bluuebunny/crossref_metadata_2025_split huggingface dataset:
L-fucose enhances anti-tumor immunity through modulation of monocyte-derived dendritic cell signaling
Authors: Burton Chase , Lester Daniel , Lau Eric 
Published: August 2023

Abstract
Although immunotherapies have emerged as standard of care for severals cancer types, their efficacy remains limited to ~30% of patients. Limited efficacy has in part been attributed to a lack of infiltrating tumor-suppressive immune cells and overabundance of immunosuppressive cell populations. To combat this problem, extensive research efforts have focused on modulating intratumoral T cells to overcome the immunosuppressive tumor microenvironment (TME). While these treatment modalities show promise in preventing intratumoral T cell exhaustion, they fail to provide robust and durable anti-tumor immune responses. For this reason, the targeting of other intratumoral immune subpopulations—particularly dendritic cells (DC)—has become a subject of increased study. The dual activation of anti-tumor and suppression of pro-tumor immune populations is expected to significantly enhance immunotherapeutic outcomes.
Recently, our group found that the dietary sugar L-fucose (L-fuc) reduces melanoma tumor growth via a CD4+ T cell-dependent mechanism. Interestingly, analysis of immune cells in the tumors of L-fuc-treated mice revealed dynamic changes in the DC compartment, predominantly in the monocyte-derived DC subset (moDC). Additionally, myeloid cells treated ± L-fucex vivo exhibited enhanced T cell stimulation, increased neutral fluorescent bait uptake, and altered cytokine production. qPCR-RT analysis revealed that L-fuc affects downstream CD209 signaling, leading to a shift towards an immunostimulatory signature. Together, these data suggest a mechanism to enhance anti-tumor immunity by increasing abundance and immunostimulatory activity of moDCs and supports the use of L-fucose as an immunotherapeutic agent.
Supported by funding from Moffitt Cancer Center Skin Cancer Sport Project
Iron-overload affects anti-tumor immune response in pancreatic cancer
Authors: Cappello Paola , Scagliotti Alessandro , Tiberi Giorgia , Autino Federica , Greco Luana , Rubbino Federica , Connor James , Laghi Luigi 
Published: August 2023

Abstract
HFE is the most frequently mutated gene in hereditary hemochromatosis, a disease in which iron accumulates in organs leading to toxicities. Worldwide, two main polymorphisms in HFE protein have been observed, namely C282Y and H63D. Even if associated to milder iron accumulations, H63D polymorphism increases the risk of cancer development and aggressiveness. As observed in pancreatic cancer patients, a great percentage of those that underwent to surgery also present H63D polymorphism but displayed a worse survival.
To better characterize how the iron accumulation affects anti-tumor response in the context of the pancreatic ductal adenocarcinoma (PDA), we crossed genetically engineered mice (GEM) that spontaneously develop PDA with mice engineered to express H67D mutation (HfeH67D), the orthologue of H63D in human. Histological analysis of GEM showed that H67D polymorphism accelerated PDA progression, increased the number of metastases, and altered the composition of infiltrating immune cells. Accordingly, these mouse models also displayed a reduced overall survival compared to controls. By exploiting OVA as a well-known antigen, we injected OVA-expressing PDA cells subcutaneously in mice carrying or not the H67D mutation to assess their ability to mount a normal memory response. OVA-expressing PDA cells grew faster in the presence of H67D polymorphism, but immunized mice displayed a reduction in tumor volume similar to the WT counterpart. HfeH67D mice displayed an increased production of anti-OVA IgG antibodies, especially IgE, but a decreased in OVA-specific IFNγ-secreting T cells.
Overall, H67D polymorphism impacts the activation of anti-tumoral immune response, which consequently worsens PDA progression.
Supported by grants from AIRC (ID22234; ID26341)
Adjuvant skewing of host immune response to vaccination and subsequent infection with influenza virus
Authors: Noureddine Moataz , Chang Lauren A , Warang Prajakta , Jangra Sonia , De Geest Bruno , García-Sastre Adolfo , Schotsaert Michael 
Published: August 2023

