bluuebunny/crossref_metadata_2025 huggingface dataset:
Integrating Family History Into the Post-Pandemic Elementary Learning Space
Authors: Alexander-Ashley Belinda M.
Published: January 2023

The COVID-19 pandemic highlighted existing inequities as a result of zero tolerance and exclusionary policies that disproportionately impacted the world's learners living in poverty, people of color, and those experiencing special challenges. Under the existing educational system, marginalized students often feel devalued and without a voice. Integrating family history and genealogy into the elementary school learning space provides a methodology and framework that focuses on the historical conditions that promote healthy dialogues and sustain discourses connecting to other historical events. The process of creating positive experiences with family history, improving the classroom environment, effectively communicating, rapport and trust building, and strengthened socio-emotional skills reduce childhood trauma. The six recommended strategies include introspection and reflection, navigating parallel time periods, valuing genealogical tools, encouraging an environment of hope, normalizing authenticity, and transforming the learning environment.
Enhancing Supply Chain Resilience Through Digital Capabilities
Authors: Umar Muazu Adeiza
Published: January 2023

The objective of the chapter was to review pertinent literature on the concepts of supply chain resilience and digital capabilities. Further reviews were undertaken to identify the supply chain digital technologies presently in use and to explore the relevance of theoretical underpinnings, where these theories or perspectives attempt to explain the relationships between the constructs. It was concluded that although the digital technologies available in the market to support supply chain resilience are sufficient, the same cannot be said for the underpinning theories, and neither are the concepts under consideration properly grounded in the literature. More work, therefore, needs to be done on the concepts and the theories.
An Overview of Fraud Applications and Software on Social Media
Authors: Hussien Nadia Mahmood , Mohialden Yasmin Makki
Published: January 2023

Fraud involves intentional deception. While the victim's rights are violated, the criminal profits from the situation. Fraud can be committed by individuals, groups, or even entire companies. The importance of social media cannot be overstated. The management of natural disasters and medical difficulties are both handled through social media. The use of social media has resulted in the establishment of a marketplace for goods and services. Because social networking has evolved into a money-making opportunity, dishonest users now cheat in many ways. Fraud on social media platforms has been on the rise. Cybercriminals exploit unethical social engineering to steal data. Indicators of fraud include excessive pressure, a broad scope, honey traps, and justification. People who are unaware of something lose both money and information. In the current environment, information technology professionals are unable to protect social media users from being scammed or spammed.
Experimental Work and Discussion of Results on Deepfakes in Stock Prices Using Sentiment Analysis and Machine Learning
Authors: Anbarasu N.  , Karuppusamy S. , Prabhu S. , Sobana P.
Published: January 2023

In the field of artificial intelligence, the employment of computer algorithms that develop themselves mechanically as they get more and more experience in experimental work and discussion of results on deepfakes is essential to the research process. It is frequently thought of as being in close proximity to artificial intelligence. Machine learning is a set of techniques that generates a model by making use of sample data, commonly referred to as training data, in order to make predictions about the future without actively attempting to do so, by separating the various types of tweets taken from the data on Twitter regarding a number of different companies. A classification algorithm was applied to the data that was collected and used in order to categorise the tweets. In addition, the OHLCV of the data from a number of different organisations is taken into account in order to improve efficiency.
Organic Nutrient Solutions for Hydrophonic Spinach (Basella alba) Production in Urban Agriculture
Authors: Dhananjani B. A. , Pakeerathan K.
Published: January 2023

Purpose: Macro and micronutrients rich vegetable and fish wastes are generated hugely in Sri Lanka, but not recycled properly into valuable byproducts. It is a serious problem and needs to be managed to make environment free from pollution. Extraction of nutrients from organic wastes can be used as liquid fertilizers for hydroponic crop production even in congested urban cities.
Research Method: Organic hydroponic medium using vegetable waste extracts (M2), fish emulsion (M3), vermi tea (M4), the mixture of M2+M3+M4, and Albert’s solution (M1-control) were prepared and tested on Spinach. The experimental setup was arranged in a CRD with four replicates. The nutritional qualities of formulated liquid fertilizers (N, P, K, Ca, Na), pH, EC, spinach growth parameters such as plant height, number of leaves per plant, leaf area, root length, weight (Fresh /Dry) were measured eight weeks after planting and subjected to ANOVA using SAS 9.1. Tukey's HSD multiple comparison test to identify the best treatment combination at P <0.05.
Findings: The results revealed that the amount of each N, P, K, Na, and Ca was significantly different between treatments (P <0.05). The highest quantity of each N (11.1mg/L), P (37.99mg/L), and K (145,5 mmol/L) was detected in M2. Whereas, Na and Ca quantities were highly significant in M4 with the values of 208.7 and 96.2 mmol/L, respectively. Plant height and root length in M1, M2 and M3 were significantly higher than those of other treatments. Fresh and dry weights were also significantly higher in M3, M1 and M2 treatments when compared with other treatments. The highest leaf number and leaf area were also recorded in M3 treatment after 8 weeks of planting.
Originality/Value: This study shows that fish emulsion solution was found to be more promising and both fish emulsion and vegetable waste hydrophonics medium can be used as alternatives to Albert’s solution.
Calibration and Validation of APSIM Millet Model for Proso millet (Panicum miliaceum L.) Accessions as a Basis for Crop Diversification
Authors: Wimalasiri E. M. , Ashfold M. J. , Walker S. , Nissanka S. P. , Karunaratne A. S.
Published: January 2023

