Liposome-based nanocarriers loaded with anthrax lethal factor and armed with anti-CD19 VHH for effectively inhibiting MAPK pathway in B cells
Autor: | Davoud Ahmadvand, S. Reza Banihashemi, Shahryar Khoshtinat Nikkhoi, Fatemeh Rahbarizadeh, Ahmad Zavaran Hosseini |
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Rok vydání: | 2021 |
Předmět: |
MAPK/ERK pathway
MAP Kinase Signaling System p38 mitogen-activated protein kinases Immunology Antigens CD19 Bacterial Toxins CD19 Immunoliposome Cell Line Tumor Neoplasms Immunology and Allergy Humans Cytotoxicity Pharmacology Antigens Bacterial biology Chemistry Kinase Pyroptosis Single-Domain Antibodies Raji cell Cell biology Liposomes biology.protein Drug Screening Assays Antitumor Nanoparticle Drug Delivery System |
Zdroj: | International immunopharmacology. 100 |
ISSN: | 1878-1705 |
Popis: | Objective One of the vital signaling pathways in cancer development and metastasis is mitogen-activated protein kinases (MAPKs). Bacillus anthracis Lethal Toxin (LT) is a potent MAPK signaling inhibitor. This toxin is comprised of two distinct domains, Lethal Factor (LF), MAPK inhibitor, and Protective Antigen (PA). To enter various cell lines, LF must be associated with the protective antigen (PA), which facilitates LF delivery. In the current study, to block MAPK signaling, LF was loaded into anti-CD19 immunoliposomes nanoparticle to deliver the cargo to Raji B cells. Methods The liposome nanoparticle was prepared using classical lipid film formation, then conjugated to anti-CD19 VHH. The binding efficiency was measured through flow cytometry. The targeted cytotoxicity of LF immunoliposome was confirmed by BrdU lymphoproliferation assay. This was followed by Real-Time PCR to assess the effect of formulation on pro-apoptotic genes. The inhibitory effect of LF on MAPK signaling was confirmed by western blot. Results Liposome nano-formulation was optimized to reach the maximum LF encapsulation and targeted delivery. Next, phosphorylation of MAPK pathway mediators like MEK1/2, P38 and JNK were inhibited following the treatment of Raji cells with LF-immunoliposome. The treatment also upregulated caspase genes, clearly illustrating cell death induced by LF through pyroptosis and caspase-dependent apoptosis. Conclusions In conclusion, anti-CD19 VHH immunoliposome was loaded with LF, a potent MAPK inhibitor targeting B cells, which curbs proliferation and ushers B cells toward apoptosis. Thus, immunoliposome presents as a versatile nanoparticle for delivery of LF to block aberrant MAPK activation. To use LF as a therapy, it would be necessary to materialize LF without PA. In the current study, PA was substituted with anti-CD19 immunoliposome to make it targeted to CD19+ while keeping the normal cells intact. |
Databáze: | OpenAIRE |
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