In silico studies of green fluorescent protein-tagged Hsp27-HPV16 E7 fusion protein and evaluation of cytokine secretion from antigen-presenting cells exposed to the fusion protein-carrying tumor-derived exosomes DOI Open Access
Fatemeh Rezaei, Arash Arashkia, Fatemeh Fotouhi

и другие.

Journal of Solid Tumors, Год журнала: 2024, Номер 14(1), С. 45 - 45

Опубликована: Дек. 15, 2024

Objective: Exosome (Exo)-based therapies have attracted considerable interest due to their potential as carriers for therapeutic molecules and capacity elicit anti-tumor immune responses. The objective of this study was engineer TC-1 tumor cell line-derived exosomes with GFP-tagged heat shock protein (Hsp) 27-human papillomavirus (HPV)16 E7 fusion evaluation cytokine secretion from antigen-presenting cells (APCs: macrophages dendritic cells) exposed the engineered in vitro.Methods: In study, different silico methods were employed evaluate Hsp27-E7 Hsp27-E7-GFP proteins vaccine candidates. Regarding data, gene subcloned into pCDH lentiviral vector production lentivirions harboring eukaryotic cells. Subsequently, transduced these isolate (i.e., Exo-Hsp27-E7-GFP) using ExoQuick-TCTM kit characterization physicochemical methods. Finally, key cytokines (IFN-γ, TNF-α, IL-10) evaluated through incubation (APCs) Exo-Hsp27-E7-GFP enzyme-linked immunosorbent assay (ELISA).Results: Our data showed that both constructs soluble non-allergenic, exhibited strong interaction TLR4. Indeed, linkage GFP did not affect properties, receptors. Moreover, western blot analysis confirmed presence isolated exosomes. could significantly enhance TNF-α IL-10 APCs compared Exo Exo-GFP, well.Conclusions: These vesicles derived demonstrate induce effective immunity, suggesting a promising strategy development cell-free

Язык: Английский

Molecular Farming for Immunization: Current Advances and Future Prospects in Plant-Produced Vaccines DOI Creative Commons
Dang-Khoa Vo, Kieu The Loan Trinh

Vaccines, Год журнала: 2025, Номер 13(2), С. 191 - 191

Опубликована: Фев. 15, 2025

Using plants as bioreactors, molecular farming has emerged a versatile and sustainable platform for producing recombinant vaccines, therapeutic proteins, industrial enzymes, nutraceuticals. This innovative approach leverages the unique advantages of plants, including scalability, cost-effectiveness, reduced risk contamination with human pathogens. Recent advancements in gene editing, transient expression systems, nanoparticle-based delivery technologies have significantly enhanced efficiency versatility plant-based systems. Particularly vaccine development, demonstrated its potential notable successes such Medicago's Covifenz COVID-19, illustrating capacity platforms to address global health emergencies rapidly. Furthermore, edible vaccines opened new avenues mainly settings low resources where cold chain used conventional logistics is challenge. However, optimization protein yield stability, complexity purification processes, regulatory hurdles are some challenges that still remain. review discusses current status development using operational mechanisms plant platforms, major applications prevention infectious diseases, developments, nanoparticle-mediated cancer vaccines. The discussion will also touch on ethical considerations, framework, future trends respect transformative plant-derived ensuring greater accessibility cost-effectiveness vaccination. field holds great promise disease area and, indeed, personalized medicine biopharmaceuticals near future.

Язык: Английский

Процитировано

1

Strategic and Technical Considerations in Manufacturing Viral Vector Vaccines for the Biomedical Advanced Research and Development Authority Threats DOI Creative Commons

Lindsay A. Parish,

Shyam Rele,

Kimberly A. Hofmeyer

и другие.

Vaccines, Год журнала: 2025, Номер 13(1), С. 73 - 73

Опубликована: Янв. 14, 2025

Over the past few decades, world has seen a considerable uptick in number of new and emerging infectious disease outbreaks. The development vaccines, vaccine technologies, platforms are critical to enhance our preparedness for biological threats prevent future pandemics. Viral vectors can be an important tool repertoire technologies available develop effective vaccines against diseases. In many instances, may needed reactive scenario, requiring than elicit rapid robust immune responses with single dose. Here, we discuss how viral vector utilized portfolio priority threats, some challenges manufacturing need strengthen live virus capabilities, opportunities capitalize on use improve sustainability Biomedical Advanced Research Development Authority’s portfolio.

Язык: Английский

Процитировано

0

CRISPR-Cas: a game-changer in vaccine development and the fight against viral infections DOI
Fayu Yang,

Saba R. Aliyari,

Zixiang Zhu

и другие.

Trends in Microbiology, Год журнала: 2025, Номер unknown

Опубликована: Март 1, 2025

Язык: Английский

Процитировано

0

Application of semi-quantitative loop-mediated isothermal amplification for gene expression study in Expi293 cells. DOI Creative Commons
Pavel A. Bobrovsky, Ekaterina Grafskaia, Daria Kharlampieva

и другие.

