bluuebunny/crossref_metadata_2025 huggingface dataset:
The Resilient System Architecture of Supply Chain Adapted to the Digital Transformation
Authors: Cui Yu , Idota Hiroki
Published: January 2023

In this chapter, firstly, the methodology and importance of supply chain resilience research are reviewed and summarized in the context of the drastic changes in today's world circumstance and the frequent occurrence of emergencies. It systematically analyzes and expounds the specific situation of how the most cutting-edge digital technologies play a role and influence in the process of digital transformation of enterprises. Moreover, from the analysis and discussion of digital technologies, the structure and the future development prospects of the metaverse, which is the new direction of digital transformation after the epidemic, are sorted out and clarified. In addition, a detailed review and category of blockchain technologies which is closely related to the metaverse are given. Based on these analyses, a new architectural model which allows supply chains to strengthen their own supply chain resilience without weakening the existing efficiency is proposed, so as to fully utilize today's digital technology and strengthen integration when confronted with crises and challenges.
Positioning Augmented Reality in Oil and Gas Maintenance Support
Authors: Sheikh Abd Jalil Sabrina Asyraf , Ismail Syuhaida , Abd Fatah Abdul Yasser
Published: January 2023

High-risk, hazardous, and harsh environments are synonymous with the oil and gas industry. Accidents are inevitable without proper maintenance for equipment installed in any asset. Therefore, maintenance is a critical activity required to operate oil and gas facilities. Existing methods consist of corrective and proactive maintenance, where operators are betting a bottom dollar on original equipment manufacturer (OEM) specialists to assess the functionality of the equipment. Nevertheless, beginning of 2020, the Coronavirus disease (COVID-19) pandemic has hindered most of the yearly turn around (TA) to maintain the assets due to travel restrictions and mandatory quarantine regulations. Therefore, this paper aims to develop a strategic plan during the COVID-19 pandemic to position augmented reality (AR) in the maintenance support of the oil and gas industry. This proposal used literature reviews related to AR and maintenance support as its research methodology. The strategies encourage remote support via suitable AR technology to reduce cost and shorter mean time to repair (MTTR).
Managing Information System Risk Audit
Authors: Susanto Heru , Daud Farizah
Published: January 2023

It is not a surprise anymore that e-Commerce has rapidly becoming a “normal” way of doing business as the world has become advanced with the technology. Now, buying and selling goods and services can be done online at any time and any place. However, there is still the concern towards the consumers, as some of them are oblivious on how their personal information will be handled and used. A total of 121 respondents were gathered to obtain inputs on how Bruneian perceived the risk of using e-commerce. The study provides an analysis of the perception of e-commerce customers in Brunei on the security and privacy issues. From the findings of the survey, managing risk will be constructed based on the perception of the local e-commerce customers.
Blockchain for Halal Food Supply Chain
Authors: Nasyiah Tsiqatun , Bahalwan Enggar , Hafish Maulana Abdul , Barraq Ricco Faris , Zulfikarijah Fien
Published: January 2023

The purpose is to analyze the growth, possibilities, and difficulties of the halal food industry's supply chain. Blockchain solutions in the halal food supply chain have become more and more well-liked in recent years. The approach is a descriptive qualitative approach. A qualitative approach is a method that generates descriptive data in the form of spoken or written words from individuals and the observed behavior to provide a complete picture. The goal of this descriptive study kind is to uncover facts, phenomena, and situations that are existent. According to research, the supply chain for halal food uses blockchain technology. The quantity of papers that have been published proves it. The purpose of this study is to review and evaluate earlier papers on using systematic literature from 2019–2022 journals.
Exploring Blockchain-Based Digital Marketing in the UAE
Authors: Arnaut Amina , Arnaut Marina , Naqshbandi M Muzamil
Published: January 2023

This chapter explores the possibilities of using blockchain technology in digital marketing practices in the UAE. The research also examines how blockchain may affect digital marketing's contemporary practices to provide recommendations to digital marketing practitioners on utilising blockchain technology. Secondary data analysis, an online survey, and a Delphi study were conducted to achieve the research objectives. This chapter corroborates a gap in the literature by analysing the secondary data. The online survey results depict users' frustrations, openness to advertisement incentives, transparent supply chains, and brand advocacy. Lastly, the Delphi study identified digital marketing frustrations and blockchain experts' insight. The research confirms that blockchain-based digital marketing could enhance user experiences and specific marketing objectives while facilitating market development or differentiation. It is recommended for marketers to study and remain open to the technology's adoption to improve their performance.
Business Optimisation Strategy for a Malaysian Aircraft Maintenance Repair Organisation Affected by the COVID-19 Pandemic
Authors: Abdul Rashid Muhamad Farouk , Ismail Syuhaida , Mohamed Mahmoud Abdelgadir
Published: January 2023

An aircraft maintenance programme is critical to maintaining aircraft airworthiness. However, during the coronavirus disease 2019 (COVID-19) pandemic, aircraft maintenance repair organization (MRO) businesses were adversely impacted, including but not limited to retrenchments, restructurings, and permanent business closure. Therefore, the aim of this chapter is to propose a business optimization strategy for Malaysian MRO affected by the COVID-19 pandemic to promote business growth. The data and methodology used to produce this proposal were derived from an analysis of numerous literature reviews that were identified as having a high impact Malaysian aviation. This chapter finds that the COVID-19 pandemic has a global impact on aviation businesses; travel restrictions imposed by the majority of countries have caused major disruptions in the aviation and aircraft maintenance businesses, as the majority of airlines have grounded their aircraft; however, there are still some aviation business opportunities as a direct consequence of the COVID-19 pandemic.
Oops... Communication Breakdowns in Mega Projects
Authors: Gideon Eric , Ismail Syuhaida , Fathi Mohamad Syazli
Published: January 2023

