bluuebunny/crossref_metadata_2025_split huggingface dataset:
Programming effective respiratory immunity to influenza with combination microparticle adjuvants
Authors: Bakke Brock , Marinaik Chandranaik , Toy Randall , Larsen Autumn , Lee Woojong , Kawaoka Yoshihiro , Roy Krishnendu , Suresh Marulasiddappa 
Published: January 2023

Abstract
T cell-mediated immunity (CMI) can provide long-lived protection following parenteral immunization, but eliciting durable and protective CMI in the mucosa remains a significant challenge. Here, using polylactic-co-glycolic acid (PLGA)-based cationic pathogen-like particles (PLPs) for delivery of TLR agonists, we have investigated novel ways of presenting vaccine components to the immune system to simulate the potent innate immune stimulating effects of pathogens. We generated micro particle sized PLPs loaded with TLR4 (glucopyranosyl lipid adjuvant, GLA) and/or TLR9 (CpG) agonists, and formulated them with and without a mucosal delivery enhancing carbomer-based nano-emulsion adjuvant (ADJ). These adjuvants delivered intranasally to mice safely elicited robust and durable influenza nucleoprotein (NP)-specific CD8+/CD4+ effector and tissue-resident memory responses in lungs and airways. PLPs delivering TLR4 versus TLR9 agonists drove phenotypically and functionally distinct populations of effector and memory T cells. Following influenza virus challenge, functional and phenotypic attributes of effector T cells unique to the individual PLP formulation were associated with disparate levels of viral control in the respiratory tract. These studies demonstrate: (1) how adjuvant formulations affect effector/memory T cell differentiation in the lungs; (2) feasibility of using PLGA-based PLPs in vaccine formulations to elicit protective T cell immunity in the respiratory tract.
Development of alternative vaccination strategies to overcome Tc1 vaccine failure caused by chronic schistosomiasis
Authors: Shollenberger Lisa M , Norwood Stephanie K , Bement Zachary J 
Published: January 2023

Abstract
It is a global public health imperative to develop vaccine strategies that produce effective cell-mediated immunity in schistosome-endemic areas, such as sub-Saharan Africa, where HIV infection is common. Vaccine delivery techniques effective in immune-suppressed populations are critical to combating infectious diseases in these already compromised groups. It is well-known that immune suppression, such as by chronic schistosome infection, can cause Tc1/Th1 type vaccines to fail. Listeria, a strong inducer of cell-mediated immunity, can overcome the Th2 biasing and immune suppression caused by schistosome infection. However, safety concerns cause reluctance among the vaccine community and the public to incorporate this vector into immunization programs. Since neither DNA nor the Listeria vaccine vector are ideal for clinical translation, and Gag is likely not the ideal HIV vaccine antigen, we are using this system as a model for enumeration of Tc1 responses (IFNγ+ CD8+ cytotoxic T lymphocytes, CTL); CTLs are likely necessary components of functional responses for next-generation vaccines targeting diseases that currently lack effective vaccines (HIV, TB, malaria). Here, we investigate why DNA vaccine vectors fail during chronic schistosomiasis when Listeria vaccine vectors can generate robust CTL responses to HIV Gag. Specifically, we are examining live bacteria as adjuvants for an episomally expressed or concurrently administered DNA vaccine for HIV Gag. This project is one step toward our overall goal, which is to generate cell-mediated vaccine immunity in schistosome-infected, immune-suppressed patients.
An Ebola virus vaccine that leverages helper T cells induced by prior BCG immunization
Authors: Ng Tony W , Wirchnianski Ariel A , Chan John , Jacobs William R , Chandran Kartik , Marzi Andrea , Porcelli Steven A 
Published: January 2023

