bluuebunny/crossref_metadata_2025_split huggingface dataset:
Controlled release of inactivated influenza virus and α-galactosylceramide from PLGA microparticles results in long-term protection against influenza infection
Authors: Wen Yuhan , Sparks Zachary , Chauhan Anuj , Nelson Corwin , Driver John 
Published: August 2023

Abstract
The objective of this study was to develop biodegradable microparticles with continuous release that can enhance and extend the long-term protection of single-dose vaccines. Several formulations of Poly (lactic-co-glycolic acid) (PLGA) microparticles were synthesized using a double emulsion solvent evaporation technique to encapsulate inactivated A/PR/8/34 (PR8) H1N1 influenza virus in combination with the invariant natural killer T (NKT) cell agonist α-galactosylceramide (α-GalCer). The most efficient controlled release particle formulations were injected into C57BL/6J mice. Additional control mice were injected with PBS alone, inactivated PR8 dissolved in PBS, or inactivated PR8 and α-GalCer dissolved in PBS. Thirty weeks after vaccination, mice were challenged with a lethal dose of live homologous virus. Mice immunized with the microparticle vaccine survived the infection and had reduced clinical disease and decreased weight loss compared to unvaccinated mice. After vaccination, mice injected microparticles presented a gradual increase production of PR8-specific IgGs that was dominated by IgG1 isotype that is usually produced by T helper (Th) 2 responses. In contrast, mice that received soluble vaccine presented a rapid antibody response that was dominated by antibodies of the Th1-associated IgG2a isotype. Collectively, these results demonstrate the potential of PLGA-based single-dose microparticle vaccines for providing long-term protection against influenza infection and potentially other pathogens.
Supported by grants from USAMRAA (W81XWH-19-1-0005)
The slow-release platform, VacSIM, shapes host immune response against influenza infection after vaccination
Authors: McCormick Anna L , Mousa Jarrod J , Harn Donald A 
Published: August 2023

Abstract
Influenza virus is a leading cause of respiratory infections known to cause global morbidity and mortality, despite the widespread use of multiple influenza vaccines. Current influenza vaccines target the hemagglutinin (HA) envelope protein responsible for the attachment and entry of influenza into host cells. However, high mutation rates during influenza replication lead to mutations that cause the HA protein to be unrecognizable by host antibodies, increasing levels of infection within both vaccinated and unvaccinated individuals. A promising approach to circumvent this problem is through the use of a slow release delivery platform which has been previously shown to induce the longevity of germinal center reactions, allowing for the generation of higher affinity antibodies and more evolved B cells. Here we show that vaccination with the slow-delivery platform, Vaccine Self-Assembling Immune Matrix (VacSIM), in combination with recombinant HA antigens, increases antibody titers against H1 and H3 HAs in vaccinated groups of mice. When mice immunized with VacSIM/H1 HA were challenged with a lethal H1N1 dose, a 25% increase in survival was seen compared to the non-VacSIM group and 100% increase against mock immunized. Similarly, when immunized with VacSIM/H3 HA and challenged with H3N2, a 1.8 log reduction in lung viral titers was seen. Further analysis of the vaccine-elicited antibodies showed that vaccination with VacSIM and HA increases HAI activity against multiple strains of influenza within both H1N1 and H3N2 subtypes. We anticipate that use of VacSIM will provide further insight into how consistent exposure through a slow delivery platform can shape the host immune response to increase protection against influenza infection.
Supported by grants from CIVIC
ISCOMs/MPLA adjuvanted SDAD protein nanoparticles induce improved immune responses and cross-protection in mice
Authors: Zhu Wandi , Park Jaeyoung , Dong Chunhong , Wei Lai , Pho Thomas , Champion Julie , Wang Baozhong 
Published: August 2023