Purpose: Proso millet (Panicum miliaceum L.) is an underutilised minor millet grown as a rain-fed crop in subsistence farming systems in tropical African and Asian countries including Sri Lanka. It is identified as a climate-resilient crop that exhibits a huge potential to diversify conventional farming systems, however few modelling studies have assessed this crop, especially in the tropics. The objective of this study is to calibrate and validate Agricultural Production Systems Simulator (APSIM) model for Proso millet.
Research Method: The millet model of APSIM that was developed based on Pearl millet was calibrated for five farmer-selected Proso millet accessions using the data gathered from two field experiments. Yield data collected from 35 farmers’ fields under different crop management practices were used to validate the model.
Findings: The observed phenology, leaf area index, above ground biomass and grain yield were not significantly different (p > 0.05) from the simulated values of the calibrated model. The observed yields (1187±336 kg ha–1) in the farmers’ fields were not significantly (p > 0.05) different from the simulated yields (1208±255 kg ha–1), suggesting a good calibration of the model.
Research Limitations: Lack of field data under different abiotic stress conditions is a limitation for further validation of the model.
Originality/ Value: APSIM millet model has not been tested for Proso millet previously. Derived genetic coefficients were successfully used to simulate Proso millet production.
Price Spread Analysis of Tezpur Litchi: A Case Study from Assam, India
Authors: Newar P. , Borah R.
Published: January 2023

Purpose: Being a geographical indication certified fruit, Tezpur litchi enjoys the consumer price difference of Indian Rupees 10 per litchi from the common varieties of litchi. Therefore, it is crucial to consider the different aspects of price spread which will make a way out to provide maximum benefit to the litchi growers. With this motive, this study aimed to analyse the price spread and identify the pattern of marketing channels that would be feasible and give profitable returns to the litchi growers.
Research Method: The empirical data were collected from 218 samples that included four channel members i.e., litchi growers, pre-harvesting contractors, retailers and final consumers who are mainly involved in the marketing and consumption of Tezpur litchi. The data were analysed using specific formulas of price spread, the share of channel members in consumer’s price and marketing efficiency.
Findings: Four patterns of marketing channels were identified for Tezpur litchi. Channel III which included litchi growers, retailers and final consumers was found the most feasible in the study area where 79% of litchi growers directly sell to retailers instead of pre-harvesting contractors. It was observed that price spread, the share of channel members in consumer’s price and marketing efficiency shared an inverse relationship with the number of mediators in the marketing channel. However, from the practical point of view, channel III is suggested to the litchi growers for a feasible and profitable return.
Research Limitations: Exact information on the distribution of Tezpur Litchi to the other states and countries was not available. Therefore, the geographical scope of the study is limited to Assam.
Originality/ Value: Understanding price spread is considered very important to cognise the efficiency of the marketing channels operating for a product. An effective marketing channel not only meets the demand of the consumers but also creates a favourable environment for the growers and intermediaries to increase production and add value to the product.
Comparative Studies of Some Biochemical Characteristics of Seminal Plasma in Wild and Cultured Male Catfish (Clarias gariepinus), Burchell, 1822 Broodstock
Authors: Yusuf Olatunji Yusuf , Jimoh Wasiu Adeyemi , Raji Lukman Oladimeji , Azeez Faidat
Published: January 2023

Purpose: In aquaculture, the most important aspect to consider is ensuring the development of valuable offspring by using high-quality gametes from fish broodstock. This research aims to compare the biochemical constituents of seminal plasma in both wild and cultured Clarias gariepinus broodstock.
Research Method: Apparently healthy adult male Clarias gariepinus (n=9, 900±300g mean weight per fish type (wild or cultured), 49±8.5cm mean total length) were collected from fishermen on the Oshin River, Nigeria for the wild samples and from a reputable fish farm in Ilorin and examined for some seminal plasma’s biochemical parameters following standard procedures. T-test statistics were used to compare the differences between classes.
Findings: Higher values of most of the semen biochemical compositions were recorded among the cultured Clarias gariepinus with significant variations (p<0.05) recorded in the mean values of total cholesterol, high-density lipoprotein-cholesterol (HDL-Cholesterol), low-density lipoprotein-cholesterol (LDLCholesterol), uric acid, Aspartate Aminotransferase (AST), total bilirubin, albumin, potassium, chloride and urea. Creatinine, glucose, sodium, and triglycerides, on the other hand, did not vary significantly (p>0.05) between cultured and wild Clarias gariepinus semen.
Originality/ Value: The current study’s findings on biochemical compositions could lead to more effective gamete management, higher yields, and improved semen suitability for short-term storage.
A Critical Systematic Review for Inhaled Corticosteroids on Lung Cancer Incidence: Not Yet Concluded Story
Authors: Lee Suh-Young , Yoon Soon Ho , Hong Hyunsook
Published: January 2023

Background: To systematically review studies on inhaled corticosteroids (ICS) and lung cancer incidence in chronic airway disease patients. Methods: We conducted electronic bibliographic searches on OVID-MEDLINE, EM- BASE, and the Cochrane Database before May 2020 to identify relevant studies. Detailed data on the study population, exposure, and outcome domains were reviewed. Results: Of 4,058 screened publications, 13 eligible studies in adults with chronic obstructive pulmonary disease (COPD) or asthma evaluated lung cancer incidence after ICS exposure. Pooled hazard ratio and odds ratio for developing lung cancer in ICS exposure were 0.81 (95% confidence interval, 0.64 to 1.02; I2=95.7%) from 10 studies and 1.02 (95% confidence interval 0.50 to 2.07; I2=94.7%) from three studies. Meta-regression failed to explain the substantial heterogeneity of pooled estimates. COPD and asthma were variously defined without spirometry in 11 studies. Regarding exposure assessment, three and 10 studies regarded ICS exposure as a time-dependent and fixed variable, respectively. Some studies assessed ICS use for the entire study period, whereas others assessed ICS use for 6 months to 2 years within or before study entry. Smoking was adjusted in four studies, and only four studies introduced 1 to 2 latency years in their main or subgroup analysis.Conclusion: Studies published to date on ICS and lung cancer incidence had heterogeneous study populations, exposures, and outcome assessments, limiting the generation of a pooled conclusion. The beneficial effect of ICS on lung cancer incidence has not yet been established, and understanding the heterogeneities will help future researchers to establish robust evidence on ICS and lung cancer incidence.
Molecular bases for inherited human complement component C6 deficiency in two unrelated individuals.
Authors: Nishizaka H , Horiuchi T , Zhu Z B , Fukumori Y , Nagasawa K , Hayashi K , Krumdieck R , Cobbs C G , Higuchi M , Yasunaga S , Niho Y , Volanakis J E
Published: January 2023

Deficiency of the sixth component of complement (C6D) is frequently associated with recurrent neisserial infections, especially meningitis caused by Neisseria meningitidis. We here report the molecular bases of C6D in two unrelated subjects, one African American (case 1) and the other Japanese (case 2). Screening all 17 exons of the C6 gene and their boundaries by exon-specific PCR/single strand conformation polymorphism demonstrated aberrant single stranded DNA fragments in exon 12 of case 1 and exon 2 of case 2. Nucleotide sequencing of the amplified DNA fragments revealed a homozygous single-base deletion (G1936) in exon 12 case 1 and a heterozygous single base deletion (C291/C292/C293/C294) in exon 2 of case 2. Both mutations resulted in frame shifts and premature termination of the C6 polypeptide. Sequence-specific oligonucleotide probe hybridization and direct sequencing of exon 12 amplified from genomic DNA further supported the homozygosity of the mutation in case 1. Case 2 is apparently compound heterozygote, but the putative mutation in the other allele of the C6 gene remains unknown. Both case 1 and case 2 were homozygous for the C6A allotype. These data indicate that at least three distinct mutational events can cause C6D, single nucleotide deletions in exons 2 and 12, and a mutation yet unidentified. Thus, similar to other complement protein deficiencies, the pathogenesis of C6D appears to be heterogeneous.
Adenosine pathway is associated with the control of immune activation and inflammation in nonprogressive SIV infection (VIR1P.1009)
Authors: He Tianyu , Brocca Cofano Egidio , Gillespie Delbert , Xu Cui Ling , Kristoff Jan , Ross Samantha , Ma Dongzhu , Haret-Richter George Sebastian , Rinaldo Charles , Jackson Edwin , Macatangay Bernard , Apetrei Cristian , Pandrea Ivona
Published: January 2023

Adenosine (ADO) is associated with immunosuppression in multiple diseases, yet its role in modulating immune activation/inflammation (IA/INFL) in HIV-1/SIV infection is unclear. We compared changes in ADO pathway-associated markers between African green monkeys (AGMs) that control SIV-related IA/INFL and pigtailed macaques (PTMs) that exhibit IA/INFL associated with AIDS progression. CD39/CD73 and CD26 (which promote ADO production or breakdown to inosine) were assessed by flow cytometry on T cells from blood, lymph nodes (LNs) and intestine of AGMs and PTMs pre and post SIV infection. ADO was measured by mass spectrometry in frozen tissues. Baseline CD39/CD73 expression on Tregs from LNs and intestine were intrinsically high in AGMs and remained high throughout SIV infection. CD26 and inosine did not change in AGMs. In the gut, basal ADO was high in AGMs and further increased early after infection. Conversely, in PTMs, CD26 expression dramatically increased in the gut early in infection, a change associated with increased inosine. CD39/CD73 only increased during late infection in PTMs. Our results suggest a key role for ADO in the control of IA/INFL in nonprogressive SIV infections and that ADO production may be counteracted by the early increase of CD26 in pathogenic HIV/SIV. Furthermore, changes of ADO levels predominately occur in the gut rather than in blood or LNs, suggesting that future studies of the ADO pathway should focus on mucosal sites of viral replication.
Targeted IL-10 locally suppresses inflammation in atherosclerotic sites
Authors: Volpatti Lisa R , Hubbell Jeffrey A
Published: January 2023

Atherosclerosis is a main contributor to cardiovascular disease, the leading cause of death in the U.S. Preventative measures include statins or dietary changes to reduce “bad cholesterol” or low-density lipoprotein (LDL) that accumulates on arterial walls to form plaques. However, these treatment options often lead to surgical interventions associated with high cost and morbidity. Although atherosclerosis is now recognized as a chronic inflammatory disease, no therapies targeting the underlying immunology are currently approved.

