bluuebunny/crossref_metadata_2025_split huggingface dataset:
The TLR4 agonist vaccine adjuvant, GLA-SE, requires canonical and atypical mechanisms of action for TH1 induction
Authors: Cauwelaert Natasha Dubois , Desbien Anthony L , Hudson Thomas E , Pine Samuel O , Reed Steven G , Coler Rhea N. , Orr Mark T 
Published: January 2023

Abstract
The Toll-like receptor 4 agonist glucopyranosyl lipid adjuvant formulated in a stable emulsion (GLA-SE) promotes strong TH1 and balanced IgG1/IgG2 responses to protein vaccine antigens. This enhanced immunity is sufficient to provide protection against many diseases including tuberculosis and leishmaniasis. To better characterize the adjuvant action it is important to understand how the different cytokines and transcription factors contribute to the initiation of immunity. In the present study using T-bet−/− and IL-12−/− mice and a blocking anti-IFNαR1 monoclonal antibody, we define mechanisms of adjuvant activity of GLA-SE. In accordance with previous studies of TLR4 agonist based adjuvants, we found that TH1 induction via GLA-SE was completely dependent upon T-bet, a key transcription factor for IFNγ production and TH1 differentiation. Consistent with this, deficiency of IL-12, a cytokine canonical to TH1 induction, ablated TH1 induction via GLA-SE. Finally we demonstrate that the innate immune response to GLA-SE, including rapid IFNγ production by memory CD8+ T cells and NK cells, was contingent on type I interferon, a cytokine group whose association with TH1 induction is contextual, and that they contributed to the adjuvant activity of GLA-SE.
CD16-IL15-CD33 Trispecific Killer Engager (TriKE) induces NK cell expansion, persistence, and myeloid blast antigen specific killing.
Authors: Felices Martin , Vallera Daniel A. , McElmurry Ron , Lenvik Alexander J.L. , Chu Sami , Todhunter Deborah , Taras Elizabeth P. , Tolar Jakub , Blazar Bruce R , Miller Jeffrey S 
Published: January 2023

Abstract
NK cell infusions induce complete remissions in 30–50% of patients with refractory AML when combined with lymphodepleting chemotherapy and IL-2. NK cell therapy is limited by lack of antigen specificity and by IL-2 mediated induction of Tregs. To overcome these limitations we developed a 161533 trispecific killer engager (TriKE) molecule containing an anti-CD16 scFv, to engage NK cells, an anti-CD33 scFv, to engage AML targets, and an IL-15 molecule. We have shown previously that a 1633 bispecific killer engager (BiKE) is capable of creating an immunologic synapse between NK cells and AML targets leading to target killing. These molecules generate higher affinity interactions than natural CD16 binding to Fc portions of therapeutic antibodies, resulting in increased NK cell functionality. Addition of IL-15 in the TriKE molecule is meant to enhance NK cell priming, proliferation, and maintenance without inducing Tregs. Cytotoxicity assays and cytokine secretion assessment of NK cells incubated with CD33+ HL-60 target cells indicated that the TriKE induced greater NK cell activation than the BiKE or no molecule control. The TriKE was able to induce potent NK cell proliferation and survival when compared to the BiKE. Interestingly, when compared to recombinant IL-15 the TriKE induced less proliferation of T cells while inducing similar proliferation of NK cells from healthy donors, indicating specificity. The TriKE was capable of rescuing NK cell function of post-transplant patients. In vivo functionality was tested in an NSG xenogeneic model engrafted with HL-60-luc targets and NK cells. TriKE treatment resulted in reduced tumor burden and NK cell maintenance at day 21, displaying the immunotherapeutic potential of the molecule.
A small-molecule RIG-I agonist functions to enhance vaccine protection against influenza A virus infection.
Authors: Hemann Emily A , Grigg John B , Wilkins Courtney R , Knoll Megan , Iadonato Shawn P , Bedard Kristin , Probst Peter , Loo Yueh-Ming , Gale Michael 
Published: January 2023