Abstract
Protective antibody responses against the Ebola virus (EBOV) are dependent on the quality of CD4+ helper T (Th) cells. We propose that BCG specific Th cells may provide a source of Th cells that can be recruited to drive antibody responses against EBOV, given that the majority of individuals residing in EBOV endemic regions have been previously vaccinated with BCG. To take advantage of pre-existing BCG Th cells, a vaccine was developed that consists of the EBOV GP fused to the mycobacteria Th cell epitope (P25) of Ag85B (P25-EBOV GP). Presentation of the P25 epitope by B cells recruited cognate interactions with P25 Th cells to promote antibody responses against the EBOV GP. Experiments in mice showed BCG priming induced a broad range of Th cells including Th1, Th2, and Tfh subsets. These P25 Th subsets provided a significant dose sparing effect on the P25-EBOV GP vaccine in the induction of EBOV GP antibodies and resulted in increased titers of high affinity neutralizing IgG1 antibodies. The induction of IgG2c antibodies only occurred in BCG primed mice, where BCG vaccination is known to be a strong inducer of Th1 responses that drive class-switching to IgG2c. Microscopy of sections from secondary lymphoid organs of BCG primed mice showed that the majority of P25 Th cells remained in extrafollicular regions outside germinal centers and were most responsible for the induction of the IgG2c class-switched antibodies that were weakly neutralizing but capable of mediating antibody-dependent cellular toxicity (ADCC). BCG priming followed by vaccination with P25-EBOV GP protected mice against lethal EBOV infection, thus suggesting a potentially useful strategy for vaccination against EBOV in populations with high prevalence of previous BCG vaccination.
Structural and functional analysis of cooperativity in a potent human antibody cocktail against Ebola virus
Authors: Gilchuk Pavlo , Murin Charles D , Milligan Jacob C , Cross Robert W , Mire Chad E , Ilinykh Philipp A , Huang Kai , Kuzmina Natalia , Altman Pilar X , Hui Sean , Gunn Bronwyn M , Bryan Aubrey L , Davidson Edgar , Doranz Benjamin J , Turner Hannah L , Alkutkar Tanwee , Flinko Robin , Orlandi Chiara , Carnahan Robert , Nargi Rachel , Bombardi Robin G , Vodzak Megan E , Li Sheng , Okoli Adaora , Ibeawuchi Morris , Ohiaeri Benjamin , Lewis George K , Alter Galit , Bukreyev Alexander , Saphire Erica Ollmann , Geisbert Thomas W , Ward Andrew B , Crowe James E 
Published: January 2023

Abstract
Structural principles underlying the composition of protective antiviral monoclonal antibody (mAb) cocktails are poorly defined. Here, we exploited antibody cooperativity to develop a therapeutic mAb cocktail against Ebola virus (EBOV). Systematic analysis of antibody repertoire in the human survivors identified a pair of potently neutralizing mAbs that cooperatively bind to the ebolavirus glycoprotein (GP). High-resolution structures revealed that in a two-antibody cocktail, molecular mimicry was a major feature of mAb/GP interactions. Broadly neutralizing mAb rEBOV-520 targeted a conserved epitope on the GP base region. MAb rEBOV-548 bound to a glycan cap epitope, possessed neutralizing and Fc-mediated effector function activities, and potentiated neutralization by rEBOV-520. Remodeling of the glycan cap structures by the cocktail enabled enhanced GP binding and virus neutralization. The cocktail demonstrated high levels of effectiveness against EBOV in nonhuman primates. These data illuminate structural principles of antibody cooperativity with implications for the design and development of antiviral immunotherapeutics.
Dendritic cell targeting anti-CD40 antibody-CD40L-HIV-1 vaccines are adjuvant intrinsic
Authors: Ceglia Valentina , Zurawski Sandy , Montes Monica , Bouteau Aurelie , Wang Zhiqing , Ellis Jerome , Kroll Mitchell , Igyarto Botond Z , Levy Yves , Zurawski Gerard 
Published: January 2023

Abstract
Soluble CD40L and agonistic anti-CD40 monoclonal antibody are adjuvants used in vaccination settings. Vaccines based on anti-CD40 antibodies fused to HIV-1 antigens are in clinical development. Studies with current anti-CD40-based dendritic cell (DC) targeting vaccines show that co-administration of an adjuvant is needed for maximal immune responses. We show that by fusing CD40L to CD40-targeting antibodies, activation of DCs concomitant with antigen uptake and processing is maximized, and this provides a context CD40-targeting vaccines with intrinsic adjuvant activity. Direct fusion of CD40L to L or H chain C-termini results in CD40 agonists with ‘superagonist’ properties. Especially on DCs, both potency and efficacy for induction of cytokine secretion and activation markers is greatly enhanced compared to known strong agonists like Pfizer’s anti-CD40 CP-870-89 antibody. This potency was maintained by anti-CD40-CD40L constructs fused to HIV-1 antigens from Gag, Nef, and Pol regions (HIV5pep). Anti-CD40-CD40L-HIV5pep preferentially expanded CD8+ T cells from HIV-1+ donor PBMCs compared to the same antibody-antigen fusions without attached CD40L. Anti-CD40-CD40L-TEα and anti-CD40-TEα both evoked robust proliferation of TEα-specific CD4+ T cells in human CD40 transgenic mice, but only anti-CD40-CD40L-TEα vaccine elicited TEα-specific CD4+ T cells producing IFNγ. Also, anti-CD40-CD40L-Env gp140 vaccine without adjuvant in human CD40 transgenic mice elicited stronger anti-Env gp140 antibody responses than anti-CD40-Env gp140 vaccine. Thus superagonist anti-CD40 antibodies directly fused to the natural ligand show great advantage in inducing immune responses without the use of an extrinsic adjuvant.
Characterization of a protective antibody against dengue virus non-structural protein 1 (NS1) reveals critical domains required for NS1-triggered pathogenesis
Authors: Wong Marcus P , Biering Scott B , Akey David L , Lo Nicholas TN , Brown W. Clay , de Sousa Francielle Tramontini Gomes , Puerta-Guardo Henry , Wang Chunling , Konwerski Jamie , Espinosa Diego , Li Jeffery , Blanc Sophie , Beatty P. Robert , Smith Janet L , Harris Eva 
Published: January 2023

