bluuebunny/crossref_metadata_2025_split huggingface dataset:
A high-sensitivity detection system for monitoring of rare antigen specific naïve and memory T helper cells (58.6)
Authors: Bacher Petra , Schink Christian , Teutschbein Janka , Kniemeyer Olaf , Assenmacher Mario , Brakhage Axel , Schmitz Jürgen , Scheffold Alexander 
Published: January 2023

Abstract
CD4+ T cells play a central role in the immune defense against pathogens or tumors but also in immunopathology, such as autoimmunity or allergy. However, due to technical limitations, comprehensive information about the frequency and phenotype of antigen-specific CD4+ T cells against many disease-relevant antigens is missing. In particular the nature of the human naïve repertoire is elusive. We have developed a new, sensitive detection system for antigen-specific T helper cells, based on a combination of magnetic pre-enrichment and multi-parametric flow cytometric analysis of CD154 (CD40L)-expressing CD4+ T cells upon in vitro activation with antigens. The high sensitivity of 1 cell out of 105 to 106 enables the direct enumeration of memory and naïve CD4+ T cells reactive against basically any recall or primary antigen. Importantly, our technology allowed us to rapidly analyze sufficient numbers of target cells to discriminate even small functional or phenotypic subpopulations within the total antigen-specific T cell pool, undetectable by conventional methods. We used this assay for a first characterization of tumor- and auto-antigen reactive CD4+ T cells from healthy subjects as well as for the in depth characterization of rare pathogen specific T cells. The possibility to visualize T cells reactive against any antigen of interest even in the naïve repertoire will open up new possibilities to advance human T cell research and to improve clinical diagnostics and prognosis.
Early detection of lymphangiogenesis by near-infrared imaging and responses to VEGF-C in a mouse full-thickness wound model (58.9)
Authors: Hall Mary , Aldrich Melissa , Robinson Holly , Kwon Sunkuk , Sevick-Muraca Eva 
Published: January 2023

Abstract
Wound healing is a multistep process involving interaction of various cell types, mediators such as cytokines, inflammation, and generation of new tissue. Both the blood and lymphatic vasculature aid in tissue repair, during which new vessels are generated by angiogenesis and lymphangiogenesis, respectively. We utilized a mouse full-thickness wound model to detect lymphangiogenesis near an incisional wound by near-infrared fluorescence (NIRF) imaging at multiple time points post-incision (PI). In addition, the systemic affects of locally administered vascular endothelial growth factor (VEGF)-C were investigated by measuring peripheral blood cytokine levels in treated and untreated animals. Lymphangiogenesis was detected at the wound site by NIRF imaging by day 7 or 8 PI and confirmed by intravital imaging, while angiogenesis was observed by day 2 or 3 PI. Interleukin (IL)-1α and IL-3 levels were significantly increased at day 7 and 21, respectively, relative to pre-incision levels found in the periphery of VEGF-C-treated mice. Also, regulated-upon-activation, normal T-cell expressed and secreted (RANTES) levels were significantly higher in VEGF-C-treated animals when compared to saline-treated mice at day 21 PI. Our findings demonstrate that NIRF imaging can be utilized to detect early lymphangiogenesis during wound healing, and local VEGF-C administration may help induce this process via increasing systemic levels of wound-healing mediators.
Neutrophil Gelatinase Associated Lipocalin (NGAL) deficiency accelerates the onset of serum autoantibodies in pristane induced lupus (60.10)
Authors: Pawar Rahul , Goilav Beatrice , Xia Yumin , Zuang Haoyang , Reeves Westley , Putterman Chaim 
Published: January 2023

Abstract
NGAL (lipocalin-2), a member of the lipocalin superfamily, is expressed by neutrophils, hepatocytes, and renal resident cells following ischemia or inflammatory conditions & is involved in innate immune responses during infection. However, its role in adaptive immunity has not been described. Previously, we had found that NGAL is instrumental in the pathogenesis of antibody mediated nephritis. To investigate a possible role for NGAL in the generation of autoreactive antibody responses, we examined its effect in an experimental lupus model. We injected pristane (0.5 ml per mouse i.p.) into wild type B6 (n=10) and NGAL-deficient (NGAL-/-) (n=10) mice and followed the course of disease. Analyzing the levels of serum autoantibodies at 3 months post injection, NGAL-/- mice had significant increases in IgG2a (p=0.007) and IgG2b (p=0.019) anti-double stranded DNA antibodies, and IgG2a (p=0.002) & IgG2b (p=0.010) anti-single stranded DNA antibodies, and IgG anti-histone antibodies (p=0.025) as compared to pristane challenged, NGAL sufficient mice. Immunoprecipitation studies showed elevated serum anti-Sm/RNP antibodies in NGAL-/- mice compared to the WT mice at 3 months post injection, with an increase in titer and epitope diversity of this response at 6 months post injection. In conclusion, NGAL delays the onset of anti-nuclear antibodies in pristane induced lupus. The mechanisms by which NGAL plays a protective role in humoral autoimmunity are under investigation.
IRF7-dependent IFNβ production in response to RANK facilitates medullary thymic epithelial cell development. (60.3)
Authors: Otero Dennis , Baker Darren , David Michael 
Published: January 2023

