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Bacteriophage-resistant Staphylococcus aureus mutant confers broad immunity against staphylococcal infection in mice.

Capparelli R, Nocerino N, Lanzetta R, Silipo A, Amoresano A, Giangrande C, Becker K, Blaiotta G, Evidente A, Cimmino A, Iannaccone M, Parlato M, Medaglia C, Roperto S, Roperto F, Ramunno L, Iannelli D - PLoS ONE (2010)

Bottom Line: Acquisition of phage-resistance altered several properties of A172, causing reduced growth rate, under-expression of numerous genes and production of capsular polysaccharide.The same vaccine was also effective when administered as an aerosol.The above results demonstrate that selection for phage-resistance can facilitate bacterial vaccine preparation.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Biotechnology, University of Naples, Portici, Naples, Italy.

ABSTRACT
In the presence of a bacteriophage (a bacteria-attacking virus) resistance is clearly beneficial to the bacteria. As expected in such conditions, resistant bacteria emerge rapidly. However, in the absence of the phage, resistant bacteria often display reduced fitness, compared to their sensitive counterparts. The present study explored the fitness cost associated with phage-resistance as an opportunity to isolate an attenuated strain of S. aureus. The phage-resistant strain A172 was isolated from the phage-sensitive strain A170 in the presence of the M(Sa) phage. Acquisition of phage-resistance altered several properties of A172, causing reduced growth rate, under-expression of numerous genes and production of capsular polysaccharide. In vivo, A172 modulated the transcription of the TNF-alpha, IFN-gamma and Il-1beta genes and, given intramuscularly, protected mice from a lethal dose of A170 (18/20). The heat-killed vaccine also afforded protection from heterologous methicillin-resistant S. aureus (MRSA) (8/10 mice) or vancomycin-intermediate S. aureus (VISA) (9/10 mice). The same vaccine was also effective when administered as an aerosol. Anti-A172 mouse antibodies, in the dose of 10 microl/mouse, protected the animals (10/10, in two independent experiments) from a lethal dose of A170. Consisting predominantly of the sugars glucose and galactose, the capsular polysaccharide of A172, given in the dose of 25 microg/mouse, also protected the mice (20/20) from a lethal dose of A170. The above results demonstrate that selection for phage-resistance can facilitate bacterial vaccine preparation.

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Related in: MedlinePlus

Pulsed field electrophoresis pattern of Sma I digests from the Staphylococcus aureus A170 strain.The A172 strain (not shown) displayed an identical pattern. M: DNA Size Standard, Lambda Ladder (Concatemers of λ cl857 Sam7) (Bio-Rad Laboratories, Hercules, CA).
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pone-0011720-g001: Pulsed field electrophoresis pattern of Sma I digests from the Staphylococcus aureus A170 strain.The A172 strain (not shown) displayed an identical pattern. M: DNA Size Standard, Lambda Ladder (Concatemers of λ cl857 Sam7) (Bio-Rad Laboratories, Hercules, CA).

Mentions: The strains A170 and A172 were positive for the exfoliative toxin B, the Panton-Valentine leukocidin, while negative for the exfoliative toxin A, toxic shock syndrome toxin 1, the leukotoxin LukELukD, the leukotoxin LukM and the staphylococcal enterotoxins-a, -b, -c, -d and –e. The strains A170 and A172 displayed also the same egc type (egc-4), the multilocus sequence type (ST 45) and spa-type (t6668) (spa-type repeats succession: 08-16-02-16-34-13-17-34-34-34-34-34, equivalent to Kreiswirth's IDs: XKAKBEMBBBBB). According to the Ridom spa-server (http://spaserver.ridom.de), t6668 represents a novel spa type. PFGE analysis displayed a close genetic relation between the parental phage MSa-sensitive strain A170 and the derivative phage-resistant strain A172, as proved by the identical PFGE pattern of the Sma I digests (Figure 1). A172 was isolated by growing the parental strain A170 in the presence of increasing concentrations of the MSa phage [10]. In addition to MSa, the strain A172 is also resistant to phages MSa1, MSa2 and MSa3. Prolonged liquid subculture for several months in the absence of phage or prolonged storage at −80°C did not alter the resistance of A172 to MSa.


