bluuebunny/crossref_metadata_2025_split huggingface dataset:
Design and Preclinical Evaluation of a Universal SARS-CoV-2 mRNA Vaccine
Authors: Langston Laura Katherine , Xu Renhuan , Zhu Huabin , Qin Jane , Jeon Ju , Xu Jiangsheng , Mou Stephanie 
Published: August 2023

Abstract
Due to the rapid mutations of severe acute respiratory syndrome coronavirus (SARS-CoV-2), an effective vaccine against SARS-CoV-2 variants is needed to prevent coronavirus disease 2019 (COVID-19). T cells, in addition to neutralizing antibodies, are an important component of naturally acquired protective immunity, and a number of studies have shown that T cells induced by natural infection or vaccination contribute significantly to protection against several viral infections including SARS-CoV-2. However, it has never been tested whether a T cell-inducing vaccine can provide significant protection against SARS-CoV-2 infection in the absence of preexisting antibodies. In this study, we designed and evaluated lipid nanoparticle (LNP) formulated mRNA vaccines that induce only T cell responses or both T cell and neutralizing antibody responses by using two mRNAs. One mRNA encodes SARS-CoV-2 Omicron Spike protein in prefusion conformation for induction of neutralizing antibodies. The other mRNA encodes over one hundred T cell epitopes (multi-T cell epitope or MTE) derived from non-Spike but conserved regions of the SARS-CoV-2. We show immunization with MTE mRNA alone protected mice from lethal challenge with the SARS-CoV-2 Delta variant or a mouse-adapted virus MA30. Immunization with both mRNAs induced the best protection with the lowest viral titer in the lung. These results demonstrate that induction of T cell responses, in the absence of preexisting antibodies, is sufficient to confer protection against severe disease, and that a vaccine containing mRNAs encoding both the Spike and MTE could be further developed as a universal SARS-CoV-2 vaccine.
Antigen presenting cell targeted T cell DNA vaccine inducing strong and specific cellular responses across multiple T cell epitopes of SARS-COV-2
Authors: Norheim Gunnstein , Kuczkowska Katarzyna , Stubsrud Elisabeth , Bjerkan Louise , Jørgensen Marthe , Husbyn Hannah , Bøthun Anne , Dürkoop Mirjam , Sekelja Monika , Namouchi Amine , Lindahl Torbjørn , Granum Stine , Chellappa Stalin , Skarshaug Renate , Klinger Mark , Ebert Peter , Osborne Edward , Fredriksen Agnete Brunsvik , Pedersen Mikkel Wandahl 
Published: August 2023

Abstract
The severe respiratory syndrome coronavirus 2 (SARS-CoV-2) causing COVID-19 has continuously evolved with successive new virus variants of concern (VOC). A key challenge is to develop vaccines that can prevent infection and/or protect against severe disease caused VOCs that evade vaccine induced Spike neutralizing antibodies. T cell responses likely contribute to the efficacy of approved vaccines against VOCs and provide the rationale for the development of T-cell based vaccines. VB10.2210 is a T-cell based vaccine that was developed using Nykode’s easily adaptable DNA plasmid (pDNA) vaccine platform. VB10.2210 pDNA encodes homodimers consisting of i) a targeting unit that binds chemokine receptors on antigen-presenting cells, ii) a dimerization unit, and iii) an antigenic unit consisting of a selection of validated immunogenic SARS-CoV-2-specific T cell epitopes. The T cell epitopes were identified by Adaptive using T cell receptor sequencing of more than 6500 samples from COVID-19 individuals representing diverse geographies. The vaccine candidate was designed with broad HLA coverage and contains a diversity of both MHCI and MHCII T-cell epitopes across multiple viral proteins and known VOCs. In vitro characterization of the VB10.2210 pDNA showed that intact protein was expressed and secreted in human cell culture. The immunogenicity of the vaccine candidate was evaluated in transgenic HLA-A2.1, C57BL/6 and BALB/c mice. Preclinical data in these three mouse models demonstrate that VB10.2210 consistently induced strong, broad, dose-dependent, and persistent T cell responses across multiple T cell epitopes. VB10.2210 is currently being evaluated for safety and immunogenicity in a Phase I clinical trial (NCT05069623).
Relatively preserved functional capacity with standard COVID-19 vaccine regimen in People Living with HIV asymptomatic or with mild immunodeficiency
Authors: Hsiao Sung-Han , Hung Chen-Yiu , Huang Chung-Guei , Chen Tse-Ching , Chang Chia-Shiang , Huang Yu-Lin , Lin Yung-Chang , Lin Chun-Yen , Dutta Avijit , Huang Ching-Tai 
Published: August 2023

