bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immunometabolic modulation by targeting UCP2 and IL-17A boost anti-tumor immunity against pancreatic cancer
Authors: Liu Chuan-Teng , Yen Hung-Rong 
Published: August 2023

Abstract
Metabolic reprograming of cytotoxic effectors by tumor microenvironment (TME) is one of the key mechanisms by which TME impairs the success of cancer immunotherapy. Therapeutic strategies that mitigate the immunosuppressive TME and selectively enhance bioenergetic fitness of cytotoxic effectors may enhance benefits of immunotherapy, particularly in TME-abundant pancreatic cancer. IL-17 is a key TME cytokine engaged in a feed-forward loop promoting the immunosuppressive milieu that facilitates cancer progression. Mitochondrial uncoupling protein 2 (UCP2) acts as an inhibitor of T cell metabolic fitness and directly promotes tumorigenesis. Therefore, we hypothesized that targeting IL-17 and UCP2 could act additionally to enhance antitumor immune response and directly inhibit pancreatic cancer growth. UCP2 deficiency regulated bioenergetic metabolism and mTOR signaling in cytotoxic T cells to enhance IFN-γ production and anti-tumor immune responses. UCP2 inhibition increased anti-tumor immunity in PBMCs and tumors of patients with PDAC. The dual inhibition of IL-17 and UCP2 boosted antitumor immunity and suppressed tumorigenesis in tumor-bearing mice. Combination of UCP2 and IL-17 blockade may offer additional complimentary and cooperative mechanisms of antitumor immunity against pancreatic cancer.
1. Ministry of Science and Technology (MOST 108-2320-B-039-024-MY3) 2. China Medical University Hospital (DMR-110-186) 3. Chinese Medicine Research Center, China Medical University from the Featured Areas Research 4. Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education (CMRC-CHM-2)
Gut microbiota modulation by high glucose diet invigorates anti-tumor immune response to glioblastoma multiforme
Authors: Kim Jaeho , Kim Hyunjin , Kang Byeong Hoon , Kim Yumin , La Jeoungwoo , Ku Keun Bon , Lee Heung Kyu 
Published: August 2023

Abstract
Most studies about the relationship between high sugar diet and tumor progression state that the pro-tumoral effects of high sugar diet are induced by metabolic diseases such as obesity and diabetes caused by long-term, excessive sugar diet. However, there are still many unknowns about the effect of high sugar diet on tumors when it lasts for a relatively short period that does not cause metabolic diseases. In this background, we find that survival rate of glioblastoma (GBM) is increased through supplement of high glucose diet (HGD) for several weeks. We find that gut microbiota modulation by HGD plays a crucial role in enhancement of anti-tumor immune response. Through single cell RNA sequencing analysis, modulation of gut microbiota by HGD results in an increase in T cell-mediated anti-tumor immune response in GBM. In addition, we find the novel cytotoxic CD4+ T cell population in GBM is increased due to synergistic action with anti PD-1 and the change occurs only through the synergistic action of anti PD-1 treatment and HGD. As a result, we identify that HGD can enhance anti-tumor immune response in GBM through changes in gut microbiota and suggest that the role of glucose diet on GBM needs to be elucidated from a different perspective.
γ-aminobutyric acid drives glioblastoma in females in an immune-dependent manner
Authors: Bayik Defne , Chin Diana , Zhou Yadi , Cheng Feixiong , Lathia Justin 
Published: August 2023

Abstract
Interaction between tumor cells and neurons drives the progression of glioblastoma (GBM), which is characterized by enhanced immunosuppressive myeloid cell accumulation. Our previous results revealed that myeloid-derived suppressor cell subsets (MDSCs) play a critical role in tumorigenesis in a sex-specific manner, with monocytic MDSCs (mMDSCs) accumulating in male tumors and granulocytic MDSCs (gMDSCs) correlating with a worse outcome in females. However, host factors governing sex differences in MDSC subset activity remained unknown. Here, we report that gMDSCs express g-aminobutyric acid (GABA) receptor and female gMDSCs respond to GABA by upregulating the antiviral innate immune pathway. Female mice treated with the GABA analogue pregabalin succumbed to disease earlier, consistent with our previous observation that gMDSCs have a central role in GBM progression in females. In contrast, pregabalin treatment had no effect in male mice with tumors or immunocompromised female mice. Assessment of the antiviral response signature revealed that GABA upregulates nitric oxide synthase 2 (NOS2) expression in gMDSCs. Correspondingly, the tumor-accelerating effect of pregabalin was diminished in NOS2 KO female mice. Collectively, these results identify a neuron-immune-cancer communication axis and indicate that GABA alters anti-tumor immune response in a sex-specific manner. These results support the future assessment of GABA pathway inhibitors as potential immunotherapy agents.
VeloSano Philanthrophic Funds
Nontypeable Haemophilus influenzae specific memory TH17 cells protect against secondary bacterial pneumonia following Influenza A virus infection
Authors: Yang Ying 
Published: August 2023

