Cystatin C Regulates the Cytotoxicity of Infection-induced Endothelial-derived β-amyloid

Authors

Ron Balczon, University of South AlabamaFollow
Kyle A. Morrow, Edward Via Virginia College of Osteopathic Medicine
Silas Leavesley, University of South AlabamaFollow
Michael Francis, University of South AlabamaFollow
Trevor C. Stevens, University of South Alabama
Ezinne Agwaramgbo, University of South Alabama
Christopher Williams, Mayo Clinic
Reece P. Stevens, University of South Alabama
Geri Langham, University of South Alabama
Sarah Voth, University of South Alabama
Eugene A. Cioffi, University of South AlabamaFollow
Susan E. Weintraub, University of Texas at San Antonio
Troy Stevens, University of South AlabamaFollow

Document Type

Article

Publication Title

FEBS Open Bio

Abstract

Infection of rat pulmonary microvascular endothelial cells with the bacterium Pseudomonas aeruginosa induces the production and release of cytotoxic oligomeric tau and beta amyloid (Aβ). Here, we characterized these cytotoxic amyloids. Cytotoxic behavior and oligomeric tau were partially resistant to digestion with proteinase K, but cytotoxicity was abolished by various denaturants including phenol, diethylpyrocarbonate (DEPC), and 1,1,1,3,3,3-hexafluoro-2-isopropanol (HFIP). Ultracentrifugation for 8 h at 150 000 g was required to remove cytotoxic activity from the supernatant. Ultracentrifugation, DEPC treatment, and immunodepletion using antibodies against Aβ also demonstrated that cytoprotective protein(s) are released from endothelial cells during P. aeruginosa infection. Mass spectrometry of endothelial cell culture media following P. aeruginosa infection allowed identification of multiple potential secreted modulators of Aβ, including cystatin C, gelsolin, and ApoJ/clusterin. Immunodepletion, co-immunoprecipitation, and ultracentrifugation determined that the cytoprotective factor released during infection of endothelial cells by P. aeruginosa is cystatin C, which appears to be in a complex with Aβ. Cytoprotective cystatin C may provide a novel therapeutic avenue for protection against the long-term consequences of infection with P. aeruginosa.

First Page

2464

Last Page

2477

DOI

10.1002/2211-5463.12997

Publication Date

11-2020

Department

College of Medicine

Comments

© 2020 The Authors.

Published by FEBS Press and John Wiley & Sons Ltd. https://doi.org/10.1002/2211-5463.12997

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Recommended Citation

Balczon, R., Morrow, K.A., Leavesley, S., Francis, C.M., Stevens, T.C., Agwaramgbo, E., Williams, C., Stevens, R.P., Langham, G., Voth, S., Cioffi, E.A., Weintraub, S.E. and Stevens, T. (2020), Cystatin C regulates the cytotoxicity of infection-induced endothelial-derived β-amyloid. FEBS Open Bio, 10: 2464-2477. https://doi.org/10.1002/2211-5463.12997