Compositions and methods for treatment of microbial infections

• US 7,329,409 B2
• Filed: 06/30/2005
• Issued: 02/12/2008
• Est. Priority Date: 02/13/2002
• Status: Expired due to Term

• Current Assignee: Immunology Laboratories, Inc.
• Original Assignee: Immunology Laboratories, Inc.
• Inventors: Balcarek, John C., Pillich, Jiri
• Primary Examiner(s): GRASER, JENNIFER E
• Application Number:
  US11/170,972
• Publication Number:
  US 20050238670A1
• Time in Patent Office: 957 Days
• Field of Search: 424/243.1, 424/234.1, 424/197.11, 435/243, 435/252.3, 435/320.1, 530/412, 530/413, 530/414, 530/415, 530/416, 530/417
• US Class Current: 424/243.1
• CPC Class Codes: A61K 2039/521 inactivated (killed)
• CPC Class Codes: A61K 2039/54 characterised by the route of administration
• CPC Class Codes: A61K 2039/543 intranasal
• CPC Class Codes: A61K 2039/545 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 2039/57 characterised by the type of response, e.g. Th1, Th2
• CPC Class Codes: A61K 2039/58 raising an immune response against a target which is not the antigen used for immunisation
• CPC Class Codes: A61K 35/741 Probiotics probiotic yeast, e.g. saccharomyces A61K36/06
• CPC Class Codes: A61K 39/02 Bacterial antigens
• CPC Class Codes: A61K 39/0266 Klebsiella
• CPC Class Codes: A61K 39/085 Staphylococcus
• CPC Class Codes: A61K 39/092 Streptococcus
• CPC Class Codes: A61K 39/104 Pseudomonadales, e.g. Pseudomonas
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 11/06 Antiasthmatics
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/00 : Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 : Antibacterial agents
• CPC Class Codes: A61P 37/04 : Immunostimulants
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 1/205 : Bacterial isolates
• CPC Class Codes: C12N 2795/00011 : Details
• CPC Class Codes: C12N 2795/00034 : Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2795/00071 : Demonstrated in vivo effect
• CPC Class Codes: C12N 7/00 : Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00
• CPC Class Codes: C12P 1/04 : by using bacteria
• CPC Class Codes: C12R 2001/22 : Klebsiella
• CPC Class Codes: C12R 2001/385 : Pseudomonas aeruginosa
• CPC Class Codes: C12R 2001/445 : Staphylococcus aureus

Abstract

The present invention relates to methods and compositions for treatment of microbial infections and for the enhancement of resistance to infection. The invention comprises administration of an effective amount of bacterial lysate compositions for the treatment of pathological conditions of microbial infections. The present invention can also be used to enhance the immune system to prevent infections by the administration of an effective amount of the compositions.

Claims

1. A method for inducing an immune response in a subject to Staphylococcus aureus infections, comprising, administering to a human or animal a composition comprising an effective amount of a bacterial lysate to induce an immune response, wherein the bacterial lysate is obtained from lysis of at least one bacterial strain by infection with at least one bacteriophage, wherein,

a) the at least one bacterial strain is a Staphylococcus aureus (S. aureus) strain deposited with the Czech Collection of Microorganisms (CCM) having accession numbers CCM 4992, CCM 4993, CCM 4994, CCM 4995, CCM 4996, CCM 4997, or CCM 4998;

and,

b) the at least one bacteriophage is a bacteriophage deposited with the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmBH (DSM) having accession number DSM 14614, DSM 14615, or DSM 14616.

2. The method of claim 1, wherein the composition is administered to a human subject.

3. The method of claim 1, wherein the composition is administered to an animal subject.

4. The method of claim 3, wherein the composition is administered to a bovine species subject.

5. The method of claim 1, wherein the composition further comprises an adjuvant.

6. The method of claim 1, wherein the bacterial lysate stimulates a metabolic burst in an in vitro assay in polymorphonuclear leukocytes in blood samples used 2 hours after collection.

7. The method of claim 1, wherein the bacterial lysate induces activation of the CD 69 positive subpopulation of CD4 positive T cells in an in vitro assay.

8. A method of preparing a biologically active fraction from a bacterial lysate, comprising,

a) forming a bacterial lysate, wherein the bacterial lysate is obtained from lysis of at least one bacterial strain by infection with at least one bacteriophage, wherein

i) the least one bacterial strain is a Staphylococcus aureus (S. aureus) strain deposited with the Czech Collection of Microorganisms (CCM) having accession numbers CCM 4992, CCM 4993, CCM 4994, CCM 4995, CCM 4996, CCM 4997, or CCM 4998;

and,

ii) the at least one bacteriophage is bacteriophage deposited with the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (DSM) having accession number DSM 14614, DSM 14615, or DSM 14616;

b) adsorbing at least a portion of the bacterial lysate to a chromatographic matrix;

c) eluting one or more fractions from the chromatographic matrix; and

,

d) identifying the one or more fractions as a biologically active fraction.

9. The method of claim 8, wherein the biologically active fraction is identified using an assay measures stimulation of the Toll-like receptor family.

10. The method of claim 8, wherein the biologically active fraction is identified using an assay that measures stimulation of the metabolic burst in polymorphonuclear leukocytes in an in vitro assay comprising blood samples used 2 hours after collection.

11. The method of claim 8, wherein the biologically active fraction is identified using an assay that measures stimulation of the metabolic burst in monocytes in an in vitro assay comprising blood samples used 2 hours after collection.

12. The method of claim 8, wherein the biologically active fraction is identified using an assay that measures the activation of a subpopulation of T cells in an in vitro assay comprising blood samples.

13. The method of claim 12, wherein the subpopulation of CD4 positive T cells is CD 69 positive.

14. The method of claim 8, wherein a chromatographic matrix is a reversed phase chromatographic resin, a cation exchange chromatographic resin, an anion exchange chromatographic resin, an affinity chromatographic resin, or a size exclusion chromatographic resin.

15. The method of claim 8, wherein a chromatographic matrix further comprises a HPLC chromatography system.

16. A method of preparing a bacterial lysate, wherein the bacterial lysate comprises,

a) growing at least one bacterial strain in culture to a concentration that is at least 1×

10⁶ cells per milliliter, wherein the least one bacterial strain is a Staphylococcus aureus (S. aureus) strain deposited with the Czech Collection of Microorganisms (CCM) having accession numbers CCM 4992, CCM 4993, CCM 4994, CCM 4995, CCM 4996, CCM 4997, or CCM 4998;

b) adding to the bacterial strain in culture sufficient amounts of at least one bacteriophage to yield a concentration of at least 1×

10⁸ phage particles per milliliter, wherein the at least one bacteriophage is a bacteriophage deposited with the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmBH (DSM) having accession number DSM 14614, DSM 14615, or DSM 14616;

c) allowing infection and lysis to proceed from 3 to 48 hours; and

,

d) sterilizing the lysate.

17. The method of claim 16, wherein sterilizing the bacterial lysate comprises filtering the lysate through a filter with a pore size less than or equal to about 0.45 μ

m.

18. The method of claim 17, wherein the bacterial lysate further comprises the filtrate.

19. The method of claim 17, wherein the bacterial lysate further comprises the retentate.

20. The method of claim 17, wherein the bacterial lysate further comprises admixing fixed ratios of the filtrate and retentate.