Furthermore, although mild, the immunized tg mice developed heart lesions containing the mononuclear cells, and such a phenotype was more apparent in females than males. As the tg mice become available on both resistant (C57Bl/6) and susceptible (A/J) backgrounds, our TCR tg system can be used as a helpful tool to dissect the complexities of antigen-specific CD4 and CD8 T cells in the causation of DCM in various experimental settings.