Communications is pivotal in daily interactions as it allows individuals to relay messages and to coordinate with another person. This applies also in mega projects where stakeholders need to communicate among each other to attain mutual understanding. Project Management Office (PMO) plays a vital role to ensure that there are seamless communications between various stakeholders as communication breakdown between stakeholders promotes losses in terms of time and cost in a project. The aim of this chapter is to examine the main failures in managing communications between stakeholders in the Advanced Metering Infrastructure (AMI) deployment project in Malaysia and discusses the potential communication management alternatives to address the communication breakdowns between the stakeholders. This chapter was developed using secondary data and information obtained from extensive reviews of relevant literature pertaining to communication breakdowns that are related to the Advanced Metering Infrastructure (AMI) deployment project in Malaysia.
Cost Reduction Strategies for Outbound Logistics Operations at an Automotive Lubricant Manufacturer in Selangor
Authors: Mursal Norzureizal , Ismail Syuhaida , Mohamed Mahmoud Abdelgadir
Published: January 2023

Effective management strategies are crucial for every company, especially in manufacturing. Nonetheless, most studies focus on cost reduction strategies in original equipment manufacturer (OEM) companies, leaving lubricant manufacturers behind. Growing customer numbers, uncertain orders, and demand to reduce total production costs are the main challenges. This chapter suggests cost reduction strategies for outbound logistics at a Selangor-based automotive lubricant manufacturer. Adopting lean manufacturing methods, non-value-added activities may be eliminated in the outbound logistics operation. This chapter will investigate the manufacturer's existing operation waste management practises. The unnecessary costs can be utilized for ongoing development where they will be discussed in this chapter for the elimination of non-value-added work and building logistics efficiency. The findings from this chapter will assist the company to reduce costs while maintaining product quality and customer satisfaction. It should also show the company's commitment in the preparation to adopt Industry 4.0.
The Effect of IT Flexibility and IT Integration on Supply Chain Agility in SMEs
Authors: Barraq Ricco Faris , Hafish Maulana Abdul , Nasyiah Tsiqatun , Bahalwan Enggar , Zulfikarijah Fien
Published: January 2023

The ability to manage supply chain flexibly is critical to its competitive advantage, especially for small and medium-sized enterprises (SMEs) that do not have the economies of scale of large companies. Examining the relationship between organizational culture, information technology (IT) capabilities, and supply chain agility leads to a conceptual framework that helps SMEs build business strategies and practices to adapt to changing markets. The ability to make a difference is one of the most important requirements for a company to be able to compete in the 21st century. Globalization, unpredictable market pressures, and rapid technological change have made companies embrace the ability to change as a necessary capability. Companies need to adapt their business activities to the dynamic changes in the market environment. Supply chain management is as important as any other functional area in seeking its integrity.
Innovative Technology Adoption of Cross-Border Runners as a Trusted Service of Information Preservation
Authors: Haziqah Auji , Susanto Heru
Published: January 2023

As the courier service market becomes more competitive and the business environment changes, courier providers are being forced to become more customer-oriented and to establish long-term relationships with their clients. Courier companies presently place a strong emphasis on the quality of their services, as well as on customer satisfaction and loyalty. Customer satisfaction serves as a fundamental assessment of a company's service delivery system, whereas customer satisfaction is a “post-consumption” experience that evaluates perceived quality with expected quality. When a provider creates a benefit for consumers, it increases the likelihood that they will continue and even increase their use of the provider's courier services. Today's courier service providers are looking for ideas on how to increase customer loyalty and increase sales. Reduced marketing expenditures, more sales, and lower operating costs all contribute to the improved profit.
Analysis of Usability Level on E-Grocery Space (Convenience Stores With Online Deliveries) In Indonesia
Authors: Restuputri Dian Palupi , Hakiki Risnaldi Nur , Mufti Wahyu Fitrianda
Published: January 2023

Mobile applications in online shopping have made buying and selling transactions easier with effortless access to transactions anywhere and anytime. Alfagift and Klikindomaret is an online shopping application from Alfagift and Klikindomaret franchised Indonesian convenience store chain. The testing method assesses user interaction with the application. For this purpose, in analyzing the Klikindomaret and Alfagift applications, the usability scale (SUS) system is used. The SUS method was considered insufficient to solve the problem, so it was strengthened by the PACMAD (People at the Center of Mobile Application Development) method. Respondents in this study amounted to 96 people. The SUS-based usability values measurement revealed unsatisfactory results and scores where the SUS score for the Klikindomaret application was 68%. Meanwhile, the SUS score for the Alfagift application was 53%. The measurement of usability value using the PACMAD method on the Klikindomaret application is a score of 55.06% for the effectiveness variable. Meanwhile, the consumer opinions regarding the Alfagift application had a score of 49.533%.
Deepfakes on Smart Devices in Stock Price Prediction Using Machine Learning
Authors: Sobana P. , Prabhu S. , Karuppusamy S. , Anbarasu N.
Published: January 2023

