bluuebunny/crossref_metadata_2025 huggingface dataset:
Using Bayesian networks with tabu algorithm to explore factors related to chronic kidney disease with mental illness: A cross-sectional study
Authors: Yuan Xiaoli , Song Wenzhu , Li Yaheng , Wang Qili , Qing Jianbo , Zhi Wenqiang , Han Huimin , Qin Zhiqi , Gong Hao , Hou Guohua , Li Yafeng
Published: August 2023

While Bayesian networks (BNs) offer a promising approach to discussing factors related to many diseases, little attention has been poured into chronic kidney disease with mental illness (KDMI) using BNs. This study aimed to explore the complex network relationships between KDMI and its related factors and to apply Bayesian reasoning for KDMI, providing a scientific reference for its prevention and treatment. Data was downloaded from the online open database of CHARLS 2018, a population-based longitudinal survey. Missing values were first imputed using Random Forest, followed by propensity score matching (PSM) for class balancing regarding KDMI. Elastic Net was then employed for variable selection from 18 variables. Afterwards, the remaining variables were included in BNs model construction. Structural learning of BNs was achieved using tabu algorithm and the parameter learning was conducted using maximum likelihood estimation. After PSM, 427 non-KDMI cases and 427 KDMI cases were included in this study. Elastic Net identified 11 variables significantly associated with KDMI. The BNs model comprised 12 nodes and 24 directed edges. The results suggested that diabetes, physical activity, education levels, sleep duration, social activity, self-report on health and asset were directly related factors for KDMI, whereas sex, age, residence and Internet access represented indirect factors for KDMI. BN model not only allows for the exploration of complex network relationships between related factors and KDMI, but also could enable KDMI risk prediction through Bayesian reasoning. This study suggests that BNs model holds great prospects in risk factor detection for KDMI.
Nietzsche and Badiou: Event, Intervention, “God is dead”
Authors: Bunta Aleš
Published: August 2023

The article draws attention to a certain multi-layered parallel between Nietzsche and Badiou’s theory of the event, which the author argues Badiou evaded by a kind of strategic relocation. The article does not focus so much on (and certainly not against) Badiou’s philosophy, but attempts to assess the possible implications of this relocation for Badiou’s interpretation of Nietzsche. In the first part of the article, the key concepts of Badiou’s account of Nietzsche are introduced, such as “archi-politics”, “antiphilosophy”, and the “event Nietzsche”. Later, it is shown that, despite some fundamental differences, there is a strong resemblance between some key concepts of Badiou’s theory of the event, especially those related to the notion of “intervention”, and some concepts found in Nietzsche. Finally, the paper focuses on the problem of the “death of God”, which is somehow absent in Badiou’s interpretation, even though the term itself resonates strongly in various aspects of Badiou’s philosophy.
Voyeurism and Exhibitionism on the Internet: The Libidinal Economy of the Spectacle of Instanternity
Authors: Kolenc Bara
Published: August 2023

Today, in the situation that we call the instantiernity of the digital age, the visual aspect of the social (and power) relations is ever more important. A majority of human interaction on internet is happening in the field of vision. In this field, human desire follows the scopic drive, which is, according to Freud, expressed in the ambivalence of voyeurism and exhibitionism. This means that voyeurism and exhibitionism are the fundamental mechanisms operating in, and structuring, the digital virtual. This topic, in a broader sense, tackles the inscription of the subject within the digital virtual spectacle, which deals with the relation between individual’s imaginary and symbolic identification, that is, between ideal ego and ego ideal. To a certain extent, this also relates to what has been marked as »pathological narcissism«. Even if the changes brought about by the digital virtual, as far as the subject is concerned, are not ontological, i.e. they do not concern its relation to being, they do concern its entry into the field of the Other, and can, because they are systemic changes, fundamentally restructure the social fabric. The bet of this article is therefore not only to try to understand the mechanisms driving the formation of subectivity within the digital virtual, but also to trace their transformative potential.
Bibliometric Analysis Examining the Adoption of Augmented Reality in Tourism Research
Authors: Tinesa Fara Prihandini  , Mochammad Bruri Triyono  , Priyanto
Published: August 2023

This study aims to identify the use of Augmented Reality technology and the trend of usingAR in tourism with time limitation from 2013 until 2022. It also aims to determine the valuablecontributions of documents, sources, authors, countries, and institutions worldwide intourism. The qualitative method using bibliometric analysis is suitable for this study becauseit can provide an overview and classify. Data was collected from the Scopus database andthen analyzed using VOSviewer software. The results show that (1) museum, (2) sustainabletourism, and (3) heritage tourism are the keywords most often used in research. The potentialresearch of AR in tourism is directed at the development of underwater tourism, the applicationof virtual museums, ecotourism, culture heritage tourism, digital heritage and destinationimage. Sustainability (Switzerland), Current Issues in Tourism, and The International Journalof Human-Computer are the journals with the most publications. Tom Dieck M. C.M.C, JungT, and Chung N are authors with a high degree of collaboration with other authors. UnitedKingdom, Spain, and Italy became the countries with the highest publications. Researchtrends and advances in augmented reality in tourism have been discussed in more detail inthis paper.
How Harsh Should the Legislation Be to Prevent Financial Crimes?
Authors: Iren Perihan , Kim Moo Sung
Published: August 2023

