bluuebunny/crossref_metadata_2025_split huggingface dataset:
Myeloid responders to programmed cell death
Authors: Messmer Michelle N , Snyder Annelise G , Gerner Michael Y , Oberst Andrew 
Published: January 2023

Abstract
Necroptosis is a programmed form of cell death that more efficiently primes CD8 T cell responses when compared to apoptosis. This efficiency has been associated with key signaling effects downstream of RIP kinase activation. However, mechanisms whereby these cell-intrinsic signals translate into enhanced adaptive immunity remain unclear. We hypothesized that dying cells would attract and engage with distinct myeloid populations based on which cell death pathway is active, and that these interactions would determine T cell priming ability. By introducing dying cells loaded with non-degradable ‘antigen’ into naïve mice, we have tracked myeloid cells that interacted with necroptotic or apoptotic cell debris up to 72 hrs after injection, corresponding to early detectable differences in T cell priming. Migratory dendritic cells equivalently took up antigen following either necroptosis or apoptosis. However, necroptosis resulted in a greater number of antigen positive macrophages and lymph node resident dendritic cells. Understanding how the immune system recognizes and responds to distinct forms of cell death will benefit investigations into cytotoxic and immunotherapeutic drug combinations.
Autophagy-based unconventional secretion of alarmin HMGB1 by keratinocytes plays a pivotal role in psoriatic skin inflammation
Authors: WANG ZHEN , ZHOU HONG , ZHENG HUAPING , TENG XIU , WEI XIAOQIONG , LI JIONG 
Published: January 2023

Abstract
The precise mechanism by which autophagy affects psoriasis is poorly understood. Here we show that keratinocytes (KCs) autophagy was activated in psoriatic lesions of patients and mice model, positively correlating with psoriatic severity, and could be inhibited by MAPK family (ERK, p38, JNK) inactivation, implying autophagy was associated with psoriatic inflammation. Indeed, impaired autophagy flux, caused by autophagy inhibitors or KC-specific deletion of Atg5, alleviated psoriasisform inflammation, demonstrating autophagy positively regulated psoriatic inflammation. We then found an autophagy-based unconventional secretory pathway (autosecretion), depended on Atg5 and GRASP55, promoted the psoriasiform KCs inflammation. Moreover, alarmin HMGB1 was more effective than other autosecretory proteins, IL-1β and IL-18, to regulate the psoriasiform cutaneous inflammation. HMGB1 neutralization in autophagy-efficient KCs vanished the psoriasiform inflammation differences between autophagy-efficient KCs and autophagy-deficient KCs. Conversely, recombinant HMGB1 almost completely restored the psoriasiform inflammation in autophagy-deficient KCs in vivo. These results suggested that HMGB1-associated autosecretion, not other intracellular autophagy pathways, played a pivotal role in cutaneous inflammation. Finally, we demonstrated that KC-specific HMGB1 deficient mice, but not DC- or myeloid cell-specific HMGB1 deficient mice, displayed attenuated psoriatic inflammation, due to the essential crosstalk between KC-specific HMGB1-associated autosecretion and γδT cells. Thus, we uncover a novel autophagy mechanism in psoriasis pathogenesis, and imply its clinical significance in psoriasis treatment.
Exploiting altered enhancer landscapes to decode pathogenic changes in gene expression of diverse hepatic macrophages
Authors: Troutman Ty Dale , Seidman Jason S. , Sakai Mashito , Gola Anita , Ouyang Zhengyu , Spann Nathanael J. , Bruni Cassi M. , Bennett Hunter , Ego Kaori , Vu BaoChau , Sun Xiaoli , Pasillas Martina , Witztum Joseph L. , Germain Ronald N. , Glass Christopher K. 
Published: January 2023