Abstract
Viral sensing by RIG-I and downstream activation of antiviral defenses along with the induction of innate immune cytokines is essential from protection against influenza A virus (IAV) infection. We have identified a novel, small-molecule RIG-I agonist, KIN1148, which binds and activates RIG-I to signal the activation of IRF3 and the innate immune response. We are developing this molecule as an adjuvant to enhance vaccination against pandemic H1N1 (pH1N1) IAV. Ex vivo treatment of dendritic cells with KIN1148 leads to their activation and maturation. We determined the ability of KIN1148 to enhance suboptimal IAV vaccine responses in vivo. Administration of KIN1148 leads enhanced protection during high dose pH1N1 infection following a single, intramuscular administration of KIN1148 with IAV vaccine. This increase in protection is accompanied by a significant reduction in virus titers, as well as lung pathology. Analysis of the immune response induced following vaccination with KIN1148 as well as challenge demonstrates an increase in chemoattractant cytokines, germinal center B cells, IAV-specific antibodies, and IAV-specific CD4 and CD8 T cells compared to vaccination alone, indicating the induction of a broad anti-IAV immune response. Together these results demonstrate that prophylactic drug targeting of the RIG-I pathway with a small molecule enhances vaccine protection and highlight the potential of KIN1148 to enhancing vaccines against RNA virus infection.
A trivalent alphavirus virus-like replicon particle vaccine induces cell-mediated responses against conserved glycoprotein regions
Authors: Cohen Courtney A , Bakken Russ , Glass Pamela J 
Published: January 2023

Abstract
Venezuelan (VEEV), eastern (EEEV) and western (WEEV) equine encephalitis viruses are highly infectious and neuroinvasive when aerosolized, making them threats for weaponization. In contrast to subcutaneous infection, studies suggest that protection from aerosol challenge is partially cell-mediated, albeit poorly understood. A trivalent (VEEV/EEEV/WEEV) virus-like replicon particle (triVRP) vaccine is protective against aerosol challenge in mice, yet the correlates of protection are unclear. To investigate, mice were vaccinated (day 0 and 28; PBS or triVRP) and spleens were harvested for analysis. Using an ex vivo splenocyte expansion assay with peptides from VEEV, EEEV and WEEV glycoprotein libraries, IFNγ-producing memory CD8+ T cells were identified. T cell-specific epitopes were mapped, and the most immunodominant were found within regions of sequence homology between strains. Thus, ongoing studies evaluate potential monovalent vaccine cross-protection, which would greatly reduce manufacturing costs. Understanding vaccine-induced responses will inform rational design and clinical evaluation.
Rapamycin adjuvant therapy did not improve the outcome of severe influenza virus infection in an experimental mouse model system
Authors: Huang Ching-Tai , Dutta Avijit , Chen Tse-Ching , Lin Chun-Yen , Lin Yung-Chang , Kao Kuo-Chin , Hung Chen-Yiu , Chang Chia-Shiang , He Yueh-Chia , Huang Yu-Lin 
Published: January 2023

Abstract
The host immune system responds to influenza virus infection for eradication of the virus but the immune response associated inflammation causes tissue injury and damage to the host as well. Severe influenza with morbidity and mortality is commonly believed to be the result of overwhelming immune response. In addition to antiviral agent, adjuvant therapy with immune modulating agent is a reasonable approach for severe influenza and has been reported to be effective in clinical series. As the immunity is also critical for virus eradication and recovery, we investigated to try to define the appropriate window for immune modulation in our murine animal model of influenza virus infection. Rapamycin treatment in a wide dose range did not alter the clinical course of severe influenza. In combination with antiviral agent Oseltamivir, rapamycin failed to offer additional benefit beyond treatment with Oseltamivir alone. Oseltamivir barely rescued the mice from severe influenza if the treatment was delayed until 48 hours after infection. Rapamycin did not provide salvage effect in this scenario. In fact, rapamycin suppressed influenza antigen specific CD4+ T cell response and impaired the virus eradication capacity. The virus burden in the lung increased with intense neutrophil infiltration and hyaline deposition in the alveoli. In conclusion, rapamycin cannot be relied upon as a salvage measure in severe influenza with progressive illness under treatment with Oseltamivir. T cell immunity is suppressed but other arms of immunity are recruited with undesirable effect on the disease outcome.
Identification of a unique lipid from Francisella tularensis as an effective therapeutic for viral mediated inflammation
Authors: Bosio Catharine M , Nardone Glenn , Broeckling Corey , Chiramel Abilash , Best Sonja , Ireland Robin 
Published: January 2023