The collapse of Enron and its auditor Arthur Andersen was one of the biggest events in US corporate history and it had long-lasting repercussions in the corporate world. Following the Enron – Arthur Andersen scandal, the US Congress passed the Sarbanes Oxley Act (SOX). The legislation was designed to strengthen existing securities regulations and enacted harsh penalties on lawbreakers. SOX was one of the first and heavy responses to corporate fraud. Since the passage of SOX, debates revolving around the costs and benefits of it have prevailed and several studies investigated its effectiveness to prevent future financial crimes. This study first reviews the chronology of the events leading to SOX, followed by a brief discussion of the rules and regulations established by SOX. The empirical evidence presented by previous studies on the effectiveness of legislation and the harsh penalty Enron's auditor had received are assessed and evaluated.
Circular Frauds and Abuses of VAT
Authors: Stipic Valentina Vinsalek
Published: August 2023

Organized tax frauds include circular frauds which are very complex in EU member states. Circular fraud is a way of the organized and systematic extraction of money from the state budget by exploiting the shortcomings of the tax system. The destination principle is based on the same tax treatment and the same market position of domestic and imported goods, but it also enables circular fraud. New member countries of the EU are struggling with circular fraud. Losses in the territory of the EU are measured in billions of euros and thus the negative effects on the budgets of those countries are growing. Tax frauds also involve countries outside Europe (UAE, Hong Kong, Russia). On the other hand, in tax havens, illegally acquired money is transferred as a part of the money laundering process. Monitoring and suppression are becoming more and more demanding because the center of gravity has moved to the domain of intangible assets due to the evolution of the internet. The regulatory bodies of the EU are trying hard to combat circular fraud and VAT abuse.
The Political Economy of Corruption
Authors: Kırşanlı Fatih
Published: August 2023

Lebanon is facing one of the most acute socioeconomic and political crises in history. While it was the financial center during the 1950s, 60s, and early 70s with its sophisticated banking sector, the civil war between 1975 and 1990 demolished this position. Beginning from 1990 until 2019, the Lebanese economy was slowly dragged into an economic collapse with depreciated Lebanese lira against foreign currencies, which was the preferred monetary policy. This chapter focuses on this collapse and the so-called Ponzi scheme, organized by the Lebanese central and commercial banks. It elucidates the leading cause of the issues (e.g., financial engineering, Beirut Explosion, political deadlock, economic turmoil), namely rampant corruption, from the perspective of political economy. It also discusses foreign interventions, the dominance of Hezbollah, and terror financing issues. Lastly, it sheds light on future research by providing policy recommendations for the Lebanese political economy.
Legal, Financial and Fiscal Measures to Support the Collection Solutions of Biowaste in Portugal
Authors: David Fátima , Rodrigues Pedro Melo , Soares Elisabete , Azevedo Tânia Cristina
Published: August 2023

The Portuguese Decree-Law No. 102-D/2020, of December 10 (PCM, 2020b), dictates the implementation of biowaste selective collection networks or the separation and recycling of biowaste at the source until December 31, 2023. This chapter aims to analyze the financial support provided to municipalities (local authorities) for implementing biowaste collection solutions, such as on public roads or through door-to-door collection, and to citizens for adopting biowaste separation and recycling practices through domestic or community composting. The authors also explore the application of the polluter-pays principle, which requires citizens to pay environmental taxes and fees to support sustainable development. Regarding this focus, the chapter highlights the legal, financial, and fiscal measures used to support biowaste collection and recycling efforts in Portugal, providing insight into the steps needed to achieve a sustainable environment while addressing the economic and social needs of the population.
Sustainable Taxi Fleet
Authors: Dinis Ana Arromba , Carvalho André , Dias Sara Luís
Published: August 2023

It is a well-known fact that the automotive sector plays a key role in a country's socio-economic development, due to the large contribution of vehicles to greenhouse gas emissions (GHG). Alternatives such as car-sharing, public transport, cycling, the transition to electric vehicles, or walking can help reduce emissions. Therefore, it becomes a sector where environmental regulation should be demanded. This research aims to contribute to the tax literature by extending previous studies on corporate taxation by evaluating green tax incentives as an effective environmental tax measure. Considering taxis as an important part of urban public transport systems, the authors set out to investigate Portuguese taxi companies that have taken advantage of tax incentives for less environmentally polluting vehicles. The authors conclude that the income tax level, the economic profit, and the number of employees, are determining factors in these firms ‘access to the tax incentives under study.
The Use of Artificial Intelligence in Education
Authors: Mupaikwa Elisha
Published: August 2023

Over the past few decades, there has been an increase in ongoing research into the use of artificial intelligence in education. The usage of artificial intelligence in educational institutions has grown during the past few years. This chapter examines how artificial intelligence is used in education. The establishment of inclusive learning environments, collaborative learning and research, social robots, student assessment, automated grading, recommender systems, and student performance prediction were identified to be the main applications of artificial intelligence in education. Lack of funding, lack of faith in the system, lack of portability of systems, infrastructure issues, ethical issues regarding data privacy and security, and a lack of experience were some of the difficulties noted. After that, the chapter included recommendations for infrastructure improvements, ICT policy changes at the national and institutional levels, curriculum revisions for teacher- and school-level preparation, and additional research.
Research on Artificial Intelligence Risk Prevention and Control System for Smart Libraries
Authors: Wang Xu , Liu Binbin
Published: August 2023