Abstract
Kupffer cells have specialized roles supporting the environment of the liver during homeostasis and disease. However, the key regulatory elements governing these behaviors are unknown. Using scRNA-seq, we found diversification of Kupffer cells and recruitment of additional macrophage subtypes during nonalcoholic steatohepatitis (NASH). A significant source of macrophage heterogeneity during NASH was traced to Cx3cr1 expressing monocytes. Further, macrophage subsets were localized in distinct niches, suggesting environmental specification as a key determinant of macrophage heterogeneity. We profiled chromatin accessibility of the major NASH associated macrophage populations to identify transcription factors governing their environmental specification. These results predict greater NFκB, RUNX, and AP1 activity in recruited hepatic macrophages compared to Kupffer cells. Surprisingly, we found minimal significant chromatin accessibility changes comparing Kupffer cells from healthy mice to mice with NASH, even though several thousand genes were differentially expressed. Instead, NASH led to altered chromatin activity, as measured by H3K27ac ChIP-seq, at Kupffer cell enhancer regions. A binding element for LXR (liver X receptor) was the top transcription factor motif identified in Kupffer cell enhancers with reduced activity during NASH. Furthermore, LXRα was required to maintain expression of a unique gene signature defining healthy Kupffer cells. Thus, our studies establish for the first time gene regulatory events controlling diverse hepatic macrophages during homeostasis and NASH.
Ptpn6 inhibits Caspase-8- and Ripk3/Mlkl-dependent inflammation
Authors: Croker Ben A , Speir Mary , Nowell Cameron J , Chen Alyce A , O’Donnell Joanne A , D’Cruz Akshay A , Bliss-Moreau Meghan , Wang Shu , Kelliher Michelle A , Hakem Razq , Gerlic Motti 
Published: January 2023

Abstract
Neutrophilic dermatoses are a group of inflammatory skin disorders characterized by sterile infiltrates of neutrophils. These syndromes include pyoderma gangrenosum (PG) and Sweet’s syndrome (SS), and they are associated with an increased incidence of inflammatory bowel disease, rheumatoid arthritis, and acute myeloid leukemia. IL-1β is detectable in skin lesions, and IL-1 neutralizing therapies have met with success in some patients. Splicing variants and promoter region deletions of protein tyrosine phosphatase-6 (PTPN6) are a feature of SS and PG. Mice lacking Ptpn6 develop a cutaneous inflammatory disease that is dependent on the IL-1 receptor, G-CSF, and neutrophils, but the source of IL-1 and the mechanisms of IL-1 release remain unclear. Here, we investigated the mechanisms controlling IL-1α/β release specifically from neutrophils (Ptpn6ΔPMN) by inhibiting Caspase-8-dependent apoptosis and Ripk1/Ripk3/Mlkl-regulated necroptosis. Loss of Ripk1 from neutrophils accelerated disease onset, whereas combined deletion of caspase-8 and either Ripk3 or Mlkl strongly protected Ptpn6ΔPMN mice. Ptpn6ΔPMN neutrophils displayed increased p38-dependent Ripk1-independent IL-1 and TNF production, and were prone to cell death. Together, these data emphasize dual functions for Ptpn6 in the negative regulation of p38 MAP kinase activation to control TNF and IL-1α/β transcription, and in maintaining Ripk1 function to prevent caspase-8- and Ripk3/Mlkl-dependent cell death and concomitant IL-1α/β release. These findings implicate neutrophils as the dominant producers of IL-1 in neutrophilic dermatoses, and identify novel therapeutic targets for the treatment of these skin disorders.
Extracellular calcium-phosphate nanoparticles activate the NLRP3 inflammasome through CaSR signaling
Authors: Jäger Elisabeth , Stäubert Claudia , Sungur Pelin , Höppener Stephanie , Strobel Sarah , Murthy Supriya , Rossol Manuela , Wagner Ulf 
Published: January 2023

Abstract
Calcium and phosphate are well known as the main minerals in bone ossification. But they also have an immunomodulatory potential as components of calciprotein particles. Such particles precipitate spontaneously in body fluids if the calcium and phosphate homeostasis is disturbed. An abnormally increased mineral load could lead to soft tissue calcification. In our study we investigated the cellular response of freshly isolated peripheral blood monocytes in serum-containing medium under precipitating (high calcium and high phosphate content) conditions. Using dynamic mass redistribution analyses with the EPIC® system and IL-1β measurements as readout, we identified phosphate as a necessary co-modulator of calcium-triggered overall cellular responses and NLRP3 inflammasome activation. Moreover, flow cytometry and ImageStream® analyses revealed the uptake of spontaneously formed nanoparticles under stimulating conditions, which depends on the activation of the G protein-coupled receptor Calcium-sensing receptor (CaSR). Our study unraveled a GPCR-mediated mechanism by which monocytes detect imbalances in calcium phosphate homeostasis in the extracellular space resulting in an enormous pro-inflammatory cellular response.
A sustained small increase in NOD1 expression promotes ligand-independent oncogenic activity
Authors: Rommereim Leah , Akhade Ajay Suresh , Dutta Bhaskar , Hutcheon Carolyn , Lounsbury Nicolas , Rostomily Clifford , Savan Ram , Fraser Iain , Germain Ronald , Subramanian Naeha 
Published: January 2023