Abstract
Francisella tularensis (FT) is a highly pathogenic bacterium which can cause acute, lethal disease following exposure to fewer than 20 bacteria. FT is a successful pathogen due to its ability to potently and rapidly suppress inflammatory responses in host cells. We previously established that crude lipid preparations from virulent FT dampen inflammation in vitro and in vivo, but did not characterize the active specie(s). Utilizing a variety of biochemical techniques, we identified one of the active lipids found in FT as a unique phosphatidylethanolamine (PE) consisting of tetracosanoyl and decanoyl acyl chains (PE2410). Similar to crude lipid, enriched PE2410 restricted production of cytokines in human and mouse cells stimulated with well-defined microbial ligands. PE2410 was also effective at dampening inflammation provoked by infection with West Nile Virus. Activity of PE2410 was dependent on presence of the host phosphatase SHP-1. Importantly, a synthetic version of the PE2410 mimicked the ability of natural PE2410 to inhibit inflammation triggered by a variety of sources. Together our data support use of PE2410 as a novel therapeutic for pathogenic inflammation triggered by viral, or other, infection.
Suppression of dendritic cell necroptosis by pandemic influenza virus
Authors: Hartmann Boris M. , Albrecht Randy A , Marjanovic Nada , Sealfon Stuart C 
Published: January 2023

Abstract
The public health risk of a new pandemic influenza virus (IAV) approaching the scope and mortality of the devastating 1918 strain makes it important to identify virus-host mechanisms unique to pandemic strains. Human dendritic cell infection by two different H1N1 seasonal IAV strains (1991 and 1999) induces widespread RNA degradation and programmed cell death beginning at 4 h after infection. Characteristic of necroptosis, the cell death showed nuclear condensation and fragmentation that was caspase independent and RIP3 kinase dependent. Notably, infection with the pandemic 1918 and 2009 H1N1 IAV strains did not induce dendritic cell RNA degradation and necroptosis. Furthermore, necroptosis induced in dendritic cells by poly I:C exposure was suppressed by concomitant infection with the pandemic 2009 IAV. These results indicate that infection with the pandemic viruses not only fails to induce the dendritic cell necroptosis seen with the seasonal IAV strains, but that it actively suppresses necroptosis. Virus chimera studies implicate the HA virus segment as mediating the necroptosis inhibition by the pandemic IAV strains. Necroptosis leads to the release of intracellular components which serve as danger associated molecular patterns (DAMP), making this a highly immunogenic cell death mechanism. Consistent with this view, we find that the suppression of virus-induced necroptosis by the pandemic IAV reduces T cell activation. These studies show that pandemic viruses are unique in suppressing a key immunological danger signal. The suppression of the generation of DAMPs from infected immune cells may contribute to the pathogenicity of pandemic H1N1 IAV viruses.
Influenza exposure history dictates cellular dynamics in CD8 T cell-mediated lung protection
Authors: Van Braeckel-Budimir Natalija , Martin Matthew D , Hartwig Stacey , Badovinac Vladimir P , Harty John T 
Published: January 2023

Abstract
Influenza-specific memory (M) CD8 T cells directed against conserved viral antigens can mediate protection against heterosubtypic influenza infections. In the light of flu seasonality, it is important to understand how multiple influenza exposures shape specific M CD8 T cells and their capacity to provide protection. To address this major knowledge gap we studied primary (1M) and secondary (2M) influenza-specific endogenous M CD8 T cell responses in mice. We show that secondary influenza infection generates large, long lasting, Ag-specific CD8 T cell responses, which exceed 1M responses by 5–20× in all tissues analyzed. To evaluate CD8 T cell-mediated protection, without the contribution of other immune mediators induced by multiple influenza exposures, we used intranasal infection with vaccinia virus expressing influenza Ag. Interestingly, despite the large difference in numbers of Ag-specific 1M and 2M CD8 T cells, both groups of mice showed similar control of infection in the lung. Importantly, initial numerical differences between 1M and 2M CD8 T cells in the lung disappear with the course of infection (days 0–3), due to superior accumulation of 1M compared to 2M CD8 T cells, as shown by Pertussis toxin-induced migration block. Thus, while protection mediated by 2M CD8 T cells relies mainly on a large number of lung-residing 2M CD8 T cells, 1M CD8 T cells display superior recruitment to the target tissue and exhibit higher proliferative capacity, allowing them to compensate for fewer M numbers at the time of re-infection. Therefore, repetitive influenza exposures have the capacity to change numerical and dynamic properties of protective M CD8 T cell responses, which should inform the design of more efficient influenza vaccines.
Antibody idiotype plays a critical role in regulating antibody-dependent cell-mediated cytotoxicity (ADCC) against influenza A virus
Authors: Miller Matthew S , He Wenqian , Tan Gene S , Mullarkey Caitlin E , Krammer Florian , Palese Peter 
Published: January 2023