Abstract
The development of a universal influenza vaccine to protect against influenza variants will be the priority goal in combating influenza virus infection. In this study, we exploited a novel type of protein nanoparticles consisting of influenza NP as the core and NA1-M2e or NA2-M2e fusion proteins as the coating antigens by using an SDAD heterobifunctional crosslinker. Intramuscular immunization of the NP/NA-M2e (core/shell) nanoparticles induced increased humoral and cellular immune responses. ISCOMs/MPLA as adjuvants further improved the nanoparticle immunogenicity and elicited stronger immune reactions conferring protection against different influenza viral challenges. The nanoparticles were delivered through intranasal route to determine the application as a mucosal vaccine. While the nanoparticles alone barely induced immune response after intranasal immunization, ISCOMs/MPLA adjuvanted nanoparticles induced strengthened antigens-specific antibodies responses, cellular responses and higher levels of mucosal IgA. Meanwhile, increased lung Trm, Brm, and alveolar macrophages, were observed from ISCOMs/MPLA adjuvanted nanoparticle group. Only the mice intranasally vaccinated with ISCOMs/MPLA adjuvanted nanoparticles fully survived during the infections, and better protection efficacy was observed when compared with other adjuvants. Our results emphasized the importance of supplementing of appropriate adjuvants to improve the immunogenicity and mucosal immune responses of vaccines in mucosal immunization. In conclusion, the ISCOMs/MPLA adjuvant combination could significantly improve the immune responses and protective efficiency of protein nanoparticles in different immunization routes.
Pandemic preparedness: Addition of TLR agonists CpG+MPLA to emulsified H5 confers protection against H5N1 challenge after a single dose
Authors: Davies Jenny E , Hwang Michael , Dollinger Emmanuel , Felgner Jiin , Jain Aarti , Nie Qing , Othy Shivashankar , Pone Egest , Davies D Huw 
Published: August 2023

Abstract
H5N1 vaccines adjuvanted in the oil-in-water emulsion MF59 are maintained in the Strategic National Stockpile for pre-pandemic preparedness. The US population are currently naïve to H5 and a prime/boost regimen will likely be needed to engender a protective response. Ideally, a rapid response to pandemic influenza would benefit from being able to induce protection from a single dose of vaccine. Here, we have used a systems immunology approach in mice to examine the effect of formulating recombinant H5 in AddaVax alone or AddaVax with CpG+MPLA. AddaVax emulsion alone does not stimulate the innate immune system, although the oil droplets do act as a nanocarrier for antigen and enhance B cell activation, presumably by BCR cross-linking. In vitro, germinal center reactions show inclusion of CpG+MPLA to the emulsion (a combination adjuvant we call IVAX-1) synergize to enhance B cell maturation and class-switching. Similarly, in vivo, the combination of H5/IVAX-1 enhances both H5-specific B and T cell responses compared to individual components. Single cell mRNA sequencing of spleen cells after a boost indicate responses to H5/CpG+MPLA and H5/IVAX-1 are more similar to each other but distinct from H5/AddaVax or H5/PBS. Unexpectedly, H5/AddaVax, H5/CpG+MPLA and H5/IVAX-1 all engendered 100% efficacy against lethal intranasal challenge of H5N1 virus, although only H5/IVAX-1 engendered 100% protection after a single dose. Our data indicate the potential for certain combination adjuvants to enhance the immunogenicity of influenza vaccines and achieve efficacy from single dose vaccination. Supported by AI160397.
NIAID Grant AI160397
Single cell and genomic profiling of PBMC-expanded NK cells in first-in-dog clinical trial of allogeneic adoptive NK cell therapy for dogs with cancer
Authors: Razmara Aryana , Farley Lauren , Harris Rayna , Judge Sean , Lammers Marshall , Dunai Cordelia , Murphy William J , Rebhun Roberty , Brown Titus , Kent Michael , Canter Robert 
Published: August 2023

Abstract
Natural killer (NK) cells are innate cytotoxic cells with a crucial role in anti-tumor responses, making them a promising candidate for adoptive immunotherapy (IT). CD5 depletion was previously used to isolate CD5dim-expressing NK cells as a source of expanded NK cells for IT, but this technique can limit the yield of the final NK product. Here, we used bulk and single cell (SC) sequencing to compare ex vivo culture conditions using standard CD5 depletion versus unmanipulated PBMCs as the source of expanded NK cells comparing transcriptomic profiles from 5 matched healthy beagle donors using both techniques and subsequently from 5 dogs with naturally occurring malignant melanoma who received allogeneic PBMC-expanded NK cells in a first-in-dog clinical trial. Notably, beagle-derived PBMC-expanded cells had upregulation of NK pathways related to activation and function compared to CD5 depleted cells, suggesting bulk PBMCs may yield a superior product. Clinically, there were no serious adverse events using PBMC-expanded allogeneic NK cells, and one dog survived 445 days post-treatment, suggesting possible clinical benefit. SC analysis of PBMCs from this dog before, during, and after treatment showed a robust NK cell population across timepoints with high expression of cytotoxicity-related genes, such as GZMB and NCR3, lasting 14 days post-NK transfer. SC data also suggested a previously uncharacterized concordance of NK and CD8 T cell gene expression profiles in canine peripheral blood. Overall, allogeneic NK transfer using PBMC-expanded NK cells appears promising in dogs with cancer, and pre-clinical and clinical data support the use of SC analysis as a valuable technique for biomarker identification and optimizing NK IT in dogs.
This study was funded by the National Institutes of Health/National Cancer Institute grant U01 CA224166-01, the 3U01CA224166-02S1 and 5R03CA252793-02 grants, and the V Foundation Victory over Cancer through the Canter Laboratory. Student support was provided by the AVMA/AVMF 2nd Opportunity Summer Research Scholarship.
A Novel Tissue Explant Culture Model For Testing of Candidate Treatments for Inflammation of the Uterus in Pigs
Authors: Giles Joanna L , Walton Robert , Wonfor Ruth , Nash Debbie 
Published: August 2023