The objective of this project is to locally suppress inflammation at atherosclerotic sites. IL-10 has been shown to suppress vascular inflammation, but its poor pharmacokinetic profile and possible side effects limit its therapeutic potential. To overcome these challenges, we engineered fusion proteins in which one side is IL-10 and the other side is an antibody fragment (Fab) that binds to protein epitopes of LDL. Thus, we specifically target IL-10 to atherosclerotic plaques.

We validated the fusion proteins in vitro, confirming Fab binding affinity with surface plasmon resonance and IL-10 bioactivity in a macrophage-like cell line. We subsequently evaluated their immunosuppressive effect in the apolipoprotein E−/− mouse model of atherosclerosis. LDL-binding Fab-IL-10 constructs significantly reduced the total number of leukocytes, macrophages, T cells, and B cells in the aorta to levels similar to wild type mice. Conversely, neither IL-10 fused to a Fab of irrelevant specificity nor native IL-10 showed any differences to untreated mice. Therefore, plaque-targeted Fab-IL-10 constructs effectively suppress local inflammation and show promise as novel atherosclerosis therapies.

Supported by NIH NHLBI T32, Chicago Immunoengineering Innovation Center
Pandemic-induced pan-academic educational response in a minority university.
Authors: Fazal Nadeem , Faridi Hafeez
Published: January 2023

Education system is extremely vulnerable to the consequences caused by COVID-19, especially the need to strengthen pharmacy education. We utilized this opportunity to rebuild education post-pandemic at Chicago State University College of Pharmacy. Of many avenues opened to development, teachers obligated to update their digital competencies. The pandemic had widened achievement gaps and exposed weaknesses in education systems we had an opportunity now to reimagine a more equitable and resilient education system that delivered a better education to all minority students. Current educational set up was built when our understanding of human learning and learners, knowledge, and skills, as well as teaching pedagogy and teachers’ expertise was quite different from present. Pandemic gave us one-time life window of opportunity to pause and update our educational needs. The purpose of this project was to support such a process of exchange of knowledge. The panic button was sounding and both educators and students were baffled by how to proceed in the wake of pandemic. We encouraged to implementing technology into science education in a way allowing both teachers and students to develop collaborative skill sets and advance adaptability. We recognized the need to support not only students but also students’ families, especially in vulnerable minority communities. We found a “spring effect” and found that pandemic-induced suppression of education bounced back and even rise above the level of past normal. Since it has involved the whole world (pandemic) the educational response should be (pan-academic) and hopefully it will serve as a solution to global problem (panacea) like (vaccine) for all.
“Heard immunity”-pandemic teaches immunology vocabulary through hearing from news and social media outlets.
Authors: Fazal Nadeem , Brehm Danielle , Alakbarova Gulgaz , Cebiyev Nihat , Meher Satish , De Herde Laura
Published: January 2023

COVID-19 surprised the world and brought with it free immunology lessons. Ever since the pandemic news broke, the public were bombarded with immunological vocabulary. Immunology as a biomedical science had never attained student interest, let alone having an overwhelming global attention among the general public. The purpose of our study was to evaluate the extent to which immunology had become a common knowledge because of the main stream media and social outlets broadcasting news. We conducted a global survey of all people, regardless of gender identity, age, education, and geographical location to ask if they knew the meanings of specific immunological terms used in mainstream news outlets and social media. The words included, but were not limited to, antigen, antibody, T-cell, B-cell, mRNA, DNA, coronavirus, quarantine, lockdown, social distancing, PCR, rapid antigen tests, outbreak, herd immunity, vaccine, viral infections, infection rate, mortality rate, immunotherapy, and immune boosting drugs, etc. When we analyzed the data, we found the following trends. 139/179 (77.7%) said ’yes’, they did learn about immunology in lockdown. 100/179 (55.9%) said they learnt about Pfizer, followed by Moderna (33.5%), BioNTech (31.8%), and AstraZeneca (29.6%). 86/179 (48%) learnt about the body’s defense system, while 20.7% of people already knew about T cells, B cells, immune therapy, and immunity. 95/179 (53.1%) of people asked learned about airborne infections, making them the most common type of respiratory infections learnt about during COVID-19. The word ‘pandemic’ was learnt by 111/179 (62%) people. Overall, COVID-19 has propelled immunology into the general public’s vocabulary and knowledge in a way that has never been done before.
Designing a successful K-12 education program aimed to prevent infectious disease
Authors: Jarvi Susan Irene , Howe Kathleen , Brewer Francis , Baker Wendy
Published: January 2023

Developing collaborative efforts to train K-12 teachers and students as community educators is a strategy that is useful in preventing disease. The current pandemic illuminates the need for a greater focus on immunology and public health in K-12 STEM/STEAM education. The components of a course design and delivery impact success. University of Hawaii, Daniel K. Inouye College of Pharmacy’s Jarvi Lab, in collaboration with non-profits and professional educators, offers a for-credit, Professional Development Education course to Hawaii K-12 teachers using a curriculum focusing on rat lungworm disease, a debilitating and sometimes lethal disease caused by the parasite Angiostrongylus cantonensis. The PDE course is place-based; learning is centered on the local environment and in the community. It is taught online, allowing for statewide participation. The lessons are research-driven and standards-based. Parasite-host interactions, and disease presentation, treatment and prevention are emphasized. Course participants are provided access to peer-reviewed papers, educational videos, and webinars, which can be used in the classroom and support diverse learning styles. Participation in live (on-line) discussions with scientists and researchers help advance teacher understanding of the scientific methodology and study designs. Hands-on activities focus on modeling, data collection and analysis, and communication skills. Citizen Science, Design Thinking, and ArcGIS story mapping are introduced. Lessons encourage student-led inquiry and teamwork. Teachers’ final portfolios reflect the knowledge gained, their reviews of the course are positive, and they report a high level of student interest.

