Extracellular vesicles derived from gram-positive bacteria, and use thereof

• US 8,969,653 B2
• Filed: 08/26/2010
• Issued: 03/03/2015
• Est. Priority Date: 09/01/2009
• Status: Active Grant

• Current Assignee: Aeon Medix Inc.
• Original Assignee: Aeon Medix Inc.
• Inventors: Gho, Yong Song, Kim, Yoon Keun, Lee, Eun Young, Hong, Sung Wook, Kim, Ji Hyun, Choi, Seng Jin
• Primary Examiner(s): Schultz, Doug
• Application Number:
  US13/394,113
• Publication Number:
  US 20120159658A1
• Time in Patent Office: 1,650 Days
• US Class Current: 800/9
• CPC Class Codes: A01K 2207/00 Modified animals
• CPC Class Codes: A01K 2267/0375 Animal model for cardiovascular diseases
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12P 1/04 by using bacteria
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: Y02P 20/582 Recycling of unreacted starting or intermediate materials

Abstract

The present application relates to extracellular vesicles (EVs) derived from gram-positive bacteria. In detail, the present application provides animal models of disease using extracellular vesicles derived from gram-positive bacteria, provides a method for screening an active candidate substance which is capable of preventing or treating diseases through the animal models of disease, provides vaccines for preventing or treating diseases caused by extracellular vesicles derived from gram-positive bacteria, and provides a method for diagnosing the causative factors of diseases caused by gram-positive bacteria using extracellular vesicles.

Claims

1. A method of causing to induce antibody production or a T cell immune response against Gram-positive bacteria, the method comprising:

isolating extracellular vesicles from Gram-positive bacteria with a cell wall, the extracellular vesicles comprising a biological substance of the Gram-positive bacteria; and

providing a composition comprising extracellular vesicles and a pharmaceutically acceptable carrier for administering the composition to a subject in need of such induction at a sublethal dose,

wherein isolating comprises;

culturing Gram-positive bacteria in a culture medium,

obtaining supernatant of the culture medium by centrifugation, and

filtering the supernatant with a filter having a pore size sufficient to isolate extracellular vesicles contained in the supernatant.

2. The method of claim 1, wherein the Gram-positive bacteria comprise bacteria of Firmicutes phylum.

3. The method of claim 1, wherein the Gram-positive bacteria comprise bacteria of a genus selected from the group consisting of Staphylococcus, Streptococcus, Enterococcus, Bacillus, Corynebacterium, Norcardia, Clostridium, Actinobacteria and Listeria.

4. The method of claim 1, wherein the Gram-positive bacteria are selected from the group consisting of Enterococcus faecalis, Staphylococcus aureus, Staphylococcus epidermidis, and Bacillus subtilis.

5. The method of claim 1, wherein the Gram-positive bacteria comprise bacteria of Mollicutes class.

6. The method of claim 1, wherein the Gram-positive bacteria comprise Mycoplasma.

7. The method of claim 1, wherein the Gram-positive bacteria comprise transformed bacteria.

8. The method of claim 1, wherein the extracellular vesicles contain a drug or an immunostimulant.

9. The method of claim 1, wherein the biological substance comprises proteins of the Gram-positive bacteria.

10. The method of claim 1, wherein the administration is selected from the group consisting of dermal administration, intravenous injection, intranasal administration, sublingual administration, intratracheal inhalation, oral administration, and intrarectal administration.

11. The method of claim 1, wherein the subject is in need of treating or inhibiting a disease selected from the group consisting of a dermal disease, a respiratory disease, a digestive disease, a genital disease, a vascular disease, a metabolic disease, a lung disease, a bone disease, and a cranial nerve disease.

12. The method of claim 1, wherein the subject is in need of treating or inhibiting skin infection, respiratory infection, urogenital infection, bone infection, central nervous system infection, and sepsis.

13. The method of claim 1, wherein the subject is in need of treating or inhibiting a disease selected from the group consisting of sepsis, Pneumonia and atopic dermatitis.

14. The method of claim 1, wherein the method causes to induce IgG antibody production.

15. The method of claim 1, wherein the method causes to induce Th1- and Th7-mediated immune response.

16. The method of claim 1, the method causes to induce production of IFN-γ

and IL-17.

17. The method of claim 1, wherein the pore size is 0.22 μ

m.