Abstract
Nontypeable Haemophilus influenzae(NTHi) is one of the most common bacteria that cause secondary bacterial pneumonia following influenza A virus (IAV) infection, leading to hospitalization and death associated with IAV infection worldwide. Current efforts to develop vaccines against NTHi infection focus on inducing antibodies but are hindered by antigenic diversity among NTHi strains. Therefore, we interrogated the contribution of the memory T-helper type 17 (TH17) response in protective immunity against IAV/NTHi co-infection. We observed that co-infected mice exhibited prolonged NTHi infection in the lungs even at mild IAV infection and impaired NTHi-specific TH17 responses compared to NTHi mono-infected mice. However, immune mice that recovered from a prior NTHi infection were protected from homologous and heterologous secondary NTHi strains following IAV infection. Our data further revealed that memory NTHi-specific TH17 cells overcame IAV-driven impairments of anti-NTHi TH17 responses and were cross-reactive with different NTHi strains. Last, mice immunized with a NTHi protein that induced a strong TH17 memory response were broadly protected against diverse NTHi strains following challenge with IAV/NTHi co-infection. These results indicate that NTHi-specific memory TH17 cells are capable of providing serotype-independent protection against co-infection and demonstrate the advantage of developing broadly protective TH17-inducing vaccines against secondary bacterial pneumonia.
Mechanistic investigations of granzyme A adjuvant effects on γ9δ2 T cells
Authors: Phelps Kathleen , Rasi Valerio , Xia Mei , Hoft Daniel 
Published: August 2023

Abstract
γδ T cells are ideal effector cells against cancer and pathogens for numerous reasons: they are found in blood, mucosa, and lymphoid organs, develop memory responses independent of MHC restriction, present antigen, produce cytokines, and are cytolytic. An obstacle for the development of biomedical interventions targeting γδ T cells is the need for an adjuvant that promotes their expansion and is suitable for human use. The adjuvant routinely used, IL-2, has multiple drawbacks limiting its use for vaccination. Our lab has shown that memory γ9δ2 T cells, induced by BCG vaccination, are expanded in vitro by M. tuberculosis, M.bovis (BCG), and our lab’s vaccine candidate 6-O-methylglucoselipopolysaccharide (mGLP), in the presence of IL-2. Additionally, the expanded γ9δ2 T cells inhibit intracellular mycobacterial growth. This inhibition is dependent on granzyme A (GzmA) production by γ9δ2 T cells, and GzmA alone can inhibit intracellular mycobacterial growth.
We recently identified that GzmA can also promote expansion of γδ T cells, without exogenous IL-2, in response to mGLP and HMBPP, a phosphoantigen that stimulates all γ9δ2 T cells. To identify γδ T cell adjuvants, in vitro expansion assays were performed using human peripheral blood mononuclear cells, dendritic cells, antigen, and adjuvant. The absolute numbers of expanded effector γδ T cells were calculated using flow cytometry after 7 days of culture. We identified that the ability of GzmA to promote γδ T cell expansion is dependent upon TLR4, CD14, and GzmA homodimerization, but independent of GzmA enzymatic activity. This new information has broad implications for vaccine design, promoting γδ T cell responses, and understanding the role of GzmA in infectious disease and cancer.
Supported by grants from NIH (R01 AI048391) and Bill & Melinda Gates Foundation (OP1118659)
Pre-exposure to mRNA-LNP inhibits adaptive immune responses and alters innate immune fitness in an inheritable fashion
Authors: Qin Zhen , Bouteau Aurélie , Herbst Christopher , Igyártó Botond Z 
Published: August 2023

