bluuebunny/crossref_metadata_2025_split huggingface dataset:
Administration of Δ8-tetrahydrocannabinol enhances lung function and attenuates neutrophiles infiltration in a preclinical model of acute respiratory distress syndrome
Authors: Aladhami Ahmed Khaleel , Wilson Kiesha Maria , Carter Tayler , Nagarkatti Prakash , Nagarkatti Mitzi 
Published: August 2023

Abstract
Acute respiratory distress syndrome ARDS, characterized by severe hypoxemia, is an inflammatory response leads to high mortality rate. Patients with ARDS, such as some COVID-19 patients, suffer from pulmonary alveolar damage and major oedema due to diffuse infiltration of immune cells. Neutrophils are predominantly elevated in the bronchoalveolar lavage fluids of ARDS patients. Our laboratory already reported that Δ9-tetrahydrocannabinol (THC) administration effectively attenuated lung inflammation, cytokine storm, and prevents mortality in ARDS. In current study, we tested the effect of Δ8-THC, a cannabinoid which is less psychoactive than Δ9-THC, in lipopolysaccharide-endotoxin (LPS)-induced model of ARDS. Our data showed that Δ8-THC exhibited improvement of pulmonary function tests and decreased bronchoconstriction index in freely conscious-tested animals accompanied with significant decreasing in immune cells infiltration, specifically neutrophiles, and decreasing of serum tumor necrosis factor alpha (TNFα) level. Interestingly, Single cell RNA sequencing analysis of pulmonary cell tSNE clusters showed that Δ8-THC dramatically reduced neutrophils clusters and dysregulates certain genes involved in movement of neutrophils and chemokines production such as CCl6, CCl9, Cxcl3, Cxcl10, and CCl5. Together, these findings suggest that Δ8-THC mitigates ARDS by preventing neutrophiles trafficking into the lungs, and thus, may serve as a therapeutic agent to treat ARDS.
This work was supported in parts by NIH grants R01ES030144, P01AT003961, P20GM103641, and R01AI123947, R01AI160896
Immunoregulatory Roles of TGF-β and IL-4 during Mouse-adapted SARS-CoV-2 Infection
Authors: Sima Solomon Taye , Puebla-Clark Lucinda , Gonzalez-Orozco Maria , Shelite Thomas R , Federman Hannah G , Huante Matthew B , Martinez-Martinez Yazmin Berenice , Dann Sara M , Endsley Mark A , Siracusa Mark C , Menachery Vineet D , Endsley Janice J , Rajsbaum Ricardo , Stephens Robin 
Published: August 2023

Abstract
Severe COVID-19 is due to a dysregulated immune response; however, the mechanisms preventing recovery of airway function are unclear. Cytokines regulate the quality of inflammation, and drive both anti-viral and disease tolerance mechanisms. There was a mostly monocytic infiltration in the lungs with alveolar thickening and consolidation 3 days post-infection with mouse-adapted SARS-CoV-2 (CMA3p20) at the peak of weight loss. Based on the cytokine profiles observed in human and animal studies, we targeted IL-4 and TGF-β as potentially regulatory cytokines, and quantified changes using flow cytometry. Neutralizing TGF-β, which regulates inflammatory cytokine production, significantly increased cellular infiltration of the lung. Neutralization of TGF-β particularly affected inflammatory monocyte (Ly6C+) recruitment, with a slight decrease in alveolar macrophages (CD11c-int). Neutralizing IL-4 prolonged weight loss to 4 dpi but visibly improved resolution of alveolar thickening on 7 dpi, suggesting that IL-4 is protecting from prolonged peak pathology, but delaying resolution. Neutralizing IL-4 also increased the fraction of polymorphonuclear cells and Tregs, as well as improving the ratio of type II to type I alveolar epithelial cells in the lungs compared to the isotype group. To begin to understand mechanisms connecting IL-4 to epithelial repair, we are studying the quality of the monocyte response and found alternatively activated macrophages with Ym1 staining in activated macrophages in the alveoli of infected mice in an IL-4 dependent manner. In order to improve therapeutics for severe COVID-19, it will be important to determine the cellular mechanisms linking regulatory cytokines to epithelial repair.
R01AI135061 (RS, LPC), R01AI166668 (RR, MGO), R01AI131634 (MCS, HGF)
IL-13 protects epithelial cells from SARS-CoV-2 infection by inhibiting early ACE2-mediated events
Authors: Pivniouk Vadim I , Pivniouk Oksana , Uhrlaub Jennifer , Molzahn Allyson , Kimura Hiroki , Ledford Julie , Kraft Monica , Nikolich-Zugich Janko , Vercelli Donata 
Published: August 2023