Supported by Hawaii State Legislature, Daniel K. Inouye College of Pharmacy
Strength of CAR signaling reveals bifurcation of T and ILC2 lineage differentiation from pluripotent stem cells
Authors: Li Suwen , Wang Chloe S , Montel-Hagen Amélie , Casero David , Lopez Shawn , Chen Ho-Chung , Zhou Olivia , Botero Carlos , Tsai Steven C , Wong Claudia , Crooks Gay M , Seet Christopher S
Published: January 2023

T cells and type 2 innate lymphoid cells (ILC2) are two closely related lymphoid lineages that share certain developmental and transcriptional programs, including a requirement for Notch and IL-7 signaling during differentiation. We recently developed an artificial thymic organoid (ATO) system that supports in vitro differentiation of human pluripotent stem cells (PSCs) to mature αβT cells. We show here that lentiviral introduction of a CD19-targeted chimeric antigen receptor (CAR) into PSCs surprisingly resulted in ILC2-biased lymphopoiesis from PSCs at the expense of T cell differentiation. PSC-derived ILC2s expressed the classical ILC2 markers CD25, CD200R, and CRTH2 as well GATA3, ID2, and TCF7, and responded to both cytokine stimuli and antigen-dependent CAR signaling by producing IL-5 and IL-13. To understand CAR-mediated lineage diversion from T to ILC2, we performed single cell RNAseq of PSC-derived hematopoietic progenitor cells before the phenotypic onset of either T or ILC2 differentiation. This revealed a gene signature of T cell receptor signaling, suggesting aberrant CAR activation at the earliest stages of lymphocyte development. We established that CAR signaling in ATOs was antigen-independent and thus likely driven by tonic signaling. We applied this finding to rationally modulate the T/ILC2 lineage decision by fine-tuning CAR signaling strength during lymphocyte development from PSCs, permitting restoration of conventional CAR-T cell differentiation or, conversely, directed differentiation of isogenic antigen-specific CAR-ILC2s. Taken together, our findings shed light on human ILC2 development and inform the applied differentiation of both ILC2s and conventional CAR-T cells from PSCs.

Supported by Tobacco-Related Disease Research Program Predoctoral Award (SL), Broad Stem Cell Research Center UCLA Fellowship (SL, CSS), CIRM Bridges Program (CB), NIH T32 (ST), NIH K08CA235525 (CSS).
Proteogenomic immune signatures delineate the landscape of pediatric acquired demyelinating syndromes
Authors: Espinoza Diego A , Mexhitaj Ina , Smiler Jacqueline , Mafra Fernanda , Da Silva Renata Pellegrino , Fadda Giulia , Yeh E Ann , Marrie Ruth Ann , Arnold Douglas L , Li Rui , Banwell Brenda , Bar-Or Amit
Published: January 2023

Approximately 20–30% of children presenting with acquired inflammatory demyelinating syndromes (ADS) have multiple sclerosis (MS). Another 30% harbor serum antibodies against myelin oligodendrocyte glycoprotein and are referred to as having MOG-associated disease (MOGAD). While MS and MOGAD can have similar features, differences in response to immune therapies point to distinct underlying immune mechanisms.

To assess potentially distinct immune mechanisms underlying MS and MOGAD, we applied proteogenomics to high quality cryopreserved peripheral blood mononuclear cells collected from patients with ADS prior to institution of immune therapy, as well as from healthy controls. CITE-Seq profiling was applied to a total of 92,716 single cells with equal contribution from 24 children (6 healthy donors; 6 with ADS but neither MS or MOGAD; 6 with MOGAD; and 6 with MS, ascertained with long-term follow-up).

Analysis revealed a pan-ADS enrichment of atypical (CD11c+) B cells compared to healthy controls. Children with MS were distinguished from children with MOGAD by MS-specific enrichments of checkpoint-molecule (TIGIT and CD137)-expressing CD8 memory T cells, STAT4+ Th1 CD4 memory T cells and CD56dim/CD16+ NK cells.

Overall, our study identifies distinct features of circulating cellular immune profiles that may serve to distinguish children with MS and MOGAD, and provides novel insights into early immune mechanisms that may be involved in each of these conditions.
T Cell Responses In Children With Asthma Against SARS-CoV-2 Correlates To Asthmatic Outcomes
Authors: Eyoh Enwono , Murrell Amelie , Werthmann Derek , Trinh Ivy , Stone Addison , Chandra Sruti , Elliott Debra , Smira Ashley , Velazquez Jalene V , Schieffelin John S , Kolls Jay K , Robinson James , Gunn Bronwyn M , Rabito Felicia , Norton Elizabeth B
Published: January 2023

