Recombinant PBP۲a/autolysin conjugate as PLGA-based nanovaccine induced humoral responses with opsonophagocytosis activity, and protection versus methicillin-resistant Staphylococcus aureus infection
Publish place: Iranian Journal of Basic Medical Sciences، Vol: 25، Issue: 4
Publish Year: 1401
نوع سند: مقاله ژورنالی
زبان: English
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JR_IJBMS-25-4_003
تاریخ نمایه سازی: 25 اردیبهشت 1401
Abstract:
Objective(s): Methicillin-resistant Staphylococcus aureus (MRSA) reasons extreme infections, can resist various conventional antimicrobial agents, and cause morbidity and mortality worldwide. Vaccination seems to help modulate MRSA infections. Nanovaccine is considered a novel strategy in vaccine technology. The primary purpose of the present study was to develop a conjugate vaccine based on recombinant PBP۲a and MRSA autolysin formulated in PLGA as a nanoparticle capable of enhancing protective responses against MRSA in the murine model.Materials and Methods: Recombinant PBP۲a and autolysin have been expressed and purified by nickel-nitrilotriacetic acid (Ni-NTA) affinity column and characterized by SDS-PAGE and western blot. PLGA was bound to recombinant proteins by using ۱-ethyl-۳-(۳-dimethylaminopropyl) carbodiimide hydrochloride (EDAC) and adipic acid dihydrazide (ADH) as a linker and spacer, respectively. Conjugation of recombinant proteins to PLGA was confirmed by the AFM assay, zeta potential, and size distribution, and its efficacy was evaluated in mice. Total IgG, IgG۱, IgG۲a, IgG۲b, and IgM titers were analyzed to assess immune responses. Lastly, the bioactivity of antibodies was tested by using the opsonophagocytosis assay. Results: Mice immunized with the r-PBP۲a-r-autolysin–PLGA nanovaccine led to increased levels of opsonic antibodies and IgG۱, IgG۲a, IgG۲b, and IgM when compared with other experimental groups. Our results confirmed that vaccination with nanovaccine could reduce the mortality rate against the sub-lethal dose of MRSA challenge. Furthermore, the nanovaccine could eliminate MRSA from the kidney of infected mice. Conclusion: This study may provide valuable insights into the protective power of the r-PBP۲a-r-autolysin–PLGA conjugate vaccine against MRSA infection.
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Authors
Setareh Haghighat
Department of Microbiology, Faculty of Advanced Science and Technology, Tehran Medical Sciences, Islamic Azad University, Tehran, Iran
Seyed Davar Siadat
Department of Mycobacteriology & Pulmonary Research, Microbiology Research Center, Pasteur Institute of Iran, Tehran, Iran
Abbas Akhavan Sepahi
Department of Microbiology, Faculty of Basic Sciences, North Tehran Branch, Islamic Azad University, Tehran, Iran
Mehdi Mahdavi
Advanced Therapy Medicinal Product (ATMP) Department, Breast Cancer Research Center, Motamed Cancer Institute, Academic Center for Education, Culture and Research(ACECR), Tehran, Iran
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