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Publications

Journal Article Epublish

PubMed: 39835181 DOI: 10.2147/IJN.S497742

Viral Mimetic Bacterial Outer Membrane Vesicles for Targeting Angiotensin-Converting Enzyme 2.

Int J Nanomedicine 20:669-684

Authors: Gna Ahn, Hyo-Won Yoon, Ju Hwan Jeong, Yang-Hoon Kim, Woo-Ri Shin, Min-Suk Song, Ji-Young Ahn

Affiliations

  • Department of Microbiology, Chungbuk National University, Cheongju, Republic of Korea. Center for Ecology and Environmental Toxicology, Chungbuk National University, Cheongju, Republic of Korea.
  • Department of Microbiology, Chungbuk National University, Cheongju, Republic of Korea.
  • Department of Microbiology, College of Medicine and Medical Research Institute, Chungbuk National University, Cheongju, Republic of Korea. Functional Biomaterial Research Center, Korea Research Institute of Bioscience and Biotechnology, Jeongeup, Republic of Korea.
  • Department of Microbiology, Chungbuk National University, Cheongju, Republic of Korea. Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA. Department of Animal Bioscience & Integrated Biotechnology, Gyeongsang National University, Jinju, Republic of Korea.
  • Department of Microbiology, College of Medicine and Medical Research Institute, Chungbuk National University, Cheongju, Republic of Korea.

Abstract

Purpose

Outer membrane vesicles (OMVs) derived from Gram-negative bacteria naturally serve as a heterologous nano-engineering platform, functioning as effective multi-use nanovesicles for diagnostics, vaccines, and treatments against pathogens. To apply refined OMVs for human theranostic applications, we developed naturally exposed receptor-binding domain (RBD) OMVs grafted with antigen 43 as a minimal modular system targeting angiotensin-converting enzyme 2 (ACE2).

Methods

We constructed -derived OMVs using the antigen 43 autotransporter system to display RBD referred to as viral mimetic Ag43β700_RBD OMVs. Based on this, Ag43β700_RBD protein were expressed onto () membrane. Artificial viral mimetic Ag43β700_RBD OMVs were fabricated by self-assembly through membrane disruption of the Ag43β700_RBD using a chemical detergent mainly containing lysozyme. Through serial centrifugation to purify fabricated OMVs, spherical Ag43β700_RBD OMVs with an average diameter of 218 nm were obtained. The confirmation of the RBD expressed on OMVs was performed using trypsin treatment.

Results

Our viral mimetic Ag43β700_RBD OMVs had an impact on the theranostic studies: (i) angiotensin-converting enzyme 2 blockade assay, (ii) enzyme-linked immunosorbent assay for the OMVs, and (iii) intracellular uptake and neutralization assay. As serodiagnostic surrogates, Ag43β700_RBD OMVs were applied to ACE2 blockade and OMVs-ELISA assay to quantify neutralization antibodies (nAbs). They reduced the robust immune response in vitro, especially IL-6 and IL-1β. Experiments in mice, Ag43β700_RBD OMVs was successfully proven to be safe and effective; they produced a detectable level of nAbs with 39-58% neutralisation and reduced viral titres in the lungs and brain without weight loss.

Conclusion

The developed viral mimetic Ag43β700_RBD OMVs may therefore be applied as a nanovesicle-theranostic platform for further emerging infectious disease-related diagnosis, vaccination, and treatment.

MeSH

Nanomedicine, Vaccines, Angiotensin-Converting Enzyme 2, Peptidyl-Dipeptidase A, Enzyme-Linked Immunosorbent Assay, Bacterial Outer Membrane, Angiotensins, Enzymes, Therapeutics, Membranes