A5013651491- Immune Cell Function and Interaction
- Immunotherapy and Immune Responses
- Toxin Mechanisms and Immunotoxins
- CAR-T cell therapy research
- T-cell and B-cell Immunology
- HIV Research and Treatment
- T-cell and Retrovirus Studies
- Herpesvirus Infections and Treatments
- SARS-CoV-2 and COVID-19 Research
- Pharmacological Effects of Natural Compounds
- Cancer, Hypoxia, and Metabolism
- Virus-based gene therapy research
- Animal Virus Infections Studies
- RNA Interference and Gene Delivery
- Angiogenesis and VEGF in Cancer
- COVID-19 Clinical Research Studies
- Cancer Research and Treatments
- Cancer Immunotherapy and Biomarkers
- Cytomegalovirus and herpesvirus research
- IL-33, ST2, and ILC Pathways
- Immune cells in cancer
- Extracellular vesicles in disease
- Viral gastroenteritis research and epidemiology
- MicroRNA in disease regulation
- Transgenic Plants and Applications
Center for Cancer Research
2010-2023
Frederick National Laboratory for Cancer Research
2013-2023
National Cancer Institute
2013-2023
Government of the United States of America
2022
National Institutes of Health
2007-2021
University of Patras
2018
National Cancer Institute
2017
Universidade do Porto
2014
Frederick Community College
2013
Institut de Recherche Vaccinale
2013
Extracellular vesicles (EV), including exosomes and microvesicles, are nano-sized intercellular communication vehicles that participate in a multitude of physiological processes. Due to their biological properties, they also promising candidates for the systemic delivery therapeutic compounds, such as cytokines, chemotherapeutic drugs, siRNAs viral vectors. However, low EV production yield rapid clearance administered by liver macrophages limit potential use vehicles. We have used...
Early responses to vaccination are important for shaping both humoral and cellular protective immunity. Dissecting innate vaccine signatures may predict immunogenicity help optimize the efficacy of mRNA other strategies. Here, we characterize cytokine chemokine 1st 2nd dose BNT162b2 (Pfizer/BioNtech) in antigen-naive previously coronavirus disease 2019 (COVID-19)-infected individuals (NCT04743388). Transient increases interleukin-15 (IL-15) interferon gamma (IFN-γ) levels early after boost...
We show that co-expression of interleukin 15 (IL-15) and IL-15 receptor alpha (IL-15Ralpha) in the same cell allows for intracellular interaction two proteins early after translation, resulting increased stability secretion both molecules as a complex. In absence co-expressed IL-15Ralpha, large portion produced is rapidly degraded immediately synthesis. Co-injection into mice IL-15Ralpha expression plasmids led to significantly levels cytokine serum well biological activity IL-15....
The development of extracellular vesicles (EV) for therapeutic applications is contingent upon the establishment reproducible, scalable, and high-throughput methods production purification clinical grade EV. Methods including ultracentrifugation (U/C), ultrafiltration, immunoprecipitation, size-exclusion chromatography (SEC) have been employed to isolate EV, each facing limitations such as efficiency, particle purity, lengthy processing time, and/or sample volume. We developed a...
Background Interleukin-15 (IL-15) promotes growth and activation of cytotoxic CD8 + T natural killer (NK) cells. Bioactive IL-15 is produced in the body as a heterodimeric cytokine, comprising receptor alpha chains (hetIL-15). Several preclinical models support antitumor activity hetIL-15 promoting its application clinical trials. Methods The from mammalian cells was tested mouse tumor (MC38 colon carcinoma TC-1 epithelial carcinoma). functional diversity immune infiltrate cytokine/chemokine...
Interleukin-15 (IL-15), a 114-amino acid cytokine related to IL-2, regulates immune homeostasis and the fate of many lymphocyte subsets. We reported that, in blood mice humans, IL-15 is present as heterodimer associated with soluble receptor α (sIL-15Rα). Here, we show efficient production this noncovalently linked but stable clonal human HEK293 cells release processed IL-15·sIL-15Rα medium. Purification sIL-15Rα polypeptides allowed identification proteolytic cleavage site IL-15Rα...
IL-12 is a 70-kDa heterodimeric cytokine composed of the p35 and p40 subunits. To maximize production from plasmid DNA, molecular steps controlling IL-12p70 biosynthesis at posttranscriptional posttranslational levels were investigated. We show that combination RNA/codon-optimized gene sequences fine-tuning relative expression two subunits within cell resulted in increased heterodimer. found subunit plays critical role enhancing stability, intracellular trafficking, export subunit. This...