The stock market is a widely used investment scheme promising high returns, but it has some risks. It is an act to forecast the future value of the stock market. The change in the stock market is explosive, and there are multiple sophisticated financial indicators. Still, the enhancement in technology provides a chance to grow constant fortune from the stock market and so helps experts to detect the most enlightening indicators to produce better predictions. Machine learning algorithms have made a magnificent effect in determining stocks precisely. This paper proposed a multiple regression algorithm for determining the future value of a stock. The first thing that was taken into account is the dataset of the companies Apple and Microsoft. Live historical data has been collected from yahoo finance. The dataset was preprocessed and tuned up for real analysis. Hence, this paper also focused on preprocessing of the raw dataset. After preprocessing the data, some forecasting measures are suggested, such as momentum, volatility, index volatility, and stock.
Machine Learning Algorithms to Detect Deepfakes Fine Tuned for Anomaly Detection of IoT
Authors: Sridhar N. , Shanmugapriya K. , Marimuthu C. N.
Published: January 2023

The internet of things (IoT) is a worldwide network of interconnected gadgets that enables devices to communicate with one another and share data in a continuous manner. Any deviation from the typical course of events is referred to as an anomaly, and it might serve as an early indicator that there is a problem. The authors differentiate themselves from previous tactics by requiring less time to identify and respond to attacks since they implement a variety of machine learning algorithms while the programme is running. This effort intends to establish a system for anomaly detection that is capable of screening IoT flaws and alerting the organization's CEO as well as the help network. The authors make use of a machine learning approach called k-nearest neighbor (KNN) in conjunction with a random forest (RF) algorithm in order to fine-tune the parameters of the spreading network. As a result, this framework improves the performance of the model without causing it to overfit.
Enhancing the Protection of Information in Digital Voting Using the Fraud Application of Blockchain Technology
Authors: Ganesan P. , Victoria D. Rosy Salomi , Chouhan Arun Singh , Saravanan D. , Baghel Rekha , Saikumar K.
Published: January 2023

Elections are conducted electronically instead of using paper ballots to cut down on mistakes and discrepancies. Recently, it has been discovered that paper-based balloting fails owing to security and privacy difficulties, and the electronic balloting approach has been recommended as a replacement. For the sake of keeping your data safe, the authors have designed and developed a hashing algorithm based on SHA-256. The blockchain's adaptability is aided by the sealing of the block concept's incorporation. Consortium blockchain technology is employed to ensure that only the election commission has access to the blockchain database, which candidates and other outside parties cannot modify. When used in the polling method, the methodology discussed in this chapter can yield reliable findings. The authors used a hashing algorithm (SHA-256), block generation, data collection, and result declaration to get to this point.
Features Manipulation of Classification and Recognition of Images Under Artificial Intelligence Using CNN Algorithm and LSTM
Authors: Priyadharshini J. , Padma E. , Prabhadevi S.
Published: January 2023

The established model provides appropriate picture pixel gaining knowledge in image detection. Additionally, it also affords an alternative solution for item tracking and predicting the usage of deep gaining knowledge of strategies. The proposed technique offers a fine overall performance in photo recognition issues or even outperforms humans in positive cases. Deep learning architectures containing dispensed techniques will become more critical as the scale of datasets increases. Then, it is important to understand which are the most green approaches to carry out distributed education, so as to maximize the throughput of the gadget, while minimizing the accuracy and model regression. This chapter explores features manipulation of classification and recognition of images under artificial intelligence using CNN algorithm and LSTM.
Techniques and Applications of Demographic Determinants of Mobile Phone Brand Switching in Attitude Towards Switching and Switching Intentions
Authors: Patel Binit
Published: January 2023

The Indian telecom industry is growing at a very rapid pace. Both networking and handset-related technologies are refining day by day and year by year. Even the routine activities are executed by the use of mobile phones. Hence, mobile phones nowadays have become a necessity and not a luxury. Switching has now become a trend. There are demographics that actually suggest the switching behaviour; hence, they need to be identified. The descriptive study was used with a total of 1502 respondents across the state of Gujarat, wherein the respondents were selected using the hybrid sampling method. A structured questionnaire was designed using different items, constructs, and dimensions found in the literature review. The study has developed a regression model that can further help in predicting the switching behaviour of mobile phone users. There are demographics that moderate the relationship between ‘switching intention' and ‘attitude towards switching'.
Narrative Approach in Qualitative Research
Authors: Rosairo Rohitha
Published: January 2023

No abstract available
Farmers’ Independence Level in the Urban Area of Subak Sembung Denpasar City, Bali Province, Indonesia
Authors: Darmawan D. P. , Arisena G. M. K. , Djelantik A. A. A. W. S. , Krisnandika A. A. K. , Utari N. K. S. , Korri N. T. L.
Published: January 2023

