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Novel Antimicrobial Peptide Mitigates Inflammation in Human Lung Cells Infected with Pseudomonas aeruginosa

Undergraduate #102
Discipline: Biological Sciences
Subcategory: Microbiology/Immunology/Virology

Robert Emeh - Alabama State University
Co-Author(s): Ejovwoke Dosunmu, Komal Vig, Shree R. Singh, and Vida A. Dennis, Alabama State University, Montgomery, AL



Pseudomonas aeruginosa is an opportunist Gram-negative pathogen that causes infection in immuno-compromised individuals, such as cystic fibrosis (CF) patients. In the airways of CF patients, P. aeruginosa induces vigorous inflammatory response that does not necessarily eliminate the pathogen, but leads to a progressive loss of lung function and diminished life expectancy. We have shown that antimicrobial proprietary peptides (AMPs) have the ability to inhibit the in vitro growth of both mucoid and non-mucoid strains of P. aeruginosa. Numerous studies now indicate that AMPs can modulate immune responses by inducing cytokines/chemokines production and inhibiting pro-inflammatory cytokine production. In this study, we investigated the anti-inflammatory properties of a proprietary antimicrobial peptide, TP359, in human A549 lung cells infected with P. aeruginosa. Hypothesis: We hypothesized that the peptide, TP359, will reduce the inflammation caused by P. aerugonisa infection. Cytokine ELISA was used to determine the anti-inflammatory effect of TP-359 by measuring pro-inflammatory cytokines Interleukin-6 (IL-6), Interleukin-8 (IL-8), and Tumor Necrosis Factor (TNF). The A549 cells were exposed to TP359 at 4 different concentrations (12.5, 25, 50, and 100 ug/mL). Three types of exposure were utilized: A549 lung cells were exposed to P.aerugonisa 1 hour before TP359 treatment, A549 lung cells were exposed to TP359 1 hour before exposure to P. aerugonisa, and simultaneous exposure of P. aerugonisa and TP359 to A549 lung cells. Our results show that TP359 reduced the levels of pro-inflammatory cytokines in a dose- and time-dependent manner at all treatment conditions. These results indicate that TP359 can modulate P.aerugonisa -induced inflammatory responses in lung cells. Our further studies will investigate the pathways necessary for induction of pro-cytokines caused by a P. aerugonisa infection. Due to its confirmed anti-microbial and suggested anti-inflammatory properties, TP359 is an attractive agent for the development of therapeutics to treat Pseudomonas infections.

Funder Acknowledgement(s): The work was supported by Minority Science and Engineering Improvement Program (PD Komal Vig, Grant number P120A150008) and NSF-CREST (PI Shree Singh, HRD 12417010).

Faculty Advisor: Vida Dennis, vdennis@alasu.edu

Role: Before I joined Dr. Vida Dennis’s laboratory, it was already determined that the TP359 peptide was an antimicrobial agent against Pseudomonas aerugonisa, so my role was to help determine if the TP359 had anti-inflammatory effects on Pseudomonas aeruginosa infected human lung cells. Working alongside Dr. Dosunmu, I performed cytokine ELISA to determine the anti-inflammatory effect of of TP-359 by measuring tumor necrosis factor (TNF), interleukin (IL)-6, -8, and -1 beta secretions in A549 cells exposed to live Pseudomonas aeruginosa. Ultimately, our results confirmed our hypothesis; the TP359 peptide did in fact possess anti-inflammatory effects on Pseudomonas aerugonisa. TP359 reduced the levels of the pro-inflammatory cytokines, TNF, IL-6 and -8, secreted from Pseudomonas aeruginosa-infec

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This material is based upon work supported by the National Science Foundation (NSF) under Grant No. DUE-1930047. Any opinions, findings, interpretations, conclusions or recommendations expressed in this material are those of its authors and do not represent the views of the AAAS Board of Directors, the Council of AAAS, AAAS’ membership or the National Science Foundation.

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