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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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Phage MSA is inhibited by N-acetyl-glucosamine (GlcNAc), the teichoic acid from A170 (A170TA) or by treatment with N-acetyl-glucosaminidase (GlcNA-ase) from Canavalia ensiformis; phage MSA is not inhibited by the teichoic acid from A172 (A172TA) or glucose.(A) GlcNAc inhibits the lysis of the A170 strain by the phage MSa, while glucose (B) does not. (C) A170TA inhibits the lysis of the A170 strain by the phage MSA, while A172TA does not. (D) The phage-sensitive strain A170 grows in the presence of the MSa phage, if pre-treated with GlcAc-ase (4 U/tube; 2 h at 37°C).
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pone-0011720-g005: Phage MSA is inhibited by N-acetyl-glucosamine (GlcNAc), the teichoic acid from A170 (A170TA) or by treatment with N-acetyl-glucosaminidase (GlcNA-ase) from Canavalia ensiformis; phage MSA is not inhibited by the teichoic acid from A172 (A172TA) or glucose.(A) GlcNAc inhibits the lysis of the A170 strain by the phage MSa, while glucose (B) does not. (C) A170TA inhibits the lysis of the A170 strain by the phage MSA, while A172TA does not. (D) The phage-sensitive strain A170 grows in the presence of the MSa phage, if pre-treated with GlcAc-ase (4 U/tube; 2 h at 37°C).

Mentions: Phages often use teichoic acids for adsorption on the cell wall of Gram positive bacteria. They can use as receptor the glucose side chains of Bacillus subtilis teichoic acids [26], [27] or the N-acetylglucosamine (GlcNAc) side chains of S. aureus [28]. To identify the receptor site of the MSa phage, the A170 and A172 strains were grown in the presence or absence of 5–20 mM GlcNAc or 5–20 mM glucose. GlcNAc inhibited the lysis of A170 by phage MSA, while glucose did not. GlcNAc inhibition was dose-dependent (Figure 5A–B). The experiment was repeated using the teichoic acid from (A170TA) or from (A172TA) as inhibitor. Since the quantity of teichoic acid that is isolated from the A170 or A172 strains varies significantly (2.4 mg from 1010 CFU A170; 0.9 mg from 1010 CFU A172), the experiment was conducted using an ample concentration range of the A170TA and A172TA reagents. Under these conditions, phage inactivation occurred with A170TA, but not with A172TA (Figure 5C). A170 bacteria were then treated with N-acetylglucosaminidase (4 U/tube; 2 h at 37°C). Enzymatic hydrolysis of GlcNAc from the A170 bacteria destroyed phage lysis capacity (Figure 5D). Finally, analysis of teichoic acids from the A170 (wild) and A172 (mutant) strains of S. aureus displayed the absence of ribitol (Figure 6A) and terminal GlcNAc (t-GlcNAc) residues (Figure 6B) from A172TA. Collectively, the experiments described above demonstrate that A172 conforms with the tendency of several bacterial species - B. subtilis [26], [27], S. aureus [28], Salmonella enterica [18], [29] - to acquire phage-resistance by altering a cell wall polysaccharide component.


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)

Phage MSA is inhibited by N-acetyl-glucosamine (GlcNAc), the teichoic acid from A170 (A170TA) or by treatment with N-acetyl-glucosaminidase (GlcNA-ase) from Canavalia ensiformis; phage MSA is not inhibited by the teichoic acid from A172 (A172TA) or glucose.(A) GlcNAc inhibits the lysis of the A170 strain by the phage MSa, while glucose (B) does not. (C) A170TA inhibits the lysis of the A170 strain by the phage MSA, while A172TA does not. (D) The phage-sensitive strain A170 grows in the presence of the MSa phage, if pre-treated with GlcAc-ase (4 U/tube; 2 h at 37°C).
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2908692&req=5

pone-0011720-g005: Phage MSA is inhibited by N-acetyl-glucosamine (GlcNAc), the teichoic acid from A170 (A170TA) or by treatment with N-acetyl-glucosaminidase (GlcNA-ase) from Canavalia ensiformis; phage MSA is not inhibited by the teichoic acid from A172 (A172TA) or glucose.(A) GlcNAc inhibits the lysis of the A170 strain by the phage MSa, while glucose (B) does not. (C) A170TA inhibits the lysis of the A170 strain by the phage MSA, while A172TA does not. (D) The phage-sensitive strain A170 grows in the presence of the MSa phage, if pre-treated with GlcAc-ase (4 U/tube; 2 h at 37°C).
Mentions: Phages often use teichoic acids for adsorption on the cell wall of Gram positive bacteria. They can use as receptor the glucose side chains of Bacillus subtilis teichoic acids [26], [27] or the N-acetylglucosamine (GlcNAc) side chains of S. aureus [28]. To identify the receptor site of the MSa phage, the A170 and A172 strains were grown in the presence or absence of 5–20 mM GlcNAc or 5–20 mM glucose. GlcNAc inhibited the lysis of A170 by phage MSA, while glucose did not. GlcNAc inhibition was dose-dependent (Figure 5A–B). The experiment was repeated using the teichoic acid from (A170TA) or from (A172TA) as inhibitor. Since the quantity of teichoic acid that is isolated from the A170 or A172 strains varies significantly (2.4 mg from 1010 CFU A170; 0.9 mg from 1010 CFU A172), the experiment was conducted using an ample concentration range of the A170TA and A172TA reagents. Under these conditions, phage inactivation occurred with A170TA, but not with A172TA (Figure 5C). A170 bacteria were then treated with N-acetylglucosaminidase (4 U/tube; 2 h at 37°C). Enzymatic hydrolysis of GlcNAc from the A170 bacteria destroyed phage lysis capacity (Figure 5D). Finally, analysis of teichoic acids from the A170 (wild) and A172 (mutant) strains of S. aureus displayed the absence of ribitol (Figure 6A) and terminal GlcNAc (t-GlcNAc) residues (Figure 6B) from A172TA. Collectively, the experiments described above demonstrate that A172 conforms with the tendency of several bacterial species - B. subtilis [26], [27], S. aureus [28], Salmonella enterica [18], [29] - to acquire phage-resistance by altering a cell wall polysaccharide component.

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