With the rapid development of artificial intelligence (AI) science and technology, people's demand for library information services and risk prevention and control construction has increased in both directions, affecting profound changes in the construction of libraries in various countries. Artificial intelligence is reshaping the management model and service model of smart libraries, and libraries must pay attention to risk prevention and control while welcoming the model change. This chapter is based on the background of artificial intelligence era. To achieve the deep synergy between artificial intelligence and smart libraries, it combines risk management and multiple synergy theory, and it proposes that the types of AI risks in smart libraries are divided into their own risks and application risks. This chapter constructs the AI risk prevention and control system for smart libraries and proposes feasible AI risk prevention and control countermeasures for smart libraries.
Building Social Capital in Contemporary Major U.S. Public Libraries
Authors: Kee Helen Chui Ling , Chan Mimi Mei Wa , Chiu Dickson K. W.
Published: August 2023

As the US is a highly ethnically diverse country and the origin of public libraries, this chapter explores how US public libraries construct social capital for the public via various services and activities. This chapter selected nine cases of US public libraries with interviews of their management for analysis of library services and activities related to empowering library users and building their social capital. The findings indicate that libraries are community meeting places connecting community members through library programs, activities, and information services as community educational institutions that empower underprivileged people and new immigrants by satisfying their information needs. The process contributes to the social development of library users and their communities, building social capital. Scant studies summarize the good practice of renowned public libraries in building social capital. This chapter contributes to understanding the good practice of US public libraries as a creator of social capital, serving as a reference for public libraries worldwide.
The Impact of Twitter Users' Characteristics on Behaviors
Authors: Uppala Vishal , Palvia Prashant , Ankem Kalyani
Published: August 2023

Researchers have focused on leadership, often overlooking followership. The notion of followership was irreversibly transformed with the advent and societal adoption of followership systems, such as Twitter. To examine such emergent systems, this paper advances a distinct form of followership: eFollowership. To understand Twitter and its users, the eFollowership concept is explicated and synthesized by adapting several followership lenses from the literature. The authors empirically examined eFollowership by assessing the roles constructed by 301 Twitter users and the relationships between these users' role-based characteristics and behaviors with partial least squares structural equation modeling (PLS-SEM). Results showed that users' voicing and empowering behaviors were significantly influenced by users' characteristics: personal sense of power, eCourage, and social capital. Users' helping behaviors were related to users' personal sense of power and social capital, but not to eCourage. Surprisingly, users' disempowering behaviors were unrelated to all three users' characteristics.
Government Supervision in Curbing the Spread of COVID-19
Authors: Kong Xiaolin , Chen Yan , Lin Zijun , Kuang Xianhua
Published: August 2023

The outbreak of coronavirus disease 2019 (COVID-19) has brought severe challenges to global economic development and government management. According to a report on the United Nations News website (news. un. org), as of May 3, 2023, the cumulative number of confirmed cases of COVID-19 worldwide has reached 765 million, and more than 6 million people have lost their lives as a result. How to evaluate the effect of government management has become a key issue. In this research, the SEIR model is used to estimate the network average degree of Chinese cities under network limitation. The results show that government management has reduced the average degree of the COVID-19 network and the prevention of first-tier cities will be more difficult. This study scientifically evaluates the government management of the epidemic and provides ideas for emergency management of infectious diseases that affect the public.
Synthetic glycovaccines elicit cross-protection against Chagas disease and tegumentary leishmaniasis in a murine co-infection model
Authors: Knight Colin D , Estevao Igor L , Portillo Susana , Farani Priscilla Silva Grijo , Montoya Alba , Heydemann Emily , Carvajal Elisa G , Vinales Irodiel , Gil Eileni R , Rodriguez Felipe , Maldonado Rosa A , Michael Katja , Almeida Igor C
Published: August 2023

Chagas disease (CD) and tegumentary leishmaniasis (TL) are two of the World Health Organization’s (WHO) top 20 neglected tropical diseases. These diseases are caused by the protozoan trypanosomatids, Trypanosoma cruzi and Leishmania spp., respectively. Although historically restricted to Central and South American, there are currently, ~7 million people infected with CD worldwide, with ~300,000 chronically infected patients in the US. Between 700,000 and 1.2 million new cases of TL occur worldwide yearly. Chemotherapeutic agents are available for the treatment of both diseases; however, they may cause serious adverse events, ultimately resulting in premature treatment interruption. Moreover, no effective vaccine for either disease is currently available. Due to the evolutionary loss of the α1,3-galactosyltransferase (α1,3GT) gene in humans and Old-World nonhuman primates, α-Gal epitopes, abundantly expressed on both Trypanosoma cruzi and Leishmania spp. cell surfaces, are highly immunogenic. We, therefore, hypothesize that a synthetic glycan-based vaccine could provide effective cross-protection against a T. cruzi and L. major co-infection. Here we use an α1,3GT-knockout (α1,3GT-KO) murine model to evaluate the efficacy of two new synthetic neoglycoproteins, NGP28b and NGP29b, containing nonreducing, terminal α-Gal and β-Galf glycotopes, respectively, as potential vaccine candidates against CD and TL. Our data indicate that both vaccine candidates induce a strong cross-protective immune response, significantly reducing parasite burden and increasing animal survival in this co-infection experimental model