Abstract
During the ongoing SARS-CoV-2 pandemic, hundreds of millions of people have been exposed to the experimental mRNA-LNP-based COVID vaccines, of which long-term immune effects are unknown. We previously showed that this vaccine platform is highly inflammatory. Its synthetic ionizable lipid component, which has a long in vivo half-life, is responsible for the induction of inflammation and adaptive immune responses. The inflammatory component’s long in vivo half-life could be a reminiscence of a chronic infection, which is known to lead to immune exhaustion and non-responsiveness. Therefore, here, we sought to determine the effects of pre-exposure to the mRNA-LNP on adaptive immune responses and innate immune fitness.
We found that pre-exposure to mRNA-LNPs or LNP alone led to long-term inhibition of the adaptive immune response, which could be overcome using standard adjuvants. On the other hand, we report that after pre-exposure to mRNA-LNPs, the resistance of mice to heterologous infections with influenza virus increased while resistance to Candida albicans decreased. The diminished resistance to Candida albicans correlated with a general decrease in blood neutrophil percentages. Interestingly, mice pre-exposed to the mRNA-LNP platform can pass down the acquired immune traits to their offspring, providing better protection against influenza.
In summary, the mRNA-LNP vaccine platform induces long-term unexpected immunological changes affecting both adaptive immune responses and heterologous protection against infections. Thus, our studies highlight the need for more research to determine this platform’s true impact on human health.
National Institute of Allergy and Infectious Diseases (https://www.niaid.nih.gov) R01AI146420 and R01AI146101
Correlates of COVID-19 mRNA vaccine humoral immune responses in men living with HIV-1
Authors: Tuttle Dylan J , Castanha Priscila M S , Nasser Amro , Wilkins Maris S , Cuff Deirdre E , Chhibbar Prabal , Sluis-Cremer Nicolas P , Das Jishnu , Mailliard Robbie B , Rinaldo Charles R , Marques Ernesto T A 
Published: August 2023

Abstract
It is important that people with HIV-1 (PWH) receive an FDA-approved vaccine for COVID-19. Because PWH are known to elicit less effective immune responses to other licensed vaccines, we assessed the humoral immune response to the novel COVID-19 mRNA vaccines in 139 fully vaccinated male participants (45 with HIV-1 [MWH] and 94 without HIV-1 [MWOH]) of the MACS/WIHS Combined Cohort Study. Levels of anti-nucleocapsid (N) and anti-spike 1 (S1) IgG and IgA were determined using ELISA and the MSD electro-chemiluminescence multiplex assay platform, to classify natural infection and vaccination status, and quantify binding IgG. Functional characterization of antibodies (Ab) was determined by measuring neutralizing Ab levels against WA, Delta, and Omicron strains, and immune complex complement activation capacity by complement component 3 (C3) deposition. We found that MWH who received two standard doses of mRNA vaccines had lower levels of vaccine-induced anti-S1 IgG and neutralizing Ab against the Washington, Delta, Omicron variants compared to MWOH, and these Ab titers were boosted in the context of natural SARS-CoV-2 infection for MWH. Interestingly, Ab in MWH promoted more potent C3 deposition. In all participants the Moderna mRNA-1273 vaccine induced significantly higher levels of binding IgG and neutralizing Abs against the variants. Importantly, preexisting immunity to the endemic human coronaviruses, participant age, and blood biochemical measurements correlated with these vaccine responses. Our data indicate that COVID-19 mRNA vaccine-induced humoral responses are impaired in MWH. Further studies are needed to define the long-term effects of this impairment on prevention and control of SARS-CoV-2 infection in MWH.
Supported by grants from NIH: Pitt AIDS Research Training (PART) (NRSA 5T32AI065380-15) to DJT, NIH (NHLBI 5U01HL146208) and OAR/NHLBI Supplement (U01HL146208-03S2/S3), MWCCS Concepts X21033 and X20124, and NIH NIAID (R01-AI167711).
Development of broadly protective COVID-19 vaccine against up-to-date variants based on adenovirus type 5/35 vector platform
Authors: Kang Chang-Yuil , Chang Soojeong , Shin Kwang-Soo , Jung In Kyung , Park Hyemin , Oh Jonghan , Park Seowoo , Shin Jieun , Kim Jong Heon 
Published: August 2023