Purpose: Subak is an organization owned by the farming community in Bali which specifically regulates traditional rice field management. Subak Sembung is one of the Subak organizations that is engaged in the production to date. The novelty of this research as well as the purpose of the research is to observe the independence of urban farmers from various sides. The purpose of this research is to increase the independence of farmers in adapting to urban situations.
Research Method: Sixty-six members of Subak Sembung participated as study participants. Data were collected by observation and interview sessions. The qualitative analysis was employed to address the first, second, and third aims of the study.
Findings: Results revealed that the majority of the farmers had a high level of independence in the aspect of cultivation tool preparation, crop production, capital, capital provider, and type of cultivated commodity. However, lack of independence was identified in the aspect of post-harvest handling, management, selfdevelopment, collaborating to provide agricultural services, and technology adoption.
Originality/ Value: We suggest delivering training, workshops, and technology adoption to improve the level of independence of farmers in adapting to urban agriculture.
Monocytic Myeloid-Derived Suppressor Cells from Tumor Tissue Are a Differentiated Cell with Limited Fate Plasticity
Authors: Calvert Ryan D , Fleet James C , Fournier Pierrick G J , Juarez Patricia , Burcham Grant N , Haverkamp Jessica M , Guise Theresa A , Ratliff Timothy L , Elzey Bennett D
Published: January 2023

Owing to ease of access and high yield, most murine myeloid-derived suppressor cell (MDSC) knowledge comes from the study of spleen-derived MDSCs rather than those isolated from the tumor. Although several studies have identified subtle differences in suppressive function between these MDSCs, a recent report demonstrated that the whole peripheral myeloid compartment poorly reflects myeloid populations found at the tumor. We confirm and extend these observations by presenting data that indicate extensive differences exist between peripheral and tumor MDSCs, suggesting that it may be inappropriate to use spleen MDSCs as surrogates for studying tumor MDSCs. Using cytospins, we observed that tumor MDSCs have undergone a morphologic shift from immature myeloid cell forms commonly seen in bone marrow (BM) and spleen MDSCs and acquired mature myeloid cell characteristics. Spleen and BM monocyte-like MDSCs (M-MDSCs) readily responded to differentiation signals for multiple myeloid cell types whereas tumor M-MDSCs had remarkably reduced cellular plasticity. At the time of isolation, M-MDSCs from BM or spleen have little to no T cell suppressive activity whereas those from the tumor possess immediate and efficient T cell suppressive function. Finally, microarray analysis revealed that the transcriptomes of tumor and spleen M-MDSCs possessed >4500 differentially expressed transcripts. We conclude that tumor M-MDSCs are more differentiated and mature, and that they are morphologically, genetically, and functionally distinct from spleen and BM M-MDSCs. These observations have important implications for the design of anti-MDSC therapies and suggest that preclinical studies using nontumor MDSCs could lead to results not applicable to tumor MDSCs.
Human Plasmacytoid Dendritic Cells Express C-Type Lectin Receptors and Attach and Respond to Aspergillus fumigatus
Authors: Maldonado Samuel D , Dai Jihong , Dutta Orchi , Hurley Harry J , Singh Sukhwinder , Gittens-Williams Lisa , Kalyoussef Evelyne , Edelblum Karen L , Rivera Amariliz , Fitzgerald-Bocarsly Patricia
Published: January 2023

Plasmacytoid dendritic cells (pDCs) have been implicated as having a role in antifungal immunity, but mechanisms of their interaction with fungi and the resulting cellular responses are not well understood. In this study, we identify the direct and indirect biological response of human pDCs to the fungal pathogen Aspergillus fumigatus and characterize the expression and regulation of antifungal receptors on the pDC surface. Results indicate pDCs do not phagocytose Aspergillus conidia, but instead bind hyphal surfaces and undergo activation and maturation via the upregulation of costimulatory and maturation markers. Measuring the expression of C-type lectin receptors dectin-1, dectin-2, dectin-3, and mannose receptor on human pDCs revealed intermediate expression of each receptor compared with monocytes. The specific dectin-1 agonist curdlan induced pDC activation and maturation in a cell-intrinsic and cell-extrinsic manner. The indirect activation of pDCs by curdlan was much stronger than direct stimulation and was mediated through cytokine production by other PBMCs. Overall, our data indicate pDCs express various C-type lectin receptors, recognize and respond to Aspergillus hyphal Ag, and serve as immune enhancers or modulators in the overarching fungal immune response.
HLA-Inception: A structure-based deep learning framework for MHC-I binding motif prediction
Authors: Wilson Eric A , Cava John , Anderson Karen , Singharoy Abhishek
Published: January 2023

The ability to accurately identify peptide ligands for a given major histocompatibility complex class I (MHC-I) molecule has immense value for targeted anticancer and antiviral therapeutics. However, the highly polymorphic nature of the MHC-I protein makes universal prediction of peptide ligands challenging due to lack of experimental data describing most MHC-I variants, and the vast number of protein variants precludes comprehensive experimental determination. Therefore, there is a need for a framework to cluster MHC-I alleles to prioritize for experimental validation as well as identify alleles with potential disease associations.

