bluuebunny/crossref_metadata_2025_split huggingface dataset:
Heterogenicity of Tissue Distribution Recognized by A panel of Monoclonal Antibodies
Authors: Zhang Hong , Lin Mongshang , Wen Chiaokai , Kim Daniel , Frey Avery , Chronopoulou Efthalia , Yang Xifeng , Ni Jessie 
Published: February 2023

Abstract
ACE2 (Angiotensin Converting Enzyme 2) is the main viral entry point for SARS-CoV and SARS-CoV-2. Human ACE2 is widely expressed on different organs but if virus attached all the organs that highly expressed ACE2 is not fully studied. Most recent finding showed that a novel short isoform of ACE2 expressed in the airway epithelium is substantially upregulated in response to interferon stimulation, but not SARS-CoV-2 infection due to this short isoform lacking SARS-CoV-2 spike high-affinity binding sites. This arouses our interest in whether ACE2 has more isoforms expressed in different tissues that directly affect different organs response to virus infection. To support understanding ACE2 expression in response to viral infection, we generated human ACE2 monoclonal antibody by immunizing rat with recombinant human ACE2 protein covering extracellular domain (Gln18-Ser740). We found that there is discrepancy in different clones on different tissues. After three rounds subcloning, four clones have been selected based on chromosomally stable IgG producing hybridoma cells. Our data showed that all four clones showed equivalent signals on ACE2 transfectants but staining signals varied on tubules of kidney and seminiferous tubules and leyding cells on testis, two clones showed medium level staining signals on intestine and colon, but only one clone showed positive staining signals on spleen vascular and red pulp sinus endothelium. In addition, different neutralization potency and Western blot detection performance were observed among the collections of clones. Our data indicated that different clones might recognize different epitopes of ACE2 and indirectly suggested that ACE2 might have different isoforms in different tissues.
Chronic air pollution exposure exaggerates obesity-induced tissue inflammation and insulin resistance by activating inflammasome
Authors: Rao Xiaoquan , Zhong Jixin 
Published: February 2023

Abstract

Background:
Air pollution has been associated with cardiometabolic diseases such as obesity and insulin resistance. However, the underlying mechanisms were not fully understood. In this study, we examined the role of NLRP3 inflammasome in chronic air pollution-induced tissue inflammation and insulin resistance.

Methods:
8 weeks old male WT and NLRP3−/− mice were exposed to filtered air (FA) or concentrated ambient particles (CAPS) in a small animal whole-body exposure system with or without high fat diet for 6 hours/day, 5 days/week, for a total of 12 weeks (n= 5–8 per group).

Results:
CAPS exposure enhanced insulin resistance in the WT mice but not in the NLRP3−/− mice. Cytokine array and ELISA assays indicated that IL-1β secretion was increased in the culture supernatant of macrophages isolated from CAPS-exposed WT mice, compared to that of FA-exposed mice. Electronic microscopy showed particulate deposites in the peritoneal macrophages isolated from the peritoneal cavity of CAPS-exposed animals. The gene expression of Nlrp3 and Il-1β, the rate-limiting factor of NLRP3 inflammasome activation, were also increased in CAPS-exposed WT mice. Deficiency of NLRP3 blocked CAPS exposure-induced lung inflammation, inflammatory gene expression, and IL-1β secretion in mice. Subsequent ex vivo experiments by culturing macrophages with particulate matters indicate that air pollution particles not only initiate the transcription of Nlrp3 and Il-1β, the first signal for inflammasome activation, but also induce the activation of caspase-1, the second signal for inflammasome activation.

Conclusions:
Our data suggest that NLRP3 inflammasome activation mediates air pollution-induced tissue inflammation and insulin resistance.
Controlling for Confounding Factors when Diagnosing Cancer from T Cell Receptor Repertoires
Authors: Ostmeyer Jared L , Christley Scott , Reuben Alexandre , Zhang Jianjun , Cowell Lindsay 
Published: February 2023