SARS CoV-2 in children or special populations post-infection has not been well studied. Asthma is a heterogenous disease marked by chronic airway inflammation; triggers include cockroaches (CRA), and other inhaled irritants. Asthma also has links to viral infections like respiratory syncytial virus (RSV). Additionally, some with moderate to severe asthma are more likely to be hospitalized with COVID. T cells play key roles in asthma and control of viruses such; however, there is limited information connecting T cell responses in people with asthma to viral infections. We aimed to evaluate T cells and antibodies in an asthma confirmed cohort. Thirty-five children aged 5–17 years were included. We measured SARS CoV-2 spike (S) and Nucleoprotein (N) plasma antibody responses or effector functions and CD4, CD8 T cells specific to CRA or peptide pools made from RSV, S and N antigen using an activation induced markers (AIM) assay. Surprisingly, in this population many CD4 and CD8 T-cell AIM responses to S, N, CRA, and RSV were significantly associated, particularly for CD8 T-cells (Spearmans r = 0.57–0.76) and the restimulation antigens CRA and RSV. N-specific CD4 AIM was the only immune measure to correlate to a recent asthma attack within a month of the blood collection visit, though anti-N antibodies, CD4 AIM to CRA, RSV or CD8 AIM to CRA, RSV, or N antigens also correlated with asthmatic outcomes (e.g., ER visits, night waking from symptoms, etc.) whereas allergen specific IgE or anti-S IgG did not. Taken together these results indicate an immunological association between viral infection and asthma, broadly allowing for the conjecture that viral infections, in particular RSV and SARS-CoV-2 could act together as possible triggers of asthma.
Aiolos supports TFH cell differentiation by antagonizing the IL-2/STAT5 signaling pathway
Authors: Read Kaitlin , Pokhrel Srijana , Jones Devin M , Saadey Abbey , Eisele Caprice , Warren Robert T , Collins Patrick , Ghoneim Hazem E , Freud Aharon , Oestreich Kenneth J
Published: January 2023

CD4+ T follicular helper (TFH) cells are critical for the generation of robust humoral immune responses, as they provide help to B cells to support the generation of both pathogen-neutralizing antibodies and long-lived plasma cell populations. To date, the mechanisms underlying TFH differentiation are incompletely understood. Here, we identify the transcription factor Aiolos as a key regulator of TFH responses. We find that Aiolos expression is increased in TFH cell populations generated during influenza infection and that Aiolos deficiency results in compromised TFH cell differentiation and B cell helper activity. Mechanistically, loss of Aiolos results in diminished expression of TFH genes, including those encoding the key transcription factors Bcl-6, TCF-1, and Tox. Conversely, expression of genes associated with IL-2/STAT5 signaling, a known antagonist of the TFH gene program, were significantly elevated in Aiolos-deficient settings. Consistent with these data, antigen-specific Aiolos-deficient effector CD4+ T cell populations generated in response to influenza infection exhibited significantly elevated IL-2Rα surface expression. Together, our findings suggest that repression of IL-2/STAT5 signaling represents a novel mechanism by which Aiolos regulates the TFH gene program. These findings are important, as they provide critical insight that may ultimately be leveraged for the development of novel TFH-focused immunotherapies and strategies to improve vaccination approaches.

Supported by a grant from the NIAID (NIH; R01 AI134972), funds through The Ohio State University College of Medicine, and funds through The Ohio State University College of Medicine Advancing Research in Infection and Immunity Fellowship Program
Antimicrobial Peptide and Lipid Response of Alveolar Type II Model Cells to Cytokines of TH17 Signature
Authors: Toscano Cindy Lizette , Wesley-Cardwell India , Abou Abbas Dana A , Dzul Janette , Martinez Erik , Porter Edith
Published: January 2023

Mycobacterium tuberculosis (Mtb) causes ~ 10 million deaths each year worldwide. The current BCG vaccine that targets TH1 cells and anti-tuberculosis drugs have limited efficacy. Novel preventative measures are in dire need. Epithelial cells in the lungs are among the first host cells to encounter inhaled Mtb. Epithelial cells produce antimicrobial peptides (AMPs) like human beta-defensins (HBDs) and antimicrobial lipids (AMLs). Microbial products such as lipopolysaccharide (LPS), IL-1β made by macrophages, and IL-17 and IL-22 secreted by TH17 cells are all known to increase production of AMPs in epithelial cells. However, their effect on the production of AMLs is unknown. We hypothesized that TH17 cytokines increase the production of AMPs and AMLs in lung alveolar type II cells, leading to increased antimycobacterial activity. As determined by immunofluorescence, IL-17 combined with IL-22 (IL-17/22) significantly increased intracellular HBD2 in A549 cells after 48 h. Using the lipid sensitive fluoroprobes Bodipy and Nile Red, we found that IL-1β combined with LPS significantly increased intracellular neutral lipids, while IL-17/22, IL-1β/LPS, and LPS alone significantly increased neutral and polar lipid secretion after 24h stimulation. To assess the antimycobacterial effect of A549 secretions, we developed a qPCR-based assay employing Mycolicibacterium smegmatis (Ms), an avirulent model organism for Mtb, bacterial cell lysis with TRI reagent and bead beating, gDNA extraction with Direct-zol™ DNA Miniprep, SYBR Green chemistry, and 16S rRNA gene targeting primers, and testing is ongoing. This research may lead to novel vaccine approaches for Mtb based on TH17-induced antimycobacterial activity of lung epithelial cells.