Abstract
Despite being a vaccine-preventable disease, seasonal influenza is responsible for thousands of hospitalizations and deaths yearly. Currently licensed quadrivalent inactivated influenza virus vaccines (QIV) aim to induce neutralizing antibodies but are poor inducers of T-cell responses, which could be potentially overcome with adjuvants. We evaluated adjuvant skewing of host-immune responses after vaccination and its persistence after infection. Hereto, we compared two squalene-based adjuvants (AddaVax, and AddaS03) and IMDQ-PC, a lymph node-draining TLR7/8 agonist. All adjuvants resulted in enhanced antibody titers and protection against challenge with H1N1 influenza virus. Lung cytokine/chemokine profiling at 5 days post-infection showed that AddaVax results in higher levels of type 2 immunity-stimulating cytokines IL-4, -5, and -13 in the lungs, which is also reflected by a higher IgG1:IgG2a ratio. Meanwhile, IMDQ-PC showed higher serological IgG2a titers after vaccination and higher levels of IFNγ in the lungs after challenge. To investigate the role of the myeloid compartment in adjuvant skewing of host responses, we examined adjuvant-mediated skewing of bone-derived macrophages in-vitro using Arginase I and iNOS as M2 or M1 markers, respectively. We also pre-licensed macrophages with IL-4 and IFNγ to evaluate adjuvant-intrinsic skewing effects. Licensing was required to induce M1 or M2 phenotypes by squalene-based adjuvants, whereas IMDQ-PC skewed towards M1 phenotype irrespective of licensing. Correlating with the observed type I immune host response, IMDQ-PC induced iNOS more robustly with IFNγ than the other adjuvants. And, AddaS03 induced the highest levels of Arginase I in the IL-4 group.
This study was also partly funded by CRIPT (Center for Research on Inuenza Pathogenesis and Transmission), a NIH NIAID funded Center of Excellence for Influenza Research and Response (CEIRR, contract number 75N93021C00014), by the NIAID funded SEM-CIVIC consortium to improve inuenza vaccines (contract number 75N93019C00051), to AG-S. By NIH/NIAID HHSN272201400008C (L.C.), and NIH/NIAID R21AI151229 and R01AI160706 (M.S.). B.G.D.G. acknowledges funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (grant N 817938).
Modulating vaccine inflammation with thermophobic trehalose glycolipid adjuvant polymers
Authors: Mancini Rock , Hendricksen Aaron , Ryan Austin , Aguilar-Carreno Hector , Ezzatpour Shahrzad 
Published: August 2023

Abstract
Adjuvants drive the efficacy of many vaccines, however, they also elicit undesirable inflammation which is commonly pyrogenic resulting in local or systemic increases in temperature. Based on this, we envisioned that inversely coupling adjuvant potency to small (~2 °C) temperature changes could be a uniquely effective approach to balance efficacy while maintaining safety and tolerability. Here we report the first Thermophobic Adjuvant Polymers (TAPs) that reversibly attenuate potency in response to temperature. TAPs consisted of co-polymers containing both trehalose glycolipid adjuvant and thermoresponsive N-isopropylacrylamide (NIPAM) subunits. In an Influenza A/California/04/2009 virus vaccine mouse model, TAPs were effective adjuvants. Mice immunized with TAPs adjuvanted vaccine exhibited increased neuraminidase neutralizing antibodies, lymph CD4+ T cells, and CD4+/44+/62L+ lung central memory T cells. Protection after viral rechallenge was comparable between TAPs and alum adjuvanted vaccines. At 37 °C in-vitro, TAPs elicited cytokine profiles comparable to other glycolipid adjuvants in the JAWSII cell line and primary CD11c+ BMDCs. Relative to 37 °C, inflammation (TNF-α and IL-6) was enhanced at 35 °C and abrogated at 39 °C. DLS and NOESY-NMR suggest that the temperature-dependent activity is due to intramolecular hydrophobic shielding between the adjuvant and NIPAM which is enhanced beyond the lower critical solution temperature. Overall, this study demonstrates TAPs are an effective influenza vaccine adjuvant and elicit inflammation which is inversely linked to temperature. We envision this approach could be extended to many other adjuvants to enhance efficacy while maintaining safety and tolerability.
Supported by grants from NIH (1R01CA234115), AAI (Careers in immunology fellowship), and the Washington Research Foundation.
SARS-CoV-2 Ancestral and Variant of Concern Breakthrough infections boost both cross-reactive cellular and humoral immune responses in mRNA-vaccinated Hamsters
Authors: Schotsaert Michael , Diego Juan Garcia-Bernalt , Singh Gagandeep , Jangra Sonia , Garcia-Sastre Adolfo , Laporte Manon , Chang Lauren A , Handrejk Kim , Rathnasinghe Raveen , Zahed Sara El , Chang Max W , Warang Prajakta , Aslam Sadaf , Benner Christopher , Krammer Florian , Mena Ignacio 
Published: August 2023