Abstract
Our publications have shown that T cell receptor (TCR) repertoires sequenced from tissue biopsies can be used to distinguish tumor from control tissue with &gt; 90% classification accuracies, suggesting that TCR repertoires can be used to diagnose cancer.
TCR repertoires sequenced from blood samples of cancer patients are now publicly available, and it is tempting to use these existing samples to develop blood-based assays for diagnosing cancer. Already, research groups including us obtained promising results. However, these results are obtained by combining cancer and control samples from mismatching studies. Without controlling for age, sex, race, and even the reduced amount of blood volume collected from cancer patients relative to controls, these exciting results could merely be an artifact of confounding factors.
In this study, we control for possible confounding factors and observe that our ability to diagnose cancer from TCR repertoires sequenced from blood does not vanish. On patient-holdout cross-validations, we achieve diagnostic accuracies of 90% for colorectal cancer (controlling for depth coverage), 84% for breast cancer (controlling for depth coverage and age), and 83% for lung cancer (controlling for age, sex, race, and depth coverage).
While it is impossible to control for all potential confounding factors like variations in geographic locations, these results are encouraging. We propose future studies that (i) collect matching controls and cases and (ii) confirm TCR repertoire sequencing provides a distinct signal for diagnosing cancer independent of other blood-based assays and therefore can improve the diagnostic performance of these other blood-based assays.
Near infrared light reduces neutrophil chemokine production during ischemia-reperfusion injury
Authors: Compton Tyler Compton , Weihrauch Dorothee , Ninomiya James T , Lochhead Robert 
Published: February 2023

Abstract

Background:
Ischemia reperfusion injury and tissue necrosis commonly follows administration of a tourniquet. Previously, we have shown that exposure to non-thermal infrared (NIR) light decreases neutrophil and macrophage mediated inflammation and tissue necrosis in a mouse hindlimb model. This study tests whether exposure to NIR light during a period of ischemia limits tissue damage by reducing secretion of chemoattractant proteins and phenotypic switching of tissue macrophages from a pro-inflammatory (M1) to a wound healing (M2) phenotype.
Methods and Results: Mouse hindlimbs received 3 hours of tourniquet-induced ischemia followed by a period of reperfusion with or without 15 or 30 minutes of NIR exposure. Hindlimbs receiving injury and NIR treatment showed a significant decrease in the neutrophil chemoattractants CXCL1 and CXCL5 by ELISA, compared with mice receiving ischemia reperfusion injury alone and control mice receiving no injury or NIR. Protein isolated from mouse hindlimbs receiving NIR treatment and injury had a significant decrease in M1 macrophage marker CD68 and significant increase in M2 macrophage marker CD206, normalized to CD14, compared with mice receiving injury alone and uninjured mice.

Conclusion:
We have shown that NIR light affects neutrophil chemokine secretion and tissue macrophage polarization in a murine hindlimb ischemia and reperfusion model. These data indicate that NIR light treatment during tourniquet administration limits inflammatory responses, reduces tissue necrosis, and promotes wound healing following removal of the tourniquet. Thus, NIR treatment may improve limb preservation following vascular injury and inform timing of tourniquet application during surgical procedures.
Itolizumab-induced antigenic modulation of CD6 inhibits T cell activity
Authors: Chu Dalena Ngoc , Ampudia Jeanette , Connelly Stephen , Ng Cherie 
Published: February 2023

Abstract
CD6 is an immunomodulatory receptor on T cells that promotes immune synapse formation, T-cell activation, and cell migration into tissues by binding activated leukocyte cell adhesion molecule (ALCAM). Excessive activation through the CD6-ALCAM pathway has been implicated in the pathogenesis of multiple autoimmune and inflammatory diseases. Hence, the ability to modulate the level of activation is beneficial to disease resolution. Itolizumab is a humanized anti-CD6 monoclonal antibody that is specific for the membrane distal domain 1 of CD6. Previously, itolizumab was believed to sterically hinder the CD6-ALCAM interaction, thereby blocking T-cell co-stimulation. Here we describe antigenic modulation as an additional mechanism whereby binding of itolizumab to CD6 induces proteolytic cleavage of the extracellular portion of CD6. Upon treatment with itolizumab, surface levels of CD6 decreased in a dose- and time-dependent manner, as monitored by flow cytometry using a noncompetitive anti-CD6 monoclonal antibody. This loss was inhibited in the presence of protease inhibitors and the decrease in surface levels of CD6 was accompanied by an increase in levels of soluble CD6 in the supernatant. Furthermore, we assessed the effect of surface levels of CD6 on the response of cells to T cell stimulation in the presence of ALCAM. CD6low cells showed reduced T-cell activity compared to CD6highcells as measured by cell surface activation makers, including CD25, CD69 and PD-1, and cytokine production. These findings demonstrate that CD6 is an important regulator of T-cell activity and that modulating surface levels of CD6 is an effective method for fine-tuning the activity of T-cells.
MLL1-COMPASS establishes developmental patterning of the memory CD8 T cell compartment and ensures regenerative immunity.
Authors: Getzler Adam , Frederick Megan A. , Diao Huitian , Milner Justin , Belanger Simon , Tsuda Shanel , Kim Jihye , Crotty Shane , Goldrath Ananda W , Pipkin Matthew E. 
Published: February 2023

