Bacteriophage and their uses

Abstract

There is provided a bacteriophage capable of lysing a P. acnes bacterium and incapable of lysing a bacterium which is not P. acnes , and which is incapable of sustaining lysogeny in a bacterium. There is also provided a pharmaceutical composition comprising such a bacteriophage.

1 . A bacteriophage capable of lysing a  P. acnes  bacterium and incapable of lysing any bacterium which is not  P. acnes , and which is incapable of sustaining lysogeny in a bacterium. 
     
     
         2 . A bacteriophage according to  claim 1  which lacks the ability to express at least one gene necessary for sustaining lysogeny. 
     
     
         3 . A bacteriophage according to  claim 2  having a genome which lacks all or part of at least one gene necessary for sustaining lysogeny. 
     
     
         4 . A bacteriophage according to  claim 1  having a genome which comprises defects in non-coding regions which affect the ability of the phage to sustain lysogeny. 
     
     
         5 . An isolated bacteriophage according to  claim 1  which is naturally occurring. 
     
     
         6 . A bacteriophage according to  claim 1  capable of lysing a plurality of strains of the  P. acnes  bacterium. 
     
     
         7 . A bacteriophage according to  claim 1  capable of lysing at least 5 strains of the  P. acnes  bacterium. 
     
     
         8 . A bacteriophage according to  claim 1  which is isolated and selected from: 103609 (Accession no. NCIMB 41350); 103672 (Accession no. NCIMB 41351); and 1894 (Accession no. NCIMB 41349). 
     
     
         9 . A bacteriophage according to  claim 1  having a genome having sequence identity of at least 88% with the genome of the bacteriophage deposited under Accession No. NCIMB 41349. 
     
     
         10 . A bacteriophage according to  claim 1  having a genome which comprises the DNA sequence of SEQ ID NO: 5, a genome having at least 88% sequence identity with the DNA sequence of SEQ ID NO:5, or a genome which comprises a functional fragment of the DNA sequence of SEQ ID NO: 5. 
     
     
         11 . A bacteriophage according to  claim 10  wherein the functional fragment is selected from within the Open Reading Frames shown in  FIG. 4 . 
     
     
         12 . A bacteriophage according to  claim 10  wherein the functional fragment comprises a DNA sequence having sequence identity of at least 95% with the DNA sequence of SEQ ID NO:8. 
     
     
         13 . A bacteriophage according to  claim 10  wherein the functional fragment comprises the DNA sequence of SEQ ID NO:8. 
     
     
         14 . An isolated bacteriophage according to  claim 1  which is 1894. 
     
     
         15 . A bacteriophage according to  claim 1  having a genome having sequence identity of at least 87% with the genome of the bacteriophage deposited under Accession No. NCIMB 41350. 
     
     
         16 . A bacteriophage according to  claim 1  having a genome which comprises the DNA sequence of SEQ ID NO: 3, a genome having at least 87% sequence identity with the DNA sequence of SEQ ID NO:3, or a genome which comprises a functional fragment of the DNA sequence of SEQ ID NO: 3. 
     
     
         17 . A bacteriophage according to  claim 16  wherein the functional fragment is selected from within the Open Reading Frames shown in  FIG. 2 . 
     
     
         18 . A bacteriophage according to  claim 16  wherein the functional fragment comprises a DNA sequence having sequence identity of at least 95% with the DNA sequence of SEQ ID NO:6. 
     
     
         19 . A bacteriophage according to  claim 16  wherein the functional fragment comprises the DNA sequence of SEQ ID NO:6. 
     
     
         20 . A bacteriophage according to  claim 1  which is 103609. 
     
     
         21 . A bacteriophage according to  claim 1  having a genome having sequence identity of at least 88% with the genome of the bacteriophage deposited under Accession No. NCIMB 41351. 
     
     
         22 . A bacteriophage according to  claim 1  having a genome which comprises the DNA sequence of SEQ ID NO: 4, a genome having at least 88% sequence identity with the DNA sequence of SEQ ID NO:4, or a genome which comprises a functional fragment of the DNA sequence of SEQ ID NO: 4. 
     
     
         23 . A bacteriophage according to  claim 22  wherein the functional fragment is selected from the Open Reading Frames shown in  FIG. 3 . 
     
     
         24 . A bacteriophage according to  claim 22  wherein the functional fragment comprises a DNA sequence having sequence identity of at least 95% with the DNA sequence of SEQ ID NO:7. 
     