Abstract
When mRNA vaccines were rolled out under EUA, it was observed that a single vaccination, already protected vaccinees from severe disease, even before neutralizing antibodies appearded. To better understand the underlying mechanism of protection, we vaccinated hamsters once with a suboptimal dose of Pfizer/BioNTech COVID-19 mRNA vaccine (5mg/animal). Four weeks post vaccination, animals were challenged with different variants of SARS-CoV-2. Vaccination resulted in antibody ELISA titers against SARS-CoV-2 spike protein, but antibody titers were too low for efficient neutralization of antigenically matching USA-WA1/2020 (WA1) or drifted variants of concern (Alpha, Beta, Delta and Mu). Nevertheless, suboptimal vaccination resulted in reduced morbidity for WA1 and Alpha-challenged animals and complete control of lung virus titers for WA1, Alpha and Delta but with breakthrough infection for the antigenically more distant Beta and Mu. Moreover, T cell responses were higher in vaccinated animals compared to unvaccinated ones at 5 days post infection, suggesting a recall of vaccine-primed T cells. Infection with different SARS-CoV-2 variants also back-boosted neutralizing antibody responses against the challenge virus and in the case of the variants of concern, against the antigenically distant but vaccine-matched WA1. Transcriptomic analysis of host immune responses to infection reflects both vaccination status and disease course, is further compared with lung pathology and suggests a role for interstitial lung macrophages in vaccine-mediated protection. In summary, protection after suboptimal vaccination against SARS-CoV-2 challenge in hamsters correlates with recall of both vaccine-induced B- and T-cell responses.
This study was also partly funded by CRIPT (Center for Research on Influenza Pathogenesis and Transmission), a NIH NIAID funded Center of Excellence for Influenza Research and Response (CEIRR, contract number 75N93021C00014) to A.G.S and M.S. and by R01 AI160706 to M.S.
The T cell receptor repertoire reflects the early dynamics of the immune response to vaccination
Authors: Sachidanandam Ravi 
Published: August 2023

Abstract
Early ascertainment of the immune response to vaccinations is important. The T cell receptor (TCR) is a promising target to achieve this, with the complete TCR repertoire in PBMCs reflecting the T cells present in an individual. To this end, we developed Tseek, an unbiased and accurate method for profiling the TCR repertoire by sequencing the TCR α and β chains and developing a suite of tools for repertoire analysis. We explored the T cell response to both the COVID-19 mRNA vaccine (n=9) and the seasonal inactivated Influenza vaccine (n=5) at several time points. Neutralizing antibody titers were also measured in the covid vaccine samples. The COVID-19 vaccine elicited a broad T cell response involving multiple expanded clones, whereas the Influenza vaccine elicited a narrower response involving fewer clones. Distinct T cell clones responded at different time points over a month, providing temporal details lacking in the antibody measurements. The physical symptoms following administration of the vaccinations were not indicative of the TCR/antibody responses while the TCR responses broadly presaged the antibody responses. Additionally, the TCR repertoire is akin to a fingerprint: donors of blood samples taken years apart could be identified from the TCR repertoire. TCR repertoire profiling shows great promise as an early and sensitive measure of the adaptive immune response to vaccination, which can help improve immunogen selection and optimize vaccine dosage and spacing between doses.
A rationally designed combined intranasal adjuvant drives robust and broad humoral, cellular, and mucosal immune responses to SARS-CoV-2
Authors: Wong Pamela T , Jangra Sonia , Landers Jeffrey J , Laghlali Gabriel , Rathnasinghe Raveen , O’Konek Jessica J , Janczak Katarzyna W , Garcia-Sastre Adolfo , Baker James R , Schotsaert Michael 
Published: August 2023

Abstract
Multiple FDA-approved SARS-CoV-2 vaccines have provided excellent protection against severe disease. Despite this, immunity can wane relatively fast, particularly in the elderly and novel viral variants capable of evading infection- and vaccination-induced immunity continue to emerge. Furthermore, while these parenteral vaccines have provided substantial protection, they have been less effective in inducing mucosal immune responses which are critical for effectively blocking viral transmission in vaccinated individuals. As mucosal immune responses are most effectively induced through infection or mucosal immunization, we have developed an intranasal, rationally designed adjuvant integrating a nanoemulsion (NE) that activates TLRs and NLRP3 with an RNA agonist of RIG-I (IVT DI). Through activating this array of receptors, we have shown that the NE/IVT adjuvant better recapitulates the innate immune activation events that occur during natural viral infection, subsequently leading to more tailored and effective downstream adaptive immune responses. The combination adjuvant with various spike protein antigens elicited robust protective humoral, mucosal, and cellular responses to SARS-CoV-2 in mice, with markedly enhanced TH1-biased cellular responses and high virus-neutralizing antibody titers towards both homologous and antigenically distant drift variants of SARS-CoV-2. These results highlight the potential of this adjuvant to improve the immune profile of a variety of SARS-CoV-2 vaccine candidates for use mucosally.
This research was supported by R01-AI160706 to P.W. and M.S, and in part by R01-DK130425 to M.S.
A SARS-CoV-2 vaccine designed for manufacturability results in unexpected potency and non-waning humoral response
Authors: Anderson Stephen , Campbell Elliot , Dobkin Julie , Osorio Louis , Kolloli Afsal , Ramasamy Santhamani , Kumar Ranjeet , Sant’Angelo Derek B , Subbian Selvakumar , Denzin Lisa 
Published: August 2023