Abstract
CD8+ T cells with stem cell-like qualities (TSTEM) provide central memory following resolved intracellular infections, and serve as progenitor cells that sustain effector-like cell development during chronic infections and tumors, both naturally and in response to immunotherapeutic PD-1 blockade. However, the chromatin regulatory factors (CRFs) and transcriptional programs that establish TSTEM cell differentiation are unclear. Using an RNA interference screen of all CRFs we discovered that Mixed Lineage Leukemia 1 (MLL1) was required in activated CD8 T cells to promote Id3-GFP reporter expression and to coordinately restrain PD-1, LAG3 and TIM3 expression. Single cell RNA-seq analysis demonstrated Kmt2a (MLL1) was preferentially expressed in P14 CD8 T cells enriched with TSTEM gene expression during lymphocytic choriomeningitis virus (LCMV) infection. P14 CD8 T cells depleted of MLL1 failed to establish TSTEM-like cells, terminally differentiated prematurely, failed to sustain responses to chronic LCMV infection and tumors, and failed to develop central memory subsets during acute LCMV infection. MLL1 preferentially bound transcriptional start sites (TSSs) in CD8 T cells, and was essential for global accumulation of H3K4me3, H4K16ac, and RNA Polymerase II CTD-phosphorylation during TCR stimulation. These results suggest that MLL1-COMPASS organizes TSS activity to establish transcriptional programs that delineate TSTEM and terminal differentiation.
Candida albicans-induced EphA2 signaling promotes kidney injury and sepsis during invasive candidiasis
Authors: Millet Nicolas , Solis Norma , Swidergall Marc 
Published: February 2023

Abstract
Pattern recognition receptors (PRRs) play key roles in fungal recognition and initiation of host immune responses. Recently, we identified that the ephrin type-A receptor 2 (EphA2) is a key β-glucan receptor on oral epithelial cells and neutrophils which orchestrates innate immune responses thereby preventing oral Candida infection. However, the contribution of this PRR in the pathogenesis of invasive candidiasis (IC) remains unknown. Although no differences in renal fungal burden could be observed, EphA2-deficient mice were more resistant to C. albicans infection suggesting that EphA2 deficiency enhances renal host tolerance to lethal fungal challenge. By testing renal cytokines, serum NGAL, a marker of renal injury, and the sepsis marker TREM-1, we found that EphA2 deficiency altered the pro- and anti-inflammatory response, reduced renal injury, and sepsis during IC. Epha2−/− mice decreased neutrophil and monocyte recruitment during IC. However, dendritic cells lacking EphA2 migrated more efficient to infected tissue. Since EphA2 is highly expressed on both, hematopoietic and stromal cells, we generated bone marrow chimeric mice. Both Epha2+/+ mice reconstituted with Epha2−/− BM and Epha2−/− mice reconstituted with Epha2+/+ BM were more resistant to IC compared to Epha2+/+ mice reconstituted with Epha2+/+BM. However, these chimeric mice were more susceptible than Epha2−/− mice reconstituted with Epha2−/− BM suggesting that EphA2 deficiency within cells of both compartments, the hematopoietic and the stromal, is required for full protection against IC. Collectively, our data highlight a detrimental role of EphA2 in promoting kidney injury and sepsis during lethal fungal infection.
Harnessing Regulatory T cells to Suppress Pathological Immune Response during Cryptococcus neoformans Meningoencephalitis
Authors: Xu Jintao , Goughenour Kristie , Zhao Ziyin , Olszewski Michal A 
Published: February 2023

