bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development of inactivated subunit influenza vaccine endowing with IgA induction and protection against heterologous strain
Authors: Bang Yoo-Jin , Kim Yun-Hee , Lee Yu-Sun , Kim Jae-Yong , Park Hyo-Jung , Ko Hae-Li , Park Sang-In , Hwang Kyung-Ah , Kim Hun , Nam Jae-Hwan 
Published: January 2023

Abstract
Vaccination is known to be the best way to prevent and control seasonal influenza infections. Among the various available influenza vaccines, an inactivated vaccine shows improved protective effects associated with greater safety. However, since the influenza virus is continuously evolving, its response to inactivated vaccines becomes increasingly difficult to predict, leading to complete or partial loss of protection against the virus. In addition, immunogenicity is lower than other types and Th2-biased immune responses have been reported. In this study, we have investigated the role of the single-stranded RNA (ssRNA) adjuvant derived from the intergenic region of internal ribosome entry site of Cricket Paralysis virus in seasonal inactivated subunit influenza vaccine (ISIV). We found that the ssRNA adjuvant stimulated balanced cellular (indicated by IgG2a, IFN-γ, IL-2, and TNF-α) and humoral (indicated by IgG1 and HI) responses, along with mucosal (indicated by IgA) immune response. Moreover, the ssRNA adjuvant formulated ISIV enhanced viral clearance and improved lung pathology after homologous and even heterologous influenza virus infections. The proportion of memory CD4+ and CD8+ T cells, important for long term immunity, was also observed to increase. Therefore, the ssRNA adjuvant formulated ISIV is effective in inducing humoral and cellular immune responses, cross protection, and long-term immunity.
Immunology of vaccine adjuvants enhancing respiratory viral vaccine efficacy
Authors: Kang Sang-Moo , Ko Eun-Ju , Lee Youri , Kim Ki-Hye 
Published: January 2023

Abstract
We investigated the adjuvant effects of MPL (TLR4 agonist) and CpG (TLR9 agonist) on promoting protection after vaccination of C57BL/6 and BALB/c mice with inactivated influenza split virus or human respiratory syncytial virus (RSV) vaccines. Adjuvant effects of combination MPL and CpG (MPL+CpG) on improving protective vaccine efficacy were evident by higher levels of protective humoral and cellular immune responses to vaccination, more effective control of lung viral replication, prevention of weight loss, and inflammatory cytokines and cellular infiltrates in vaccinated mice after virus challenge. Vaccine-enhanced respiratory disease, a well-known phenomenon due to atypical immune priming by RSV vaccination after challenge, could be prevented by MPL + CpG adjuvants in RSV vaccination, shaping toward Th1 and IgG2a isotype immune responses. In contrast, aluminum hydroxide and squalene oil-in-water adjuvants (AddaVax) were not effective in conferring vaccine-induced protection against influenza and RSV in BALB/c mice after vaccination and challenge. Combination of MPL and CpG was found to exhibit distinct effects on modulating inflammatory micro-environment with cytokines, chemokines, and innate infiltrates in vivo within a day as well as stimulating dendritic cells in vitro to secrete IL-12p70 and TNF-α cytokines. In summary, the findings suggest that a combination adjuvant of different TLR agonists exhibits a unique pattern of innate and adaptive immune responses, contributing to protection by inactivated influenza split virus and subunit RSV vaccination.
Safety, immunogenicity, and protective efficacy of a recombinant vaccine against RSV: Evidence from murine and bovine models
Authors: Diaz Fabian E , Guerra-Maupome Mariana , Soto Jorge A , Gálvez Nicolás M S , Ríos Mariana , McGill Jodi L , Kalergis Alexis M 
Published: January 2023

