The immunomodulatory activity of Staphylococcus aureus products derived from biofilm and planktonic cultures

Abstract

Biofilms are probably one of the most common structures formed by microorganisms in various environments. The higher resistance of such microbial communities to stress conditions, including antibiotics and host immune response, is recently extensively studied. However, the weak activity of phagocytic cells against microbial biofilm is not yet fully understood and explained. The aim of this study was: (1) a qualitative and quantitative comparison of cell components/products released from Staphylococcus aureus biofilm or planktonic cultures, (2) evaluation of the influence of such cell components/products on murine leukocytes secretory function. For this, mouse peritoneal leukocytes were stimulated with biofilm or planktonic staphylococcal cultures or their acellular filtrates, and then the production of cytokines (TNF-α, IL-6, IL-10, MCP-1 and MIP-1α), hemolytic activity and staphylokinase (SAK) production was determined. It was found that similar staphylococcal components/products possessing the immunomodulatory properties, were present in both, biofilm and planktonic filtrates. Moreover, these compounds were similarly active in the stimulation of TNF-α and MCP-1 release from leukocytes. The hemolytic activity and SAK release by planktonic and biofilm cultures were also comparable. What is interesting, stronger stimulatory activity of biofilm-derived components/products of clinical S. aureus strains in the case of MIP-1α, IL-6 and IL-10 was noticed. On the other hand, taking into consideration the reference strains, MIP-1α production was enhanced by "planktonic filtrates". Thus, in our study it was proved, first of all, that biofilm is not a structure fully separated from the external environment. Second, the influence of these S. aureus constituents/metabolites on leukocytes seems to be more strain-dependent than culture phenotype-dependent. The lack of one common profile of biofilm and planktonic S. aureus cultures/filtrates biological activity indicates that the disturbances in cytokines' production could not be the only reason for the so-called "frustrated phagocytosis", connected with enhanced biofilm resistance.

Fig. 1

The presence of Hla in the filtrates obtained from planktonic (PL) or biofilm (B) S. aureus cultures. 1 Sa-8325-4 PL, 2 Sa-8325-4 B, 3 molecular mass standard, 4 Sa-Wood 46 PL, 5 Sa-Wood 46 B, 6 Sa-α11 PL, 7 Sa-α11 B, 8 Sa-α21 B, 9 Hla standard, 20 μg/ml (Sigma, Germany), 10 negative control (TSB medium)

Fig. 2

MIP-1α (a) and MCP-1 (b) production by mouse PMN after 2 and 24 h stimulation with bacterial filtrates obtained from planktonic (PL) or biofilm (B) cultures of S. aureus strains (8325-4, Wood 46, Sa-α11, Sa-α21. c Unstimulated control cells. All data are the means ± SD. *A significant differences (p < 0.02) in chemokines’ concentration compared to control