Abstract
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) omicron strain has evolved into a highly divergent variant with several sub-lineages. These new emerging variants could impair the protective efficacy of COVID-19 vaccines based on the ancestral spike, causing breakthrough infections and re-infections. Therefore, it is imperative to develop new multivalent vaccines that can provide broad protection against currently prevalent new variants. We previously developed AdCLD-CoV19-1 OMI, an Ad5/35 platform based non-replicating recombinant adenoviral vector that encodes the spike protein of BA.1, and currently performing clinical studies. We also constructed vaccines against emerging subvariants such as BA.5, BA.2.75, XBB, BQ.1.1, and BN.1 using the Ad5/35 platform and evaluated their immunogenicity by sera from vaccinated mice. To develop the bivalent vaccine, we selected a combination of BA.5 and BA.2.75 vaccines by antigenic cartography map based on cross-neutralizing activities. We found that a bivalent vaccine could induce broadly neutralizing antibodies against the predominant circulating strains and improved cross-neutralization capacity. These data suggest that the bivalent vaccine broadens immunity against current prevalent omicron subvariants. We further investigate combinations of next-generation multivalent vaccines to confer broad protection against new subvariants. Based on the results of preclinical studies, clinical trials of the polyvalent vaccines will be conducted.
Supported by grant from the Bio & Medical Technology Development Program of the National Research Foundation (NRF) funded by the Korean government (MSIT) (2020M3A9I2107463)
Targeting antigens to phosphatidylserine triggers rapid T- and B-cell responses
Authors: Hu Wenbo , Flaskamp Lavinia , Kranich Jan , Brocker Thomas 
Published: August 2023

Abstract
Phosphatidylserine (PS) is a phospholipid that is present in each cell membrane. In intact cells, PS is kept intracellularly, in dying cells, extracellular vesicles (EVs) and activated platelets, PS flips outside where it can be accessed by PS-binding reagents, such as Milk fat globule-EGF factor 8 (MFG-E8). In previous studies, we have established in vivo use of fluorescent MFG-E8 to label and characterize EVs. Here, we measured the antigen-specific immune responses elicited by targeting antigen (Ag) to PS+ structures, presumably EVs via MFG-E8 as a novel vaccination strategy. In contrast to non-targeted conventional Ag, PS-targeting by fusing Ag to MFG-E8 caused accumulation in the marginal zone of lymphoid follicles soon after injection. Hours later, Ag-MFG-E8 was deposited on the follicular dendritic cell network without the need of previous primary immunization. Mice immunized with PS-targeted Ag/Alum induced long-lasting germinal centers (GCs) and significantly more GC B cells. Furthermore, it resulted in an approx. 13,000-fold increase in Ag-specific IgG-levels and strongly accelerated antibody class switch recombination as compared to immunization with conventional Ag/Alum. Ag targeted to PS also induced more efficient CD8 T cell activation and IFN-gamma production post immunization. Therefore, targeting Ag to EVs by harnessing the PS-binding properties of MFG-E8 might turn out to be of advantage for the induction of very rapid and long-lasting immune responses.
This project was supported by the Deutsche Forschungsgemeinschaft (DFG, German Research foundation) – Project-ID 210592381 - SFB 1054 B03 to Thomas Brocker and Z02 to Jan Kranich.
Stabilization of N-terminal cysteines in HPLCHRMS quality control of peptide pools
Authors: Kreuzer Oliver J , Weller Michael G , Bosc-Bierne Gaby 
Published: August 2023

Abstract
Synthetic peptide pools are used in experimental and clinical immunology, for example, cancer immunotherapy. The analytical characterization is challenging due to the similarity of the peptides, usually only a pre-characterization of the single peptides is performed. However, regular quality control of the peptide mix would be highly desirable. Therefore, a high performance liquid chromatography-high resolution mass spectrometry (HPLCHRMS) method for quality control of a peptide pool was developed. Peptides were synthesized and purified to &gt; 90% each. The lyophilized single peptides were combined to a peptide pool containing 32 peptides and analyzed by HPLC-HRMS. Quantification is carried out UV detection, and identification is obtained by high resolution mass spectrometry. Before separation on a capillary reversed-phase column, alkylation of thiols was performed. Best results were obtained with a linear gradient from 96% of solvent A (water with 0.05% TFA), 4% of solvent B (acetonitrile with 0.04% TFA) to 56% of solvent A, 44% of solvent B in 100 minutes. Without sample preparation, peptides with N-terminal cysteine will yield almost no UV signal. This surprising effect could be eliminated using iodoacetic acid (IAC) as an alkylation reagent. The amount of IAC and reductant (Tris(2-carboxyethyl)phosphine (TCEP)) was carefully optimized to avoid overalkylation. By a combination of capillary HPLC and HRMS, a method could be developed for the quality control of complex synthetic peptide pools. In contrast to the analysis of single peptides, analyzing complex peptide pools requires the stabilization of cysteine-containing peptides. Good results were obtained using iodoacetic acid as an alkylation reagent.
Peptide-pulsed MHC class II mutant dendritic cell vaccine has superior efficacy in a murine tumor model
Authors: Seishima Noriko , Olkhanud Purevdorj B , Becker William , Maeng Hoyoung , Lopez-Lago Miguel , Wiseman Charles , Williams William V , Berzofsky Jay A 
Published: August 2023