Abstract
Central nervous systems (CNS) infections are marked by high mortality or frequently lasting neurologic disorders in the survivors. Cryptococcus neoformans meningoencephalitis (CM) is one of the leading causes of infectious neurological death. Many CM patients develop severe neurological symptoms despite evidence of ongoing immune response and fungal eradication, suggesting a dysregulated pathological host response. Our objective is to test whether and how regulatory T cells (Treg) maintain immune balance in CM and whether they could be used therapeutically to limit inflammatory CNS damage.
Using a murine CM model, we found Foxp3+ Treg massively accumulate in the infected brain. They acquire a Th1-like phenotype, expressing transcription factor T-bet, IFN-γ, and chemokine receptor CXCR3. This phenotype is critical for Treg accumulation in the CM brain. Treg depletion using anti-CD25 antibody results in deteriorated neurological symptoms and increased mouse mortality. Mechanistically, we found that brain-infiltrating Treg express immune suppressive molecules, such as IL-10 and tissue repair molecules, such as Amphiregulin compared to effector T cells. Single-cell RNA seq analysis showed broad cross-talk between brain Tregs with infiltrating T cell, monocyte-derived cells, and brain resident glial cells. Treg depletion resulted in increased activation of pathological T cells and monocyte-derived cells, promoting brain immunopathology during CM. Selectively enhancing Treg through low-dose IL-2 treatment improves mice survival and neurological function readouts. Taken together, our findings suggest that Treg limit pathological CNS inflammation and may provide the therapeutic opportunity for CM patients.
Characterization of beta2-adrenergic receptor homozygous knockout stress mouse model during Chlamydia muridarum genital infection
Authors: Belay Tesfaye 
Published: February 2023

Abstract
The relationship of stress in chlamydia genital disease pathogenesis and its influence on the host immune response remains unknown. Beta2-adrenergic receptor(β2-AR), the major receptor of the stress hormone, norepinephrine (NE), is known to impair the function of immune cells. This study was undertaken to determine whether cold-induced stress (CIS) results in differential weight loss, stress hormone level production, and gene expression of β-AR subtypes in the β2-AR knockout (KO) and wildtype (WT) C57BL/J6 mice. CIS was induced by immersing mice in cold water for 5 minutes daily for 21 days, as per previously established protocols in our lab. CIS in 21 days resulted in average body or spleen weight loss of 1.88 or 0.018 gm, respectively, to β2-AR KO and 1.7 or 0.015 gm to WT C57BL/6J compared to their respective non-stressed group. Furthermore, β2-AR deficiency lead to a significant enhancement in C. muridarum shedding from days 9–15 following intravaginal infection, but no statistically significant difference between the stressed (4000 IFU/mL) and non-stressed (4200 IFU/ml) of β2-AR KO was obtained. β2-AR OK and WT stressed mice showed about 65–70 ng/mL production of NE; however, the intensity of C. muridarum infection was low in the KO than the WT (p &lt;0.05). Relative fold-change in mRNA in splenic CD4+ T cells of stressed β2-AR OK was non-detectable. However, a slight upregulation of β1- and β3-AR was observed primarily in CD4+ T cells of β2-AR KO mice. Overall, the β2-AR and the WT C57BL/6J are related in many ways, and we predict to have similar physiological conditions suitable to compare pathogenies. Immunity, histopathology, and infertility.
The purinergic receptor P2RX7 promotes the accumulation of effector CD4+ T cells in the lung parenchyma during hypervirulent mycobacterial infection
Authors: Carvalho Igor Santiao , Bonfim Caio César Barbosa , de Almeida Santos Gislane , Melo Bruno M , Hirata Mario H , Alves-Filho José Carlos C , Alvarez José M , Lassounskaia Elena L , Silva Henrique Borges da , D‘Império-Lima Maria Regina 
Published: February 2023

Abstract
CD4+ T cells are crucial to control pulmonary tuberculosis, but also favor uncontrolled inflammation in severe disease. Effective CD4+ T cell response to tuberculosis is related to tissue location: Lung-resident CD4+ T cells display higher effector function in comparison to circulating cells. Among the factors favoring T cell residency in non-lymphoid tissues, extracellular ATP sensing via P2RX7 is crucial for the generation of virus-specific resident memory CD8+ T cells. Yet, its role in CD4+ T cell non-lymphoid tissue residency has not been defined. Moreover, mycobacterial infection induces high extracellular ATP levels due to lung tissue damage, which is increased in response to hypervirulent strains. Here, we used P2rx7 ablation models to test the role of P2RX7 in CD4+T cell function in tuberculosis. In response to the hypervirulent Mycobacterium bovis MP287/03 strain, lung-resident WT CD4+ T cells express higher P2RX7 levels compared to circulating CD4+ T cells. Compared to WT mice, P2RX7-deficient (P2rx7−/−) mice display reduced numbers of antigen-specific (I-AbESAT-64-17) and IFN-γ-producing CD4+ T cells in the lung parenchyma. Using cell adoptive transfers, while recipient CD4-deficient mice transferred with CD4+ WT T cells are protected, mice that received CD4+P2rx7−/− T cells develop severe tuberculosis. Mechanistically, CD4+P2rx7−/− T cells have poorer accumulation and IFN-γ production in the lung parenchyma; moreover, they show reduced proliferation in situ. This defect in proliferation is exclusively observed in the lung, with no differences observed in draining lymph nodes. Together, our results show that P2RX7 promote the accumulation of protective CD4+ T cells in the lung during severe tuberculosis.
Mucosal LTA1 Elicits Lung Th17 cells with reduced plasticity towards Th1 cells and resistance to FK506.
Authors: Yang Haoran , Iwanaga Naoki , Norton Elizabeth B. , Kolls Jay K 
Published: February 2023