Abstract
Severe Acute Respiratory Syndrome coronavirus-2 (SARS-CoV-2) infection is mediated by binding of viral spike (S) protein to membrane ACE2 on host epithelial cells, preferentially in the airway. Asthmatics with type 2 (T2) inflammation are reportedly less susceptible to disease, but the underlying mechanisms are unclear.
We recently showed that ACE2 expression is decreased in airway epithelial cells from T2 asthmatics and is consistently suppressed in human primary epithelial cells cultured with IL-13 (PMID: 32422146). To dissect IL-13/SARS-CoV-2 interactions, we assessed the effects of IL-13 on discrete events in SARS-CoV-2 infection. S1 protein binding to green monkey kidney Vero E6, human lung Calu-3 epithelial cells and HEK293T cells stably transfected with human ACE2 (hACE2-HEK293T) was measured by flow cytometry with His-tagged S1 protein. S protein-mediated viral entry into Vero E6 and hACE2-HEK293T cells was assessed using S protein-pseudotyped lentiviral particles carrying a GFP reporter. ACE2 transcription in Vero E6, Calu-3 and primary human airway epithelial cells was measured by RT-qPCR. ACE2 expression on Vero E6, Calu-3 and hACE2-HTK293T cells was evaluated by flow cytometry. USA-WA1/2020 SARS-CoV-2 was used to infect Calu-3 cells.
IL-13 strongly inhibited S protein binding, cell entry, and infection of epithelial cells by SARS-CoV-2. These effects resulted at least in part from IL-13-dependent inhibition of ACE2 expression because IL-13 did not affect S protein binding to, and S protein-pseudotyped lentivirus entry into, hACE2-HEK293T cells that do not downregulate ACE2 in response to IL-13. We suggest that IL-13 protects epithelial cells from SARS-CoV-2 infection by inhibiting early ACE2-mediated events.
Supported by grant from NIAID: U19 AI125357
Understanding of SARS-CoV-2 pathogenicity in COVID-19 cynomolgus macaque model and consideration of its reinfection
Authors: Urano Emiko , Okamura Tomotaka , Yasutomi Yasuhiro 
Published: August 2023

Abstract
Although rapid vaccination has been progressing in the world as a countermeasure against COVID-19, the pandemic of SARS-CoV-2 is still raging worldwide due to the continuous emergence of several variants. We have established the COVID-19 cynomolgus macaque (CM) model using healthy young CMs and advanced-age CMs and the differences observed between the pathologies of young and elderly macaques were similar that those seen in humans.
In the present study, we analyzed viral dynamics and pathology of SARS-CoV-2 variants of concern (VOCs) from Wuhan to Omicrons using the COVID-19 CM model. Delta infection displayed a greater virulence and Omicron infection showed a lesser virulence. Elucidation of how adaptive immunity in patients is induced by SARS-CoV-2 natural infection is critical for establishing a vaccine strategy, serologic therapies, public health control and prediction of infection. Therefore, we conducted the prolonged rechallenge analysis using the COVID-19 CM model reinoculated with either the same or a different strain of SARS-CoV-2 that was used for the initial infection. Re-infection was confirmed between different virus strains and PCR re-positivity was also observed between same VOC strain in some cases. Anti-RBD antibodies against Wuhan, Alpha, and Delta, were well induced by each VOC infection, but not against Omicrons. Antigen-specific cellular responses are also currently being analyzed.
Overall, studies using COVID-19 CM models should contribute to the elucidation of the complicated pathophysiology of COVID-19. In addition, COVID-19 CM model should be useful for understand the virus evolution and to consider vaccine strategy to overcome the COVID-19 pandemic.
AMED JP21fk0108414 AMED JP223fa727002 JSPS 21K19395
Peering into the future of SARS-CoV-2 evolution using a computational model of the SARS-CoV-2 serum antibody repertoire
Authors: Cohen-Lavi Liel , Koren Eilay , Yegorov Yan , Sacharen Sinai , Burkovitz Anat , Hertz Tomer 
Published: August 2023

