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Development & Production of various Probiotic products

Project ID: 1.1.1-MKI-2018-00029
Project title: Development of heat killed Lactobacillus containing immunobiotics to attenuate chemotherapy induced systemic inflammatory response syndrome

Project promoter: Fermentia Mikrobiológiai Kft
Project partners: Eötvös Lóránd Tudományegyetem, Természettudományi kar, Mikrobiológiai Tanszék
Donor partners: Dynea AS. Korlátolt Felelősségű Társaság

Project summary:

Probiotics are widely spread as nutrient supplement in the therapy of different inflammatory bowel diseases associated with diarrhoea.

In the last decade accumulating body of evidences prove that probiotic microbes may be applied successfully far beyond than the gut diseases. Lactobacillus strains have been reported to apply successfully in the case of allergic skin diseases, like atopic dermatitis and psoriasis, moreover in other studies lactobacilli proved to be protective in different heart ischaemic models.

Lactobacillus treatment resulted in the decrease of both tissue necrosis and remodelling in heart ischaemia.

Probiotics exert their protective effects in endotoxaemia also via reducing the synthesis of pro-inflammatory cytokines (TNF, IL-6, IL-8) and oxidative tissue injury in liver through increasing synthesis of reduced glutathione.

Fermentia intends to develop a probiotic to attenuate chemotherapy induced injury of intestinal epithelium associated with mucositis, which leads to endotoxemia. These processes may cause increased systemic inflammatory cytokine production (systemic inflammatory response syndrome, SIRS which may resulted in multi organ failure.

Our goal is to develop a probiotics which effectively attenuate intestinal mucositis and systemic inflammatory processes caused by endotoxemia.

Pro-inflammatory (TNF, IL-6) and anti-inflammatory (IL-10) cytokine release of human intestinal epithelial cell line will be tested after 5-fluorouracil and LPS exposure, with or without Lactobacillus treatment. Cell viability will be determined also with TTC reduction test. Bcl2, and Bax gene expression will be determined with qPCR in order to examine pro- and anti-apoptotic processes.

At in vivo phase animals will be treated with 5-fluorouracil and LPS with and without Lactobacillus treatment and serum cytokine assay (TNF, IL-6, IL-10) will be performed. Some amino acid transferases eg. (alanine-amino transferease ALT), serum urea and kreatinin (indicating tissue injury). Moreover kidney function will be determined as well.

In this stage of our project we select those strains which most effectively attenuate the LPS and 5-fluorouracil induced proinflammatory cytokine release, ALT activity and improve kidney function.

The third phase of our work is to optimize the fermentation parameters of selected Lactobacillus strains in shake flask fermentations and in 10L fermentors. Examined parameters will be the rotation speed, medium composition and determination of the optimal pH value. We will continue the scale up procedures up to 1000 litres volume.

At the end we will carry out the formulation experiments. We will compare the effectiveness of lyophilized powder in capsule and the inactivated bacterium suspension in physiological salt solution.

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