Abstract
People living with HIV (PLWH) are at a higher risk of sever disease with SARS-CoV-2 virus infection. COVID-19 vaccines are effective in most PLWH, with significant reduction in the risk of severe disease or death when the standard 2-shot regimen is taken. However, suboptimal immune response to the standard course of vaccination is a concern for PLWH, especially for those with moderate to severe immunodeficiency. An additional dose is recommended as part of the extended primary series. In Taiwan, this additional dose is endorsed even for asymptomatic or mild immunodeficiency with CD4 counts &gt;200/mm3 and suppressed virology. We aimed to investigate the immune responses with the additional dose in PLWH asymptomatic or with mild immunodeficiency. We collected peripheral blood mononuclear cells (PBMC) from 72 PLWH asymptomatic or with mild immunodeficiency and from 362 non-HIV subjects, after two and three shots respectively. Two shots elicited lower responses in PLWH, although the difference was statistically insignificant. They have comparable levels of neutralizing and anti-S antibodies and comparable effector CD4+ and CD8+ T cell responses. The 3rd shot boosted the SARS-CoV-2 immunity significantly more in PLWH compared to the non-HIV people. Upon in vitro stimulation with extracted Wuhan strain SARS-CoV-2 proteins, CD8+ T cells from PLWH after 3 shots has more durable effector responses than the non-HIV controls with extended time of stimulation. This subtle difference between PLWH and non-HIV people implied immune exhaustion with two shots in non-HIV people. Slightly compromised immunity in PLWH indeed preserved the functional capacity for further response to the 3rd shot or natural infection.
The Research Center for Emerging Viral Infections receives support from The Featured Areas Research Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education (MOE), Taiwan and NSTC 111-2634-F-182-001 from the National Science and Technology Council, Taiwan. This work was supported by Projects CMRPG3M1811 (CTH) from the Medical Research Project Fund, Chang Gung Memorial Hospital, Taiwan.
Adjuvant nanocellulose with different physicochemical property can induce immune adjuvant effects without posing cytotoxicity
Authors: Wang Lingzhen 
Published: August 2023

Abstract

Objective
To analyze the in vitro effectiveness of cellulose nanocrystals (CNC) in inducing pro-inflammatory and immune adjuvant effects in THP-1 cells, a monocyte cell line isolated from the peripheral blood of an acute monocytic leukemia patient.

Methods
THP-1 cells were counted, and their viability was calculated followed by the quantification of IL-1 β production.

Results
We observed the following: 1) CNCs were not cytotoxic to the cell lines, and 2) a 280 nm length of CNC induced the strongest immune response within our selected size spectrum.

Conclusion
This study provides a preliminary demonstration of the optimal length range CNCs that could induce most adjuvant effects without posing cytotoxicity.

This research didn’t receive funding support.
Investigating the potential of silver nanocubes and gold nanocages as protein subunit vaccine carriers
Authors: Yavuz Emine , Walters Adam , Chudasama Bhavnesh , Han Shunping , Qin Yue , Al-Jamal Khuloud 
Published: August 2023

Abstract
Metal nanoparticles have been exploited as vaccine delivery platforms for various diseases. In particular, Ag and Au NPs of different size and shape have been shown to promote delivery of antigens and act as efficient adjuvants. Yet, there is limited description of cuboidal Ag nanocubes (AgNCs) and Au nanocages (AuNCs) as antigen carriers for vaccination. Here, we first investigated the intrinsic protein binding ability of AgNCs and AuNCs, using ovalbumin (OVA) as a model. Next, the effect of AgNCs and AuNCs on bone marrow-derived dendritic cells (BMDCs) in terms of cytotoxicity and maturation was assessed. Finally, we investigated humoral and cellular immune responses of AgNCs-OVA and AuNCs-OVA following intramuscular immunization and their prophylactic effects in B16F10-OVA mice tumor model. In terms of OVA loading efficiency, AgNCs were more potent than AuNCs, despite their non-porous form. Both cubic nanomaterials were found not to induce BMDCs maturation in vitro or DCs maturation in spleen. Following the administration of nanovaccine doses, serum IgG responses were comparable between groups. However, there were significant alterations in relative frequencies of lymphocyte sub-populations. Overall, AgNCs-OVA and AuNCs-OVA immunization via intramuscular route did not alter tumour growth. This study has demonstrated the intrinsic immunomodulatory properties of AgNCs and AuNCs, preceding their potential use as nanocarriers.
Supported by grants from British Council (Newton Fund, 337313), Wellcome Trust (WT103913), and TUBITAK.
Elucidating the Immunogenic Mechanisms of Merck’s Lipid Nanoparticle (LNP) Adjuvants
Authors: Sullivan Nicole L , Secore Susan , Bass Antonia , Wolf Mary , Dangizer Andrew , Thambi Nithya , Bhattacharya Shashwati , Rodriguez Silveria , O’Donnell Gregory L , Bett Andrew L 
Published: August 2023