Bacteriophage-resistant Staphylococcus aureus mutant confers broad immunity against staphylococcal infection in mice.

Capparelli R, Nocerino N, Lanzetta R, Silipo A, Amoresano A, Giangrande C, Becker K, Blaiotta G, Evidente A, Cimmino A, Iannaccone M, Parlato M, Medaglia C, Roperto S, Roperto F, Ramunno L, Iannelli D - PLoS ONE (2010)

Pulsed field electrophoresis pattern of Sma I digests from the Staphylococcus aureus A170 strain.The A172 strain (not shown) displayed an identical pattern. M: DNA Size Standard, Lambda Ladder (Concatemers of λ cl857 Sam7) (Bio-Rad Laboratories, Hercules, CA).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2908692&req=5

pone-0011720-g001: Pulsed field electrophoresis pattern of Sma I digests from the Staphylococcus aureus A170 strain.The A172 strain (not shown) displayed an identical pattern. M: DNA Size Standard, Lambda Ladder (Concatemers of λ cl857 Sam7) (Bio-Rad Laboratories, Hercules, CA).
Mentions: The strains A170 and A172 were positive for the exfoliative toxin B, the Panton-Valentine leukocidin, while negative for the exfoliative toxin A, toxic shock syndrome toxin 1, the leukotoxin LukELukD, the leukotoxin LukM and the staphylococcal enterotoxins-a, -b, -c, -d and –e. The strains A170 and A172 displayed also the same egc type (egc-4), the multilocus sequence type (ST 45) and spa-type (t6668) (spa-type repeats succession: 08-16-02-16-34-13-17-34-34-34-34-34, equivalent to Kreiswirth's IDs: XKAKBEMBBBBB). According to the Ridom spa-server (http://spaserver.ridom.de), t6668 represents a novel spa type. PFGE analysis displayed a close genetic relation between the parental phage MSa-sensitive strain A170 and the derivative phage-resistant strain A172, as proved by the identical PFGE pattern of the Sma I digests (Figure 1). A172 was isolated by growing the parental strain A170 in the presence of increasing concentrations of the MSa phage [10]. In addition to MSa, the strain A172 is also resistant to phages MSa1, MSa2 and MSa3. Prolonged liquid subculture for several months in the absence of phage or prolonged storage at −80°C did not alter the resistance of A172 to MSa.

Bottom Line: Acquisition of phage-resistance altered several properties of A172, causing reduced growth rate, under-expression of numerous genes and production of capsular polysaccharide.The same vaccine was also effective when administered as an aerosol.The above results demonstrate that selection for phage-resistance can facilitate bacterial vaccine preparation.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Biotechnology, University of Naples, Portici, Naples, Italy.

ABSTRACT
In the presence of a bacteriophage (a bacteria-attacking virus) resistance is clearly beneficial to the bacteria. As expected in such conditions, resistant bacteria emerge rapidly. However, in the absence of the phage, resistant bacteria often display reduced fitness, compared to their sensitive counterparts. The present study explored the fitness cost associated with phage-resistance as an opportunity to isolate an attenuated strain of S. aureus. The phage-resistant strain A172 was isolated from the phage-sensitive strain A170 in the presence of the M(Sa) phage. Acquisition of phage-resistance altered several properties of A172, causing reduced growth rate, under-expression of numerous genes and production of capsular polysaccharide. In vivo, A172 modulated the transcription of the TNF-alpha, IFN-gamma and Il-1beta genes and, given intramuscularly, protected mice from a lethal dose of A170 (18/20). The heat-killed vaccine also afforded protection from heterologous methicillin-resistant S. aureus (MRSA) (8/10 mice) or vancomycin-intermediate S. aureus (VISA) (9/10 mice). The same vaccine was also effective when administered as an aerosol. Anti-A172 mouse antibodies, in the dose of 10 microl/mouse, protected the animals (10/10, in two independent experiments) from a lethal dose of A170. Consisting predominantly of the sugars glucose and galactose, the capsular polysaccharide of A172, given in the dose of 25 microg/mouse, also protected the mice (20/20) from a lethal dose of A170. The above results demonstrate that selection for phage-resistance can facilitate bacterial vaccine preparation.

Show MeSH
Related in: MedlinePlus