This study was partially supported by a grant NIH-U54MD007592 (to Robert A. Kirken (PI), ICA (Co-I)), from the National Institute on Minority Health and Health Disparities (NIMHD). We would like to thank the staff of the Border Biomedical Research Center (BBRC) Biomolecule Analysis Omics Unit (BAOU). BAOU was supported by the NIMHD grant 5U54MD007592. CDK was supported by the UT System LSAMP program which is funded by the National Science Foundation grant number HRD-1202008.
Induction of multivalent and balanced T cell responses by modular nanoshell vaccine technology
Authors: Huang Pinghan , Tsai Hsiao-Han , Tat Vivian , Hsu Jason C , Aleksandra Drelich , Chen Hui-Wen , Hu Che-Ming Jack , Tseng Chien-Te
Published: August 2023

After the outbreak of COVID-19, cellular immunity has been shown to provide protection especially in breakthrough infection and B cell-deficient patients. Furthermore, durable cellular immunity targets conserved epitopes among SARS-CoV-2 variants or different coronavirus then inspiring an attractive strategy for vaccine development. However, how to simultaneously induce balanced T cell responses against multiple epitopes still needs to be elucidated. In this study, we sought to design a T cell-based vaccine that can adjust valency against different T cell epitopes. We selected seven SARS-CoV-2 specific short peptides as antigens, including I-Ab/H-2Kb/H-2Db restricted epitopes. Furthermore, we used PLGA nanoparticle as vaccine platform to achieve the precise loading of the indicated peptides. Mice were immunized with different combinations of peptides and splenocytes were used to access epitope specific-IFN gamma expressing CD8+ or CD4+ T cell responses. First, we confirmed the robust immunogenicity of each peptide. We then combined an equal amount of indicated epitope peptides and accessed the immunogenicity of each epitope in the combination. The results showed peptides compete with each other to induce T cell responses regardless of MHC allele and MHC class. In addition, dominant epitopes significantly suppressed the immunogenicity of the subdominant epitopes. Finally, by optimizing the antigen dose, nanoparticle vaccine induced more balanced T responses against all seven epitopes. Taking advantage of nanoparticle, our results reveal a viable strategy to induce multivalent T cell response. Furthermore, the result suggests that competition of T cell epitopes is one of the factors controlling cellular immunity.
Single-Chain B cell receptor mice as a novel tool for antibody generation and exploring B cell signaling
Authors: Borbet Timothy C , Zaldana Kimberly , Ilmain Juliana K , Chan Kun-Wei , Luo Christina C , Bryan Arielle M , Kirilov Milen , Kong Xiang-Peng , Bhabha Gira , Ekiert Damian C , Weng Wei , Sheiffele Paul , Koralov Sergei B
Published: August 2023

Conventional antibodies consist of a pair of heavy and light dimers, while heavy-chain only antibodies (HCAbs), like those found in camelid and shark species, lack a domain critical for light chain binding and develop without a light chain. HCAbs have unique physiochemical properties that confer advantages: they are smaller, have greater accessibility to their target antigens, higher hydrophilicity, higher tissue penetrance, and greater conformational stability under environmental stresses compared to conventional Abs. They are also highly amendable to biochemical engineering. We have generated a mouse model, that can produce HCAbs from the endogenous murine heavy chain locus. We have shown that these mice undergo efficient B cell development in the bone marrow and generate a diverse repertoire of B cells that populate all peripheral immune compartments. Single-cell sequencing of B cells revealed some biases in VH and JH usage, but highlighted the tremendous diversity of rearrangements that give rise to B cells bearing HCAbs. With these mice, we have been able to generate antigen specific HCAbs to the SARS-CoV2 spike protein that have a KD in the nM range, measured via bio-layer interferometry. Further, the IgG3dCH1 mice provide us with a novel model to understand how single domain B cell receptors (BCRs) function in comparison to conventional heavy/light chain BCRs. We have observed single-chain BCRs have distinct transcriptional profiles by single-cell sequencing and support higher plasma cell differentiation in vitro and in vivo. These data highlight the utility of our small animal platform for the generation of clinically relevant HCAbs, and as a model to study the biology of HCAb encoding B cells which remain poorly understood.

Bernard Levine Postdoctoral Fellowship, R43AI136141
Targeting CD70 using CAR NK cells to enhance NK cells cytolytic effect against osteosarcoma
Authors: Rav Emily Simon , Anjier Ariana , Acharya Sunil , Basar Rafet , Wang Yifei , Daw Najat , Rezvani Katy , Gopalakrishnan Vidya , Gorlick Richard , Kleinerman Eugenie S , Gordon Nancy
Published: August 2023

Background

Cure rates for osteosarcoma (OS) have not improved in >30 years and no new effectivetherapies have been identified for metastatic disease. Natural killer (NK) cells have become an attractive tool for cancer immunotherapy. Efficay is improved by direct tumor targeting introducing a specific chimeric antigen receptor (CAR). Emerging knowledge suggests CD70 to be a viable candidate as expression is higher in lung metastases and it has been correlated with tumor escape from immune surveillance.