Abstract
Lipid nanoparticles (LNPs) have been shown to be a safe and potent platform for vaccine delivery. When added to subunit proteins, they have the potential to induce strong immune responses capable of generating long-lasting immunity. Merck has developed a variety of LNPs utilizing different cationic lipid components including LNP A, which has a long half-life in vivo, and LNP B, which has a shorter half-life in vivo and is more biodegradable. Protein subunit vaccines containing LNP A have been shown to generate robust antigen-specific humoral and cellular responses with some dose dependent reactogenicity while vaccines formulated with LNP B elicit lower immunogenicity and increased tolerability. To gain insight into the innate signaling pathways utilized by these LNPs, we performed an immunogenicity study in MyD88 knockout mice utilizing RSV F subunit protein vaccine plus LNP A or B. Wild type mice immunized with LNP A vaccines produced significantly higher levels of pro-inflammatory and neutrophil recruitment serum cytokines than the MyD88 knockout mice. However, with LNP B, MyD88 deficiency had no effect on cytokine expression. Humoral and cellular immune readouts are ongoing. Next, we stimulated whole human blood with the LNPs to compare at cytokine production. Stimulation with LNP A produced cytokine significant upregulation of monocyte and neutrophil-specific cytokines at 24 hours while stimulation with LNP B had no effect on cytokine production. Finally, we are evaluating differential gene expression to gain insight into the cellular responses of leukocytes to stimulation with the LNPs. Together, these experiments will help mechanistically determine how these fundamentally different LNPs can activate immune responses.
Modulating IL-1β and its receptors shapes spike-specific CD4+ T cell responses to mRNA vaccination
Authors: Park Hong-Jai , Shin Min Sun , Shin Junghee J , Kim Hyoungsu , Kang Byunghyun , Par-Young Jennefer , Unlu Serhan , Afinogenova Yuliya , Catanzaro Jason , Young Juan , Kim Minhyung , Lee Sang Jin , You Sungyong , Racke Michael K , Bucala Richard , Kang Insoo 
Published: August 2023

Abstract
Augmenting and sustaining a durable immunity provided by COVID-19 mRNA vaccines is important. Thus, we characterized S protein-specific CD4+ T cells in healthy individuals who received COVID-19 mRNA vaccines utilizing single cell RNA-seq and mass cytometry. In humans, S protein-specific CD4+ T cells highly expressed IL-1 receptor (R) 1 and its decoy receptor IL-1R2. IL-1b promoted IFN-g expression by S protein-stimulated CD4+ T cells, which was furthered by adding anti-IL-1R2-blocking antibodies, supporting the functional implications of IL-1R1 and IL-1R2. Following the 2nd dose of vaccination, IL-1R1 expression increased while IL-1R2 expression decreased in S protein-specific CD4+ T cells. The expression levels of such cytokine receptors correlated with S-specific antibody production. Our findings provide novel insight into the role of IL-1β and its receptor system in developing COVID-19 mRNA vaccine-induced CD4+ T cell immunity.
Supported by grants from NIH (1R01AG056728 and R01AG055362) and Quest Diagnostics.
Serological assessment of the durability of vaccine-mediated protection against SARS-CoV-2 infection
Authors: Bates John T , Lirette Seth T , Farmer Andrew P , Bierdeman Michael A , Seyfarth Kristina B , Montgomery Denise D , Burnett Grace C , Ederer Dallas R , Marshall Gailen D 
Published: August 2023