Abstract
The human Respiratory Syncytial virus (RSV) is a leading cause of lower respiratory tract infection in infants worldwide. Despite its health burden, there are no licensed vaccines to prevent human RSV-caused disease. In mice, immunization with a single subcutaneous dose of 3 × 105 CFUs of a recombinant BCG expressing hRSV N protein (rBCG-N-hRSV) is safe, and induces robust, long-lived humoral and cellular Th1/Th17 immunity, which prevents lung damage after an intranasal hRSV challenge. Vaccinated mice display reduced weight loss, less neutrophil airway infiltration, and reduced lung viral loads, when compared to unimmunized controls. To further explore the safety, immunogenicity and efficacy of this formulation, we carried out a vaccination and challenge study on a bovine RSV - neonatal calf infection model. Five day old calves received a subcutaneous dose of the rBCG-N-hRSV vaccine and a booster vaccination after 14 days. On day 28, calves were inoculated via aerosol challenge with ~104 TCID50/mL of BRSV strain 375. Clinical data was registered daily until euthanasia on day 7 post infection. Virus-specific IgA and neutralization activity increased in vaccinated calves at 7dpi, and the animals mounted a robust virus-specific IFN-γ/IL-17 response observed after PBMC antigen recall assays. Following bRSV challenge, we observed reduced clinical disease compared to unvaccinated control calves, without signs of vaccine-enhanced disease. Reduction trends in viral shedding and lung viral load were observed in vaccinated animals, as well as a significantly reduced neutrophil infiltration in BAL. These results suggest that the rBCG-N-hRSV vaccine would be safe for neonatal use and support further investigation in clinical studies.
Chitosan adjuvantation improves protection elicited by single, low dose influenza A virus recombinant protein vaccination
Authors: Lampe Anna T , Farris Eric J , Ballweg Matthew D , Pannier Angela K , Brown Deborah M 
Published: January 2023

Abstract
Continued development of effective vaccines requires investigation of new adjuvants that are able to direct and enhance immune responses. Adjuvants have potential to allow for fine-tuning of immune responses against pathogens for which vaccines are suboptimal, or non-existent. Despite this capability, few adjuvants are currently used in human vaccines. Biomaterials provide a pool of potential adjuvants that have been vetted for biocompatibility and adverse effects. One such material is chitosan (CS), a linear, positively charged, partially deacylated derivative of chitin. Varying molecular weights of CS were examined for their ability to improve influenza A virus vaccination. It was observed that mice receiving CS during single, low dose protein vaccination experienced significantly less weight loss after homologous challenge than mice receiving protein alone. CS elicited enhanced protection from weight loss was accompanied by elevated IgG and a trend of increased IgG1. When T cell activation was examined two days after challenge, CS adjuvantation resulted in no significant changes in IFN-γ or IL-4 production in the DLN, however CS induced IL-4 production by CD4 T cells in the lung at day five. This effect appeared to be dependent on the molecular weight of the CS. Additionally, CS adjuvantation resulted in elevated CD69+/CD103+ CD4 T cells in the lung two days after homologous challenge, potentially mediating improved protection against infection. Further, we observed enhanced in vitro cytokine production and IRF pathway activation in antigen presenting cells treated with CS. Taken together, these results suggest that CS is able to act as a vaccine adjuvant and may be used to instruct immune responses and improve vaccine efficacy.
PD1 blockade augments adoptive T cell therapy via endogenous T cells rather than direct enhancement of transferred T cells
Authors: Davies John S , Karimipour Farrah , Zhang Ling , Serna Carylinda , Hinrichs Christian 
Published: January 2023

Abstract
Adoptive T cell therapy (ACT) has yielded promising clinical results but improvements are needed. Cell therapy efficacy may be improved by blockade of the inhibitory receptor PD1. Independent T cell models demonstrated cooperation between ACT and α-PD1. The cooperative effect of α-PD1 was abolished in three separate hosts devoid of polyclonal T cells (Rag1-KO, Trac-KO and irrelevant TCRtg hosts). Moreover, endogenous CD4 and CD8 T cells in the tumor increased and transferred T cells decreased in number following α-PD1 treatment. Efficacy of ACT was not improved when transferred cells were engineered with PD1-targeting miRNA or over-expression of PD1 without the cytoplasmic signaling domain. However, ACT with PD1-KO T cells was improved with the combination of α-PD1. Additionally, transferred T cells did not respond to α-PD1 when they were repeatedly stimulated until high co-expression of PD1+LAG3+TIM3+ (85% vs 34%) prior to transfer, when they were allowed prolonged in vivo tumor engagement prior to α-PD1 administration, or when they were targeting tumors with enforced expression of PDL1. Lastly, α-PD1 therapy showed no efficacy during the treatment of less immunogenic tumors, engineered with ubiquitin-tagged antigen constructs rather than RFP-antigen constructs. This finding established that PD1-sensitive endogenous T cells targeted immunogenic antigens within RFP. The role of endogenous T cells was further corroborated by TCR clonotype skewing when α-PD1 was administered. These data demonstrated that blocking the PD1 axis did not directly benefit adoptively transferred T cells. Rather, the benefit of α-PD1 was dependent on the presence of endogenous T cells specific for immunogenic tumor antigens.
Modulation of thymic architecture and T cell function by Mesenchymal stromal cell extracellular vesicles in a model of neonatal hyperoxia
Authors: Reis Monica , Willis Gareth R. , Fernandez-Gonzalez Angeles , Yeung Vincent , Taglauer Elizabeth , Liu Xianlan , Mitsialis S. Alex , Kourembanas Stella 
Published: January 2023