Abstract
The generation of strain-specific neutralizing antibodies against influenza A virus is known to confer potent protection against homologous infections. Recently, elicitation of broadly-neutralizing antibodies which target the conserved hemagglutinin stalk domain have emerged as a promising “universal” influenza virus vaccine strategy. The ability of these antibodies to elicit Fc-dependent effector functions, such as antibody-dependent cell-mediated cytotoxicity, has emerged as an important mechanism through which protection is achieved in vivo. However, the way in which Fc-dependent effector functions are regulated by polyclonal influenza-binding antibody idiotypes in vivo has never been defined. Here, we demonstrate that complex interactions among viral glycoprotein-binding antibody idiotypes regulate the magnitude of antibody-dependent cell-mediated cytotoxicity induction. We propose that antibody idiotype serves as a critical determinant in the activation of ADCC. This is the first demonstration that cross-talk among antibody idiotypes is important for the regulation of Fc-dependent effector functions. This phenomenon will have major implications for not only the development of universal influenza virus vaccines, but also in any scenario wherein Fc-dependent effector functions are engaged in the context of polyclonal responses containing multiple antibody idiotypes.
ADAMTS4 modulates lung tissue repair following lethal influenza A infection in mice
Authors: Boyd David F , Sanders Catherine J , Bajracharya Resha , Diercks Alan H , Thomas Paul G 
Published: January 2023

Abstract
Lung tissue damage is a major contributor to morbidity and mortality following infection with influenza A virus. Tissue repair, including remodeling of the extracellular matrix (ECM), is a critical process in recovery from infection. A Disintegrin and Metalloproteinase with Thrombospondin motifs (ADAMTS) family proteins are secreted zinc metalloendopeptidases and important mediators of ECM remodeling. The role of ADAMTS proteins in lung tissue repair following influenza A infection remains unclear. To investigate the regulation of ADAMTS genes in the context of influenza A infection, we performed a microarray analysis of whole lung homogenates harvested at 24 hours post-infection from B6 mice that were uninfected or infected with mouse-adapted influenza A PR8. In the context of infection, several ADAMTS genes, including ADAMTS4, −6, and −9 exhibited a ≥ 3-fold increase in gene expression compared to uninfected lungs. ADAMTS-4 exhibited the most dramatic increase in gene expression (&gt;100-fold) at this early timepoint. We further investigated the role of ADAMTS4 in survival and recovery from influenza A infection by infecting wild-type (WT) and ADAMTS4 −/− knockout B6 mice with a lethal dose of influenza A PR8. Interestingly, ADAMTS4 −/− mice exhibited significantly improved survival compared to WT littermates (log-rank test, p= 0.04) with 75% vs. 41% survival at 15 days post-infection, despite no observable difference in weight loss. Consistent with a role in tissue repair and recovery, differences in survival were observed after 9 days post-infection. These results raise the possibility that increased expression of ADAMTS4 following influenza A infection modulates the tissue repair process and contributes to lung pathology.
SP-R210 (Myo18A) enhances influenza A virus pathogenicity by alveolar macrophages and dendritic cells in vivo
Authors: Fino Kristin K , Yang Linlin , Hu Sanmei , Halstead Scott E , DiAngelo Susan L , Silveyra Patricia , Chroneos Zissis C 
Published: January 2023