To address this challenge, we have developed a deep convolutional neural network, HLA-Inception, capable of predicting MHC-I peptide binding motif using data derived from the structure of the MHC-I binding pocket. By approaching this problem from a 3-dimensional perspective, we can fully consider the impact of sidechain arrangement and topology of the MHC-I binding pocket on peptide binding motif, which is not inherently captured by the popular protein sequence-based approaches. Through a combination of homology modeling and biophysical simulations, we created protein structure models for all full-length HLA-ABC alleles. The topology and interaction forces within the MHC-I binding pocket were accounted for by solving the 3-dimensional electrostatic potential near the surface of the protein. HLA-Inception was then trained on all MHC-I alleles with known MHC-I binding motifs and applied to the full set of MHC-I models. We found that predicted peptide binding motifs fell into distinct and well-defined clusters which maintained known peptide binding and disease associations.
The monocyte cell surface as a novel site of autoantigen generation in Rheumatoid Arthritis
Authors: Thomas Mekha A , Naik Pooja , Wang Hong , Jang Yura , Johnson Tory P , Curran Ashley M , Crawford Jonathan D , Jahanbani Shaghayegh , Robinson William H , Na Chan Hyun , Darrah Erika
Published: January 2023

Citrullination is recognized as a key pathogenic process in rheumatoid arthritis (RA), as evidenced by the formation of anti-citrullinated protein antibodies (APCAs) in the majority of patients; however, the mechanisms that result in citrullinated autoantigen generation are not fully understood. Although the citrullinating enzyme peptidylarginine deiminase IV (PAD4) is predominantly expressed by neutrophils and monocytes, the contribution of monocytes to the citrullinated autoantigen pool has been underexplored. In this study, we utilized multiple complementary methods including flow cytometry, immunofluorescence, and transmission electron microscopy, which revealed a predominantly extranuclear localization of PAD4 in monocytes with a fraction present on the cell surface. Surface PAD4 was enzymatically active and citrullinated both extracellular fibrinogen and endogenous surface proteins in a calcium dose–dependent manner. In addition, human monoclonal ACPAs cloned from patients with RA recognized fibrinogen citrullinated by monocyte-surface PAD4. Mass spectrometry analysis of citrullinated proteins from the cell surface fraction revealed CD11b to be a novel PAD4 substrate. Citrullinated CD11b was recognized by autoantibodies in 60% of ACPA+ RA patients compared to 6% of healthy controls (p=0.0021) and 0% of ACPA− RA patients (p≤0.001). Taken together, our study demonstrates that PAD4 is expressed on the surface of monocytes in an enzymatically active state that renders the monocyte surface a novel site of citrullinated autoantigen generation in RA.

Supported by funds received from Bristol-Myers Squibb.
Plasma-derived exosomes as biomarkers for therapy monitoring in head and neck cancer patients treated with surgery and (chemo)radiotherapy
Authors: Theodorakis Marie-Nicole , Lotfi Ramin , Whiteside Theresa L , Laban Simon , Hofmann Linda
Published: January 2023

Background

Head and neck squamous cell carcinomas (HNSCC) are highly immunosuppressive and aggressive malignancies. Exosomes are small extracellular vesicles that mediate intercellular communication. HNSCC patients have a significantly higher exosome load compared to healthy donors. Here, we evaluated the potential of plasma-derived exosomes as biomarkers for therapy monitoring in advanced HNSCC treated with conventional therapy.

Methods

Plasma from 17 HNSCC patients was collected before, during, and after treatment by surgery with adjuvant (chemo)radiation and at time of recurrence. Total exosomal protein (TEP) of exosomes was measured to estimate exosome load. Flow cytometry was used to evaluate relative fluorescence intensity (RFI) of tumor-associated and immunoregulatory proteins on exosomes. Effects of exosomes on CD8+ T cells and adenosine production by CD39+ T cells was assessed by flow cytometry and mass spectrometry.

Results

TEP levels and tumor-derived exosomes varied during and after therapy with an overall decrease in the tumor-free follow up but with an increase at time of recurrence. RFI values of immune checkpoints on exosomes, their potential for T cell inhibition, and adenosine production changed during and after therapy. Recurrent patients showed a different immunoinhibitory profile compared to disease-free patients. Exosomal PD-L1 emerged as the earliest discriminator for treatment failure and disease-free survival.

Conclusions

Plasma-derived exosomes from HNSCC patients emerge as promising non-invasive biomarkers for early detection of treatment failure. These exosomes can accurately inform about changes in patients’ immune status and may alert of impeding recurrence.
Old mice and elderly humans make increased autoimmune antibodies, inflammation and SASP from aged-increased B cells (ABCs in mice and DN in humans) which are hypermetabolic
Authors: Blomberg Bonnie , Diaz Alain , Romero Maria , Frasca Daniela
Published: January 2023

Immune mechanisms of human and murine diseases of aging include generation of chronic inflammation and autoimmunity. We and others have associated aging in mice and humans with increased chronic inflammation, reduced vaccine response and an increase in a B lymphocyte subset referred to as Age-associated B cells (ABCs) in mice and DN (double negative) in humans. Immune cell function is dependent on metabolic pathways and to date understudied. A. In this work we sorted splenic ABCs (CD19+AA4.1-CD21-CD23−) from young (3–4 months) and old (18–22 months) C57BL/6 mice showing an increased percentage in old and increased RNA expression of SASP (senescence-associated secretory phenotype) pro-inflammatory cytokines (TNFa, IL-6), proinflammatory micro-RNAs (miRs 155 and 16) and cell cycle regulators p16INK4 and p21Waf1 in old ABCs. We also show here old B cells produce more autoimmune IgG antibodies (to MDA, malondialdehyde and ADA, adipocyte-derived antigens), ABCs more than FO (follicular), and ABCs from old more than those from young. Total B cells and ABCs from old mice also have a higher metabolic profile compared with those of young mice. B. The frequency of DN cells is increased in human elderly as well as those younger with obesity. Plasma of obese had more autoantibodies to dsDNA, MDA, and ADA and the DN B cells showed higher levels of IA (Immune activation) markers and transcription factors, e.g. tbx21 (TBET) associated with autoimmunity. We have recently found that elderly human B cells are, like murine, hypermetabolic and are discovering possible mechanisms, such as fatty acids.