Abstract
Small genetically-determined differences in transcription are implicated in complex disease but the mechanisms by which small changes in gene expression impact the origin of complex disease are unknown. Recent GWAS fine-mapping studies show that many causal alleles for complex immune-related diseases lie in non-coding regions of genes, generating expression quantitative trait loci (eQTLs), with the extent of expression variation between susceptible and resistant genotypes often in the 1.5–3 fold range. Concurrently, systems-level studies across several cancer types suggest a regulatory organization in which disease-promoting genomic mutations lie upstream of functional master regulator proteins, a change in whose expression by 1.5–3 fold is responsible for propagating a tumor cell state. Here we show that a persistent small (~1.5 fold) increase in expression of the innate sensor NOD1 (Nucleotide-binding Oligomerization Domain-containing protein 1) precipitates large cancer-promoting changes in cell state in the absence of ligand-driven activity. A ~1.2–1.4-fold increase in NOD1 protein concentration by loss of microRNA regulation sensitizes cells to ligand-induced inflammation with an additional slight increase leading to ligand-independent NOD1 activation that upregulates the expression of proto-oncogenes and is linked to poor prognosis in gastric cancer. Our data show that tight expression regulation of NOD1 prevents this sensor from exceeding a physiological switching checkpoint that promotes persistent inflammation and oncogene expression and reveal the impact of a single small quantitative change in cell state on cancer.
γδ T cell activation in response to a keratinocyte wound signature
Authors: Johnson Margarete Daisy , Higginson Cody , Tekkam Srinivas , Crooke Stephen , Lloyd Jessica , Witherden Deborah , Finn M.G. , Havran Wendy L 
Published: January 2023

Abstract
Epidermal resident γδ T cells are key players in the wound healing response, providing growth factors and cytokines that promote keratinocyte proliferation following recognition of damaged self. These cells are known to be unresponsive in chronic wounds and as such represent a novel target for new wound healing therapies. In this study we seek to modulate the γδ T cell response in chronic wounds through the identification, characterization, and delivery of keratinocyte-expressed proteins that comprise a “wound signature” allowing γδ T cells to recognize and respond to epithelial damage. We have identified a set of target genes though RNA-seq analysis which are upregulated following wounding in mice and correspond to membrane-bound and secreted proteins available for interactions with neighboring cells. Candidate gene expression was measured following wounding of human skin tissue, and ICAM1, CRISPLD2, and HSPA8 were shown to also be significantly upregulated in human skin wound samples. Hspa8 and ICAM-1 have been validated through in vitro assays to activate γδ T cells, and these proteins have been chosen for in vivo delivery. In order to deliver these candidate proteins into in vivo wound models we have developed a hydrogel-based delivery system with a highly tunable time frame of release. This system improves the retention of protein cargo at the wounds site post-wounding in mice and is being used to administer our candidate proteins into mouse models of non-healing wounds to assess effects on acceleration of tissue repair.
Tracking the in vivo history and fate of IL-2 producing CD8 T cells during memory differentiation
Authors: Yuzefpolskiy Yevgeniy , Sarkar Surojit , Kalia Vandana 
Published: January 2023

Abstract
IL-2 is a key driver of T cell responses critical for the conversion of resting antigen specific T cells into glycolytic powerhouses that undergo massive expansion and elicit cytolytic function to clear infections. Recent studies have used ex vivo antigen challenge combined with somatic IL-2 deletion to show that both paracrine and autocrine IL-2 signals are indispensable for the formation of highly functional memory CD8 T cells. Despite this critical role of autocrine IL-2 for memory T cell fate, the in vivo history and fate of autocrine IL-2 producing CD8 T cells remains unknown. We generated a mouse model in which autocrine IL-2 production leads to transcription of TdTomato, leading to an indelibly marked T cell. Following acute infection, a proportion of antigen specific CD8 T cells made autocrine IL-2, however, more than half of the T cell response had no history of IL-2 production. Furthermore, contrary to their ex vivo functionality, memory precursor and terminal effector T cells had similar history of in vivo IL-2 production. Despite autocrine IL-2 production being a benchmark of highly functional memory T cells, we observed no increase in the ex vivo polyfunctionality of T cells that produced in vivo IL-2 during infection. Finally, while strong TCR engagement is critical for the recruitment of robust T cell responses, we saw that in vivo autocrine IL-2 production is more effectively elicited by lower affinity TCR signals. Thus, we see that while proportion of T cells rely on autocrine IL-2 during infection, the ability of a CD8 T cell to recruit autocrine IL-2 function heavily depends on the avidity of the TCR to the peptide-MHC complex. Together these data provide the first insights into the in vivo function of IL-2 in antigen specific CD8 T cells.
Delineating the functions of RIPK1 in vivo through CRISPR/Cas9-mediated gene editing in mice
Authors: Zhang Xuhua , Dowling John P. , Zhang Jianke 
Published: January 2023