Abstract
The signal transducer and activator of transcription 1 (STAT1) is an essential part of signaling cascades induced by type I and II interferons. Deficiency of STAT1 results in heightened susceptibility to infections and autoimmunity in mouse and humans. While the role of IFNγ in T cell development is well known, the role of type I interferons in thymic development has largely remained unexplored. Here we show that STAT1 and IFNAR-deficient mice harbor a defect in medullary compartment characterized by a significant reduction in self-antigen presenting, AIRE expressing medullary thymic epithelial cells. Using a novel reporter mouse we demonstrate that constitutive IFNAR signaling is taking place in the thymic medulla in the absence of any infection or inflammatory signals. In addition, we found that RANK stimulation in thymic stromal cells results in the upregulation of IFNβ which in turn is inhibits RANK signaling thereby facilitating AIRE expression. Finally, we show that IRF7 is required for the induction of IFNβ by RANKL, and that IRF7-, but not IRF3-deficient mice display a defect in thymic architecture and medullary thymic epithelial cell differentiation similar to that observed in IFNAR- or STAT1-deficient animals. These results illustrate that a spatially and temporally coordinated crosstalk between the RANKL/RANK signaling pathway and the IRF7/IFNβ/IFNAR/STAT1 axis is essential for the differentiation of AIRE-expressing medullary thymic epithelial cells.
p21 selectively regulates the activation of autoimmune of lpr memory T cells and attenuates autoimmunity (60.5)
Authors: Weber Kathrin , Vázquez-Mateo Cristina , Daszkiewicz Lidia , Balomenos Dimitrios 
Published: January 2023

Abstract
Fas drives activated T cell apoptosis in vitro, but lymphadenopathy and lupus in Fas-deficient (lpr) mice cannot be explained only on the basis of the lpr T cell apoptotic defect. Unexplained also remains the hyperproliferation of T cells in lpr mice, including CD4+, CD8+ and CD4-CD8-TCR+ cells. We investigate these questions by generating T cell-specific p21-transgenic and p21-/- BL/6 lpr mice. p21, a cell cycle inhibitor and a lupus-like autoimmunity suppressor, regulates memory T cell but not naïve T cell expansion. p21 overexpression in lpr T cells did not affect primary T cell responses, but profoundly diminished lpr T cell expansion upon repeated activation, without affecting the apoptosis defect of lpr T cells. Importantly, p21 overexpression restrains lpr T cell memory expansion and IFN-gamma and IL-17 production, resulting in reduced autoantibody production, glomerulonephritis and lymphadenopathy in lpr mice. In p21-/- lpr mice, lack of p21 enhanced the activation and accumulation of memory T cells, which produced higher levels of IFN-gamma and IL-17. Lack of p21 led to severe glomerulonephritis and death in BL/6 lpr mice, which normally develop non-lethal mild autoimmunity. Overall, we conclude that independently of their lpr apoptotic defect, T cell hyperactivation and autoimmunity development can be controlled by p21. Thus, analysis of the role of p21 in T cell immunity may be of therapeutic value for both Lupus and the Autoimmune Lymphoproliferative Syndrome.
Stressor-enhanced colitis is abrogated in mice deficient in CCL2 (61.6)
Authors: Mackos Amy , Galley Jeffrey , Parry Nicola , Eubank Tim , Bailey Michael 
Published: January 2023