Abstract
Virus-specific neutralizing antibodies are generally accepted as a correlate of protection against infection, though many questions remain about which components of the immune response protect against SARS-CoV-2 infection. We longitudinally measured Spike receptor binding domain (RBD)-specific serum IgG and nucleocapsid protein (NP)-specific serum IgG in a human cohort immunized with the Pfizer BNT162b2 vaccine. Serum samples were collected and stored at −80 °C. Antibody titers were quantitated using an endpoint dilution ELISA and reported as inverse titers. A four-fold or greater increase in NP-specific antibody titers was used as serological marker of clinical infection at some point prior to the sample collection. Using the RBD-specific IgG titer measured in participants’ serum collected prior to seroconversion for NP, we were able to calculate a protective threshold titer for RBD-specific IgG. We calculated the rate of decay of antibodies in participants who did not exhibit a four-fold increase in NP-specific IgG, i.e. had no serological evidence of infection, and found that, on average, the RBD-specific IgG response waned below the protective threshold by four months following vaccination. These data highlight the need for a more durable immune response to vaccination which may require different adjuvants, dosages, or vaccination schedules.
This work was supported by The Robert M. Hearin Foundation, The Bower Foundation, and James L. Barksdale.
Heterozygosity for Crohn’s Disease Risk Allele of Atg16L1 Protects from Salmonella Infection
Authors: Yao Xiaomin , Rudensky Eugene , Martin Patricia , Torres Victor , Cadwell Ken 
Published: August 2023

Abstract
Autophagy is an evolutionarily conserved pathway for the formation and trafficking of lipid vesicles in response to stress, including inflammation and microbial infection. A missense variant of the essential autophagy gene Atg16L1 (rs2241880, T300A) is carried by a majority of the human population and is linked to Crohn’s Disease (CD). Previous studies found that T300A enhances proteolytic cleavage and degradation of Atg16L1, leading to dysregulated cytokine expression and impaired control of microbial infection in knock-in mice. However, these studies have focused on homozygous animals while the plurality of the human population is heterozygous for the allele. Here we demonstrate that heterozygosity for T300A protects mice from Salmonella infection. We find the variant allele results in altered complex formation independent of cleavage, allowing for enhanced inflammasome activation but impaired conjugation of Atg8 to single membranes (CASM). In vivo, one copy of T300A is sufficient to elevate the production of inflammatory cytokines, which is required for protection from Salmonella infection. Meanwhile, mice deficient in CASM genes are susceptible to Salmonella infection and this pathway is shown to be impaired in homozygous but not heterozygous mice. Finally, we demonstrate that heterozygosity in human macrophages also results in elevated cytokine expression, while maintaining functional non-canonical autophagy. These results clarify the altered autophagy function of the CD risk allele Atg16L1T300A and suggest it can be beneficial to heterozygous carriers, providing a potential explanation for its prevalence in the human population.
NIH R01 AI121244 F31 HL149238
SPATIOTEMPORAL REGULATION OF RP105 SUBCELLULAR LOCALIZATION SHAPES ENDOSOMAL TLR4 SIGNALING
Authors: Mathmann Carmen , Schultz Thomas , Ongtengco Cherica Felize J , Dalton Emma , Domínguez Cadena Leslie C , Zamoshnikova Alina , Stow Jennifer L , Blumenthal Antje 
Published: August 2023

Abstract
Innate immune sensing of microbial and endogenous ligands by Toll-like receptor (TLR) 4 drives inflammation and antimicrobial mechanisms. While critical for host defense against pathogens, unchecked TLR4 activation causes organ damage and chronic disease. Molecular mechanisms that drive versus turn off TLR4 activity are being pursued as potential targets for preventing and treating infections and inflammation-driven pathologies. Mechanisms of the temporal and spatial regulation of TLR4 signaling originating from the cell surface are well established. In contrast, the regulation of TLR4 activation originating from endosomal compartments is not well known. Here we show that the TLR-related receptor, Radioprotective 105 kDa (RP105), localizes to TLR4-containing endosomal compartments in murine macrophages. RP105 has been proposed as a negative regulator of TLR4-MyD88 signaling originating from the cell surface, but whether it modulates endosomal TLR4 signaling is unknown. Our data shows that RP105-deficiency reduces LPS- and E. coli-induced type I IFN responses, suggesting that RP105 facilitates endosomal TLR4-TRIF signaling. We employed high-resolution microscopy and AI-guided quantitative analyses to interrogate the spatiotemporal relationship between RP105 and endosomal compartments. Cellular activation led to a transient reduction of RP105 from endosomal compartments. Our results suggest TLR4 activation via TRIF signaling mediates this process, but not MyD88. Moreover, our data identify phosphatidylinositol 3-kinase p110-delta and Bruton’s tyrosine kinase as critical regulators of RP105 endosomal localization. Collectively, our data identify RP105 as a positive regulator of endosomal TLR4 signaling.
AAI Careers in Immunology Fellowship, 2022–2023
Mkp-1 deficiency strengthens immune defense against fungal infection
Authors: Liu Yusen , Mager Carli E , Shelton Evan D , Crowell Sara A , Tang Juan , Kim Victoria Y , Nelin Leif D , Zhang Jian 
Published: August 2023