Abstract
The global emergence of SARS-CoV-2 has provided a wealth of immunological data, including more than 600 solved structures of SARS-CoV-2 Spike antibodies (Abs) available in the Protein Data Bank. We reasoned that this data could be used to generate a computational model of the serum Ab repertoire. We extracted a set of 275 non-redundant Abs, and clustered them using an unsupervised clustering method based on their contact positions with Spike. We identified 9 unique Ab clusters that together summarize the major binding modes of Abs to the Spike protein. To characterize each cluster, we mined 65 published papers extracting detailed information on each Ab. We then demonstrated Spike Ab contacts could be predicted based on the Ab’s CDRs sequence. Next, we developed a computational method for predicting immune escape. We extracted the set of all Spike mutations in SARS-CoV-2 sequences from GISAID. To assess the effects of single mutations on immune escape, we computed the ΔΔG for each Ab-mutation pair and constructed an aggregated escape score for each mutation. Our predicted escape scores were positively correlated with published experimental measurements of Ab escape. We used our escape score to rank novel viral variants and are dynamically ranking strains and novel variants of interest as they appear. To further validate its predictive power, we chose a set of predicted Ab-escaping mutations and are testing their escape from Abs in human sera samples. Taken together, our analysis demonstrates the feasibility of generating a computational model of the serum Ab repertoire, which may be used for the rapid characterization of novel Abs, and to computationally estimate the effects of mutations on immune escape from Ab recognition.
Supposed by the Israel Science Foundation (ISF) grant no. 2683/21, and the NIH\NIAID CEIRR under contract 75N9302100016, St Jude Children’s Research Hospital.
A new mouse model expressing human ACE2, TMPRSS2, and FCGRT for evaluating human antibody responses to SARS-CoV-2
Authors: Ana-Sosa-Batiz Fernanda , Verma Shailendra Kumar , Shafee Norazizah , Miller Robyn , Conner Chris , Hastie Kathryn M , Timis Julia , Maule Erin , Nguyen Michael N , Saphire Erica Ollman , Shresta Sujan 
Published: August 2023

Abstract
Protection against viruses is most often measured by levels of plasma binding and neutralizing antibodies (Abs) in vitro, which do not capture the full picture of Ab immunity, including Fc functions. The continuing evolution of SARS-CoV-2 underscores the need to understand protection mediated by Abs. We generated triple knock-in (TKI) mice expressing human ACE2, TMPRSS2, and FCGRT in place of murine genes. A human anti-SARS-CoV-2 IgG1 with a half-life extending mutation but not wild-type IgG1 exhibited long half-life and protected against BA.2 infection in lungs, thereby validating our TKI mice for evaluating human Ab clearance and function. We next evaluated protective efficacy of plasma samples from vaccinated San Diegans against Delta or BA.5 by performing passive transfer experiments. Treatment with pooled plasma from vaccination plus infection but not triple vaccination individuals cleared infectious Delta in lungs of TKI mice, although Delta-binding and -neutralizing Ab titers in vitro were similar between the two groups of people. In contrast, neither group’s plasma impacted lung viral burden upon BA.5 challenge. Thus, Abs elicited by vaccination plus infection but not triple vaccination reduce Delta infection in this mouse model, and Abs from both groups are ineffective against BA.5. These results are consistent with epidemiological data, and show that in vitro Ab titers do not predict in vivo protection. Our TKI mouse model represents an important tool to test efficacy of human monoclonal Abs against SARS-CoV-2/related CoVs, predict human polyclonal Ab-mediated protection against newly emerging variants/CoVs, and dissect mechanisms of Ab-mediated protection in vivo. Results will aid development of pan CoV vaccines.
Phenotypical and Functional Characterization of Unswitched Memory B cells verses Switched during SARS-CoV-2 Infection
Authors: Castleman Moriah J , Santos Adriana Luna , Lesteberg Kelsey , Maloney James , Janssen William , Mould Kara , Beckham J David , Pelanda Roberta , Torres Raul M 
Published: August 2023

