The inhibition of microbial adhesion in the presence of immune compounds on the human cell culture model

Abstract number: 1135_25

Afinogenova A.G., Afinogenov G.E.

Objectives:  

To comparatively evaluate the influence of the commercial preparations of normal human immunoglobulin (Russia), Intraglobin and Pentaglobin (Biotest, Germany) on the clinical strain S. aureus adhesion to the human embryonic skin fibroblasts in culture.

Methods:  

The clinical isolate of S. aureus with the highest adhesive capacity was selected in the model of it's co-incubation with the target cells in culture. The inoculums was prepared at a baseline concentration of 108 cfu/ml after 24 h grown on meat-peptone agar. The antiadhesive activity of the normal human immunoglobulin (NHI), Intraglobin and Pentaglobin at a baseline concentration of 20 mg/ml was assessed. The estimation of the influence of the agents under investigation on the bacterial adhesion process was performed on the above mentioned model of co-incubation of microorganism and the target-cells in culture as well as using 1 h pre-incubation with agents to be tested. Also the experiments were held in the presence Protein A from the staphylococcus cell wall as well as clostridial collagenasum and animal hyaluronidasum. The intensity of bacterial adhesion and antiadhesive properties of used compounds were expressed as following indices: Index of adhesion (IA) calculated as mean number of attached microbes per one eukaryotic cell; Percent of affected cells of the monolayer (AC%); Microbial contamination of the monolayer (MC) obtained as a product of IA•AC%; Percent of inhibition of bacterial adhesion (II%) calculated from the figures of IA with respect to the control.

Results:  

All preparations revealed more or less significant inherent antiadhesive activity in vitro, which became apparent in the reduction of all indices characterizing colonization of target-cells. Most effective proved to be the preparations with 1 hour pre-incubation with clinical isolate of S. aureus which protected fibroblasts against the bacterial adhesion in more than 92% in this case.

Conclusion:  

The protective effect of NHI and Intraglobin disappeared with addition of Protein A, collagenasum or hyaluronidasum to nutrient medium but remained the same for Pentaglobin. According to its specific polymeric structure and prospective form as well as content of IgM Pentaglobin possesses much more antiadhesive activity in the presence of Protein A, collagenasum or hyaluronidasum. In the presence of Protein A and enzymes IgM makes the antiadhesive activity of Pentaglobin irreversible.