Supported by grants from NIH (AG32576, AG059719, and AG023717)
CD8 T cell exhaustion is reduced in subjects with autoimmune-associated DR4 risk alleles
Authors: Long Sarah Alice , Jones Britta , Wall Valerie , Muir Virginia , Ylescupidez Alyssa , Uchtenhagen Hannes , Linsley Peter S , Buckner Jane H
Published: January 2023

Exhausted CD8 T cells (TEX) are associated with worse outcome in cancer and better outcome in autoimmunity. However, factors contributing to reduced TEX in the context of autoimmunity are not well understood. Here, we identify co-expression of TIGIT and KLRG1 as a broad identifier of human TEX: TIGIT+KLRG1+ CD8 memory T cells share EOMES signature genes in healthy controls (HC), T1D and Rheumatoid Arthritis (RA) subjects, are increased with age and in chronic viral-specific cells, and are hyporesponsive (reduced proliferation and cytokine production) in vitro. We applied this inclusive, sample sparing and high throughput measure of TEX to larger longitudinal and autoimmune cohorts. Consistent with a genetic determinant, TEX are stable within HC and T1D individuals (Interclass correlation coefficient=92.0 and 83.7%, respectively), but display high interindividual variation (range 0.37–28.47 of CD8). In a large cohort of HC and RA subjects (n>100/cohort), lower levels of EOMES modules and TEX were associated with the HLA Class II DR4 autoimmune risk allele, regardless of disease status. While the DR4 locus is complex, these findings suggest that TEX may be indirectly influenced by APC or CD4 T cell help. Moreover, therapeutic modulation of TEX may differ in DR4 risk and non risk subjects, with implications for precision medicine.

Supported by grants from the NIH (R01 AI141952, R21 5R21AR073508-02) and JDRF.
Interrogation of STAT3 activation in patients with polyarticular juvenile arthritis
Authors: Wood Stephanie Leigh , Branch Justin , Vasquez Pricilla , DeGuzman Marietta M , Brown Amanda , Sagcal Gironella A Carmela , Singla Saimun , Ramirez Andrea , Vogel Tiphanie P
Published: January 2023

Juvenile idiopathic arthritis (JIA) can cause pain and disability, but early disease control improves outcomes. The polyarticular JIA (polyJIA) subset is clinically similar to adult rheumatoid arthritis (RA), yet its pathogenesis remains incompletely understood. A pathway of interest in polyJIA is STAT3 signaling. STAT3 is a transcription factor critical to development of inflammatory Th17 cells and modulation of regulatory T cells. Elevated Th17s and activated STAT3 have been shown in RA. We aimed to profile T cell subsets and STAT3 activation in polyJIA.

Blood from 13 patients with polyJIA was collected at initial diagnosis and serially until remission. Peripheral blood mononuclear cells were isolated and T cell subsets and STAT activation (phosphorylation) were evaluated using flow cytometry.

Regulatory T cells from patients with polyJIA were found at similar levels as pediatric controls. However, treatment naïve polyJIA patients had increased Th17s compared to pediatric controls (0.15% v 0.44%, p=0.037). We identified IL-17 and IFN-γ dual+ helper T cells (non-classical Th1s) appearing in post-treatment samples from polyJIA patients. After stimulation (IL-6, IL-27 or IFN-α), a subset of polyJIA patients had highly activated STAT3, but not STAT1 or STAT5, in CD3+CD4+ T cells.

A better understanding of the mechanisms underlying polyJIA is needed for more efficient and effective clinical care. Although multiple therapeutics to treat JIA exist, a biomarker-driven approach to treatment does not. Patients with polyJIA have increased Th17s and a subset have highly activated STAT3. These immunologic features may help identify patients that would benefit from upfront, tailored treatment with therapeutics targeting this pathway.
Blockade of dendritic cell glutaminolysis induces allergic asthma desensitization via suppression of Tfh13 polarization
Authors: Tharakan Anuj , Liu Jinze , Cowart Ashley , Martin Rebecca
Published: January 2023

Allergic asthma is a pulmonary disease characterized by airway hyperresponsiveness (AhR) in response to inhaled allergens. AhR is largely mediated by allergen-specific (Ag-specific) IgE induced mast cell (MC) degranulation which drives inflammation of the airways following allergen exposure. Ag-specific IgE production requires T follicular helper 13 cells (Tfh13s) which must first be polarized by dendritic cells (DCs). The mechanisms by which DCs polarize Tfh13s, however, are unclear. We hypothesized that the Tfh13 inducing allergen Alternaria would induce a unique metabolic response compared to non-Tfh13 inducing stimuli. We found, by untargeted metabolomic analysis, that Alternaria induced accumulation of the glutamine-derived TCA cycle metabolite α-ketoglutarate compared to LPS stimulated or unstimulated DCs. Glutamine is converted to α-ketoglutarate by glutaminase (Gls), so we predicted that inhibition of Gls may suppress Tfh13 polarization by DCs. Indeed, we found that treatment with the Gls inhibitor, CB-839, reduced Tfh13 polarization and increased T follicular regulatory cells following Alternaria exposure. This finding was recapitulated in a DC adoptive transfer model, indicating that this mechanism is DC intrinsic. We next tested if CB-839 causes allergen desensitization in a model of allergic asthma. Mice were intranasally sensitized and challenged with Alternaria and intranasally treated with CB-839 or vehicle control prior to challenge. CB-839 treated mice exhibited reduced AhR, Ag-specific IgE, and airway MC degranulation. These data demonstrate that DC glutaminolysis is required for Tfh13 polarization, and that Gls inhibitors may promote allergen desensitization in allergic asthma.