Abstract
Memory B cells are comprised of unswitched (CD27+IgD+) and switched (CD27+IgD-) subsets. The origin and function of unswitched human memory B cells remains controversial, whereas switched memory B cells are primed to respond to recurrent infection. Unswitched memory B cells were described to be reduced in frequency with severe SARS-CoV2 infection and here we report on their activation status, BCR functionality, and contribution to virally-induced cytokine production. Analyses of whole blood from healthy individuals, people immunized against SARS-CoV2, and those with mild or severe SARS-CoV2 infection, confirm a loss of unswitched memory B cells during severe SARS-CoV2 infection and demonstrate this loss is associated with increased levels of systemic TNFα. We further report an increased frequency of ‘IgD+’ only memory B cells that correlate with an increase in IgG autoantibody levels (anti-chromatin, anti-cardiolipin, anti-smith) in severe SARS-CoV2 infection. Unswitched and switched memory B cells from severe SARS-CoV2 infection displayed evidence of heightened activation with concomitant reduction in expression of the inhibitory receptor CD72. Functionally, both populations of memory B cells from severe SARS-COV2 infection harbored a signaling competent BCR that displayed enhanced activity in the unswitched population. Finally, we demonstrate that B cells from mild SARS-CoV2 infection secrete pro-inflammatory cytokines IL-6 and TNFα. Importantly, unswitched memory B cells were a major producer of IL-6 and switched memory B cells were a major producer of TNFα in response to viral TLR ligands. Together these data suggest that B cells contribute to the inflammatory milieu during viral infection.
Supported by grants from NIH R01 AI124474 (RP) and R01 AI136534 (RMT).
Pre-existing SARS-CoV-2-reactive CD4+ T cells are established and highly functional in early childhood
Authors: Olofsson Anna , Humbert Marion , Niessl Julia , Hodcroft Emma , Gao Yu , Sohlberg Ebba , Malmberg Karl-Johan , Lund-Johansen Fritjof , Björkhem-Bergman Linda , Jenmalm Maria C , Ljunggren Hans-Gustaf , Buggert Marcus , Karlsson Annika C 
Published: August 2023

Abstract
The pre-existing SARS-CoV-2-reactive T cells identified in unexposed individuals are likely induced by prior infections with seasonal coronaviruses and may modulate disease outcomes. By assessing humoral and cellular immunity in paired samples from SARS-CoV-2 unexposed children at ages 2 and 6 (n=16), and SARS-CoV-2 seronegative adults (n=94, ages 26–83), this study aims to elucidate how age influences the immune responses against coronaviruses.
Using flow cytometry, the phenotype and function of seasonal coronavirus OC43-specific and pre-existing SARS-CoV-2-reactive memory (m) CD4+ T cells were characterized after a 10h stimulation with viral peptides. The frequency of SARS-CoV-2-reactive mCD4+ T cells were closely linked with the OC43-specific responses in all age groups. However, a positive response to either OC43- or SARS-CoV-2-derived peptides was more common at age 6 than in adults. Similarly, the functional capacity, defined by IFN-γ, IL-2 and TNF expression, of both OC43-specific and SARS-CoV-2-reactive mCD4+ T cells peaked at age 6 and then declined with age. Age-dependent phenotypic markers were also identified, including CD226, TIGIT and CD38. Lastly, in contrast to Rhinovirus, EBV, Influenza and other coronaviruses, OC43 seroprevalence did not increase between ages 2 and 6, or from childhood to adulthood, further establishing OC43 as a childhood infection.
Our results show that memory responses to coronaviruses develop at a young age and exhibit age-dependent qualities, findings which may aid in future vaccine development. In addition, the high prevalence and functional capacity of pre-existing SARS-CoV-2-reactive mCD4+ T cells at age 6 may contribute to the low incidence of severe COVID-19 in this age group.
Autoantibodies produced by SARS-CoV-2 infection trigger damage to the lacrimal and salivary glands and lead to the development of Sjögren’s Disease
Authors: Shen Yiran , Voigt Alexandria , Goranova Laura , Abed Mehdi , Kleiner David , Maldonado Jose , Beach Margaret , Pelayo Eileen , Chiorini John , Craft William , Ostrov David A , Ramiya Vijay , Warner Blake , Nguyen Cuong 
Published: August 2023