Objective

To assess whether using CD70 CAR-NK cells we can enhance trafficking, homing and proliferation of NK cells and therefore therapeutic efficacy against OS.

Methods

We analyzed the cBioportal database to verify CD70 expression in OS. Surface expression of CD70 on human OS cell lines and normal osteoblasts was determined by flow cytometry. CD70 CAR constructs were sequenced, verified and transduced into NK cells. CD70 CAR NK and control NK cells cytolytic activity against OS cells was evaluated by IncuCyte. In vitro cytokine release was measured upon CD70 CAR NK/NK cells exposure to OS cells (IsoPlexis).

Results

Database analysis confirmed OS CD70 gene amplification. Flow cytometry showed variable expression of CD70 on OS cells and no expression on human osteoblasts. There was an increase in CD70 CAR NK percent lysis against OS cells at the highest effector:target ratios compared to control NK cells. We found a predominant release in the chemoattractive/stimulatory cytokines upon OS cells exposure to CD70 CAR NK cells.

Conclusion

CD70 has immunotherapy potential against OS. CD70 CAR NK cells have increased cytolytic activity against OS cells in vitro. Cytolytic activity may be influenced by the release of specific cytokines.
CD4 depletion enhances the efficacy of Super2 IL-33 armored CAR T cell against solid tumors and leads memory formation of both endogenous and CAR T cells
Authors: Mohamed Asmaa , Ferry Shannon Lynn , Huang Yina H
Published: August 2023

Chimeric antigen receptor (CAR) T cells exhibit tremendous efficacy in patients with relapsed and recurrent liquid cancers. However, CAR T cells fail to control solid tumors because of their inability to infiltrate the suppressive tumor environment and their poor persistence in vivo. The development of 4th generation armored CAR T cells that employ direct tumor killing and release cytokines that can modulate the immune response to enhance tissue/tumor infiltration may improve CAR T cell persistence. Our lab has developed Super2 and IL-33 (S233) armored CAR T cells that significantly delayed B16F10 melanoma tumor growth and enhanced tumor infiltration of CARs and endogenous immune cells. Combination therapy of S233 CAR T Cells with anti-CTLA4 or anti-CD4 further enhanced this phenotype. S233 CAR T cells and anti-CD4 combination therapy promoted Melanoma-associated vitiligo (MAV), which correlates with development of tumor protective Tissue Resident Memory (TRM) cells. To assess whether CAR T cells differentiate into memory T cells, we treated tumor-bearing mice with S233 CAR T Cells with or without anti-CD4, resected their residual tumors and tracked memory T cell responses. We found that S233 CAR T cells persisted longer in the tumor draining lymph node and developed accelerated MAV when combined with anti-CD4. Both CAR T cells and endogenous T cells differentiated into central and effector memory but only endogenous T cells in the CD4 depleted group became TRM cells. While these findings provide insights on strategies to enhance long-term persistence of CAR T cells, additional studies are needed to induce CAR TRM cells and assess the durability of anti-tumor responses.

NIH grant T32-AI007363 NIH grant P30-CA023108
Persistent activation of TNFR1 on spinal cord interneurons contributes to immune dysfunction after spinal cord injury in rodents
Authors: Martynyuk Tetyana , Bracchi-Ricard Valerie , Ricard Jerome , Tom Veronica J , Bethea John R
Published: August 2023

Spinal cord injury (SCI) is a devastating condition with 250,000 to 500,000 new cases globally each year. Respiratory infections, e.g., pneumonia and influenza, are the leading cause of death after SCI. For instance, an individual with SCI is 37 times more likely to become infected and hospitalized from the flu. Yet, the field has a poor understanding of how immune dysfunction arises after trauma to the central nervous system (CNS) and how altered neuro-immune communication impacts an individual’s outcome to infection. In humans and rodents, SCI leads to maladaptive changes in the spinal-sympathetic reflex (SSR) circuit which is crucial to sympathetic function. Soluble Tumor Necrosis Factor (sTNF) is a pro-inflammatory cytokine that is elevated in the CNS after SCI and remains elevated for several months after injury. We recently demonstrated that blocking TNFR1 activation in the CNS, using XPro1595, decreased the maladaptive excitability of SpINs in the SSR circuit leading to improved immune response to infection after SCI. We hypothesize that hyper-excitability of glutamatergic SpINs after SCI due to persistent TNFR1 activation contributes to immune dysfunction. To test this, we utilized viral mediated targeted knock-down of TNFR1 on excitatory SpINs, and demonstrated that inhibiting TNFR1 on excitatory interneurons contributes to improved immune function, including an increase in virus-specific T cells and decrease in viral load remaining in the lung after infection.