Abstract
Therapies based on mesenchymal stem/stromal cell extracellular vesicles (MEx) have shown promise in experimental models of bronchopulmonary dysplasia (BPD), a disease of disrupted lung development that can result from neonatal exposure to hyperoxia. The effects of oxygen exposure on other developing organs in the neonatal period remain poorly understood and whether MEx treatment may modify these responses is not known. Our research aimed at understanding the effects of neonatal exposure to hyperoxia and the immunomodulatory properties of MEx on thymic architecture and T cell development. Using a mouse model of neonatal hyperoxia, we showed that, in addition to lung injury resulting in BPD, there was involution of the thymic medulla with altered T cell function. Administration of a single dose of MEx resulted in preservation of thymic architecture, and restoration of normal thymic function through modulation of the thymic medullary antigen presentation network and T cell phenotypes. Consequently, MEx reinstated the mechanisms of thymic-driven central tolerance that are perturbed upon hyperoxia exposure, as demonstrated by increased FoxP3+ regulatory T cell generation and decreased T cell autoreactivity. Overall, our study demonstrates that MEx represent a promising cell-free therapeutic option to restore thymic architecture and ensure the development of central tolerance by preventing thymic injury associated with neonatal hyperoxia exposure.
TLR enhanced GVAX elicits tumor-specific tissue resident memory T cells independent of T cell priming
Authors: Korrer Michael J , Taylor David R , Kim Young Jun 
Published: January 2023

Abstract
GVAX, a genetically modified whole cell vaccine, is thought to work by recruiting and activating APCs which then traffic to the draining lymph node and elicit tumor-specific T cells. GVAX has little therapeutic benefit as a monotherapy, but when combined with a Toll-like receptor (TLR) 4 adjuvant, TLR Enhanced GVAX (TEGVAX) significantly reduces tumor burden in mouse models. Paradoxically, the increased therapeutic benefit of TEGVAX corresponds with a decrease in delivery of tumor antigen to the dLN suggesting a different mechanism of tumor suppression than GVAX.
To determine if TEGVAX alters the priming of CD8 T cells, we performed longitudinal studies of OT-1 CD8 T cell proliferation in vivo. GVAX induced rapid proliferation of OT-1 cells, whereas TEGVAX failed to induce any. We then determined if TEGVAX required myeloid or NK cells to function. TEGVAX reduced B16-mOVA tumor burden in NK depleted, but not myeloid cell depleted mice. To determine if T cell circulation from the dLN to the tumor was necessary, we administered FTY720, which sequesters circulating T cells in lymph nodes, prior to vaccination. TEGVAX significantly reduced B16-mOVA tumor growth even in the presence of FTY720 treatment, suggesting T cell priming was not required. Immune phenotyping of TEGVAX treated B16 tumors and showed the presence of activated tumor infiltrating tissue-resident memory (Trm) T cells.
Our results demonstrate that combining a TLR4 agonist with GVAX completely alters the immune response to GVAX vaccination. TEGVAX primes naïve T cells poorly and does not require T cell circulation to reduce tumor growth. These findings suggest that TEGVAX functions by eliciting a tumor-specific Trm T cell response independent of lymph node priming.
Cytokine driven effector functions of CD8 cells: application to protease-activated cytokine therapy for cancer
Authors: Norville Karli , Skrombolas Denise , Ferry Shannon , Corson Nancy , Frelinger John G. 
Published: January 2023

Abstract
Cytokines are potent immune mediators and play a crucial role in the differentiation and activation of effector CD8 T cells which are critical in anti-tumor responses. Here we tested the ability of IL-2, IL-12, and IL-18 to drive expression of gamma interferon (IFNg) in the absence of intentional antigen stimulation. We show in vitro that spleen cells from both BALB/c and C57BL/6 mice produced large amounts of IFNg and downstream mediators in response to these cytokines individually or synergistically in combination. Moreover, CD8 cells purified from Colon38 tumors can be similarly activated, demonstrating that they are not permanently inactivated or exhausted. These data illustrate innate-like function of CD8 cells, including those in the tumor microenvironment (TME). To address concerns of severe toxicities that can accompany systemic delivery of cytokines, we designed protease-activated cytokine fusion proteins (FPs) which exploit the overexpression of proteases in tumors to release active cytokine in the TME. Using gene transfection techniques, we show that an IL-2FP with a matrix metalloproteinase 2, 9 cleavage sequence can be activated in the TME and reduce tumor growth. In contrast, a FP with a scrambled non-cleavable sequence does not reduce tumor growth, illustrating the specificity of cleavage of the FP in vivo. Additionally, we demonstrate that systemic expression of the IL-2FP elicits little systemic toxicity in striking contrast to the delivery of free IL-2. Collectively, these data support the development of cytokine fusions proteins as a new class of therapeutics to promote anti-tumor effectors with limited detrimental cytokine side effects.
The impact of glycoyslation on filovirus vaccine immunogenicity in mice
Authors: Clarke Elizabeth C , Bradfute Steven 
Published: January 2023