Abstract
Alveolar macrophages (AMs) are critical for the host response to severe influenza A virus (IAV) infection; IAV is known to cause abortive infection in AMs that is thought to initiate the lung’s immune response to the virus. Our recent studies indicate that the surfactant protein A receptor SP-R210 mediates IAV entry in AMs. Based on these findings, we asked whether SP-R210 modulates the host response to IAV infection.
To address this question, we used SP-R210 floxed mice to delete SP-R210 in CD11c-Cre (dendritic cells and AMs) or Clec9A-Cre (CD103+ dendritic cells) expressing cells. Mice were infected intranasally with a sublethal dose of 1,000 FFU IAV PR8 strain and morbidity was monitored daily by body weight. We found that disruption of SP-R210 significantly reduced susceptibility to infection in both CD11c-CRE/SPR210+/− and Clec9A-Cre/SP-R210+/− as indicated by accelerated body weight recovery 10 days after infection compared to control littermates. The increased recovery of SP-R210-deficient mice was accompanied by a faster decrease in viral titer 7 to 14 days post infection (dpi) as well as increased numbers of CD4+ and CD8+ T cells 14 dpi. Furthermore, flow cytometry analysis revealed significantly increased number of CD11b+Ly6C++Ly6G- and CD11b+SiglecF+ macrophages and AMs in SP-R210-deficient mice, respectively. Consistent with these results, real time qPCR showed persistent suppression of the AM marker SiglecF in control littermates whereas inhibition of SiglecF expression in SP-R210-deficient mice was transient returning to near normal levels by 14 dpi. Taken together, these findings support the model that IAV causes depletion of AMs through SP-R210 disrupting restoration of immune homeostasis after IAV infection.
Decreased polyfunctionality and PD-1 elevation of CD8+ T cells in sepsis patients with human cytomegalovirus reactivation
Authors: Choi Young Joon , Kim Sun Bean , Kim Jong Hoon , Park Su-Hyung , Park Moo Suk , Kim June Myung , Han Sang Hoon , Shin Eui-Cheol 
Published: January 2023

Abstract
Human cytomegalovirus (HCMV) establishes a lifelong chronic latent infection and often reactivates in immunocompromised patients. Additionally, HCMV reactivates in patients with sepsis or other critical illnesses, particularly in patients with poor prognoses. However, the immunological characteristics of sepsis patients with HCMV reactivation have not been elucidated. In the present study, we examined T cell responses in severe sepsis patients with and without HCMV reactivation. First, we found that PD-1 expression was significantly increased in the total CD8+ T cell population in sepsis patients with HCMV reactivation, indicating CD8+ T cell exhaustion in these patients. Next, we studied HCMV pp65-specific T cell functions by intracellular cytokine staining for IFN-γ, TNF-α, and MIP-1β and CD107a staining. When we analyzed the data for each single function, we found no difference between the patient groups. However, the relative frequency of polyfunctional CD8+ T cells was significantly decreased in sepsis patients with HCMV reactivation and inversely correlated with the relative frequency of PD-1+ cells in the CD8+ T cell population. Herein, we demonstrate that HCMV reactivation in severe sepsis patients is associated with CD8+ T cell exhaustion and impaired polyfunctionality of CD8+ T cells.
The Herpes Simplex Virus LAT Gene is Associated with a Broader Repertoire of Virus-Specific Exhausted CD8+ T Cells Retained Within The Trigeminal Ganglia of Latently Infected HLA Transgenic Rabbits
Authors: Benmohamed Lbachir , Srivastava Ruchi , Khan Arif A 
Published: January 2023

Abstract
Persistent pathogens, such as herpes simplex virus type 1 (HSV-1), have evolved a variety of immune evasion strategies to avoid being detected and destroyed by the host’s immune system. A dynamic cross talk appears to occur between the HSV-1 Latency-Associated Transcript (LAT), the only viral gene that is abundantly transcribed during latency, and the CD8+ T cells that reside in HSV-1 latently infected human and rabbit trigeminal ganglia (TG). The reactivation phenotype of TG that are latently infected with wild type HSV-1 (i.e. LAT(+) TG) is significantly higher than TG latently infected with LAT null mutant (i.e. LAT(−) TG). Whether LAT promotes virus reactivation by selectively shaping a unique repertoire of HSV-specific CD8+ T cells retained in LAT(+) TG is unknown. In the present study, we assessed the frequency, function, and exhaustion status of TG-resident CD8+ T cells specific to 40 different epitopes derived from HSV-1 gB, gD, VP11/12 and VP13/14 proteins, in Human Leukocyte Antigen (HLA-A*0201) transgenic rabbits infected ocularly with LAT(+) vs. LAT(−) virus. Compared to CD8+ T cells from LAT(−) TG, CD8+ T cells from LAT(+) TG: (i) recognized a broader selection of non-overlapping HSV-1 epitopes; (ii) expressed higher levels of PD-1, TIM-3 and CTLA-4 markers of exhaustion; and (iii) produced less TNF-a, IFN-g, and GzmB. These results suggest a novel immune evasion mechanism by which the HSV-1 LAT may contribute to the shaping of a broader repertoire of exhausted HSV-specific CD8+ T cells in latently infected TG, thus, allowing for increased viral reactivation.