This work was funded by VCU’s National Center for Advancing Translational Sciences (UL1TR002649) and NIAID (R56AI139658).
E3 Ubiquitin Ligase Cbl-b Inhibits Type 2 Innate Lymphoid Cell Development by Targeting ST2 for Ubiquitination
Authors: Guo Hui , Huang Ling , Waldstein Kody , Varga Steven M , Zhang Jian
Published: January 2023

Type 2 innate lymphoid cells (ILC2) play a crucial role in driving allergic airway inflammation, but the regulation of ILC2 development is not fully understood. Previously we have shown that E3 ubiquitin ligase Cbl-b negatively regulates Th2 response, but whether Cbl-b also regulates ILC2-mediated type 2 immunity remains to be defined. In this study, we found that Rag1−/−Cblb−/− mice are hypersensitive to inhalation of A. Fumigatus, a fungus that causes allergic asthma, as revealed by heightened airway hyperresponsiveness (AHR), increased airway inflammation, and mucus production. This hypersensitivity is associated with increased frequencies of ILC2 and eosinophils, but reduced frequency of alveolar macrophages in the lungs. Consistent with this, type 2 cytokines are also increased in the lung homogenates of Rag1−/−Cblb−/− mice. At the molecular level, ST2 undergoes ubiquitination and proteasome-mediated degradation upon IL-33 stimulation, which is abrogated in cells expressing Cbl-b C373A mutant. Therefore, our data indicate that Cbl-b inhibits the development of ILC2 and ILC2-mediated airway inflammation by targeting ST2 for ubiquitination.

Supported by NIH R01 AI090901, AI121196, and AI123253
PD-1 agonist modulates ILC2 metabolism and ameliorates airway hyperreactivity
Authors: Helou Georges Doumet , Shafiei-Jahani Pedram , Lo Richard , Howard Emily Diane , Hurrell Benjamin P , Galle-Treger Lauriane , Painter Jacob Dean , Lewis Gavin , Soroosh Pejman , Sharpe Arlene H , Akbari Omid
Published: January 2023

Allergic asthma is a chronic inflammatory disease characterized by airway hyperreactivity (AHR) and type-2 immune response. Type-2 innate lymphoid cells (ILC2s) mimic T helper 2 (Th2) cells in cytokine secretion and are among the first pulmonary immune responders to the allergen-induced alarmins. Programmed cell death protein-1 (PD-1) is known as an immune checkpoint equipped with tyrosine-based inhibitory motifs in the cytoplasmic tail. Since PD-1 is associated with immune regulation in many inflammatory diseases, the objective of this study was to investigate the role of PD-1 in the initiation and development of AHR. Lung function tests, RNA sequencing, flow cytometry, targeted metabolomic assays, and adoptive transfer experiments were principally used to explore the role of PD-1 in AHR mouse models and in human ILC2s. Using IL-33 and Alternaria alternata models, we have demonstrated that PD-1 knockout mice develop a higher AHR and lung inflammation as compared to control wild-type mice. In particular, PD-1 is highly inducible on lung ILC2s and downregulates their effector functions. Moreover, PD-1 controls glycolysis and methionine catabolism, limiting, therefore, the proliferation of activated pulmonary ILC2s. In line with mice data, PD-1 is inducible and functional in human ILC2s in response to IL-33. To confirm the translational relevance of our findings, we tested a novel human PD-1 agonist in vitro and in a humanized mouse model of AHR. Interestingly, the PD-1 agonist decreases human ILC2 activation and is able to dampen AHR and lung inflammation. Altogether this study reveals the protective role of PD-1 as regulators of ILC2s and highlights for the first time the therapeutic potential of PD-1 agonists in allergic asthma.

Supported by grants from NIH: R01 ES025786, R01 ES021801, R01 HL144790, and R21 AI109059 (O.A.)
Immune determinants of cardiometabolic risk in pre-existing type-2 diabetes (T2D) severe COVID-19 patients
Authors: Gunasena Manuja Gayashan Bandara , Wijewantha Yasasvi , Bowman Emily , Kumar Amendra , Weragalaarachchi Krishanthi , Furay Jerra , Skladany Tatum , Liu Shan-lu , Vilgelm Anna , Bednash Joseph , Kasturiratna Dhanuja , Demberg Thorsten , Funderburg Nicholas , Liyanage Namal
Published: January 2023

COVID-19, the disease caused by SARS-CoV-2, has led to a global public health emergency. Severity of disease course may be related to a dysregulated immune response and pre-existing health conditions. Recent studies have demonstrated that SARS-CoV-2 infection may directly or indirectly lead to an increase in cardiometabolic complications in patients with pre-existing type-2 diabetes mellites (T2DM) when compare to non-DM patients. A lack of mechanistic and systematic studies on how SARS-CoV-2 infection related immune responses may contribute to increase risk of cardiometabolic complications in pre-existing T2DM patients, hinder early risk identification and therapeutic interventions. Thus, in this study we investigate the biomarkers of cardiometabolic risk in non-DM and T2DM, severe COVID-19 patients admitted to the Intensive care unit.