Abstract

Objective:
Based on our previous work, we identified a vaccine composed of the A1 domain of heat-labile toxin from E. coli (LTA1) adjuvant and Outer membrane protein (Omp) X from K2 strain of Klebsiella pneumoniae (K. pneumoniae), which can elicit serotype independent protection against K1 strain by generating lung Th1 and Th17 cells. To better understand the characteristics of these Th17 cells, we investigated the TCR repertoire, plasticity, and resistance to FK506.

Methods:
Naïve or immunized lung cells were collected and stained with Anti-Mouse TCR Vβ Screening Panel for FACS. Naïve CD4+ cells were isolated from spleens and cultured under Th17 differentiation conditions in vitro. Immunized lung Th17 cells were purified. To assess plasticity towards Th1, cells were cultured w/wo IL-12 and analyzed by FACS.
To assess resistance to calcineurin inhibitors, the naïve/immunized CD4+ cells were treated w/wo FK506 both in vivo and ex vivo, analyzed by FACS.
Results and Conclusions: The LTA1-OmpX elicited CD4+ cells were oligoclonal with skewing towards TCRVβ14+ cells.
After incubation with IL-12, 57.73 ± 3.9% of the in vitro differentiated Th17 cells showed plasticity towards Th1 cells whereas in immunized lung Th17 cells only showed 6.25 ± 1.6%. Lung Th17 cells also showed substantially reduced STAT4 phosphorylation and surface IL-12rβ2 expression compared to in vitro Th17 cells.
Furthermore, lung Th17 cell IL-17A expression was resistant to FK506 treatment both in vivo and ex vivo, which potentially indicated that this vaccine strategy may confer protection in the setting of transplant immunosuppression.
Complex immunogenomic control of serum thyroid hormone levels in fetuses and piglets challenged with Porcine Reproductive and Respiratory Syndrome Virus
Authors: Van Goor Angelica G , Pasternak Alex , Walugembe Muhammed , Hamonic Glenn , Dekkers Jack , Harding John C. S. , Lunney Joan K 
Published: February 2023

Abstract
Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) causes respiratory disease in piglets and reproductive disease in sows. However, the interaction between host immunity and hormone homeostasis is not completely understood but piglet and fetal serum thyroid hormone (i.e., T3 and T4) levels decrease rapidly in response to PRRSV infection. Our objective was to estimate genetic parameters, identify QTL, and uncover nearby candidate genes for T3 and/or T4 levels of piglets and fetuses challenged with PRRSV2. Serum from 5-week old pigs (N=1792) at 11 days post infection (dpi) with PRRSV2 were assayed for T3 levels (piglet_T3). While serum from fetuses (N=1267) at 12 or 21 dpi from sows (N=150) challenged with PRRSV2 in late gestation were assayed for T3 (fetal_T3) and T4 (fetal_T4) levels. Animals were genotyped for 60K or 650K SNPs, heritabilities were estimated and genome wide association studies were performed for each trait separately. We found all three traits to be moderately heritable (18–28%) and the significant QTL (N=37) across several chromosomes collectively explained 24–34% of the genetic variation. A shared QTL identified for all traits on chromosome 6 contained the IL12RB2 and IFI44 genes. The largest QTL identified for both piglet_T3 and fetal_T3 on chromosome 5 contained IL17RA. A large QTL identified for piglet_T3 on chromosome 7 contained 9 MHC genes (e.g., SLA-DMA, SLA-DOA, SLA-DQA, SLA-DRA, and MHC-IA7). In conclusion, our data support the concept that there are complex interactions between thyroid hormone levels and multiple immunogenetic pathways during PRRSV infection, and that there is potential to select for pleiotropic QTL to simultaneously improve host immunity for reproductive and respiratory PRRS.