     
         25 . A bacteriophage according to  claim 22  wherein the functional fragment comprises the DNA sequence of SEQ ID NO:7. 
     
     
         26 . A bacteriophage according to  claim 1  which is 103672. 
     
     
         27 . A bacteriophage according to  claim 1  having a genome which does not comprise the DNA sequence of SEQ ID NO:9. 
     
     
         28 . A bacteriophage according to  claim 1  having a genome which does not comprise the DNA sequence of SEQ ID NO:10. 
     
     
         29 . A bacteriophage according to  claim 1  having a genome which does not comprise the DNA sequence of SEQ ID NO:11. 
     
     
         30 . A bacteriophage according to  claim 1  having a genome which does not comprise the DNA sequence of SEQ ID NO:12. 
     
     
         31 . A bacteriophage according to  claim 1  having a genome which does not comprise the DNA sequence of SEQ ID NO:13. 
     
     
         32 . A bacteriophage according to  claim 1  modified to comprise a marker molecule. 
     
     
         33 . An isolated polynucleotide having the nucleotide sequence of the genome of a bacteriophage according to  claim 1 . 
     
     
         34 . An isolated polynucleotide comprising the nucleotide sequence of SEQ ID NO:3 or of a functional fragment of the DNA sequence of SEQ ID NO:3, wherein a bacteriophage comprising the polynucleotide is a bacteriophage according to  claim 1 . 
     
     
         35 . An isolated polynucleotide having sequence identity of at least 87% with the DNA sequence of SEQ ID NO:3, wherein a bacteriophage comprising the polynucleotide is a bacteriophage according to  claim 1 . 
     
     
         36 . An isolated polynucleotide according to  claim 34  wherein the functional fragment is selected from within the Open Reading Frames shown in  FIG. 2 . 
     
     
         37 . An isolated polynucleotide according to  claim 34  wherein the functional fragment comprises a DNA sequence having sequence identity of at least 95% with the DNA sequence of SEQ ID NO:6. 
     
     
         38 . An isolated polynucleotide according to  claim 36  wherein the functional fragment comprises the DNA sequence of SEQ ID NO:6. 
     
     
         39 . An isolated polynucleotide comprising the nucleotide sequence of SEQ ID NO:4 or of a functional fragment of the DNA sequence of SEQ ID NO:4, wherein a bacteriophage comprising the polynucleotide is a bacteriophage according to  claim 1 . 
     
     
         40 . An isolated polynucleotide having sequence identity of at least 88% with the DNA sequence of SEQ ID NO:4, wherein a bacteriophage comprising the polynucleotide is a bacteriophage according to  claim 1 . 
     
     
         41 . An isolated polynucleotide according to  claim 39  wherein the functional fragment is selected from within the Open Reading Frames shown in FIG. 
     
     
         42 . An isolated polynucleotide according to  claim 39  wherein the functional fragment comprises a DNA sequence having sequence identity of at least 95% with the DNA sequence of SEQ ID NO:7. 
     
     
         43 . An isolated polynucleotide according to  claim 41  wherein the functional fragment comprises the DNA sequence of SEQ ID NO:7. 
     
     
         44 . An isolated polynucleotide comprising the nucleotide sequence of SEQ ID NO:5 or of a functional fragment of the DNA sequence of SEQ ID NO:5, wherein a bacteriophage comprising the polynucleotide is a bacteriophage according to  claim 1 . 
     
     
         45 . An isolated polynucleotide having sequence identity of at least 88% with the DNA sequence of SEQ ID NO:5, wherein a bacteriophage comprising the polynucleotide is a bacteriophage according to  claim 1 . 
     
     
         46 . An isolated polynucleotide according to  claim 44  wherein the functional fragment is selected from within the Open Reading Frames shown in  FIG. 4 . 
     
     
         47 . An isolated polynucleotide according to  claim 44  wherein the functional fragment comprises a DNA sequence having sequence identity of at least 95% with the DNA sequence of SEQ ID NO:8. 
     
     
         48 . An isolated polynucleotide according to  claim 46  wherein the functional fragment comprises the DNA sequence of SEQ ID NO:8. 
     
     
         49 . An isolated polynucleotide according to  claim 33  having a nucleic acid sequence which does not comprise that of SEQ ID NO:9. 
     
     
         50 . An isolated polynucleotide according to  claim 33  having a nucleic acid sequence which does not comprise that of SEQ ID NO:10. 
     