Abstract

Background
Endometritis is an inflammatory condition of the uterus leading to complications with reproductive organs, miscarriage and infertility.
The first-line treatment of endometritis in pigs are antibiotics, which do not always resolve inflammation and contribute to antibiotic resistance. There is an urgent need for the development of alternative treatments for endometritis. Here we propose and validate the first porcine tissue explant model for uterine inflammation and test naturally derived compounds for their effectiveness as anti-inflammatory therapies in porcine endometrial tissue.

Methods
Uterine endometrial explants were harvested from uteri of sows sent to slaughter. Tissue biopsies were collected at follicular and luteal phases of the oestrous cycle. Biopsies were washed and these tissue ‘explants’ were cultured. Explants were treated at 24 hr with oxytocin (to stimulate PGF2α production) or LPS (to mimic infection and induce inflammation. Explants were retained for RT-PCR analysis. Supernatants were harvested and concentrations of markers were measured by ELISA and Luminex.

Results
The data generated thus far show a dose dependent response of IL-6 secretion to LPS stimulation, which demonstrates the validity of our model. Inflammation can be stimulated in an in vitro setting to mimic porcine endometritis and we present data to show the change in cytokine levels in response to treatment with naturally derived compounds.

Conclusions
This data shows that inflammation can be stimulated in an in vitro setting to mimic porcine endometritis This model is a tool for testing of novel compounds and their potential anti-inflammatory effects on the porcine endometrium.

Supported by an Innovate UK Grant (Project Reference: 98015) (https://gtr.ukri.org/projects?ref=98015)
Benefits of Being Big: How a T cell response can occur as rapidly in a human as a mouse
Authors: Cannon Judy L , Moses Melanie E , Ferdous Jannatul , Fricke Matthew 
Published: August 2023

Abstract
The adaptive immune response must be fast in order to protect against exponentially growing novel pathogens. A remarkable property of the immune system is that despite a human which is approximately 2500 times larger than a mouse, the human immune response is initiated as fast as that of mice. This is in contrast to most biological processes slowing systematically with body mass. We have now collected data comparing the sizes of immune organs including spleen and lymph nodes across a large number of species including mice, rat, rabbit, macques, human, pigs, camels, cattle, and elephants. We mathematically demonstrate that while spleens scale similarly to other organs such as the hearts and kidneys, the size and number of lymph nodes scale differently than typical organ size. Using a novel computational modeling approach to predict the time that the first T cell is likely to encounter a dendritic cell presenting cognate antigen after infection, we show that the size and number of lymph nodes determine the timing of the adaptive immune response and enable even very large animals to respond quickly to pathogen infection. Combined with empirical scaling of lymph node volume, this suggests that the initiation of T cell adaptive immunity in lymph nodes occurs faster in larger animals like humans, compared to small laboratory animals which are the focus of many immunological and vaccine studies.
UNM Center for Advanced Research Computing, supported in part by NSF, for high performance computing resources, as well as the James S. McDonnell Foundation and NSF awards 2030037 and 2020247 for funding.
Computational prediction of highly conserved CD8+ and CD4+ T cell epitopes in bluetongue virus: A promising data for developing broad-spectrum bluetongue vaccine
Authors: Umeshappa Channakeshava Sokke , Kolla Harish Babu , Singh Karam Pal , Karakach Tobias , Nanjundappa Roopa Hebbandi , Mertens Peter Paul Clement 
Published: August 2023