Supported by grants from NIH (R01 NS111761)

Supported by grants from NIH (R01 NS111761)
IL-12 alters T-cell metabolism to augment anti-tumor immunotherapy
Authors: Mills Stephanie Ann , Dwyer Connor , Quinn Kristen , Oberholtzer Nathan , Chakraborty Paramita , Mehrotra Shikhar
Published: August 2023

To better limit tumor growth, adoptive T cell therapy (ACT), a cutting-edge new strategy for treating cancer, permits patients’ T cells to be modified. Long-lived central memory cells (Tcm), a specific subset of T cells, have been demonstrated through studies conducted in our group and others to be more effective at controlling tumor burden. Powerful inflammatory cytokine IL-12 has been extensively studied for its roles in the interaction of innate and adaptive immunity; however, therapeutic potential has not been optimized. We hypothesized that ex vivo administration of IL-12 may improve metabolic fitness and anti-tumor capabilities of T cells. Using a B16 murine melanoma model and T cells that are reactive to the melanoma epitope gp100, we found ex-vivo conditioning with IL-12 increased stem cell memory (Tscm) and central memory (Tcm) T cell populations. Additionally, we found unique metabolic signatures and proliferation patterns were dependent on IL-12 duration. We also observed enhanced engraftment and tumor control for up to 60 days after adoptively transferring these cells into animals with subcutaneous melanoma tumors. These metabolically conditioned T cells showed improved mitochondrial mass and function, elevated cytokine output, and perforin expression. Aside from switching from glycolysis to oxidative phosphorylation, these cells were also able to continue producing cytokines even when glucose or pyruvate levels dropped. Together, these findings imply that IL-12 modifies T cells’ reliance on glucose for energy while preserving their anti-tumor ability, potentially amplifying their ability to fight tumors in nutrient deprived microenvironments. IL-12 could therefore have a large impact on future ACT strategies.

R01 CA250458, R01 CA236379
Blockade of IL-18 signaling downregulates homing of effector CD4+ T cells to the gut mucosa and provides protection from experimental IBD
Authors: Leber Andrew J , Hontecillas Raquel , Juni Nuria Tubau , Fitch Sarah N , Bassaganya-Riera Josep
Published: August 2023

The immunological complexity of inflammatory bowel disease (IBD) results in persistent intestinal inflammation in millions of patients. As an epithelial cell-derived cytokine essential in the differentiation of CD4+ T helper cells, interleukin 18 (IL-18) is emerging as a therapeutic target for IBD. When naïve CD4+ T cells are stimulated with IL-18, the production of IFNγ and differentiation of Th1 cells increase. IL18R1−/− CD4+ T cells produce less IFNγ and inflammatory cytokines compared to WT. To examine the viability of targeting IL-18 in vivo, colitis was induced in WT, IL18−/−, and IL18R1−/− mice by DSS. In both knockouts, lower proportions of IFNγ- and IL17-producing CD4+ T cells were observed in the colon along with lesser overall severity relative to WT. To further investigate, we examined the interplay of antigen presenting cells and CD4+ T cells in vitro. IL18R1−/− Th1 cells had decreased α4β7 and GPR15 expression when co-cultured with WT BMDCs. Meanwhile, IL18R1−/− Tregs had decreased IFNγ and CCR9 expression when co-cultured with WT BMDCs that were stimulated with LPS and retinoic acid, suggesting that IL-18 may play a role in the differential gut homing of CD4+ T cell subsets. When CD4+ T cells are transferred to Rag2−/−, mice receiving IL18R1−/− T cells possess significantly lower proportions of α4β7+ Th1 cells while the proportion of Tregs in the colon are increased. Treatment of mice with DSS or Mdr1a−/− colitis with an oral IL-18 inhibitor (20 mg/kg) resulted in decreased Th1 cells and neutrophils, lower histological scores and improved disease severity. From converging data, we deduce that inhibition of IL-18 may alleviate exacerbated immune responses associated with IBD, yielding a favorable therapeutic.
Evaluating IL-27 production at the maternal-fetal interface and its role in innate antiviral immune defense during pregnancy
Authors: Barksdale Briah , Merlino Madeline S , Jurado Kellie Ann
Published: August 2023

During pregnancy, it is critical to protect the womb and developing fetus from encountered pathogens. A way to enact protection is through localized immune cells. Immune cells communicate via specialized effector molecules called cytokines. Interleukin-27 (IL-27) is a cytokine that can initiate a pro-inflammatory or anti-inflammatory response, dependent on other microenvironmental queues. Interestingly, IL-27 and its receptor (IL27RA) are known to be expressed in the placenta throughout pregnancy. This research aims to mechanistically describe the functional role of IL-27 at the maternal-fetal interface. To accomplish this aim, we will use human trophoblast organoids to determine the kinetics of IL-27 by trophoblast cells. Additionally, we will use a murine model of pregnancy to assess IL-27 production and localization at the maternal-fetal interface in individual concepti. Localized production will be visualized using immunohistochemistry of implantation site cross-sections from an IL-27 reporter (p28-GFP) mouse. Our second aim is to determine the impact of IL-27 signaling on viral replication in our model systems using Zika virus as a model pathogen. We will determine the contribution of IL-27 to antiviral activities by assessing viral burden through transcriptional and immunofluorescence readouts. We hypothesize that IL-27, through autocrine signaling, induces constitutive antiviral programming that directly restricts viral replication in trophoblasts due to its ability to induce STAT1 signaling. Overall, this project will elucidate the presence and functionality of IL-27 at the maternal-fetal interface within murine and human models.

