Shiga Toxin 2a Binds to Complement Components C3b and C5 and Upregulates Their Gene Expression in Human Cell Lines
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Shiga Toxin 2a Binds to Complement Components C3b and C5 and Upregulates Their Gene Expression in Human Cell Lines. / Kellnerová, Sára; Chatterjee, Sneha; Bayarri-Olmos, Rafael; Justesen, Louise; Talasz, Heribert; Posch, Wilfried; Kenno, Samyr; Garred, Peter; Orth-Höller, Dorothea; Grasse, Marco; Würzner, Reinhard.
In: Toxins, Vol. 13, No. 1, 2021.Research output: Contribution to journal › Journal article › Research › peer-review
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T1 - Shiga Toxin 2a Binds to Complement Components C3b and C5 and Upregulates Their Gene Expression in Human Cell Lines
AU - Kellnerová, Sára
AU - Chatterjee, Sneha
AU - Bayarri-Olmos, Rafael
AU - Justesen, Louise
AU - Talasz, Heribert
AU - Posch, Wilfried
AU - Kenno, Samyr
AU - Garred, Peter
AU - Orth-Höller, Dorothea
AU - Grasse, Marco
AU - Würzner, Reinhard
PY - 2021
Y1 - 2021
N2 - Enterohemorrhagic Escherichia coli (EHEC) infections can cause EHEC-associated hemolytic uremic syndrome (eHUS) via its main virulent factor, Shiga toxins (Stxs). Complement has been reported to be involved in the progression of eHUS. The aim of this study was to investigate the interactions of the most effective subtype of the toxin, Stx2a, with pivotal complement proteins C3b and C5. The study further examined the effect of Stx2a stimulation on the transcription and synthesis of these complement proteins in human target cell lines. Binding of Stx2a to C3b and C5 was evaluated by ELISA. Kidney and gut cell lines (HK-2 and HCT-8) were stimulated with varied concentrations of Stx2a. Subsequent evaluation of complement gene transcription was studied by real-time PCR (qPCR), and ELISAs and Western blots were performed to examine protein synthesis of C3 and C5 in supernatants and lysates of stimulated HK-2 cells. Stx2a showed a specific binding to C3b and C5. Gene transcription of C3 and C5 was upregulated with increasing concentrations of Stx2a in both cell lines, but protein synthesis was not. This study demonstrates the binding of Stx2a to complement proteins C3b and C5, which could potentially be involved in regulating complement during eHUS infection, supporting further investigations into elucidating the role of complement in eHUS pathogenesis.
AB - Enterohemorrhagic Escherichia coli (EHEC) infections can cause EHEC-associated hemolytic uremic syndrome (eHUS) via its main virulent factor, Shiga toxins (Stxs). Complement has been reported to be involved in the progression of eHUS. The aim of this study was to investigate the interactions of the most effective subtype of the toxin, Stx2a, with pivotal complement proteins C3b and C5. The study further examined the effect of Stx2a stimulation on the transcription and synthesis of these complement proteins in human target cell lines. Binding of Stx2a to C3b and C5 was evaluated by ELISA. Kidney and gut cell lines (HK-2 and HCT-8) were stimulated with varied concentrations of Stx2a. Subsequent evaluation of complement gene transcription was studied by real-time PCR (qPCR), and ELISAs and Western blots were performed to examine protein synthesis of C3 and C5 in supernatants and lysates of stimulated HK-2 cells. Stx2a showed a specific binding to C3b and C5. Gene transcription of C3 and C5 was upregulated with increasing concentrations of Stx2a in both cell lines, but protein synthesis was not. This study demonstrates the binding of Stx2a to complement proteins C3b and C5, which could potentially be involved in regulating complement during eHUS infection, supporting further investigations into elucidating the role of complement in eHUS pathogenesis.
KW - C3
KW - C5
KW - HCT-8
KW - hemolytic uremic syndrome
KW - HK-2
KW - intracellular complement
KW - Shiga toxin 2a
U2 - 10.3390/toxins13010008
DO - 10.3390/toxins13010008
M3 - Journal article
C2 - 33374102
AN - SCOPUS:85099114677
VL - 13
JO - Toxins
JF - Toxins
SN - 2072-6651
IS - 1
ER -
ID: 282602488