Abstract
Bluetongue (BT) is an economically important arboviral disease of sheep, cattle, goats, and wild ruminants, particularly in America and Europe. However, it has remained uncontrolled due to the evolution of &gt;32 serologically distinct BT virus (BTV) serotypes and the lack of broad-spectrum vaccines. While outer VP2 and VP5 proteins, involved in cell penetration, are less conserved, certain core BTV proteins NS1, NS2, NS3, and VP7 have higher amino acid conservancy. Here, using in silico epitope mapping and multiple sequence alignment, we analyzed all the antigenic BTV proteins for the presence of conserved T cell epitopes recognized by different mammals. We find that mouse Major Histocompatibility Complex-I (MHC-I) conserved epitopes are present in NS1, NS3, and VP7, and MHC-II-conserved epitopes in NS1, NS2, NS3, and VP7; and in bovines, Bovine Leukocyte antigen class-I and II-(BoLA-I & II)-conserved epitopes present in NS1, NS2, NS3, and VP7 proteins. The presence of these conserved epitopes from NS1, NS2, NS3, and VP7 proteins of BTV in the selected mammals, including bovine that is closely related to sheep, we believe they are also likely to be presented by MHCs of the primary natural host sheep. Thus, harnessing this knowledge in developing a pan-BTV vaccine would confer broad-spectrum protection against all the BTV serotypes in sheep and bovines.
We are grateful for start-up funding by the Department of Microbiology and Immunology, Faculty of Medicine, Dalhousie University. PPCM wishes to acknowledge support from EU H20:20 grant ‘PALE-Blu’ (project number 727393-2).
Effect of estrogen signaling in CD8 T cell mediated antiviral response
Authors: Mithila Farha J , Hasan Mohammad , Pacia Jessica , Beura Lalit 
Published: August 2023

Abstract
Estrogen, the major female sex hormone has been linked with immune response disparity between males and females. However, the exact mechanism by which estrogen impacts our ability to fight viral infection and development of immunologic memory remains unknown. CD8 T cells represent the primary protective mechanism against viruses and cancer. Here we aim to understand how CD8 T cell mediated antiviral immune response is regulated by estrogen. Systemic LCMV infection in ovariectomized female mice generated lower virus-specific memory CD8 T cells compared to sham surgery controls. To check if this reduction in memory response is CD8 T cell intrinsic, I generated a transgenic virus-specific estrogen receptor-α deficient CD8 mouse line (ESR1 KO). Using a co-adoptive transfer model, I compared the differentiation of ESR1 KO and control CD8 T cells in both systemic LCMV and local influenza infection. The lack of estrogen signaling resulted in reduced abundance of effector CD8 T cells in the context of both infections. ESR1 KO CD8 T cells also displayed reduced cytotoxic ability. In vitro studies revealed that the CD8 T cell expansion was primarily impacted by the lack of estrogen signaling, whereas activation and cellular apoptosis remain unchanged. Altogether, this suggests a crucial role of sex steroid estrogen in initial effector CD8 T cell differentiation, that has significant ramification for generation of protective immune responses in pathological or physiological estrogen deficiency.
Supported by grants from Searle Scholars Foundation, NIH 2P20GM109035, NIH P20GM121298, NIH T32HL134625, NIH T32GM136566
ASXL1 disruption preserves CD8 T cell exhaustion-progenitor epigenetic programming during chronic stimulation
Authors: Kang Tae Gun , Lan Xin , Alli Shanta , O’Connell Casey L , Gronbaek Kirsten , Jones Peter , Han Jin-Hwan , Spatz Marianne L , Youngblood Ben A , Zebley Caitlin C 
Published: August 2023