Genes and Cells, Год журнала: 2024, Номер unknown

Опубликована: Дек. 1, 2024

BACKGROUND: Mammalian cell cultures play a key role in the pharmaceutical industry, requiring constant monitoring of conditions during fermentation. In addition to physical-chemical parameters, it is important evaluate internal state cells, for which gene expression analysis used. Currently, quantitative reverse transcription polymerase chain reaction (qPCR) dominant method. However, loop-mediated isothermal amplification (LAMP) technique also attracts attention due its high specificity, sensitivity and rate. LAMP becoming promising tool rapid expression, especially under limited biological material or large volume samples. AIM: Development quantifying level target genes human culture Expi293. METHODS: For LAMP, recombinant fragment Bacillus stearothermophilus DNA (Bst-pol) was obtained, purified, optimal were determined. SYBR Green I LUCS13 used as intercalating dyes. The parameters different concentrations enzyme dye analyzed. Standard kits qPCR. Both methods compared when analyzing IGF 1, FGF2 EIF3i lines with an increased these genes. RESULTS: It has been shown that using provides classic S-shape signal accumulation curve while causes artifacts. concentration Bst-pol 40 ng/µl. When comparing two methods, found greater sensitivity, allows determining accuracy comparable qPCR, demonstrating shorter time (up 35 minutes). CONCLUSION: Although PCR remains main method assessing offers number advantages make attractive alternative various biotechnological purposes. Due speed, ease execution accessibility, well valuable monitoring.

Язык: Английский

Процитировано

1

Chikungunya virus release is reduced by TIM-1 receptors through binding of envelope phosphatidylserine DOI
Judith Mary Reyes Ballista,

Ashley J. Hoover,

Joseph Thomas Noble

и другие.

Journal of Virology, Год журнала: 2024, Номер 98(8)

Опубликована: Июль 15, 2024

ABSTRACT T-cell immunoglobin and mucin domain protein-1 (TIM-1) mediates entry of chikungunya virus (CHIKV) into some mammalian cells through the interaction with envelope phospholipids. While this enhances entry, TIM-1 has been shown to tether newly formed HIV Ebola particles, limiting their efficient release. In study, we investigate ability surface receptors such as sequester budded virions on infected cells. We established a luminescence reporter system produce viral particles that integrate nano-luciferase easily quantify particles. found host significantly reduced CHIKV release efficiency in comparison other factors. Removal cell direct cellular knock-out or altering lipid distribution enhanced Over course infection, was able counteract tethering effect by gradually decreasing levels process mediated nonstructural protein 2. This study highlights importance phosphatidylserine mediating not only but also its could aid developing lines capable vaccine production. IMPORTANCE Chikungunya is an enveloped alphavirus transmitted bites infectious mosquitoes. Infection results development fever, joint pain, arthralgia can become chronic last for months after infection. Prevention disease still highly focused vector control strategies. December 2023, new live attenuated against approved FDA. aimed factors involved release, better understand CHIKV’s efficiently infect spread among wide variety lines. abrogate exit information be beneficial maximizing particle production laboratory settings during manufacturing.

Язык: Английский

Процитировано

0

Serum-Free Media Formulation Using Marine Microalgae Extracts and Growth Factor Cocktails for Madin-Darby Canine Kidney and Vero Cell Cultures DOI Open Access

Areumi Park,

Yeon-Ji Lee,

Eunyoung Jo

и другие.

International Journal of Molecular Sciences, Год журнала: 2024, Номер 25(18), С. 9881 - 9881

Опубликована: Сен. 12, 2024

The development of serum-free media (SFM) is critical to advance cell culture techniques used in viral vaccine production and address the ethical concerns contamination risks associated with fetal bovine serum (FBS). This study evaluated effects marine microalgal extracts growth factor cocktails on activity Madin-Darby canine kidney (MDCK) Vero cells. Five species were used: Spirulina platensis (SP), Dunaliella salina (DS), Haematococcus pluvialis (HP), Nannochloropsis (NS), Tetraselmis sp. (TS). DS SP significantly increased proliferation rate both MDCK had a 149.56% 195.50% cells, respectively, compared that medium (SFM). Notably, superoxide dismutase (SOD) activity, which was 118.61% cells 130.08% for DS, 108.72% 125.63% SP, indicating reduction intracellular oxidative stress. Marine extracts, especially are feasible alternatives FBS as they promote proliferation, ensure safety, supply essential nutrients while reducing

Язык: Английский

Процитировано

0

In silico studies of green fluorescent protein-tagged Hsp27-HPV16 E7 fusion protein and evaluation of cytokine secretion from antigen-presenting cells exposed to the fusion protein-carrying tumor-derived exosomes DOI Open Access
Fatemeh Rezaei, Arash Arashkia, Fatemeh Fotouhi

и другие.

Journal of Solid Tumors, Год журнала: 2024, Номер 14(1), С. 45 - 45

Опубликована: Дек. 15, 2024

Objective: Exosome (Exo)-based therapies have attracted considerable interest due to their potential as carriers for therapeutic molecules and capacity elicit anti-tumor immune responses. The objective of this study was engineer TC-1 tumor cell line-derived exosomes with GFP-tagged heat shock protein (Hsp) 27-human papillomavirus (HPV)16 E7 fusion evaluation cytokine secretion from antigen-presenting cells (APCs: macrophages dendritic cells) exposed the engineered in vitro.Methods: In study, different silico methods were employed evaluate Hsp27-E7 Hsp27-E7-GFP proteins vaccine candidates. Regarding data, gene subcloned into pCDH lentiviral vector production lentivirions harboring eukaryotic cells. Subsequently, transduced these isolate (i.e., Exo-Hsp27-E7-GFP) using ExoQuick-TCTM kit characterization physicochemical methods. Finally, key cytokines (IFN-γ, TNF-α, IL-10) evaluated through incubation (APCs) Exo-Hsp27-E7-GFP enzyme-linked immunosorbent assay (ELISA).Results: Our data showed that both constructs soluble non-allergenic, exhibited strong interaction TLR4. Indeed, linkage GFP did not affect properties, receptors. Moreover, western blot analysis confirmed presence isolated exosomes. could significantly enhance TNF-α IL-10 APCs compared Exo Exo-GFP, well.Conclusions: These vesicles derived demonstrate induce effective immunity, suggesting a promising strategy development cell-free

Язык: Английский

Процитировано

0