This research was supported by NIH R25GM061331, NIH T34GM08228, and CalStateLA Provost’s Research Restart Grant.
Time course scRNAseq analysis on host mucosal immune responses to influenza virus infection during aging
Authors: Wu Yue , Tang Jinyi , Gao Xiaochen , Li Chaofan , Zhu Bibo , Zhang Ruixuan , Fain Cori , Ayasoufi Katayoun , Johnson Aaron J , Dong Haidong , Sun Jie
Published: January 2023

Aging has been shown to be one of the major risk factors for host morbidity and mortality following infections with respiratory viruses including influenza virus and SARS-CoV-2. The unique immune properties of the aged hosts not only influence pathogen clearance and tissue damage responses, but also modulate the pathogenesis of the long-term sequalae post acute viral clearance. Currently, the dynamics and features of mucosal immune responses following acute respiratory viral infections in the aged hosts remain to be fully elucidated. Here, we characterized the lung cellular and molecular profiles following influenza virus infection in young (~3-month-old) or aged (~24-month-old) mice with a chronological manner from acute to chronic phase. Using DNA-oligo-conjugated CD45 antibody and i.v. labeling, we were able to distinguish the unique responses of circulating vs lung-resident immune cells to influenza virus infection during aging. These responses were further validated by spectral flow cytometry analysis of the lung innate and adaptive immune cells. Further data analysis and integration are warranted for this unique large data set detailing the single cell molecular profiles of mucosal responses to influenza virus infection comparing young and aged hosts.
Epithelial-derived oxysterol production tunes intestinal IgA secretion against commensals and enteric pathogen in tissue
Authors: Ceglia Simona , Berthelette Alyssa , Howley Kelsey , Li Yun , Yiew Nicole K H , Xu Ying , Brink Robert Adrian , Cyster Jason G , Hooper Lora V , Randolph Gwendalyn J , Reboldi Andrea
Published: January 2023

Immunoglobulin A (IgA) secretion by plasma cells (PCs), terminally differentiated B cells residing in the intestinal lamina propria, assures microbiome homeostasis and protects the host against enteric infections. However, whether exposure to diet-derived and commensal-derived signals instruct tissue resident PCs effector function and dynamically shape IgA immune responses at the mucosal barrier remain largely uninvestigated. Here, we demonstrated that intestinal epithelial cells (IECs) integrate luminal input to produce 7α,25-dihydroxycholesterol (7α,25-HC), a cholesterol metabolite (oxysterol) with immunomodulatory functions. In IECs, both cholesterol uptake via NPC1L1 and commensal recognition via MyD88 controlled oxysterol production. Mice lacking the oxysterol enzyme CH25H specifically in IECs abolished 7α,25-HC production and allowed to study oxysterol generation and activity in the small intestine. Inability of IECs to generate 7α,25-HC enhanced IgA secretion by PCs in the gut, suggesting that oxysterol negatively regulate humoral response at the mucosal barriers. Mechanistically, we showed that intestinal PCs sensed 7α,25-HC via the chemoattractant receptor GPR183 to position in the lamina propria tissue. This IEC-PC axis was rapidly modulated by cholesterol dietary content and tuned Salmonella-specific IgA response.

Our finding revealed a new mechanism linking dietary cholesterol and humoral immune responses centered around PC localization for efficient mucosal protection.

Supported by grants from Kenneth Rainin Foundation, Innovator Award, Charles H. Hood Foundation Child Health Research Awards Program, The Leukemia and Lymphoma Society New Idea Award and the Multiple Myeloma Research Fellowship, NIH (AI40098) and The American Association of Immunologists Careers in Immunology Fellowship Program.
Increased tryptase expression in HαT is associated with upregulation of epithelia-derived MRGPRX2 agonists and MRGPRX2+ MCs in the GI mucosa
Authors: Glover Sarah C , Williams Haley , Pride Yilianys , Owings Anna H , Robinson Tanya , Lyons Jonathan , Deepak Vishwa , Ali Hydar
Published: January 2023

Hereditary α-tryptasemia (HαT) is a genetic trait characterized by elevated serum tryptase and increased copy number of the TPSAB1 gene. It is associated with flushing, itching, GI symptoms and anaphylaxis. HαT gut epithelia undergo increased rates of pyroptosis, and HαT gut mast cells (MCs) are increased in number with highly expressed FCER1 and CD203C. Gut MCs express Mas-related G protein–coupled receptor-X2 (MRGPRX2). Studies have clearly shown that this receptor contributes to host defense, mediates pseudo-allergy, and is involved in a number of atopic skin conditions. Herein, we sought to evaluate the impact of MRGPRX2 in individuals with HαT and GI symptoms. We observed increased numbers of MRGPRX2+ mast cells in HαT versus non-HαT gastrointestinal (GI) biopsies (colon->duodenum->stomach->Ileum). mRNA expression of tryptase and MRGPRX2 in intestine of HαT increased ~6-fold and 3-fold respectively over non-HαT biopsies. Epithelial derived agonists of MGPRX2 including human beta defensin (HBD) 2 and adrenomedullin (PAMP12) mRNA expression were also found to be upregulated ~2-fold and ~3-folds in HαT over non-HαT biopsies. MC protease carboxydeptidase-3 was also increased. Taken together, this data suggests that in addition to upregulation of MRGPRX2 in MCs, increased tryptase expression in HαT leads to upregulation of MRGPRX2 agonists and modifiers in gut epithelial cells, nociceptors and MCs, and results in GI symptoms.

