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. 2022 Sep 9;10(9):2245.
doi: 10.3390/biomedicines10092245.

Efficacy of Selected Live Biotherapeutic Candidates to Inhibit the Interaction of an Adhesive-Invasive Escherichia coli Strain with Caco-2, HT29-MTX Cells and Their Co-Culture

Bronwyn Smit  1 , Chiemeka C Chinaka  1 , Albert A Scott  1 , Kirsten Gaiduschek  1 , Eva Hatje  1   2 , Anna Kuballa  3 , Samantha Coulson  4 , Wayne Finlayson  4 , Mohammad Katouli  1
Affiliations

Efficacy of Selected Live Biotherapeutic Candidates to Inhibit the Interaction of an Adhesive-Invasive Escherichia coli Strain with Caco-2, HT29-MTX Cells and Their Co-Culture

Bronwyn Smit et al. Biomedicines. .

Abstract

Adherent-invasive Escherichia coli (AIEC) has been implicated as a microbiological factor in the pathogenesis of inflammatory bowel disease (IBD). We evaluated the ability of six live biotherapeutic products (LBPs) to inhibit the interaction of an AIEC strain to three cell lines representing human gut epithelium. Co-inoculation of LBPs with AIEC showed a reduction in adhesion (up to 73%) and invasion of AIEC (up to 89%). Pre-inoculation of LBPs in HT-29-MTX and Caco-2 cells before challenging with AIEC further reduced the adhesion and invasion of the AIEC, with three LBPs showing significantly (p < 0.0001) higher efficiency in reducing the adhesion of AIEC. In co-inoculation experiments, the highest reduction in adhesion (73%) of AIEC was observed in HT-29-MTX cells, whereas the highest reduction in invasion (89%) was seen in HT-29-MTX and the co-culture of cells. Pre-inoculation of LBPs further reduced the invasion of AIEC with highest reduction (97%) observed in co-culture of cells. Our results indicated that whilst there were differences in the efficacy of LBPs, they all reduced interaction of AIEC with cell lines representing gut epithelium. Their efficiency was higher when they were pre-inoculated onto the cells, suggesting their potential as candidates for alleviating pathogenesis of AIEC in patients with IBD.

Keywords: AIEC; Caco-2:HT-29-MTX; adhesion; invasion; live biotherapeutic products.

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Conflict of interest statement

The authors declare that there is no conflict of interest with the organization that sponsored this research and publications arising from this research. Servatus Biopharmaceuticals provided the LBP strains and had input towards the study design and final report. However, the company was not involved in data collection, analysis or interpretation of the results.

Figures

Figure 1
Figure 1
Colonization of HT-29-MTX, Caco-2 and co-culture of Caco-2/HT-29-MTX cells by AIEC alone formula image, AIEC when co-inoculated with the LBPs formula image, AIEC in the presence of pre-inoculated LBPs formula image and negative control [NC] formula image. * Significant differences between the percentages of cells showing adhesion with AIEC before addition of LBPs strains in co-inoculation or pre-inoculation experiments; p ≤ 0.0001 (HT-29-MTX), p ≤ 0.0001 (Caco-2), p ≤ 0.0001 (Co-culture). ** Significant difference between co-inoculation and pre-inoculation for SVT 06B1 (p = 0.03) in Caco-2.
Figure 2
Figure 2
The number of AIEC cells adhering per cell on HT-29-MTX, Caco-2 and co-culture of Caco-2/HT-29-MTX cells. AIEC alone formula image, AIEC when co-inoculated with the LBPs formula image, AIEC when cells were pre-inoculated with LBPs formula image and negative control [NC] formula image. Significant differences (*) were observed before and after the addition of LBP strains (co-inoculation or pre-inoculation); p < 0.0001 (HT-29-MTX), p < 0.0001 (Caco-2), p < 0.0001 (Co-culture). ** Significant difference between co-inoculation and pre-inoculation for SVT 05P2 (p = 0.049) and SVT 06B1 (p = 0.04) in Caco-2, and SVT 08Z1 (p < 0.0001) in HT-29-MTX.
Figure 3
Figure 3
The number of cells showing invasion of AIEC on HT-29-MTX, Caco-2 and co-culture cells. Invading AIEC alone formula image, AIEC when co-inoculated with the LBPs formula image, AIEC in the presence of pre-inoculated LPBs formula image and negative control [NC] formula image. p < 0.0001 (HT-29-MTX), * p < 0.0001 (Caco-2), * p < 0.0001 (Co-culture). ** Significant difference between co-inoculation and pre-inoculation for SVT 05P2 (p = 0.0003) in Caco-2.
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