Abstract
Despite the availability of a number of COVID-19 vaccines, there still exists a large unmet need for vaccines which provide durable immunity and protection against emerging SARS-CoV-2 variants, and for vaccines with lower manufacturing costs and reduced logistical requirements for distribution. We have developed a novel protein component vaccine candidate, MT-001, based on a fragment of the SARS-CoV-2 spike protein surrounding the receptor binding domain (RBD). Mice vaccinated with a 2-dose prime-boost regimen of MT-001 adjuvanted with alum and CpG exhibited peak anti-spike IgG ELISA titers comparable to those reported for mRNA vaccines from Pfizer/BioNTech (BNT162b2) and Moderna (mRNA-1273), and, in contrast to the mRNA vaccines, these titers were found to be durable with midpoint GMTs in the range of 105 – 106 up to 12 months post-vaccination. Syrian golden hamsters vaccinated with a similar 2-dose regimen of MT-001 exhibited undetectable levels of SARS-CoV-2 in lung tissue four days after intranasal challenge with SARS-CoV-2/US-WA1. In mice, the results showed an increased breadth of the response, with significant neutralization titers against the Omicron BA.1 variant at this timepoint.
These attributes make MT-001 a compelling candidate for a next-generation COVID-19 vaccine. MT-001 could be particularly valuable as an annual booster for individuals with diverse histories of vaccination, SARS-CoV-2 infection, and/or predispositions resulting in an immunocompromised state. MT-001 was designed for manufacturability and ease of distribution, and we demonstrate that these attributes are not inconsistent with a highly immunogenic vaccine that confers durable and broad immunity to SARS-CoV-2 and its emerging variants.
Supported by grants from Rutgers University CCRP2 and the New Jersey Health Foundation. In-kind support from CEPI is also gratefully acknowledged.
MEF2C is a critical regulator of human NK cell metabolism
Authors: Li Joey , Senthilkumar Vignesh , Shah Siya , Padilla Eddie , Riggan Luke , Ball Andrea , Matulionis Nedas , Divakaruni Ajit , Christofk Heather , O’Sullivan Timothy 
Published: August 2023

Abstract
Natural killer (NK) cells are critical for the first line of defense against viral infection and malignancy. While the transcriptional and epigenetic regulation of activated NK cells is well characterized in mice, these processes are not yet fully understood in human NK cells. We performed a targeted CRISPR/Cas9 ribonucleoprotein (RNP) transcription factor screen in mature primary human NK cells to identify putative transcription factors required for optimal effector function. The transcription factor myocyte enhancer factor 2C (MEF2C) emerged as a previously unidentified regulator of NK cell homeostasis, cytokine production, and cytotoxicity. MEF2C-deficient NK cells activated with IL-2 and IL-15 displayed impaired proliferation, degranulation, and production of granzyme B. CRISPR-mediated deletion of MEF2C resulted in disrupted glycolysis and oxidative phosphorylation using both SCENITH and Seahorse extracellular flux analysis. Liquid chromatography/mass spectrometry (LC/MS)-based metabolomics of MEF2C-deficient NK cells revealed reductions in the metabolites glyceraldehyde-3-phosphate and a-ketoglutarate/succinate, accompanied by increased accumulation of lipid metabolism products including phosphorylethanolamine and saturated long-chain fatty acids. In vivo, CRISPR/Cas9 RNP-mediated disruption of MEF2C expression in mouse NK cells resulted in decreased expansion during mouse cytomegalovirus infection. Together, these studies reveal MEF2C as a novel regulator of NK cell effector function through control of multiple critical cell-intrinsic metabolic pathways.
Macrophage recognition of non-self MHCI triggers signaling pathways directing differentiation and initiating tissue repair
Authors: Warunek Jordan , Mathews Lisa , Oberbarnscheidt Martin H , Lakkis Fadi G , Turnquist Heth R 
Published: August 2023