Abstract
Differentiation of CD8 T cells into a terminally exhausted state is a major hurdle limiting efficacy of both immune checkpoint blockade and adoptive cellular immunotherapy approaches. Prior studies describing disruption of the epigenetic regulators DNMT3A and TET2 in CD8 T cells have revealed a critical role of DNA methylation in repressing the proliferative capacity and effector potential of CD8 T cells during exhaustion. Disruption of these genes has also been noted to preserve the proliferative capacity of hematopoietic stem cells (HSCs), resulting in clonal hematopoiesis (CH) and establishing the broader concept that these epigenetic regulators control cellular developmental potential. In addition to DNMT3A and TET2, the histone modifier ASXL1 is a critical regulator controlling CH. Here we report that disruption of ASXL1 in CD8 T cells preserved a stem-like phenotype denoted by increased TCF1 expression, IFN-γ production during chronic infection. At the chronic stage of infection, ASXL1 KO CD8 T cells retained a heightened ability to undergo a proliferative burst during PD-L1 blockade. Genome-wide epigenetic profiling demonstrated that disruption of ASXL1 resulted in inhibition of DNA methylation accompanied by changes in chromatin accessibility. These data document crosstalk between histone modifications and DNA methylation in regulating the transition of stem-like progenitor T cells into terminally exhausted T cells. Collectively, these data demonstrate that ASXL1 is a novel epigenetic regulator controlling development of exhausted CD8 T cells and represents a novel immunotherapeutic target for improving responses to T cell-based cancer immunotherapy.
Pannexin-1 channels promotes CD8+ T cell effector and memory responses through distinct metabolic pathways
Authors: Banuelos Alma , Vardam-Kaur Trupti , Gabaldon-Parish Maria , Zhou Maggie , Wanhainen Kelsey Marie , van Dijk Sarah , de Gois Macedo Bruna , Salgado Caio Loureiro , Santiago-Carvalho Igor , Peng Changwei , Jameson Stephen , Da Silva Henrique Borges 
Published: August 2023

Abstract
Sensing of extracellular metabolites controls CD8+ T cell function. Their accumulation can occur through export by specialized molecules, such as the release channel Pannexin-1 (Panx1). Here, we report that T cell-specific Panx1 is needed for efficient CD8+ T cell responses to viral infection and cancer. Panx1 favors both the expansion of effector CD8+ T cells and the survival of memory CD8+ T cells. Our data suggests that, while memory CD8+ T cells require Panx1 for mitochondrial function, Panx1 promotes the glycolytic pathway in effector CD8+ T cells. Panx1 promotes memory CD8+ T cell survival together with the eATP sensor P2RX7. However, Panx1 induces effector CD8+ T cells independently of eATP. Rather, we found an unexpected link between Panx1, lactate export and the activation of effector CD8+ T cells. Therefore, Panx1 regulates effector and memory CD8+ T cells through export of distinct metabolites and by engaging different intracellular pathways.
CpG therapeutic efficacy in a murine model of metastatic Lymphangioleiomyomatosis (LAM) is mediated by plasmacytoid dendritic cell and T cell immune responses
Authors: Amosu Mayowa , McCright Jacob , Moore Kaitlyn , Maisel Katharina 
Published: August 2023

Abstract

Introduction
LAM is a slow growing, metastasizing neoplasm affecting women of reproductive age, marked by abnormal growth of smooth muscle-like cells leading to cystic lung destruction. LAM has hallmarks of cancer, like expression of immune checkpoint receptors. Effective cancer therapies promote robust T cell responses through activated dendritic cells (DCs). Anti-cancer strategies, like toll like receptor (TLR) activation and immune checkpoint inhibition could work as LAM therapy. In this study, we examine the effect of CpG, a TLR9 agonist, on the DC and T cell responses in murine LAM.

Methods
We used a mouse model of metastatic LAM to determine survival after biweekly intranasal CpG therapy (10μg/ 5μg) with/ without systemic α-PD-1, rapamycin, or α-CD317 therapy. We used ELISA to measure the cytokine profile and flow cytometry to quantify cell populations between CpG-treated and untreated LAM lungs.

Results
We found that CpG treatment enhances median survival from 32 to 60 days in murine LAM (p &lt;0.0001). Efficacy of CpG treatment in LAM is facilitated by plasmacytoid DCs. pDC depletion in CpG-treated mice decreases survival from 60 to 52 days (p=0.028). CpG-treated LAM lungs also produce more IFN-α (p=0.031). CpG-treated LAM lungs have increased IFN-γ (p= 0.012) and IL-12 (p= 0.005), cytokines secreted by cytotoxic T cells.
CpG-treatment is synergistic with α-PD-1 checkpoint inhibition (p=0.004) and can be administered with rapamycin without adverse effects on median survival.

Conclusions
In LAM, CpG therapy is mediated by pDC dependent immune responses and cytotoxic T cells. This suggests that adjuvant immunotherapy, like CpG, may offer new treatment options for LAM compatible with the current standard of care, rapamycin.

The LAM Foundation Research Grant Award