Abstract
Sjögren’s Disease (SjD) is a chronic, systemic autoimmune disease characterized by lymphocytic infiltration and the development of dry eyes and mouth as the result of the secretory dysfunction of the lacrimal and salivary glands. In recent years, infectious agents have been shown to be associated with SjD. Several studies indicated that SARS-CoV-2 infection induces autoantibodies production, patients also experienced various autoimmune disorders. This study used SARS-CoV-2 WA1/2020 to infect hACE2 mice to investigate the relationship between SARS-CoV-2 infection and SjD. Infected mice showed dry eyes, decreased salivary flow, lymphocytic infiltration of the lacrimal and salivary glands, elevated antinuclear antibodies (ANA), specifically anti-SSB/La, and apoptotic cells near the acinar cells. Notably, counting of apoptotic cells showed a significantly higher number in the lacrimal glands of mice infected with a low viral load than high viral load. We confirmed with clinical data that we observed the elevation of SjD-specific autoantibodies (ANA, anti-SSB/Ro52, and anti-SSA/La) in sera from COVID-19 patients. Salivary gland biopsies of patients diagnosed with SjD several months after SARS-CoV-2 infection also showed lymphocytic infiltration and fibrosis. Lastly, to identify whether monoclonal antibodies produced by COVID-19 patients are reactive against nuclear antigens, antibodies tested for greater antiviral binding capacity were shown to be more responsive. Overall, by observing SjD-like phenotypes in mouse models and patients, our study confirms a direct pathogenic role of SARS-CoV-2 in SjD. It also provides a different pilot for the study of the mechanisms of autoimmune responses induced by viral infections.
This research was funded by the National Institutes of Health (NIH), National Institute of Dental and Craniofacial Research (NIDCR), Division of Extramural Research (DE028544, PI-Nguyen; DE028544-02S1, PI-Nguyen), and the NIH/NIDCR Division of Intramural Research (Z01-DE000704, PI-Warner; Z01-DE00695, PI-Chiorini).
Prevalence and complement activation of anti-lymphocyte IgM antibodies in hospitalized COVID-19 patients
Authors: Perez-Diez Ainhoa , Liu Xiangdong , Calderon Stephanie , Bennett Ashlynn , Lisco Andrea , Kellog Anela , Galindo Frances , Memoli Matthew , Rocco Joseph M , Laidlaw Elizabeth , Sneller Michael C , Manion Maura , Sereti Irini 
Published: August 2023

Abstract
Recent studies have shown that a proportion of COVID-19 patients develop auto-antibodies against a wide range of secreted and membrane proteins, some of which are expressed by lymphocytes. It is unclear, however, if and what proportion of patients might develop anti-lymphocyte antibodies (ALAb) and whether they might have cytotoxic function. We systematically characterized the presence, isotype and lytic function of ALAb in a cohort of 85 COVID-19 patients during hospital admission from May 2020 to June 2022. As controls we analyzed 20 healthy donors (HD) and seven Flu infected volunteers. Patients were assigned into mild (N=65), or moderate/severe (N=20) groups, based on oxygen requirements, and 37 patients were followed up after recovery. We found that 27% of COVID-19 patients had IgM ALAb (binding CD4, CD8 and/or B cells), compared to 15% of the control populations. IgM ALAb prevalence was independent of disease severity and persisted during early convalescence. Sera from some of the IgM ALAb positive COVID-19 patients, and not from the controls, induced complement dependent cytotoxicity (CDC) of HD PBMCs, mostly targeting B cells. Complement activation was corroborated by detecting complement deposition ex vivo on lymphocytes from 70% of COVID-19 acutely infected patients. The level of C3b deposition on CD4 T cells inversely correlated with CD4 numbers in blood of moderate/severe patients. Our findings indicate that the antibody mediated auto-immune response found in COVID-19 patients includes targeting of lymphocytes with potential contribution to the observed lymphopenia.
This work was supported by the Divisions of Intramural Research of NIAID and NIBIB of the NIH
Type I Innate Lymphoid Cells Sense and Proliferate in Response to Distal Viral Infection
Authors: Wise-Dent Olivia , Shannon John P , Reynoso Glennys , Lujan Ramon A , Hickman Heather D 
Published: August 2023

Abstract
Innate lymphoid cells (ILCs) are a diverse group of tissue-resident lymphocytes that lack antigen-specific receptors commonly found on adaptive immune cells. While many different types of ILCs have been described, only group 1 ILCs (ILC1s) are known to exhibit marked antiviral activity. We recently described ILC1-mediated protection of the oral mucosa, a critical barrier tissue and site of continued pathogen exposure. Based on these studies, we hypothesized that these innate antiviral cells might also respond to inflammation induced by a distal viral infection. To address this, we used a common mouse model of vaccinia virus (VACV) infection of the peritoneal cavity, which spreads locally but does not result in infection of the oral mucosa. Confocal microscopy of frozen sections of the lip demonstrated a marked increase in the number and density of oral mucosal ILC1s in response to distal infection. Further, ILC1s remained sustainably elevated for weeks following the initial infection. Ongoing experiments will assess the ability of this enhanced layer of ILC1s to provide heightened antiviral immunity in the oral mucosa and determine the timeframes in which this occurs. Together, our data suggest that the ability of ILC1s to globally sense infection and enhance protection in response to distal infection may represent a previously unappreciated mechanism of trained immunity in the tissue.