     
         51 . An isolated polynucleotide according to  claim 33  having a nucleic acid sequence which does not comprise that of SEQ ID NO:11. 
     
     
         52 . An isolated polynucleotide according to  claim 33  having a nucleic acid sequence which does not comprise that of SEQ ID NO:12. 
     
     
         53 . An isolated polynucleotide according to  claim 33  having a nucleic acid sequence which does not comprise that of SEQ ID NO:13. 
     
     
         54 . An isolated polynucleotide having a nucleic acid sequence which is complementary to that of the polynucleotide defined in  claim 33 . 
     
     
         55 . An isolated polynucleotide comprising the polynucleotide according to  claim 33  and a nucleic acid sequence encoding a marker molecule. 
     
     
         56 . An isolated polypeptide having an amino acid sequence encoded by the polynucleotide according to  claim 33 . 
     
     
         57 . A composition comprising at least one bacteriophage according to  claim 1  and an adjuvant, carrier or vehicle. 
     
     
         58 . A composition according to  claim 57  comprising two or more different bacteriophage. 
     
     
         59 . A composition according to  claim 58  wherein each of the two or more bacteriophage is a bacteriophage as defined above. 
     
     
         60 . A composition comprising an isolated polynucleotide according to  claim 33  and/or an isolated polypeptide as defined above and an adjuvant, carrier or vehicle. 
     
     
         61 . A composition according to  claim 57  in a form suitable for oral, intravenous or topical administration. 
     
     
         62 . A composition according to  claim 57  which further comprises at least one further agent selected from antibiotics, anti-comedonals, anti- P. acnes  agents, anti-inflammatories and anti-seborrhoeics. 
     
     
         63 . A composition according to  claim 57  for use in the prevention or treatment of acne and wherein each at least one bacteriophage is present in an effective amount. 
     
     
         64 . A composition according to  claim 57  for use in the prevention or treatment of acne and wherein the isolated polynucleotide and/or the isolated polypeptide is present in an effective amount. 
     
     
         65 . A method of preventing or treating acne comprising administering an effective amount of at least one bacteriophage according to  claim 1  to an individual in need of such prevention or treatment. 
     
     
         66 . A method of preventing or treating acne comprising administering an effective amount of an isolated polynucleotide according to  claim 33  and/or an isolated polypeptide as defined above and/or of a composition as defined above to an individual in need of such prevention or treatment. 
     
     
         67 . A method of improving the appearance of an individual comprising administering to the individual an effective amount of at least one bacteriophage according to  claim 1 . 
     
     
         68 . A method of improving the appearance of an individual comprising administering to the individual an effective amount of an isolated polynucleotide according to  claim 33  and/or an isolated polypeptide as defined above and/or of a composition as defined above. 
     
     
         69 . A method for isolating a bacteriophage capable of lysing a  P. acnes  bacterium, incapable of lysing any bacterium which is not  P. acnes  and is incapable of sustaining lysogeny in a bacterium, comprising:
 a) obtaining a sample of bacteria from a skin surface:   b) isolating from the sample bacteriophage which lyse propionibacteria;   c) testing the bacteriophage to determine if it is capable of lysing at least one  P. acnes  strain;   d) testing the bacteriophage to determine if it is capable of lysing non- P. acnes  bacterial strains;   e) testing the bacteriophage to determine whether it is capable of sustaining lysogeny in a  P. acnes  strain;   f) isolating a bacteriophage which has been shown in steps (c), (d) and (e) to be capable of lysing a  P. acnes  bacterium, incapable of lysing any bacterium which is not  P. acnes , and incapable of sustaining lysogeny in a bacterium.   
     
     
         70 . A method for identifying a bacteriophage which is capable of lysing a  P. acnes  bacterium, incapable of lysing any bacterium which is not  P. acnes  and incapable of sustaining lysogeny in a bacterium, comprising:
 a) exposing a  P. acnes  bacterium to the bacteriophage and determining that the bacterium is lysed;   b) exposing at least three different species of bacteria which are not  P. acnes  bacteria to the bacteriophage and determining that the bacteria are not lysed;   c) determining that the bacteriophage is not capable of sustaining lysogeny in a bacteria.   
     
     
         71 . A bacteriophage isolated using the method according to  claim 69  or identified using the method as defined above. 
     
     
         72 . A bacteriophage obtainable using the method according to  claim 69  or identifiable using a method as defined above.