[Pseudomonas aeruginosa infection in the hospital environment. 2. Pigment production and serotype of Pseudomonas aeruginosa isolated from clinical specimens].
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The effect of tobramycin and polyvalent corpuscular Ps. aeruginosa vaccine on the infectious process in mice with grade III burns inoculated with Ps. aeruginosa 1312 was studied. The highest percentage of the survival (100 per cent) among the animals was observed, when the vaccine was applied locally every day for 7 days. With the use of tobramycin administered intramuscularly for 2 times 95 per cent of the animals survived. When the vaccine was administered subcutaneously, 96.6 per cent of the animals survived. Morphological investigation of the organs and tissues of the mice showed that the vaccine applications to the infected burns promoted rapid elimination of microorganisms in the wounds and prevented development of sepsis due to Ps. aeruginosa. At the same time early epithelization of the wounds was observed (by the 4th-7th day). Intramuscular injections of tobramycin and subcutaneous injection of vaccine also prevented development of sepsis due to Ps. aeruginosa and protected the animals from death. Still no epithelization of the wounds by that period was observed. Microscopic examinations revealed necrosis of the epiderma and derma elements and microbial swarms on the skin surface.
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Newly developed P. aeruginosa vaccine has been shown to be safe and apyrogenic for experimental animals. Immunization with the vaccine in a single injection of 0.5 ml has been found to ensure the protection of 80--98% of mice from lethal infection caused by virulent vaccine strains, with the exception of P. aeruginosa strain No. 1311, for 9 weeks. Immunity to P. aeruginosa strain No. 1311 develops only by day 56 after vaccination. No sharp correlation between the specific agglutinin level and the degree of protective effect induced by the immunization of animals with the polyvalent vaccine has been established. The vaccine has been shown to possess high immunogenicity in respect to clinical P. aeruginosa strains belonging to different serotypes (homo- and heterological vaccine strains).
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Partially purified water-soluble cellular protein antigens have been obtained from 2 newly isolated P. aeruginosa strains and 1 museum P. aeruginosa strain, belonging to immunotypes 2, 3 and 7, by the method of preparative ultracentrifugation. Such trivalent P. aeruginosa vaccine (PV) has proved to be effective in direct and cross experiments of the active protection of mice. The method of ultrafiltration has been used to prepare monovalent PV from a newly isolated strain belonging to immunotype 3/7. This monovalent PV has been found to stimulate immunity to infection with homologous or heterologous P. aeruginosa strains in mice. The comparison of the results obtained in the study of PV prepared by the methods of ultracentrifugation and ultrafiltration suggests that both these methods for the isolation and purification of P. aeruginosa protective protein antigens are equally effective.
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Elafin is a skin-derived serine-protease inhibitor. It is thought to be important to prevent human leukocyte elastase-mediated tissue damage and might play an important role in maintaining the integrity of the human epidermis. Recent studies have provided evidence for an antimicrobial activity of elafin against P. aeruginosa. As gram-negative infections typically occur in barrier-disrupted skin we were interested to determine whether supernatants of the gram-negative bacteria P. aeruginosa and Escherichia coli were capable of inducing elafin expression. Supernatants of various P. aeruginosa strains stimulated elafin mRNA-expression and protein release, whereas supernatants of E. coli did not induce elafin expression. In non-differentiated cells the relative increase of elafin mRNA was much higher (100-fold) than in differentiated cells (sixfold), although the latter exhibited higher constitutive mRNA-expression (150-fold). However, concentrations of secreted elafin were similar in differentiated and non-differentiated cells after stimulation. We could not confirm a bactericidal effect against P. aeruginosa as described previously but observed that its growth was inhibited as demonstrated for different strains in liquid cultures. Growth of E. coli was not affected by elafin. In conclusion, the data presented in this paper suggest that elafin represents an innate immune response factor induced by secreted products of P. aeruginosa. Besides its elastase inhibitory potency elafin is an antimicrobial agent against P. aeruginosa.