Abstract
Fungal infections are a serious threat to people with a compromised immune system. The recognition of fungal pathogens relies on pathways mediated by TLRs and C-type lectin receptors. Activation of these TLR and C-type lectin receptor pathways converge on both NF-κB and MAPKs to initiate cytokine and chemokine production, leading to phagocyte recruitment and pathogen elimination. Mkp-1 is a negative feedback regulator of the p38 and JNK MAPKs. To understand the role of Mkp-1 in immune defense against fungal pathogens, we assessed the effects of Mkp-1 deficiency on the immune response to C. albicans, the most common fungal pathogen. Contrary to our previous observation with E. coli or bacterial products such as LPS, Mkp1−/− mice were protected from a lethal dose of C. albicans. In response to C. albicans challenge, Mkp1−/− mice produced greater amounts of TNF-α, had lower kidney fungal burdens and smaller fungal lesions. In wildtype mice large number of neutrophils were found in the kidneys, forming clusters surrounding fungal lesions. In contrast, far less neutrophils were found in the kidneys of Mkp-1−/− mice, although these neutrophils appeared to express greater levels of neutrophil elastase and form tighter clusters surrounding the fungi. Upon stimulation with heat-killed C. albicans yeasts and hyphae Mkp-1−/− bone marrow-derived macrophages (BMDM) also produced greater amounts of TNF-α and had higher p38 and JNK activities than did WT BMDM. TNF-α production in both wildtype and Mkp-1−/− BMDM in response to heat-killed C. albicans was attenuated by the pharmacological inhibitors of MEK1, JNK, p38, and Syk. Taken together, our results suggest that during fungal infection Mkp-1 acts to hamper the immune response against fungal pathogens.
*The studies were supported by NIH grants (AI113930 and AI124029 to Y.L., and AI121196 and AI123253 to J.Z.).
Aromatic amino acid metabolites influence interferon signaling and Influenza infectivity
Authors: Anand Gautam , Clark-Dinovo Colin , Perry Alexandra M , St. Raymond Emma , Sakleshpur Sonia , Steed Ashley L 
Published: August 2023

Abstract
The gut microbiota extensively metabolizes the three aromatic amino acids. Previously we found that a tyrosine metabolite can alter type I interferon (IFN) signaling. Herein we screened 17 metabolites of these amino acids for effects on IFN signaling alone and with LPS, IFN-β, and Influenza utilizing IFN reporter human lung epithelial cells (A549) and monocytes (THP1). While the tryptophan metabolites reduced IFN signaling in both cell types, tyrosine and phenylalanine metabolites had varied effects, depending on cell type. The cells were also treated with pooled metabolites from each family and a composite treatment with all three family metabolites. The composite treatment reduced IFN signaling in both cell types, suggesting a tryptophan metabolite effect dominance. Strikingly, when pretreated with composite treatment and infected with influenza, cells had reduced viral recovery and IFN signaling in both cell types. RNA sequencing validated earlier observation of reduced viral loads and IFN signaling. Gene silencing utilizing single siRNAs for upregulated genes identified (EGR2, ATP6V0D2, SPOCK1, and IL31RA) did not completely abrogate the repressed IFN signaling. However, these targets demonstrated linkage to TGF-beta signaling, and treatment with a TGF-beta inhibitor and gene silencing of all four gene targets led to a significant reversal of IFN signaling suppression. Our in vivo work demonstrated that intranasal administration of these metabolites prior to influenza infection leads to reduced animal weight loss, viral titers, and histological inflammation. Our work implicates that microbial metabolites alter IFN signaling through TGF-beta signaling and can promote beneficial outcomes during influenza infection.
The study was supported by grant from NIH (K08 AI135097-02) and Burroughs Wellcome Fund, awarded to Dr. Ashley L. Steed