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Study of potential anti-adherence compounds in a lung cell model as a means for the prophylaxis of bacterial infections in cystic fibrosis
Áčová, Andrea ; Hodek, Petr (advisor) ; Nosková, Libuše (referee)
5 Abstract Cystic fibrosis (CF) is an insidious genetic disease with autosomal-recessive inheritance. The impaired function of the CFTR protein, caused by a mutation in the eponymous gene, results in a wide range of symptoms, the most serious being the effect on the respiratory system. The main impact on the respiratory system is the appearance of thick mucus, which contributes to the accumulation of bacterial cells in the patient's lungs. Life-threatening pathogens include Burkholderia cenocepacia (BC) and Pseudomonas aeruginosa (PA). These bacteria produce several virulence factors such as BC2L-A or BC2L-C lectins in BC and PA-IIL in PA. The virulence factors allow bacteria to bind via protein-saccharide interactions to lung cells with altered glycosylation, which is seen in the CF patient. This diploma thesis focuses on the above-mentioned lectin interaction and the inhibitory effect of hen IgY antibodies or trivalent fucosylated glycoclusters on the adherence of BC (strain ST-32, CCM 7291) / PA (strain PAK, ST 1763) to lung epithelial cells of the CuFi-1 (from a CF patient) / NuLi-1 (from a healthy donor). The possible influence of glycoclusters on PA (PA-lux strain) cell viability was studied, but no significant effect was observed. To verify the expression of the BC2L-C lectin, it was necessary to...
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Study of glycocluster prophylaxis on bacterium adherence model
Áčová, Andrea ; Hodek, Petr (advisor) ; Kubíčková, Božena (referee)
Cystic fibrosis (CF), as autosomal-recessive inherited disease caused by mutation in CFTR gene, induces an ion channel disorder which triggers big amount of CF patients' health problems. The gravest are complications associated with the respiratory system. Accumulation of thick mucus and altered glycosylation in lung lead to increased adherence of bacterial cells, mainly Pseudomonas aeruginosa (PA) and Burkholderia cepacia complex, to lung epithelial cells of CF patients. The presented thesis, focusing on one of the virulence factors of PA, the PA-IIL lectin, studies the adherence of the PA control strain (PAK; ST 1763) under the influence of anti-PA-IIL chicken IgY antibodies and multivalent glycoconjugates based on fucose, to lung epithelial cells collected from healthy person (NuLi-1) and CF patient (CuFi-1). Before the adherence testing, the presence of PA-IIL in PAK culture was checked by "Western Blotting" followed by immunodetection. For quantitative evaluation of the adherence testing, both lung epithelial and bacterial cells were first fluorescently labeled with PKH dyes. The results of spectrofluorimetric measurement after the adherence testing were also compared to microscopic observation of PAK adherence. Different fractions of chicken anti-PA-IIL antibodies from egg yolks showed...
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