Abstract
Dengue virus (DENV) is a medically important, mosquito-borne flavivirus that infects up to 390 million people annually. A signature of severe DENV infection is vascular leak, which can lead to shock and organ failure. We and others have shown that secreted DENV non-structural protein 1 (NS1) triggers endothelial cell (EC) hyperpermeability and vascular leak. Antibodies (Abs) targeting NS1 prevent NS1-induced EC hyperpermeability in vitro and are protective against lethal DENV challenge, as shown by vaccination of mice against NS1 and passive transfer of α-NS1 polyclonal serum. How α-NS1 Abs protect against pathogenesis, however, remains obscure. Here we characterize possible mechanisms behind the protective ability of an NS1-specific monoclonal Ab, 2B7. We show that 2B7 abrogates NS1 binding to ECs and EC hyperpermeability in vitro and inhibits vascular leak and mortality in vivo. Guided by crystal structures of NS1 complexed to a 2B7 Fab fragment, we mutated the amino acids on NS1 that comprise the NS1:2B7 interface. Intriguingly, while these contact sites were critical for the interaction between 2B7 and NS1, they were dispensable for NS1:EC binding. The structures and mutagenesis results suggest that 2B7 abrogates NS1 binding indirectly by sterically hindering the binding domain on NS1 from interacting with ECs. The mutagenesis also identified a key residue on NS1 that is necessary for inducing EC hyperpermeability without affecting NS1:EC binding, suggesting that 2B7 may also block an additional step in NS1 pathogenesis. This work provides mechanistic insight into the blockade of DENV NS1-mediated pathogenesis by α-NS1 Abs and identifies critical domains that serve as targets for drugs and vaccine-induced α-NS1 Ab responses.
Adjuvanted bacteriophage vaccine targeting Pseudomonas aeruginosa
Authors: Román-Cruz Valery Cristina , Miller Shannon , Buhl Cassandra , Schooner Roman , Walsh Lois , Secor Patrick , Burkhart David J , Evans Jay T 
Published: January 2023

Abstract
Pseudomonas aeruginosa (Pa) is a Gram-negative bacterium which is associated with chronic diabetic wounds as well as potentially deadly lung infections in cystic fibrosis patients. Approximately 51,000 cases of Pa infections occur annually; of these cases, 13% are resistant to antibiotics which results in the deaths of ~400 individuals every year. Diversity between Pa strains presents a challenge in developing a vaccine or therapeutic capable of resolving infections caused by different strains. Pf phage (a filamentous bacteriophage) has been found in Pa from chronic diabetic wound isolates and has been show to increase virulence of Pa. Recent findings by our team indicate that a Pf phage-targeted vaccine or monoclonal antibody provide protection from the establishment of Pa infection in mice. To further improve this innovative new vaccine targeting Pa, adjuvant systems were used to enhance humoral response to the vaccine. A consensus peptide from Pf Phage coat protein was conjugated to the carrier protein CRM197 and combined with novel adjuvants and delivery systems. Selected adjuvants significantly enhanced humoral immunity to the Pf Phage peptide in a dose-dependent manner. Combination adjuvant systems were also used to further enhance antigen-specific immunity to Pf Phage and cell-mediated immunity to the carrier protein. The development of an adjuvanted Pf phage vaccine to protect against Pa infections is a new and innovative strategy with the potential to protect patients at risk for opportunistic bacterial infections.
Vaccine-Elicited T cells are Important for Intravenous BCG Mediated Protection in a Rhesus macaque Model of Tuberculosis
Authors: Zeppa Joseph John , Darrah Patricia A , Sutton Matthew , Pokkali Supriya , Scanga Chuck A , Maiello Pauline , Lin Philana L , Roederer Mario , Seder Robert A , Flynn JoAnne L 
Published: January 2023