Abstract
Exposing mice to a social stressor called social disruption (SDR) has been reported to increase CD11b+Ly-6ChiCCR2+ inflammatory monocytes in the circulation and spleen. This study was designed to test the hypothesis that stressor-induced exacerbation of pathogen-induced colitis is dependent upon CCL2-mediated recruitment of inflammatory monocytes. Mice deficient in CCL2 (i.e., CCL2-/-), a chemokine involved in the recruitment of CCR2+ monocytes, and wild type C57BL/6 mice were challenged with the colonic pathogen Citrobacter rodentium during exposure to SDR. Exposing mice to SDR, which consists of intermale aggression for 2 hrs a night on 6 consecutive nights, during challenge with C.rodentium significantly increased pathogen-induced colitis. This increase was marked by an enhanced accumulation of F4/80+ colonic monocytes/macrophages, increased pathogen burden and pathogen-induced colonic histopathology, and increased inflammatory cytokine and chemokine gene expression in comparison to C. rodentium-challenged non-stressed control. These stressor effects were dependent upon CCL2, since exposing CCL2-/- mice to SDR did not lead to an increased accumulation of colonic monocytes/macrophages in the colon and did not significantly increase gene expression or pathogen-induced histopathology. Together, these data illustrate the deleterious effects of stressor-enhanced recruitment of inflammatory monocytes during bacterial challenge.
Modulation of TNFα and TGFβ1 in botox treated rats with burn wounds (62.2)
Authors: Jurjus Abdo , Abdallah Hajj Hussein Inaya , El-Khatib Arij , Dali-Balta Nour , Shaalan Tareq , Watfa William , Atiyeh Bechara , Jurjus Rosalyn 
Published: January 2023

Abstract
Abstract Many attempts have been made to improve wound healing using growth factors and a large array of medications. This study reexamined the effect of Botox on second degree burn wound in rats. Histological alterations and temporal evolution of two cytokines TGFβ1, and TNF-α were assessed in presence and absence of Botox. Fifty adult Sprague Dawley rats were used; one group was injected 10 units of Botox into wound. Daily observation and transepidermal water loss were assessed. Punch biopsies were taken on days 0, 3, 8, 14 and 28 for RNA extraction and PCR, and for routine light microscopy. Botox treated rats showed a delayed inflammation, necrosis, and earlier crust detachment compared to controls. On day 14, a better and smoother epithelialization was noted in the Botox groups. Finally, by day 28, the wound surface area was smaller and the quality of the skin was better with more fibroblasts in the regenerated dermis of Botox group, which expressed 2-4 times more TGFβ1. It peaked on days 3 and 8 and then decreased to almost half the control group by day 28. On the other hand, the TNF-α peaked immediately after burn to almost 20 times the control, then decreased between 2.72 to 5.5 times during the rest of the experiment very much like TGFβ1, or lower. Botox could possibly play an alleviating role and have a positive effect on burn wound healing.
Distinct regulatory networks control B-lymphoid specification and lineage commitment. (63.1)
Authors: Sigvardsson Mikael , Zandi Sasan , Åhsberg Josefine , Qian Hong , Stjenrberg Jenny 
Published: January 2023

Abstract
The development of B-lymphoid cells from hematopoetic stem cells in the bone marrow involves an orchestrated interplay between transcription factor networks and external signals. In order to better understand B-lymphocyte lineage restriction, we have investigated molecular and functional properties in early B-lineage cells from Pax-5 and Notch-1 deficient animals carrying a B-lineage restricted reporter gene, allowing us to identify B-lineage specified progenitors independently of conventional surface markers. Pax-5 deficiency resulted in a dramatic increase in the frequency of B-cell progenitors marked by expression of a λ5 (Igll1) promoter controlled reporter gene. Gene expression analysis of ex vivo isolated cells revealed that Pax-5 deficient cells expressed several B-cell restricted genes, however, they displayed a high degree of plasticity for development into other lymphoid lineages at a single cell level. Even though the Pax-5 deficient progenitors expressed higher levels of Notch-1 and respond to delta-signaling in vitro, these cells expressed lower levels of the Notch-1 target gene Deltex-1, arguing for a minor role of Notch signaling in B-cell commitment in the BM. By comparison of gene expression patterns in ex vivo isolated Pax-5 and Ebf-1 deficient progenitors, it was possible to identify a set of B-cell restricted genes dependent of Ebf-1 but not Pax-5, revealing that B-cell specification and commitment is controlled by distinct regulatory networks.
Loss of MyD88 promotes the persistence of skin lesions in filaggrin deficient mice (64.1)
Authors: Leisten Sabine , Oyoshi Michiko , Bartnikas Lisa , Geha Raif 
Published: January 2023