Abstract
Injury resolution requires a coordinated immune-mediated response executed by macrophages (MΦs). Yet, how MΦs convert local stimuli into effective repair is poorly understood. Furthermore, organ transplants (Tx) will fail over time due to fibrosis, despite immunosuppressants targeting adaptive immune cells. We hypothesize this is due to dysregulated MΦ-mediated tissue repair of grafts. Donor MHCI can be recognized by monocytes and MΦs via paired-immunoglobulin-like receptor-A (PIR-A), but if non-self MHCI recognition disrupts repair is an unanswered question. To test this, Wildtype (WT), Rag2−/−γc−/−, and Pira−/− C57BL/6 (H-2b) MΦ differentiation was assessed after exposure to allogeneic (BALB/c; H-2d; allo) or syngeneic (syn) materials using flow cytometry or phosphoflow. The function of syn- and allo-exposed MΦs were compared with live-cell imaging of wound healing assays. Allo materials generated Ly6chiCD86hi MΦs in both WT and Rag2−/−γc−/− recipients in vivo. Recombinant H2-Dd alone behaved similarly in vitro. Interestingly, Ly6chi CD86hi MΦ differentiation in response to non-self MHCI was not PIR-A mediated, but by Ly49d-triggered PLCγ2 activation of NF-κB and mTOR coupled with increased glycolysis. Moreover, MΦ allo exposure profoundly amplified MΦ repair in comparison to other well characterized MΦ subsets (i.e. LPS/IFNγ or IL-4 stimulated). We identify an unappreciated relationship between innate allorecognition and MΦ repair. Specifically, MΦs recognize non-self MHCI via Ly49d, and triggering this novel pathway mediates MΦ differentiation in phenotypically pro-inflammatory subsets with amplified repair functions. Such a mechanism would contribute to Tx graft fibrosis and related pathology over time.
Supported by grants from NIH (5T32AI074490-14, R01HL122489)
Bacterial blockade of host protein synthesis triggers cell death in dendritic cells to restrict Legionella pneumophila infection
Authors: Vazquez Marrero Victor R , Zhang Jenna , Doerner Jessica , Pollock Tzvi , Shin Sunny 
Published: August 2023

Abstract
Legionella pneumophila is a facultative intracellular bacterial pathogen that is responsible for a pneumonia called Legionnaire’s disease. Legionella invades cells by using a type IV secretion system that delivers effector proteins into the host cell cytosol to facilitate its intracellular survival and replication. Macrophages serve as the primary replicative niche for Legionella. In contrast, dendritic cells (DCs) have been shown to rapidly restrict Legionella infection by activating cell death. The host and bacterial factors that induce this response remain elusive. We found that DCs undergo caspase-dependent cell death and that this response is triggered by a subset of secreted effectors that block host protein synthesis. Specifically, we show that these effectors decrease the levels of pro-survival proteins in DCs, thus leaving cells poised to undergo cell death. Altogether, our data show that pro-survival molecules in DCs serve as guard proteins that detect pathogenic activities to limit bacterial replication.
Supported by a National Science Foundation Graduate Research Fellowship Program (DGE-2236662)
A cell reporter system to examine toll-like receptor ligands in channel catfish
Authors: Felch Kristianna Lee , Bengtén Eva , Wilson Melanie 
Published: August 2023

Abstract
Toll-like receptors (TLR) are a family of innate immune receptors that recognize microbe- or danger-associated molecular patterns (MAMPs or DAMPs). TLR ligand binding results in the recruitment of adaptor proteins, which initiate signaling cascades that lead to the activation of NF-κB and interferon regulatory factors (IRFs). Importantly, this signaling initiates both innate and adaptive immune responses. Channel catfish is an important aquacultural species, and the catfish genome encodes 20 TLRs. In contrast, mammalian genomes encode from 10 to 13 TLRs. Our hypothesis is that fish, due to their aquatic environment, express additional TLRs that provide protection from pathogens and pollutants. Our goal is to gain a functional understanding of catfish TLRs by identifying their ligands. To screen potential ligands, we use the HEK Dual-null cell line, which is engineered to not express endogenous TLRs. This cell line is stably transfected with a SEAP reporter gene under the control of NF-κB. Using this model, it was confirmed that TLR7 is activated by imiquimod, a synthetic ssRNA analog. In addition, we have identified the full repertoire of IFN genes in the catfish genome. This allows us to assess products downstream of TLR activation by qPCR. Taken together, this system permits us to study the TLR signaling pathways that drive catfish immune responses. In the future, once TLR signaling is better understood, specific ligands can be incorporated into vaccines as adjuvants to direct favorable immune responses and protect farmed and domesticated animal species.
Supported by pre-doctoral fellowship USDA-NIFA-AFRI 2022-67011-36580 and grant NSF-IOS 1656419