Using high-dimensional flowcytometry and immune biomarker assays, we investigated functional and phenotypic changes in immune subsets in whole blood and plasma biomarkers of cardiovascular disease in healthy donors (n=17), T2DM severe-COVID-19 patients (n=10), non-diabetic severe-COVID-19 patients (n=10) admitted to the OSU medical center’s intensive care unit.

We found neutrophils and Intermediate monocytes (ITM) were significantly higher in the T2DM group compared to non T2DM patients. However, activated (HLA-DR+) NKT-like cells and NKG2A+ CD56 Dim CD16+ NK cells, were significantly lower in the T2DM-COVID-19+ group. Interestingly, LBP, FABP4, sCD14, IL-1b, RANTES and MIP-1a were significantly higher in the COVID-19 T2DM patients.

In this study, we identify core immune signatures that may predict increased cardiovascular disease risk in T2DM patients who had severe COVID-19.

Supported by PI’s startup funding
Selective IRAK4 degradation, not kinase inhibition, blocks TLR-activated NF-Kb and p38 Signaling leading to broad cytokine inhibition
Authors: Skouras Stephanie M , Massa Virginia , Lurier Emily , Hubeau Cedric , Wang Amy , Zheng Xiaozhang , Sharma Kirti , Walter Dirk , Browne Chris , Mayo Michele , Fitzgerald Michael , McDonald Alice , Gollob Jared , Mainolfi Nello , Slavin Anthony , Campbell Veronica
Published: January 2023

IL-1R/TLR activation plays a central role in the pathophysiology of multiple autoimmune and inflammatory diseases via the Myddosome that is dependent on both the kinase and scaffolding functions of Interleukin-1 receptor associated kinase 4 (IRAK4). Therefore, hetero-bifunctional molecules that selectively target IRAK4 for degradation and elimination by the ubiquitin proteasome pathway have the greatest potential to block IL-1R/TLR signaling, including NF-kb activation and cytokine production. To interrogate downstream signaling, phosphorylation events were monitored in monocytes and B cells following TLR7/8 or TLR9 activation, respectively. IRAK4 degradation, but not kinase inhibition, inhibited NF-kb p65 activation and phosphorylation of p38 pathway members by TLR agonists in both cell types. PBMCs pretreated with an IRAK4 degrader and then stimulated with the TLR7/8 agonist, R848, exhibited significantly broader inhibition of cytokines (IL-6, TNFa, IL-8 and IL-1b) compared to those pretreated with a selective IRAK4 kinase inhibitor. Compound washout experiments demonstrated a sustained effect of IRAK4 degrader on both target pharmacodynamics and cytokine inhibition that was differentiated from IRAK4 kinase inhibition. Overall, through removal of both scaffolding and kinase functions, IRAK4 degradation demonstrated greater activity compared to kinase inhibition across multiple TLR stimuli and cell types. These data highlight the potential for IRAK4 degraders to block multiple TLR signaling pathways across different immune cell types and thereby impact TLR/IL-1R-driven inflammatory and autoimmune diseases.
Dynamic changes in human single-cell transcriptional signatures during fatal sepsis
Authors: Qiu Xinru , Li Jiang , Bonenfant Jeff , Jaroszewski Lukasz , Mittal Aarti , Klein Walter , Godzik Adam , Li Jiang G
Published: January 2023

Systemic infections, especially in patients with chronic diseases, may result in sepsis: an explosive immune response that can lead to multisystem organ failure with a high mortality rate. Patients with similar clinical phenotypes or sepsis biomarker expression upon diagnosis may have different outcomes, suggesting that the dynamics of sepsis is critical in disease progression. A within-subject study of patients with Gram-negative bacterial sepsis with surviving and fatal outcomes was designed and single-cell transcriptomic analyses of peripheral blood mononuclear cells (PBMC) collected during the critical period between sepsis diagnosis and 6 h were performed. The single-cell observations in the study are consistent with trends from public datasets but also identify dynamic effects in individual cell subsets that change within hours. It is shown that platelet and erythroid precursor responses are drivers of fatal sepsis, with transcriptional signatures that are shared with severe COVID-19 disease. It is also shown that hypoxic stress is a driving factor in immune and metabolic dysfunction of monocytes and erythroid precursors. Last, the data support CD52 as a prognostic biomarker and therapeutic target for sepsis as its expression dynamically increases in lymphocytes and correlates with improved sepsis outcomes. In conclusion, this study describes the first single-cell study that analyzed short-term temporal changes in the immune cell populations and their characteristics in surviving or fatal sepsis. Tracking temporal expression changes in specific cell types could lead to more accurate predictions of sepsis outcomes and identify molecular biomarkers and pathways that could be therapeutically controlled.

This research was supported by the UCR School of Medicine (to A.G. and M.G.N.), the Dean Innovation Fund (to J.B. and M.G.N.), and the National Institutes of Health (NIAID, R21AI37830, and R01AI153195 to M.G.N.). The data from this study were generated at the UC San Diego IGM Genomics Center utilizing an Illumina NovaSeq 6000 that was purchased with funding from a National Institutes of Health SIG grant (#S10 OD026929).