This work was supported by National Institutes of Health grants R01-AI124182, R01-AI143185 and R01-AI149487 to Hydar Ali.
Genetically knock out of PSGL-1 increases CD19-CAR-T cells persistence in mouse pre-B-ALL disease model
Authors: Mavuluri Jayadev
Published: January 2023

Chimeric antigen receptor T cell therapy is reported as a fifth pillar of cancer therapy. CAR-T therapy has success against DLBCL’s. Although it is an effective cancer treatment, CAR-T cells still faces challenges like persistence and exhaustion. P selectin glycoprotein ligand 1 (PSGL-1) is a mucin-like selectin counter receptor on T-cells that binds to P-selectin, E-selectin, and L-selectin on endothelial cells. In our study, we observed CRISPR-Cas9 knockout of P-Selectin Glycoprotein Ligand-1(PSGL-1) in human Pan-T cells showed increased persistence by enhancing the anti-apoptotic pathway and further reduces the exhaustion by reduced expression of PD-1, TIM3 and LAG3. Hence PSGL-1 can be used as a potential target in the reprogramming of CAR T-cells and successful CAR T-cell therapy might be achieved with a smaller number of infusions to the patients. Our preclinical study in B6 mouse model, pre-B-ALL disease model clearly shows PSGL-1 KO CD19-CAR-T cells, effectively clears the tumors, persist for longer time by maintain the central memory phenotype and having lower exhaustion profile. GSEA pathway enrichment analysis from RNA seq of PSGL-1 KO CD19 CAR-T cells shows PSGL-1 KO CD19-CAR-T cells has central memory phenotype and lower exhaustion profile, further confirmed by immunophenotyping these PSGL1-KO CD19-CAR-T cells. From our study, it is evident that PSGL-1 KO CAR-T cells can overcome the limitations of CAR-T cells hurdles like maintaining the central memory phenotype and lower exhaustion In summary, our study is clear evident that PSGL-1 KO CAR-T cells can be used as universal CAR-T system to target all kind tumors not limiting to hematological cancers.
Leveraging mouse genetics to generate heavy-chain only antibodies for therapeutic application
Authors: Borbet Timothy C , Perault Andrew , Ilmain Juliana , Chan Kun-Wei , Luo Christina C , Bryan Arielle M , Kirilov Milen , Wang Wei , Sheiffele Paul , Kong Xiang-Peng , Ekiert Damian , Torres Victor , Koralov Sergei B
Published: January 2023

Conventional antibodies (Abs) consist of two heavy and two light immunoglobulin chains, while heavy-chain only antibodies (HCAbs), like those found in camelid and shark species, are unique because they lack a light chain binding partner. The unique structure of these HCAbs impart advantageous properties to these smaller Abs, including greater accessibility to their target antigens, higher hydrophilicity, and greater conformational stability under environmental stresses. We have generated a mouse model, the IgG3dCH1 strain, 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. While single-cell sequencing analysis of B cells revealed some biases in VH and JH usage, it also highlighted the tremendous diversity of rearrangements that give rise to B cells bearing these single chain antibodies. We’ve been able to show that upon immunization, IgG3dCH1 animals are protected from lethal bacterial challenge with a pathogenic strain of Enterobacter hormaechei and generate pathogen specific IgG. Further, immunization with SARS-CoV2 stabilized prefusion spike protein elicited HCAbs with a KD in the nM range, measured via bio-layer interferometry. These data support this mouse being utilized as a platform for the generation of pathogen-specific HCAbs that can be further engineered for testing and application for translational outcomes.

Supported by grants from NIH (T32 AI007180, R43 AI136141).
Discovery of potently-neutralizing antibodies against SARS-CoV-2 using Barcode Enabled Antigen Mapping (BEAM)
Authors: Stubbington Michael J T , Adams Bruce , Reyes Daniel , Royall Ariel , Song Maengseok , Marrache Sean , Shahi Payam , Tsai FuNien , Finnegan Peter , Vollbrecht Thomas , Khadilkar Tanmayee , Jaffee David , Ramenani Ravi , McDonnell Wyatt
Published: January 2023

Humans who have recovered from infectious disease possess a memory B cell pool that contains highly-potent antibodies against the cleared pathogenic agent. This pool is a valuable source of therapeutic antibodies but identifying these requires a method that can screen a large number of cells rapidly to identify promising candidates.

We used Barcode Enabled Antigen Mapping (BEAM) to screen 100 million peripheral blood mononuclear cells (PBMCs) from a donor who had recovered from COVID-19. This method labels antigens of interest with unique reporter oligonucleotides and uses these labeled antigens to stain lymphocytes according to the binding specificity of their antigen receptors. Individual antigen-bound cells are then captured and analysed using the 10x Genomics Single Cell Immune Profiling Solution. This allows us to identify the antigen-specific cells while also generating transcriptomic profiles and natively paired, heavy and light chain full-length B-cell receptor sequences from each cell.

We used BEAM to identify B cells that bound to the spike protein of SARS-CoV-2 and discovered 222 antibodies with high binding affinity confirmed by SPR. The majority of these antibodies bound to multiple variants of concern with some also recognising endemic coronaviruses. We then investigated the potential for these antibodies to neutralize live SARS-CoV-2 and confirmed that 55 exhibited potent neutralization activity. Finally, we performed epitope binning and discovered that the collection of antibodies bound to multiple different epitopes on the spike protein.

Compared with hybridoma or other approaches, single cell methods have the potential to discover therapeutic antibodies more rapidly and with a higher diversity of leads.