Abstract
Autologous dendritic cell (DC) vaccines with tumor antigens have been used clinically with limited therapeutic effects. Semi-allogeneic DC-based immunotherapy is still controversial, but it can be an alternative source and more attractive than autologous DC vaccines because the “off-the-shelf” DCs can be used for multiple patients without lengthy individual manufacturing time and may provide additional “allogeneic help”. This study aims to compare efficacy of syngeneic DC vaccines and semi-allogeneic DC vaccines and determine whether a therapeutic semi-allogeneic DC vaccine is more efficacious in tumor suppression. Female C57BL/6 mice were inoculated subcutaneously with human papillomavirus E6 and E7-expressing TC-1 cells. Syngeneic bone marrow dendritic cells (BMDCs) were generated from C57BL/6 and semi-allogeneic BMDCs were generated from two mouse strains, B6.C-H2-Kbm1/ByJ and B6(C)-H2-Ab1bm12/KhEgJ which had limited point mutations in the MHC class I H2-Kb allele or H2-IAb MHC class II allele, respectively. Each BMDC was pulsed with H-2Db-restricted E743–77 peptide and matured before injection. The mice received 4 intradermal injections of syngeneic or one of the semi-allogeneic E7-pulsed BMDC vaccines starting 8–9 days after the TC-1 implantation. Compared with saline control, the MHC class I mutant BMDC vaccine had efficacy similar to the syngeneic BMDC vaccine in suppressing TC-1 tumor growth. However, the MHC class II mutant BMDC vaccine had efficacy significantly superior to that of the other BMDC vaccines. Thus, MHC class II semi-allogeneic BMDCs may be more effective than syngeneic DC-based cancer vaccines, presumably because the class II alloantigens induce additional T cell help for anti-tumor immunity.
Elucidating the cellular mechanisms underlying adoptive cell transfer in models of glioma
Authors: Figg John William , Francis Connor , Ji Dan , Dean Bayli DiVita , Reid Alexandra , Falceto-Font Laura , Hilferty David , Mitchell Duane , Flores Catherine 
Published: August 2023

Abstract
Despite advances in solid tumor biology and oncology, pediatric and adult primary brain cancer and metastasis to the brain harbor dismal prognosis and poor outcomes. One avenue to overcome these clinical challenges is through developing robust immunotherapy to better target the tumor. Our group has previously shown that adoptive cellular therapy (ACT) provides therapeutic benefit against central nervous system (CNS) malignancies, namely medulloblastoma, brain stem glioma and glioblastoma. In this study, we characterized cell cycling differences of immune cellular sub-populations in peripheral lymphoid and tumor tissue compartments in vivo. ACT was conducted in orthotopic KR158 luciferase tumor-bearing mice, a syngeneic murine glioma model. Following completion of ACT, bromodeoxyuridine (BRDU) was administered to mice 12-hours prior to tissue collection and flow cytometry was utilized to measure the level of BRDU incorporation. 7-aminoactinomycin D was used in conjunction with BRDU to provide insight into all phases of the cell cycle. We observed significant differences in cycling populations of myeloid derived suppressor cells (MDSC) and T cells. In mice receiving ACT, MDSCs found in secondary lymphoid tissues were significantly more proliferative. In the tumor microenvironment (TME), proliferating MDSCs were significantly reduced compared to mice that did not receive treatment. Frequencies of proliferating CD4+ and CD8+ T-cells were significantly increased in cervical lymph node and TME, but reduced in spleen. These findings suggest that ACT changes immune cycling and migration dynamics in lymphoid tissue and TME to potentially promote antitumor T-cell responses and minimize tumor-mediated immunosuppression.
5R01NS112315-04