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Streptococcus pneumoniae exerts oxidative stress, subverts antioxidant signaling and autophagy in human corneal epithelial cells that is alleviated by tert-Butylhydroquinone

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Abstract

Streptococcus pneumoniae is one of the leading causes of bacterial keratitis in the developing world and globally. In the current study, we have determined oxidative stress as pathogenesis of S. pneumoniae infection in corneal tissues and human corneal epithelial cells (HCEC) and explored host immune response of HCEC towards S. pneumoniae. We also determined whether treatment with tert-Butylhydroquinone (tBHQ), a Nrf2 inducer, could alleviate oxidative stress and reduce bacterial cytotoxicity in these cells. Oxidative stress was determined in corneal tissues of patients and HCEC by immunohistochemistry and immunofluorescence analysis, respectively. The expression of antioxidant genes, cytokines and antimicrobial peptides was determined by quantitative PCR. Infection of HCEC by S. pneumoniae was determined by colony-forming units. The autophagy and cell death were determined by fluorescence microscopy. The phosphorylation of signaling proteins was evaluated by immunoblot analysis. S. pneumoniae induced oxidative stress during corneal infections and inhibited antioxidant signaling pathways and immune responses like autophagy. tBHQ aided in restoring Nrf2 activation, reduced reactive oxygen species generation and prevented cytotoxicity and cell death in S. pneumoniae-infected HCEC. tBHQ also induced autophagy in a Nrf2-dependent manner and reduced bacterial survival in HCEC. Increased expression of antimicrobial peptides by tBHQ might have contributed to a reduction of bacterial load and cytotoxicity, as exemplified in LL-37 depleted corneal epithelial cells exposed to S. pneumoniae compared to control siRNA-transfected cells. tBHQ mediates alleviation of oxidative stress induced by S. pneumoniae by activating Nrf2-mediated antioxidant signaling in corneal epithelial cells. tBHQ also enhances expression of antimicrobial peptides in corneal cells and aids in inhibition of bacterial survival and cytotoxicity of HCEC.

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Acknowledgements

The authors sincerely acknowledge Hyderabad Eye Research Foundation, LVPEI for support. This study was supported by in parts by grants from SERB-DST, Govt. of India, (Grant No. EMR/2016/001514) and DBT, Govt. of India, (BT/PR32404/MED/30/2136/2019) to S.R, and student fellowship from UGC, Govt. of India to P.S. The authors duly acknowledge the help of Pathology Department, LVPEI. Excellent technical assistance from Bharathi Bhogapurapu and Apurwa Samarth is duly acknowledged.

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  1. Prof. Brien Holden Eye Research Centre, LV Prasad Eye Institute, Hyderabad, 500034, India

    Prerana Sharma & Sanhita Roy

  2. Department of Animal Biology, University of Hyderabad, Hyderabad, India

    Prerana Sharma

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Sharma, P., Roy, S. Streptococcus pneumoniae exerts oxidative stress, subverts antioxidant signaling and autophagy in human corneal epithelial cells that is alleviated by tert-Butylhydroquinone. Med Microbiol Immunol 211, 119–132 (2022). https://doi.org/10.1007/s00430-022-00731-y

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