Abstract
Flaky tail (ft/ft) mice have a skin barrier defect due to a loss-of-function mutation in the gene for filaggrin. They spontaneously develop eczematous skin lesions and are a model for human atopic dermatitis. We tested the hypothesis that the disrupted skin barrier in these mice facilitates the entry of environmental antigens, including microbial antigens, which are recognized by Toll-like receptors (TLRs) and may trigger the development of skin lesions. To examine the role of TLR signaling in the development of eczematous skin lesions in ft/ft mice, we created MyD88KO-ft/ft mice. Unexpectedly, the MyD88KO-ft/ft mice developed persistent skin lesions over the course of 32 weeks, while the ft/ft mice developed relapsing lesions throughout the study period. Lesional skin of both strains was characterized by epidermal thickening, infiltrating eosinophils and neutrophils as well as high mRNA levels of IL-17, IL-23 and CXCL2. These results indicate that MyD88 may be a negative regulator of skin inflammation in a mouse model of filaggrin deficiency.
Peripheral Foxp3+ regulatory T cell development in response to self-peptides (65.2)
Authors: Weissler Katherine , Bedoya Felipe , Kropf Elizabeth , Garcia Victoria , Caton Andrew 
Published: January 2023

Abstract
It is generally accepted that developing thymocytes can become Foxp3+ regulatory T cells (Tregs) through recognition of self-antigens during thymic selection. Tregs can also develop from conventional CD4+ T cells responding to exogenously administered peptides (including food antigens) in vivo, and via TGF-β signaling during activation in vitro, but the processes by which Treg induction may occur in response to self-peptides in the periphery remain unclear. We are examining this question by transferring conventional CD4+CD25-Foxp3- T cells specific for influenza virus hemagglutinin (HA) into mice that express varying amounts of the HA molecule as a self-antigen (HA Tg mice). We find that conventional CD4+ T cells can convert into Foxp3+ Tregs upon recognition of a self-antigen in the lymph nodes and spleens, and that accumulation of HA-specific Tregs is greater in mice expressing HA at lower levels than it is in mice expressing relatively higher levels of HA. Conversion occurs most efficiently at sites draining mucosal surfaces, in agreement with published studies demonstrating the existence of specialized subsets of dendritic cells at these locations that promote Treg induction. We also find that the extent of Treg induction in response to a self-antigen is essentially unaffected by acute viral infection. These studies are defining parameters that determine the ability of self-peptides to induce the formation of Foxp3+ Tregs in the periphery.
Novel Role for Btk in the Induction of Classically Activated M1 Macrophages (65.9)
Authors: Ni Gabhann Joan , Wynne Claire , Smith Siobhan , Jefferies Caroline 
Published: January 2023

Abstract
This study defines a critical role for Btk in regulating TLR-mediated APC activation and differentiation. Using Btk-deficient mice we examined the effects of LPS in regulating macrophage activity and function. Reduced levels of pro-inflammatory cytokines IL-12, IL-18 and IFN-γ were observed in Btk-deficient mice and ex vivo generated macrophages and dendritic cells (DCs) following acute LPS administration, whilst enhanced production of the anti-inflammatory cytokine IL-10 was observed. In response to diverse stimuli specific populations of macrophages are induced such that macrophage activation can be either pro-inflammatory or anti-inflammatory. Exposure to bacteria and IFN-γ results in the generation of classically activated (M1) macrophages which are microbicidal and secrete pro-inflammatory cytokines, whereas IL-10 is a key cytokine produced by alternatively activated immunomodulatory (M2) macrophages. Given the immunosuppressive environment observed in the absence of Btk we investigated macrophage population induction following treatment of APCs with either polarizing cocktails or with TLR4. Comparison of gene induction profiles for WT or Btk-deficient APCs revealed an inability of Btk deficient macrophages to become M1 effector macrophages. The predominance of M2 macrophages when Btk is absent is of crucial importance when considering therapeutic inhibition of Btk for the treatment of cancer or autoimmunity given the role of M2 macrophages in driving these pathologies.
Low phosphorylation of STAT-3 on interleukin-21 pathway in common variable immunodeficiency (66.2)
Authors: Alcántara Julio , Berrón Laura , Mogica Dolores , Espinosa Sara , Espinosa Francisco , Santos Leopoldo 
Published: January 2023

