Nanostructured recombinant cytokines: A highly stable alternative to short-lived prophylactics

Abstract

Cytokines have been widely used as adjuvants and therapeutic agents in treatments of human diseases. Despite their recognized potential as drugs, the medical use of cytokines has considerable drawbacks, mainly related to their low stability and short half-life. Such intrinsic limitations imply the administration of high doses, often prompting toxicity, undesirable side effects and greater production costs. Here, we describe a new category of mechanically stable nanostructured cytokines (TNFα and CCL4/MIP-1β) that resist harsh physicochemical conditions in vitro (pH and temperature), while maintaining functionality. These bio-functional materials are produced in recombinant cell factories through cost-effective and fully scalable processes. Notably, we demonstrate their prophylactic potential in vivo showing they protect zebrafish from a lethal infection by Pseudomonas aeruginosa.

Bacterial Inclusion Bodies: Discovering Their Better Half

“…IB protein release can be verified by the physiological effect that the IB-derived soluble protein exerts in different cellular models in vitro [47,65], showing a biological impact such as reconstruction of the cytoskeleton (keratin 14) [69], cell growth recovery (dihydrofolate reductase, leukemia inhibitory factor, fibroblast growth factor) [47], reduced oxidative stress (catalase) [47] or anti-apoptotic rescuing (Hsp70) [47] and in vivo as immunostimulant (cytokines) [70]. Oral delivery of IBs in mice and intraperitoneal or intratumoral injection in zebrafish and in mice does not result in detectable signs of local or systemic toxicity [47,70].…”

“…Oral delivery of IBs in mice and intraperitoneal or intratumoral injection in zebrafish and in mice does not result in detectable signs of local or systemic toxicity [47,70].…”

“…Tumor necrosis factor (TNF)- IBs, orally administered to zebrafish, were able to establish close contact with relevant sections of the gut for further IB-derived immune protection from bacterial infections [70]. Functional IBs, when delivering intracellular proteins with potential healing value can be regarded as a novel type of nanostructured therapeutic agent or nanopill.…”

Trends in recombinant protein use in animal production

“…Nevertheless, IBs have never been explored as protein-based nanoparticles for animal reproduction, enhancers of feed efficiency, or treatment purposes, with only one exception. A recent article described for the first time that IBs formed by cytokines can successfully be used as a prophylactic measure, showing that zebra fish treated with IBs are protected against a lethal infection [192]. In the same way that cytokines have been successfully produced as IBs for prevention purposes in fish, other proteins of interest (hormones, enzymes, and antibodies, among others) or other animal species could also be explored, unfolding enormous possibilities in this field.…”

Improving Biomaterials Imaging for Nanotechnology: Rapid Methods for Protein Localization at Ultrastructural Level

“…Specifically, natural and engineered peptides and proteins are among the most suitable building blocks for nanoparticle generation due to their full biocompatibility and degradability at physiological conditions and versatility to be engineered and produced in single step by inexpensive biological processes . Moreover, protein‐based materials can be tuned by modifying both production conditions and genetic cell background, to generate a wide spectrum of protein nanoparticles with different composition, structure, size, and shape adapted to specific uses in animal and human medicine . During the last years, peptide‐ and protein‐based biomaterials have emerged as convenient nano‐ and micro‐carriers fully compatible with cells, tissues, and entire organisms .…”