Abstract
Our laboratories have shown that intravenous (IV) vaccination with Bacille Calmette-Guerin (BCG; 5×107 CFU) provides robust protection against low-dose Mycobacterium tuberculosis (Mtb) infection in rhesus macaques (9/10 protected [&lt;50 thoracic Mtb CFU]; 100,000-fold reduction compared to intradermal [ID] BCG). IV BCG also resulted in a 100-fold increase in mycobacterium--specific T cells in the airways of animals compared to ID or aerosol administration. To investigate the role of T cells in this protective model, we employed two methodologies: 1. Vaccination with decreasing doses of BCG to stimulate lower-magnitude T cell responses and; 2. Antibody-mediated CD4 and CD8a T cell depletion post-vaccination but pre-infection. Animals were vaccinated for 6 months followed by a 2–3 month low-dose Mtb challenge. Surprisingly, even at low doses of BCG (0.4–2 × 105), a subset of animals was protected, with a range of outcomes across the doses. In the second study, depletion was successful in blood, airways and tissue. Interestingly, depletion of either T cell subset resulted in an intermediate total thoracic CFU phenotype. In both studies, correlates of immunity will be assessed comparing protection (CFU) to BAL and tissue resident immune responses. Our data supports that sterilizing protection is observed across the BCG IV dose spectrum and that T cells are important, but are likely not the only factors involved in this protection. Our findings are critical for identifying correlates of protection and generating a safe and effective vaccine against TB.
Identifying mediators of STING-dependent TB vaccine immunity
Authors: Jong Robyn MY , Van Dis Erik , Baltodano Alexander , Nguyenla Xammy , Blackshire Bianca , Magee Kiran , Stanley Sarah 
Published: January 2023

Abstract
Tuberculosis (TB) is the top infectious killer in the world. The only current licensed vaccine for TB, Mycobacterium bovis Bacille Calmette-Guérin (BCG), has not proven reliably effective against adult pulmonary TB in all studies. It furthermore cannot be administered safely to immunocompromised individuals as it is a live attenuated vaccine. Protein subunit vaccines offer increased safety over live attenuated vaccines. Many recently developed protein subunit TB vaccines use toll-like receptor-stimulating adjuvants that induce potent inflammatory responses, but it is uncertain whether they can induce long-lasting memory protection against an intracellular bacterium such as TB.
Cyclic-di-nucleotides are an alternative adjuvant that target the cytosolic immunosurveillance pathway mediated by STING. Recently, a STING-activating cyclic-di-nucleotide (CDN)-adjuvanted TB protein subunit vaccine was shown to elicit an effective memory T cell response against TB challenge in mice. Intriguingly, intranasal delivery of the vaccine provided superior protection compared to subcutaneous administration of the CDN-adjuvanted vaccine or BCG. Protection induced by mucosal administration correlates with a potent Th17 response rather than the Th1-dominated CD4+ T cell response induced by subcutaneous vaccination. Protection is partially dependent on IL-17 and type I interferon, while the transcription factor STAT6 is dispensable. The Th17 response and protection both precedes Th1 influx into the lungs and also persists after the Th1 response wanes. We are currently determining whether the transcription factor IRF3 and cytokine IFNγ are also mediators of Th17 memory induction and protection against TB infection.
Pairwise interactions govern the effects of complex immune adjuvant combinations
Authors: Pandey Surya , Gruenbaum Adam , Kanashova Tamara , Mertins Philipp , Cluzel Philippe , Chevrier Nicolas 
Published: January 2023

Abstract
The immune system decides upon actions in response to complex combinations of microbial inputs. Dozens of innate immune sensing pathways can collectively respond to combinations of microbial and immunomodulatory agents such as adjuvants. We currently lack a clear understanding for the combinatorial logic that governs how the interplay between higher-order combinations of microbial or adjuvant signals shape immune responses. Here, using in vitro coculture experiments and statistical analyses, we uncover an intrinsic property of pathogen-sensing pathways whereby the effects of higher-order combinations of stimuli on immune responses can be explained by single and pairwise effects. Surprisingly, this property of combinatorial innate sensing is mechanistically rooted in the molecular organization underlying pathogen-sensing pathways. Indeed, we find that the outputs of mouse and human innate immune dendritic cells upon triplet stimulations are dictated by single and pairwise effects at the levels of transcription, chromatin and protein secretion. Furthermore, we exploit this simplifying property of combinatorial pathogen sensing to develop and characterize cell-based immunotherapies with potent anti-tumor properties in mouse models using combinations of adjuvants. We conclude that the processing of complex mixtures of microbial or adjuvant inputs by immune cells is governed by pairwise effects, which suggests a path towards the rationale selection of adjuvant combinations to alter immunity.
Promoting anti-tumor immunity via bispecific T cell engaging cytokine (biteokine) therapy
Authors: Do Priscilla , Perdue Lacey A. , Chyong Andy , Henry Curtis J. , Porter Christopher C. , Dreaden Erik C. 
Published: January 2023

Abstract
Bispecific T cell engager (BiTE) therapy bypasses the need for natural recognition of tumor cells by cytotoxic T cells, inducing potent anti-tumor immunity. Currently, the FDA approved BiTE, Blinatumomab (Blin), provides an alternative to standard chemotherapy in patients with relapsed/refractory or MRD+ Acute Lymphoblastic Leukemia (ALL) and is in clinical trials for other B cell malignancies. Blin targets malignant B cells using the antigen binding domain of an αCD19 antibody linked to the antigen binding domain of a T cell-activating αCD3 antibody. Despite progress with Blin clinically, complete response is 39–69%, and a significant proportion of patients are refractory or relapse. To improve this outcome, we have engineered a nanoparticle-based assembly and screening approach to capitalize on cytokine-enhancement of BiTE activity. We found that IL-12 significantly improves Blin-induced B-ALL cell lysis in vitro and, using nanoparticles capable of displaying ~142 antibodies/particle, we generated 28 unique formulations with varying densities and ratios of antibodies to target CD19, activate CD3, and localize IL-12 in a non-neutralizing fashion. We screened these particles in parallel using a co-culture assay for target cell lysis and T cell expansion by flow cytometry. Through regression modeling of these data, we can select-for and optimize architectures that maximize target cell lysis and T cell proliferation. Our preliminary results indicate that optimally lytic architectures favor high αCD3 to αCD19 ratios and are improved linearly with increasing IL-12. This work seeks to innovate and enhance bispecific T cell engager therapy with a modular platform that can be used broadly in the treatment of cancer and autoimmunity.
Tri-functional T-cell engagers target immune checkpoint inhibitors PD-1 and TIGIT to enhance CD8 effector T-cell functions in chronically SHIV-infected rhesus macaques
Authors: Promsote Wanwisa , Ambrozak David , Almasri Cassandra , Liu Catherine , DeMouth Megan , Hataye Jason , Pegu Amarendra , Mascola John R. , Koup Richard 
Published: January 2023

Abstract
Checkpoint inhibitors, such as PD-1 and TIGIT, are crucial dysregulators of CD8 T cell function during chronic HIV/SIV infections. Importantly, these checkpoint inhibitors are highly expressed on the surface of CD4 T cells that harbor latent HIV. We have previously demonstrated that an anti-HIV/anti-CD3 bispecific T-cell engager (BiTE) can be used to redirect follicular CD8 (fCD8) T cells to kill HIV infected cells in vitro. Here, we hypothesize that adding an extra specificity to target PD-1 or TIGIT to the BiTE will further enhance the functional activities of CD8 T cells and simultaneously lower the threshold for reactivation of HIV latently infected cells. By using HIV-infected cell lines and primary CD4 T cells isolated from chronically SHIV-infected lymph nodes (LN) as target cells in in vitro and ex vivo killing assays, we found that trifunctional anti-HIV/anti-CD3-anti-TIGIT or anti-HIV/anti-CD3-anti-PD1 increased (1) T-cell specific activation, (2) antigen-specific CD107a degranulation, levels of granzyme B, cytokine/chemokine release by CD8 T cells, and (3) the killing capability of functionally compromised CD8 T cells when compared to the BiTE. When administered at 100 mcg/kg subcutaneously to chronically SHIV-infected animals, we demonstrated that anti-HIV/anti-CD3-anti-TIGIT could safely be used to augment activation and expansion of CD8 effector T cells that further resulted in a better elimination of the SHIV-infected cells as shown by the reduction in cell-associated SHIV gag DNA and RNA. Together, our results indicate that trifunctional T cell engagers targeting checkpoint inhibitors may serve as novel immunotherapeutic strategies to eliminate infected cells in HIV infected individuals.