B cell follicles in secondary lymphoid tissues represent an immune privileged sanctuary for AIDS viruses, part because cytotoxic CD8 + T cells are mostly excluded from entering the that harbor infected follicular helper (T FH ) cells.We studied effects of native heterodimeric IL-15 (hetIL-15) treatment on uninfected rhesus macaques and had spontaneously controlled SHIV infection to low levels chronic viremia.hetIL-15 increased effector lymphocytes with high granzyme content blood, mucosal...
Apolipoprotein B mRNA-editing enzyme, catalytic polypeptide-like 3G (APOBEC3G), a human cytidine deaminase, is potent inhibitor of HIV replication. To explore possible role this protein in modulating vivo susceptibility to infection, we analyzed APOBEC3G expression HIV-exposed seronegative individuals, HIV-seropositive patients, and healthy control subjects. The results showed that the significantly increased peripheral blood mononuclear cells (PBMCs)--mainly CD14(+) cells--and cervical...
Abstract The recent approval of bevacizumab (Avastin), a humanized anti–vascular endothelial growth factor (VEGF) monoclonal antibody, in combination with chemotherapy for the treatment patients metastatic colorectal cancer, has provided proof principle efficacy antiangiogenic strategies cancer therapy. activity is primarily attributed to its ability inhibit cell survival. Whether anti-VEGF may also have direct effect on survival poorly understood. We show that serum-starved colon cells...
Background NIZ985 is a recombinant heterodimer of physiologically active interleukin (IL-)15 and IL-15 receptor alpha. In preclinical models, promotes cytotoxic lymphocyte proliferation, killing function, organ/tumor infiltration, with resultant anticancer effects. this first-in-human study, we assessed the safety, pharmacokinetics, immune effects in patients metastatic or unresectable solid tumors. Methods Single agent dose escalation data are reported from phase I escalation/expansion...
Immunotherapy has emerged as a viable approach in cancer therapy, with cytokines being of great interest. Interleukin IL-15 (IL-15), cytokine that supports cytotoxic immune cells, been successfully tested an anti-cancer and anti-metastatic agent, but combinations conventional chemotherapy surgery protocols have not extensively studied. We produced heterodimeric (hetIL-15), which shown anti-tumor efficacy several murine models is evaluated clinical trials for metastatic cancers. In this...
Optimized plasmid DNAs encoding the majority of SIVmac239 proteins and delivered by electroporation (EP) elicited strong immune responses in rhesus macaques. Vaccination decreased viremia both acute chronic phases infection after challenge with pathogenic SIVmac251. Two groups macaques were vaccinated DNA plasmids producing different antigen forms, "native" "modified," inducing distinct responses. Both showed significantly lower during phase infection, whereas group immunized native antigens...
We employed a dose-escalation regimen in rhesus macaques to deliver glycosylated IL-7, cytokine critical for development and maintenance of T lymphocytes. IL-7 increased proliferation survival cells triggered several chemokines cytokines. Induction CXCL13 lymph nodes (LNs) led remarkable increase B the LNs, germinal center follicular helper elevated IL-21 levels suggesting an follicle activity. Transcriptomics analysis showed induction IRF-7 Flt3L, which was linked frequency circulating...
The two known isoforms of IL-15 contain either a long signal peptide (LSP) or short (SSP), and are produced by alternatively spliced transcripts. It has been proposed that SSP remains exclusively intracellular, its function is unclear. In this study, we show that, similar to LSP IL-15, the stabilized secreted efficiently upon coexpression IL-15Ralpha. Coinjection IL-15- IL-15Ralpha-expressing plasmids into mice resulted in increased plasma levels bioactive heterodimeric mobilization...
Purpose: Adoptive cell transfer (ACT) is a promising immunotherapeutic approach for cancer. Host lymphodepletion associated with favorable ACT therapy outcomes, but it may cause detrimental effects in humans. We tested the hypothesis that IL15 administration enhances absence of lymphodepletion. previously showed bioactive vivo comprises stable complex chain receptor alpha (IL15Rα), termed heterodimeric (hetIL15).Experimental Design: evaluated combination regimen + hetIL15 by transferring...
Immunogenicity of HIV-1 mRNA vaccine regimens was analyzed in a non-human primate animal model. Rhesus macaques immunized with lipid nanoparticle (mRNA/LNP) formulation expressing Gag and conserved regions (CE) as immunogens developed robust, durable antibody responses but low adaptive T-cell responses. Augmentation the dose resulted modest increases vaccine-induced cellular immunity, no difference humoral The gag mRNA/lipid (LNP) provided suboptimal priming T cell for heterologous DNA...
Efficient expression vectors for interleukin 15 (IL-15) were developed combining RNA/codon optimization and modification of the IL-15 native long signal peptide. These changes resulted in elevated cytoplasmic levels optimized mRNA more than 100-fold improved production secreted human protein. Similar modifications have also led to greatly increased rhesus macaque murine production. Comparison different heterologous secretory signals showed that tissue plasminogen activator is most efficient...