Abstract
RIPK1 is originally identified as a protein associated with Fas and TNFR1, which can trigger apoptosis. However, initial studies indicated that RIPK1 is dispensable for apoptotic signaling. Instead, cells lacking RIPK1 are defective in TNFR1-induced activation of NF-kB, a critical player involved in pro-survival signaling. Recent studies indicate that a certain form of necrosis-like death, necroptosis, requires RIPK1. In contrast, the Fas-associated death domain (FADD) protein appears to play an obligatory role in apoptosis induced by Fas and TNFR1. Fadd−/− mice die in utero due to RIPK1 mediated necroptosis. More recently, studies including ours demonstrated that only when both FADD and RIPK3 were deleted will Ripk1−/− mice survive to adulthood. This result indicates perinatal lethality in Ripk1−/− mice is due to not only FADD-mediated apoptosis but also RIPK3-dependent necrosis. Interestingly, we found that in adult T cells RIPK1 only suppresses apoptosis. To facilitate the dissection of the multiple functions of RIPK1, we performed in vivo gene editing using the CRISPR/Cas9 technology. This allows selective interruption of one functional domain/motif of RIPK1. Using these novel mouse models, our data reveals a previously unappreciated function for RIPK1 in promoting apoptosis.
The NF-kappa B regulator Bcl-3 is critical factor for memory CD8+ T cell generation
Authors: Jaiswal Hemant , Siebenlist Ulrich 
Published: January 2023

Abstract
Bcl-3 is an atypical member of the IκB family. Unlike classical IκBs, Bcl-3 is not degraded in response to activation signals, but instead its expression is often induced and it acts in the nucleus to modulate transcription of NF-κB target genes by associating with p50/NF-κB1 and p52/NF-κB2 homodimers. The latter complexes do not trans-activate autonomously, and association with Bcl-3 may promote or inhibit transcription in a highly gene- and context-dependent manner. Bcl-3 has been shown in mouse models to have important biological roles in promoting adaptive and innate immune responses to infectious agents as well as autoimmunity; however, the mechanisms have not been clearly defined. Here we investigate the role of Bcl-3 in primary and secondary T cell responses to lymphocytic choriomeningitis virus (LCMV) infection. Acutely infected mixed bone marrow chimeric mice generated from WT and Bcl-3 KO mice had defective LCMV antigen-specific CD8+ T cell recall responses in the absence of Bcl-3 upon challenge with LCMV (clone 13), despite having comparable primary CD8+ T cell responses at day 8 post primary infection (acute). Bcl-3 deficient CD8+ T cells instead exhibited significantly reduced proportions of LCMV epitope specific cells after 8 weeks of acute infection. Single cell RNA sequencing analysis of sorted LCMV epitope (GP33) specific CD8+ T cells at day 8 after acute infection showed a reduced proportion of memory precursor effector cells (MPECs) in the absence of Bcl-3. This suggests a crucial role of Bcl-3 in memory CD8+ T cell generation by regulating the formation or maintenance of MPECs during the primary response.
Signaling from individual Opioid Receptors differentially modulates the functional and phenotypic potential of resting and activated human CD8+ T cells
Authors: Mazahery Claire , Valadkhan Saba , Levine Alan D 
Published: January 2023

