Dataset Information

Production of a Granulysin-Based, Tn-Targeted Cytolytic Immunotoxin Using Pulsed Electric Field Technology.

ABSTRACT: Granulysin is a protein present in the granules of human cytotoxic T lymphocytes (CTL) and natural killer (NK) cells, with cytolytic activity against microbes and tumors. Previous work demonstrated the therapeutic effect of the intratumoral injection of recombinant granulysin and of the systemic injection of an immunotoxin between granulysin and the anti-carcinoembryonic antigen single-chain Fv antibody fragment MFE23, which were produced in the yeast Pichia pastoris. In the present work, we developed a second immunotoxin combining granulysin and the anti-Tn antigen single-chain Fv antibody fragment SM3, that could have a broader application in tumor treatment than our previous immunotoxin. In addition, we optimized a method based on electroporation by pulsed electric field (PEF) to extract the remaining intracellular protein from yeast, augmenting the production and purificiation yield. The immunotoxin specifically recognized the Tn antigen on the cell surface. We also compared the thermal stability and the cytotoxic potential of the extracellular and intracellular immunotoxins on Tn-expressing human cell lines, showing that they were similar. Moreover, the bioactivity of both immunotoxins against several Tn+ cell lines was higher than that of granulysin alone.

SUBMITTER: Guerrero-Ochoa P

PROVIDER: S-EPMC7503585 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Production of a Granulysin-Based, Tn-Targeted Cytolytic Immunotoxin Using Pulsed Electric Field Technology.

Guerrero-Ochoa Patricia P Aguilar-Machado Diederich D Ibáñez-Pérez Raquel R Macías-León Javier J Hurtado-Guerrero Ramón R Raso Javier J Anel Alberto A

International journal of molecular sciences 20200826 17

Granulysin is a protein present in the granules of human cytotoxic T lymphocytes (CTL) and natural killer (NK) cells, with cytolytic activity against microbes and tumors. Previous work demonstrated the therapeutic effect of the intratumoral injection of recombinant granulysin and of the systemic injection of an immunotoxin between granulysin and the anti-carcinoembryonic antigen single-chain Fv antibody fragment MFE23, which were produced in the yeast <i>Pichia pastoris</i>. In the present work, ...[more]

PMID: 32859066

Similar Datasets

Project description:Chemo-immunotherapy uses combined systemic therapies for resectable and unresectable tumors. This approach is gaining clinical momentum, but survival increases leave considerable room for improvement. A novel form of Pulsed Electric Field (PEF) ablation combines focal tissue destruction with immune activation in preclinical settings. The PEFs induce lethal cell damage without requiring thermal processes, leaving cellular proteins intact. This affords PEF a favorable safety profile, improved antigenicity, and significant immunostimulatory damage-associated molecular pattern release compared to other focal therapies. Preclinical investigations demonstrate a combinatorial benefit of PEF with immunostimulation. This study evaluates whether this proprietary PEF therapy induces an immunostimulatory effect sufficient to augment systemic neoadjuvant chemotherapy and immunotherapy to reverse metastatic disease in an immune-cold murine tumor model. To determine whether PEF improves a neoadjuvant chemo-immunotherapy standard-of-care, partial PEF ablation was delivered to orthotopically inoculated 4T1 metastatic tumors in addition to combinations of cisplatin chemotherapy and/or αPD-1 immunotherapy, followed by resection. In addition, to determine whether PEF combined with chemo-immunotherapy improves local and metastatic response in unresectable populations, partial PEF ablation was added to chemo-immunotherapy in mice that did not receive resection. Blood cytokines and flow cytometry evaluated immune response. Partial PEF ablation generates an immunostimulatory tumor microenvironment, increases systemic immune cell populations, slows tumor growth, and prolongs survival relative to neoadjuvant systemic therapies-alone. These data suggest the addition of this proprietary PEF locoregional therapy may synergize with systemic standard-of-care paradigms to improve outcomes with potential or demonstrated metastatic disease in both resectable and unresectable patient cohorts.

Project description:Extra virgin olive oil (EVOO) is a cornerstone of the Mediterranean diet. Many studies have highlighted its crucial preventive role against cardiovascular disease, neurodegenerative disorders, metabolic syndrome and cancer, with these effects being due to the synergistic anti-inflammatory and antioxidant activities of minor components, such as polyphenols and tocols. The aim of the present study is to implement new technologies for olive oil mills and develop an efficient large-sized industrial process for the continuous extraction of healthier EVOOs that are enriched with these bioactive compounds. Non-thermal technologies, namely ultrasound (US) and pulsed electric field (PEF), have been tested, separately and in combination, to eliminate the need for traditional malaxation. There is extensive literature to support the efficacy of ultrasound-assisted extraction (UAE) and PEF treatments in EVOO production. A newly designed US device and a PEF industrial chamber have been combined into a single, integrated continuous-flow setup, the performance of which in the extraction of EVOO from green Coratina olives has been evaluated herein. Extraction yields, physico-chemical and organoleptic characteristics, and polyphenol and tocol contents were monitored throughout the trials, and the last three were measured at accelerated aging times (AAT) of 15 and 30 days. The US and combined US-PEF processes not only increased daily oil production (ton/day, by nearly 45%), but also eliminated the need for kneading during malaxation, resulting in significant energy savings (approximately 35%). In addition, these innovations enriched the resulting EVOO with nutritionally relevant minor components (8-12% polyphenols, 3-5% tocols), thereby elevating its quality and market value, as well as overall stability. The introduction of continuous-flow US and PEF technologies is a remarkable innovation for the EVOO industry, as they offer benefits to both producers and consumers. The EVOO resulting from non-thermal continuous-flow production meets the growing demand for healthier, nutrient-enriched products.

Dataset Information

Production of a Granulysin-Based, Tn-Targeted Cytolytic Immunotoxin Using Pulsed Electric Field Technology.

Publications

Production of a Granulysin-Based, Tn-Targeted Cytolytic Immunotoxin Using Pulsed Electric Field Technology.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets