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Cephalosporium curvulum lectin causes mycotic keratitis by initiating infection through MyD88 dependent cellular proliferation and apoptosis in human corneal epithelial cells

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Abstract

Physiological role of a core fucose specific lectin from Cephalosporium curvulum isolated from mycotic keratitis patient in mediating pathogenesis was reported earlier. CSL has opposite effects on HCECs, at the initiation of infection when lectin concentration is low, CSL induces proinflammatory response and at higher concentration it inhibits growth as the infection progresses. Here we delineate detailed mechanism of opposing effects of CSL by confirming the binding of CSL and anti TLR 2 and 4 antibodies to TLRs 2 and 4 purified from HCECs using Galectin-3 Sepharose 4B column. Further, the expression of signaling proteins were monitored by Western blotting and apoptosis assay. At concentration of 0.3 µg/ml, CSL induced the activation of TLR-2,-4 and adapter protein MyD88. CSL also induced the expression of transcription factors NFkB, C-Jun and proinflammatory cytokines like interleukins -6 and -8 essential in maintaining cell proliferation. In contrast at higher concentrations i.e. 5 µg/ml CSL induces apoptotic effect as evidenced by increase in early and late apoptotic population as demonstrated by Annexin V-PI assay. Western blotting revealed that CSL treated HCECs at higher concentration lead to MyD88 dependent expression of apoptotic proteins like FADD, Caspase -8 and -3. All these results are in line with and substantiate our earlier results that indeed CSL is involved in mediating host pathogen interactions by interacting with cell surface TLRs, activating downstream signaling pathways leading to pathogenesis. Findings are of clinical significance in developing carbohydrate based therapeutic strategy to control infection and the disease.

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Acknowledgements

SRI thanks the Council of Scientific & Industrial Research (CSIR, No. 27 (0350)/19/EMR-II dated 17th May 2019) and N Jagadeesh thanks the CSIR for fellowship under CSIR-SRF (No. 09/101(0056)/2019/EMR-I).

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Authors and Affiliations

  1. Department of Studies in Biochemistry, Karnatak University, Dharwad, 580003, India

    Narasimhappagari Jagadeesh, Shivakumar Belur, Suhas Ballal & Shashikala R. Inamdar

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

    Sanhita Roy

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  1. Narasimhappagari Jagadeesh
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  2. Shivakumar Belur
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  3. Suhas Ballal
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Correspondence to Shashikala R. Inamdar.

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Jagadeesh, N., Belur, S., Ballal, S. et al. Cephalosporium curvulum lectin causes mycotic keratitis by initiating infection through MyD88 dependent cellular proliferation and apoptosis in human corneal epithelial cells. Glycoconj J 38, 509–516 (2021). https://doi.org/10.1007/s10719-021-10004-0

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  • DOI: https://doi.org/10.1007/s10719-021-10004-0

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