Supported by grants from Pew Charitable Trusts Biomedical Scholars Program and Burroughs Wellcome fund
A novel heterodimer composed of IL-12p40 and CD5L promotes a unique cytokine profile from T cells
Authors: Hill Elizabeth M , Gerber Allison J , Fink Marc Y , Keegan Achsah Dorsey , Singh Nevil J
Published: August 2023

The IL-12 family of cytokines (IL-12, IL-23, IL-27, and IL-35) are heterodimers composed of α and β subunits. Both IL-12 and IL-23 utilize IL-12p40 (p40) as their β subunit yet exert distinct proinflammatory outcomes on the T cell response. Upon immune stimulation, the p40 monomer is also secreted from innate immune cells without its binding partners p35 and p19. In order to examine if p40 functions by interacting with additional partners, we generated a library of p40 binders using immunoprecipitation and mass spectrometry. Of the 19 potential binding partners identified biochemically, CD5L, a protein characterized for its role in regulating macrophages was the top hit. We generated a panel of dimer-specific monoclonal antibodies recognizing the p40-CD5L heterodimer. Using this reagent, we found that p40CD5L was present in the serum of both C57BL/6 and BALB/c mice at baseline and increased in the BAL of mice after allergic sensitization. Adoptive transfer and restimulation of transgenic T cells with recombinant p40CD5L resulted in significantly increased production of IL-4 and IL-10 – a unique cytokine profile that has not been identified with other p40-containing cytokines. From these results, we suggest that p40CD5L may function as a novel innate heterodimeric cytokine potentially promoting IL-4 and IL-10 responses.

Supported by grants from NIH (R21AI166330-02)
In Vitro Expansion of Antigen-Specific CD8+ T Cells from Human PBMCs Using Artificial T-cell Stimulating Microparticles
Authors: Sim Hajin , Livingston Natalie , Schneck Jonathan P
Published: August 2023

While current T cell-based immunotherapies have shown moderate success, clinical translation of these methods has been hindered by biological variability, limited efficacy, and high expense. Here, we create a novel T cell stimulating platform for antigen-specific CD8+ T cell activation and expansion, eliminating the extensive ex vivo expansion steps and thus making it accessible for many more cancer patients. The scaffold is formed using thiol-modified hyaluronic acid cross-linked with PEGDA, magnetic dextran nanoparticles, as well as DTT-reduced MHC-Ig dimer HLA-A*02 (A2) and anti-CD28. The gel is passed through a mesh device to form microparticles (MPs), which are then passively loaded with peptides (here, CMV and MART1) for ease of multiple expansions. They were plated with human PBMCs thawed overnight in culture media supplemented with human AB serum and a cocktail of cytokines (IL-1b, IL-2, IL-4, IL-6, and IFNγ). On days 7 and 14, cells were harvested and analyzed for proliferation of antigen-specific cells. After fourteen days of culturing PBMCs with A2 MPs, we could detect a significant expansion of antigen-specific T cells via flow cytometry. When stimulated with peptide-pulsed target cells for an intracellular cytokine assay, they expressed one or more of IFNγ, TNFα, and IL-2, demonstrating their polyfunctionality. We also observed a greater population of memory T cells, which indicates that these cells may persist longer in the body. We have developed a biomaterial-based T cell activation platform that provides critical signals, mimicking the lymph nodes. These MPs are effective in expanding antigen-specific T cells from human PBMCs, and these cells have robust killing of target splenocytes.
Design and Optimization of a Novel Proteomimetic Polymer Nanoplatform for the Co-delivery of Antigens with Adjuvants
Authors: Wang Max Mu , Choi MiRan , Zhang Bin , Gianneschi Nathan
Published: August 2023

We describe here a novel nanoplatform called the Protein-Like Polymer (PLP) with unique characteristics allowing for sustained delivery of tumor antigens in conjunction with STING agonists.

Methods

A library of PLP formulations were synthesized via ROMP and characterized. Cell uptake/functional assays were conducted with payload-specific T Cells. Ability of PLPs to co-deliver adjuvants was tested by electrostatically coupling 2′3′ cGAMP, forming stable nanostructures.

Results

Conjugating antigens to the polymer backbone using a cleavable disulfide linkage (reduces in APCs) resulted in increased endosomal localization and T cell proliferation/activation. Incorporating OEG side chains reduced enzymatic degradation while increasing immunogenicity and APC uptake. Increasing the density of side chains further improved vaccine efficacy and resistance to proteolysis. Antigen-PLP conjugates is effective only when paired with cells from their cognate system, demonstrating payload-specificity. Mice bearing established B16F10 tumors treated with gp100-PLPs resulted in increased survival time and reduced tumor burden with corresponding changes in immune cell profiles. Notably, STING-PLP complexes showed significantly smaller tumors at day 14 and allowed for subcutaneous administration of 2′3′ cGAMP. Studies looking at multiplexing pools of neoantigens onto one PLP and generation of immunological memory are ongoing.

Conclusion

The modularity of the PLP platform allows for complex nano-architectures including systems capable of delivering challenging compounds, ie small molecule STING agonists, subcutaneously through electrostatic coupling, highlighting its potential to revolutionize cancer vaccinology.