Abstract
Filoviruses are highly virulent pathogens which cause severe hemorrhagic fevers, including Ebola virus disease (EVD). There is currently an ongoing large outbreak in the Democratic Republic of Congo, and while investigational vaccines have been used in trials during the outbreak, there are no licensed vaccines to EVD, or the related Marburg virus disease (MVD). The glycoproteins of filoviruses are the only virally expressed proteins on the virion surface and are required for receptor binding and as such, they are the main candidate vaccine antigen. A major component of the glycoprotein are the N- and O-linked glycans, but these glycans vary in number, distribution and type depending on the cell type producing the glycoprotein. Both mammalian and insect cell lines are commonly used to produce filovirus proteins used in vaccine development, but produce proteins with very different glycosylation. Here, we describe the impact of the filovirus glycoprotein glycans on the immunogenicity of the transmembrane-deleted glycoprotein (GPdTM) vaccine in mice using insect-produced, mammalian-produced, and deglycosylated filovirus glycoproteins. Results suggest differences in antibody titer and T cell epitopes induced between glycoproteins with different glycosylation patterns. Studies are underway to test how these changes affect survival against live virus challenge with mouse-adapted Ebola and Marburg.
Unique regulation of the DNA methylation machinery in natural hosts contributes to CD4 gene repression and SIV disease non-progression
Authors: Mudd Joseph C , Lai Stephen , Shah Sanjana , Rahmberg Andrew R , Flynn Jacob , Perikins Molly R , Ransier Amy , Starke Carly E , Douek Danny C , Hirsch Vanessa M , Cameron Mark J , Brenchley Jason M 
Published: January 2023

Abstract
African green monkeys (AGMs) are natural hosts of Simian immunodeficiency virus (SIV) that post-thymically downregulate CD4 to maintain a large population of CD4−CD8aa+ virus-resistant T cells which retain T-helper functionality. AGMs can become aviremic and apparently cured of SIV by down-regulating CD4 to completion. To understand the mechanisms of this process, purified CD4+ T cells from four AGMs, closely-related Patas monkeys, and rhesus macaques were stimulated with SEB for 5 days and RNAseq was performed on divided cells induced to downregulate CD4 (AGM, Patas) and those that maintain CD4 expression (rhesus). 1,917 DEGs were revealed to be common among divided, CD4-downregulated AGM and Patas T cells, yet unique from divided rhesus CD4+ T cells. Genes well-known to be regulated in natural hosts were selectively present in this dataset, including CD4, CD8A, and CXCR6 (p= 1.27e−27, 2.68e−5, 6.72e−15, respectively). Pathway analysis of DEGs revealed proteins involved in DNA methylation to be enriched in CD4-downregulated AGM and Patas T cells (p=0.013). Inhibition of DNA methyltransferases (DNMT) with 5-aza-2 deoxycitidine inhibited CD4 downregulation in AGM CD4+ T cells induced to divide in vitro (p=0.005), indicating CD4 can be pharmacologically manipulated in natural hosts. Cytosine residues within the AGM CD4 promoter region became methylated during CD4 downregulation in vitro (p= &lt; 0.0001), and were stably inherited in AGM CD4−CD8aa+ T cells sorted directly ex vivo. These results suggest AGMs employ epigenetic mechanisms to durably silence the CD4 gene. Manipulation of these mechanisms could provide avenues for modulating SIV/HIV-1 entry receptor expression in hosts that become progressively HIV-1/SIV-infected.
The novel innate immune-antagonistic effects of the multifunctional ectromelia virus C15 protein
Authors: Peauroi Elise , Forsyth Katherine S , Eisenlohr Laurence C. 
Published: January 2023