Abstract
Introduction: Common variable immunodeficiency (CVID) is a heterogeneous disorder characterized by recurrent infections and defective immunoglobulin production. Interleukin-21 (IL-21) is expressed by activated CD4+ T cells. Its receptor (IL-21R) is expressed on T, B, dendritic cells, macrophages and NK cells. The role of IL-21 in B cell differentiation make this an attractive candidate as a potential cause of CVID. Objective: To evaluate IL-21, IL-21R and its pathway in patients with CVID in naïve and memory B cells populations. Methods: Six patients with established diagnosis of CVID, according to European Society for Immunodeficiencies criteria, were included. CVID patients were previously analyzed to discard known genetics causes of CVID (ICOS, CD40L, TACI, CD19). To evaluate the pathway of IL-21R, PBMCs were stimulated with rhIL-21 for 15 minutes and phosphorylation of STAT-3 was evaluated by flow cytometry. Results: The patients show consistent low phosphorylation of STAT-3 after the stimulation with rhIL-21 for 15 minutes. IL-21 was equally expressed by PBMC from patients and healthy controls. Patients showed over expression of IL-21R on T and B cells. Conclusion: An unexpected discover in IL-21R signaling was the low phosphorylation of STAT-3 in CVID patients. This suggested that the production and regulation of STAT-3 may have an important role in B cell activation. So far, we have found no evidence of defects in IL-21, IL21R.
TLR4 and alveolar macrophages mediate lung ischemia reperfusion (IR) sterile inflammatory injury in a mouse model of ventilated lung ischemia (67.6)
Authors: Prakash Arun , Mesa Kailin , Hellman Judith 
Published: January 2023

Abstract
Background: Ischemia reperfusion (IR) injury is sterile inflammatory process associated with diverse clinical situations, for example, hemorrhage followed by resuscitation, transient embolic events, and organ transplantation. IR injury can result in lung dysfunction whether the IR occurs in the lung itself or in a remote organ. Increasingly it appears that receptors and pathways of the innate immune system are involved in recognizing sterile inflammation and overlap considerably with those involved in recognition and response to pathogens and pathogen-derived products. Methods: We used a mouse surgical model of transient unilateral pulmonary artery occlusion to the left lung without bronchial involvement to create IR injury and tracked injury in vivo and ex vivo. Additionally, we mimicked IR injury in vitro by transiently depriving cells of nutrients. Results: In the mouse ventilated lung IR model we demonstrate an upregulation of inflammatory markers and recruitment of neutrophils that is characteristic of IR injury. We show that the early inflammatory response to IR requires TLR4. Furthermore, we demonstrate in vitro that cooperativity and cross-talk exists between macrophages and endothelial cells resulting in an augmented inflammatory response to nutritional deprivation and that in vivo macrophages are indeed required for early sensing of IR injury. Remarkably, we find that selective depletion of alveolar macrophages rendered mice resistant to lung IR injury.
The local immune response in a mouse model of acute Clostridium difficile infection (67.8)
Authors: Sadighi Akha Amir , Theriot Casey , McDermott Andrew , Reeves Angela , Young Vincent , Huffnagle Gary 
Published: January 2023

Abstract
The current report presents our initial findings on the immune response to C. difficile infection (CDI) in an acute mouse model. 5-8 week old male C57BL/6 mice were either left untreated or received a cocktail of 5 antibiotics in drinking water for 3 days followed by a single intra-peritoneal dose of clindamycin. The antibiotic-treated mice were then infected with 105 CFUs of the C. difficile strain VPI10463 and sacrificed 42 hours later. Flow-cytometric study of the spleens, mesenteric lymph nodes (MLN), colons and ceca of the infected mice showed a significant increase in the numbers of B cells, CD4 and CD8 T cells, FOXP3+ CD4 cells, NK cells and neutrophils in the colons and ceca, as well as cells of the monocyte/macrophage lineage and dendritic cells in the ceca of the infected mice. The spleens and MLN of the infected mice did not display such an increase. A substantial fraction of the recruited neutrophils in the colons and ceca of the infected mice displayed upregulated levels of CD11b, whereas a similar fraction of CD4 and CD8 cells in the colons and ceca of uninfected and infected mice showed an upregulated level of CD69. Multiplex qPCR analysis of colonic and cecal tissue from the infected mice showed a significant increase in the mRNA levels of numerous neutrophil-attracting chemokines and a number of cytokines produced in an innate immune response, but no indication of T cell polarization, thereby collectively underscoring the innate, local nature of the response.