Supported by grants from the NIH (5F30CA257519-03)
Mechanisms underlying focused ultrasound-induced immunogenicity in melanoma
Authors: Pelka Sandra , Bandyopadhyay Sanmay , Pandey Sanjay , Guha Chandan , Macian Fernando
Published: August 2023

Advanced melanoma is an aggressive skin cancer with low survival rates. While the anti-tumor response relies on dendritic cells (DCs) to process tumor antigens, migrate to lymph nodes, and activate T-cells, within the tumor microenvironment (TME), DC activation is impeded. Subsequent immature DCs are a major mechanism underlying the ineffective T-cell response. We have shown that low-intensity focused ultrasound (LOFU), a novel therapy that delivers mechanical and thermal stresses to the tumor site, may enhance DC function in a B16 melanoma model. We hypothesize that LOFU enhances the dendritic cell-mediated anti-tumor response by inducing changes in tumor cell and dendritic cell autophagy and increasing pro-immunogenic factors in the TME. Autophagy is a lysosome-dependent process of cellular content degradation also implicated in the regulation of multivesicular body formation/degradation, and thus in the secretion of extracellular vesicles. Autophagy may subsequently modify the cargo of tumor-derived vesicles in a pro-immunogenic manner. We have found that LOFU treatment leads to increased LC3-II flux in melanoma cells and identified 82 proteins within melanoma-derived vesicles that are significantly enriched/reduced with LOFU, with enriched proteins partaking in processes such as antigen processing and intracellular trafficking. DCs pulsed with vesicles from LOFU-treated melanoma cells show enhanced activation of CD4+ and CD8+ T-cells, suggesting that LOFU may promote a DC-mediated anti-tumor response. Further understanding of DC involvement in the anti-tumor response will provide novel targets for the development of immunotherapies to improve survival and quality of life in advanced melanoma patients.

Supported by grant from the NIH (R01 CA226861)
Immunostimulatory effects of nanosecond pulsed electric fields
Authors: Muratori Claudia , Mazzarda Flavia , Chittams-Miles Alexandra , Vernier P Thomas
Published: August 2023

Intense pulsed electric fields (PEF) are increasingly employed for tumor ablation, including inoperable tumors, bleeding metastases, and tumors unresponsive to alternative treatments. PEF disrupt the membrane barrier of tumor cells leading to irreversible changes in homeostasis and cell death. Using different tumor models, we found that treatment with nanosecond duration pulsed electric fields (nsPEF) enhance the host antitumor immune response. The strongest effect was measured in mice bearing CT26 colon carcinoma tumors where the local treatment with nsPEF protected 78% of the animals from a second tumor cell challenge. We hypothesize that nsPEF-treated tumors act as an in-situ cancer vaccine by initiating innate immunity while molding and sustaining adaptive immunity. Indeed, we provide evidence that the damage created by these short electrical stimuli is sensed by the innate immune platform known as inflammasome. Upon assembly, the inflammasome induces proinflammatory cytokine processing and a form of cell death known as pyroptosis. We found that nsPEF triggered pyroptosis in macrophages accompanied by caspase-1 activation and release of IL-1β. Concurrently, to sustain adaptive immunity, nsPEF in tumor cells activate persistent ER-stress accompanied by phosphorylation of translation initiation factor 2α (eIF2α): a biomarker of immunogenic cell death. Tumor antigen-release from nsPEF-treated tumor cells comes with the release of damage-associated molecular patterns such as ATP, HMGB1 and calreticulin. Understanding and manipulating the cross talk between innate and adaptive immunity after nsPEF is expected to improve the immunotherapeutic response triggered by these treatments.

Pulse Biosciences, Inc.
Heterologous prime-boost viral vector cancer vaccines have protective effects against a syngeneic mouse model of glioblastoma
Authors: Steffke Emily E , Noblecourt Laurine , Almeida Vinnycius Pereira , Wicki Amanda , McAuliffe James , Kohanbash Gary , Okada Hideho , Leung Carol , Terabe Masaki , Van den Eynde Benoit
Published: August 2023

Glioblastomas are immunologically “cold” tumors with intense immunosuppressive myeloid cell- and sparse cytotoxic T-cell infiltration. Therefore, viral vector vaccines may be a promising approach to boost the induction of glioblastoma-targeted T cells. It has been previously shown that heterologous prime-boost vaccination with chimpanzee-derived adenovirus ChAdOx1 and modified vaccinia Ankara (MVA) vectors can induce a high magnitude of CD8+ T cells specific for cancer-associated antigens and have therapeutic effects against mouse models of cancer. Therefore, we aimed to evaluate whether using ChAdOx1 and MVA vaccines targeting model antigen P1A, a mouse equivalent to human tumor-associated Melanoma Antigen GenE (MAGE)-type antigens, could have protective effects against a P1A-transfected BGL-1 glioblastoma model. Mice were vaccinated with ChAdOx1/MVA-P1A vaccines, then challenged subcutaneously with P1A-expressing BGL-1 cells. Prophylactic vaccination with ChAdOx1/MVA-P1A significantly prolonged the survival of syngeneic mice subcutaneously challenged with P1A-expressing BGL-1 tumors. Furthermore, different vaccination schedules significantly impacted the magnitude of antigen-specific CD8+ T-cell responses and protective efficacy. Future studies will investigate the protective effects of vaccination against intracranial P1A-expressing BGl-1 tumors.

Supported by grants from the Ludwig Institute for Cancer Research, University of Oxford; the NIH Intramural Program (ZIA BC011877), the NIH Oxford-Cambridge Scholars Program, and the Marshall Scholarship Program.