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IgG binds Escherichia coli serine protease EspP and protects mice from E. coli O157:H7 infection

ABSTRACT: The effect of pooled immunoglobulins (IgG) on E. coli O157:H7 colonization and the course of disease in an EHEC mouse model was investigated showing an improved survival and decreased intestinal and renal pathology. Treatment was given after inoculation thereby corresponding to the clinical setting. In vitro studies identified E. coli serine protease EspP as the E. coli O157:H7 protein that IgG bound to, via the Fc fragment, in both murine and human IgG preparations, and blocked its enzymatic activity. EspP is a virulence factor previously shown to promote colonic cell injury and the uptake of Shiga toxin by intestinal cells. The results suggest that IgG in commercial preparations binds to EspP protecting the host from E. coli O157:H7 infection and could potentially be beneficial in patients.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Escherichia Coli

SUBMITTER: Charlotte Welinder

LAB HEAD: Diana Ora Karpman

PROVIDER: PXD029563 | Pride | 2022-02-08

REPOSITORIES: Pride

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Project description:Two lineages of enterohemorrhagic (EHEC) Escherichia coli O157:H7 (EDL933, Stx1+ and Stx2+) and 86-24 (Stx2+) were investigated in regards to biofilm formation on an abiotic surface. Strikingly, EDL933 strain formed a robust biofilm while 86-24 strain formed no biofilm on either a polystyrene plate or a polyethylene tube. To identify the genetic mechanisms of different biofilm formation in two EHEC strains, DNA microarrays were first performed and phenotypic assays were followed. In the comparison of the EDL933 strain versus 86-24 strain, genes (csgBAC and csgDEFG) involved in curli biosynthesis were significantly induced while genes (trpLEDCB and mtr) involved in indole signaling were repressed. Additionally, a dozen of phage genes were differentially present between two strains. Curli assays using a Congo red plate and scanning electron microscopy corroborate the microarray data as the EDL 933 strain produces a large amount of curli, while 86-24 forms much less curli. Also, the indole production in the EDL933 was 2-times lower than that of 86-24. It was known that curli formation positively regulates and indole negatively regulates biofilm formation of EHEC. Hence, it appears that less curli formation and high indole production in the 86-24 strain are majorly responsible for no biofilm formation. For the microarray experiments, E. coli O157:H7 EDL933 and 86-24 were inoculated in 25 ml of LB in 250 ml shake flasks with overnight cultures that were diluted 1:100. Cells were shaken at 250 rpm and 37°C for an absorbance of 4.0 at 600 nm. Cells were immediately chilled with dry ice and 95% ethanol (to prevent RNA degradation) for 30 sec before centrifugation in 50 ml centrifuge tubes at 13,000 g for 2 min; cell pellets were frozen immediately with dry ice and stored -80°C. RNA was isolated using Qiagen RNeasy mini Kit (Valencia, CA, USA). RNA quality was assessed by Agilent 2100 bioanalyser using the RNA 6000 Nano Chip (Agilent Technologies, Amstelveen, The Netherlands), and quantity was determined by ND-1000 Spectrophotometer (NanoDrop Technologies, Inc., DE, USA).

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IgG binds Escherichia coli serine protease EspP and protects mice from E. coli O157:H7 infection

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