Abstract
The success of poxviruses as pathogens depends upon their extensive antagonism of host immune responses by a large arsenal of immunomodulatory proteins. The C15 protein of ectromelia virus (ECTV, the agent of mousepox) is the largest of the ECTV immunomodulatory proteins and is a member of a well-conserved poxviral family previously studied as inhibitors of T cell activation. We have recently determined that C15 also facilitates early viral spread in vivo, suggesting a second non-adaptive function of C15. Accordingly, we sought to further investigate this new function and identify the cellular target. We first found that C15 maintains its replication-promoting effect in RAG KO but not RAG IL2RG double KO mice, implying the targeting of NK cells. Nevertheless, the impact of C15 is drastically less in RAG KO than WT mice, suggesting that C15 also targets a RAG-dependent immune component. Further studies in various immune deficient strains implicate type I and III interferons, perforin and the NK gene complex in this novel function. The results prompt further in vitro and in vivo studies into the impact of C15 on NK cell and γδ T cell function, as the RAG-independent and -dependent targets respectively, and lead us to propose that, unique among currently identified viral virulence factors, C15 selectively antagonizes both the adaptive and innate immune responses.
HIV-1 Replication is Metabolically Regulated in an Ex Vivo Human Tonsil Histoculture Model of Infection
Authors: Newcombe Kevin Luna , Nixon Douglas F. , Furler Robert Lance 
Published: January 2023

Abstract
Current in vitro HIV-1 infection models use CD4+ T cell lines, primary CD4+ T cell subsets, or PBMC. However, these lack the 3-dimensional structure of lymphoid tissues, where HIV-1 replication primarily occurs. We use an ex vivo tonsil histoculture model of HIV-1 infection to measure the effects of metabolism on HIV-1 replication kinetics. Histocultures were infected with a CCR5-tropic HIV-1 isolate, ADA, in the presence or absence of inhibitors of glycolysis, oxidative phosphorylation, glutaminolysis, or fatty acid oxidation (2-Deoxy-D-glucose (2-DG), Oligomycin, 6-Diazo-5-Oxo-L-Norleucine (DON), and Etomoxir, respectively). HIV-1 replication was measured using p24 ELISA every 3–4 days for a minimum of 3 weeks. Four tonsil sections (~1 mm3 each) were placed on a collagen support in each well of a 6-well plate. 2-DG and Oligomycin significantly decreased HIV-1 production compared to untreated in this system. However, while DON and Etomoxir initially suppressed HIV-1 replication, viral production increased over the course of the experiments. Following T cell activation, increased aerobic glycolysis and oxidative phosphorylation occurs to meet the biomolecule and energy needs of a growing and/or proliferating cell. Inhibiting these pathways in our ex vivo histoculture model significantly reduced HIV-1 production. Next, we will test if these metabolic inhibitors can drive latency in our model and compare our approach to other published cytokine-driven in vitro models.
The replication-competent HIV reservoir is detectable in circulating monocytes and is increased in ART-suppressed chronic HIV-infected individuals with cognitive impairment
Authors: Mitchell Brooks I. , Laws Elizabeth I. , Souza Scott A. , Paul Robert H. , Milne Cris I.P. , Nakamoto Beau K. , Shikuma Cecilia M. , Ndhlovu Lishomwa C. 
Published: January 2023

Abstract

Background
Measuring replication-competent HIV (rcHIV) in monocytes (MO) in the setting of antiretroviral therapy (ART)-suppression, remains a topic of intense interest, particularly due to the continued presence of cognitive (cog.) decline, as well as in our pursuit for an HIV cure.

Methods
PBMCs from ART-suppressed chronic HIV-infected donors, whom had available cog. performance scores, were cell sorted using flow cytometry to obtain ultra pure MO and CD4 T cells confirmed by subsequent flow analyses. Using a novel modified TZM-bl co-culture assay, frequencies of cells producing rcHIV (IUPM) were calculated. Total p24 associated with rcHIV produced by MO or CD4 T cells was quantified using a relative light unit to rcHIV-associated p24 standard curve. The infectious potential (IP) was calculated as a ratio of total rcHIV-associated p24 and IUPM. Non-parametric tests assessed group comparisons.

Results
All HIV-infected donors in cog. impaired (n=7) and cog. normal (n=9) groups were male, on ART &gt; 3 months, and virally-suppressed. IUPMs in MO were detectable in 11 of 16 donors (median IUPM=1.32). IUPMs in CD4 T cells were detectable in 13 of 16 donors (median IUPM=7.22) and were higher as compared to MO (p=0.004). Median IP was slightly higher in MO as compared to CD4 T cells (0.443 vs. 0.104, p=0.271). Cog. impaired donors had higher IUPMs in MO as compared to cog. normal donors (4.43 vs. 0.31, p=0.015). IP between cog. groups were similar for both cellular compartments.

Conclusions
Despite ART-suppression, circulating MO are capable of producing rcHIV and may contribute to cog. impairment, as well as pose a challenge for HIV remission efforts. Monitoring and targeting HIV persistence in MO during ART-suppression are warranted.