Abstract
Opioid use disorder is a worldwide epidemic. Endogenous opioids are released at sites of injury, and their cognate GPCR opioid receptors (OR) are expressed on immune cells; drugs of misuse appropriate these ORs. Conflicting published data show that opioid users have elevated risk of infection, opioids activate innate immune cells, and opioids attenuate murine T cell autoimmunity models. Intravenous drug use transmits blood borne pathogens, particularly viruses, making the study of CD8+ T cells pertinent to the crisis. Sequencing studies on leukocytes from heroin users show an altered transcriptome and a higher frequency of a SNP in a gene regulating long noncoding RNA. We demonstrate functional ORs (μ-, κ-, δ-subclasses) on human CD8+ T cells and employed unbiased analyses (flow cytometry with tSNE and transcriptome mapping) to study the effects of DPDPE (δ-OR agonist) and morphine (μ-OR biased agonist) on phenotype and activation status. Both agonists decrease activation markers on resting T cells, with δ-OR affecting naïve cells and μ-OR affecting naïve and memory cells. Pre-exposure to opioids changes the response to T cell receptor stimulation. Within 24 h, pre-exposure to either OR agonist decreases CD3, CD69, and CD25 expression. At 48 h, pre-exposure to μ-OR agonist downregulates CCR7 and CD27, while δ-OR agonist sustains CD3 expression. Regulation does not appear to be mediated by signaling cascades or calcium flux, suggesting a transcriptional mechanism: opioids dramatically modify the protein-coding and non-coding transcriptome of resting and activated T cells, with hundreds of non-coding RNAs showing increased expression. Selective engagement of OR represents a possible pathway to fine tune an immune response.
Immunology as a basis for STEM related teaching that is culturally appropriate and engages underrepresented High School students
Authors: Sika-Paotonu Dianne , Cockburn Alana 
Published: January 2023

Abstract
Indigenous Māori and Pacific Peoples in New Zealand (NZ) remain overrepresented both in adverse health statistics and education outcomes. To improve education and health outcomes for Māori and Pacific communities, various strategies and interventions have been proposed. This includes efforts to increase the Māori and Pacific health workforce populations figures more reflective of the make-up of the NZ population. These health careers require participation in science and health related subject matter throughout high school and at higher education learning levels. Māori and Pacific students remain underrepresented amongst these groups. The purpose of this work was to support improved interest and participation in science related learning that was Immunologically focused, amongst high school students from underrepresented groups that included Māori and Pacific students. A visit was carried out by a Biomedical Scientist to an all-girls high school class comprised of students who were of predominantly Māori and Pacific heritage to teach and share about Immunology research work that was Cancer based. The female Biomedical Scientist was of Pacific heritage and was an important consideration in the application of the role modelling concept for the Pacific students within this interaction. Verbal and written feedback highlighted the session was well-received and enjoyed by those in attendance. An important observation was the positive impact and enthusiasm of the students in response to the emphasis on the Immunologically focused Cancer based examples. This work demonstrated the importance of cultural considerations blended with scientific knowledge communication and interactions in the classroom.
The new paradigm shift in microbiology and immunology education in Azerbaijan Medical University Baku Azerbaijan
Authors: Fazal Nadeem , Ali Shamsideen , Sheriff Dauda , Brewer David , Childs Porscha 
Published: January 2023

Abstract
Medical Schools in USA and the rest of western world are launching their first major redesign of its early curriculum since the 1980s, integrating multiple disciplines into single courses and introducing earlier clinical immersion and flipped classrooms. It will require medical students to learn more actively, rather than cram and memorize material, and that it seeks to reflect how medicine has changed over the last four decades. Medical curricular change is part of a broader shift in medical education from asking students to memorize material to emphasizing its application. The new paradigm is driven by new innovations in medicine as well as new needs in medicine. Azerbaijan is a central Asian country at the cross roads of Western and Eastern cultures. As a former member of the Soviet Union, Azerbaijan science and education developed in last century on the basis of German-Russian model. However, in recent years, scientific, medical, and education fields are also drawing on American, British, French and Turkish models in developing new research/education standards. In this study we used innovative teaching techniques and modern web-based tools to advance student’s learning and comprehension. These parameters were evaluated in proposing a new paradigm shift in medical curriculum design at Azerbaijan Medical University via discussions, dialogues and planning, envision and strategize future efforts to slowly mold towards short-term and long-term goals. A pilot online course in Medical Microbiology and Immunology was tested for students registered in medicine, dental, pharmacy and public health faculties. Data from student and faculty surveys showed a marked acceptance of the need for a curricular redesign and welcoming paradigm shift.