PROCESSED MICROBIAL EXTRACELLULAR VESICLES

1 . A pharmaceutical composition comprising isolated processed microbial extracellular vesicles (pmEVs).

2 . The pharmaceutical composition of claim 1 , wherein at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% of the microbial-derived content of the pharmaceutical composition is pmEVs.

3 . The pharmaceutical composition of claim 1 or claim 2 for use in the treatment of a disease via immune suppression.

4 . The pharmaceutical composition of claim 1 or claim 2 for use in the treatment of a disease via immune activation.

5 . The pharmaceutical composition of claim 1 or claim 2 for use in the treatment of a disease via activation or enhancement of one or more immune responses in the subject.

6 . The pharmaceutical composition of claim 1 or claim 2 for use in the treatment of a disease via promotion of immune suppression in the subject.

7 . The pharmaceutical composition of any one of claims 2 to 6 , wherein the disease is a cancer, an autoimmune disease, an inflammatory disease, or a metabolic disease.

8 . The pharmaceutical composition of any one of claims 1 to 7 , comprising a therapeutically effective amount of the pmEVs.

9 . The pharmaceutical composition of any one of claims 1 to 8 , wherein the composition activates innate antigen presenting cells.

10 . The pharmaceutical composition of any one of claims 1 to 9 , wherein the composition has one or more beneficial immune effects outside the gastrointestinal tract when orally administered.

11 . The pharmaceutical composition of any one of claims 1 to 10 , wherein the composition modulates immune effects outside the gastrointestinal tract in the subject when orally administered.

12 . The pharmaceutical composition of any one of claims 1 to 11 , wherein the composition comprises pmEVs from one strain of bacteria.

13 . The pharmaceutical composition of any one of claims 1 to 12 , wherein the pmEVs are lyophilized (e.g., the lyophilized product further comprises a pharmaceutically acceptable excipient).

14 . The pharmaceutical composition of any one of claims 1 to 13 , wherein the pmEVs are gamma irradiated.

15 . The pharmaceutical composition of any one of claims 1 to 14 , wherein the pmEVs are UV irradiated.

16 . The pharmaceutical composition of any one of claims 1 to 15 , wherein the pmEVs are heat inactivated.

17 . The pharmaceutical composition of claim 16 , wherein the pmEVs are heat inactivated at about 50° C. for two hours or at about 90° C. for two hours.

18 . The pharmaceutical composition of any one of claims 1 to 17 , wherein the pmEVs are acid treated.

19 . The pharmaceutical composition of any one of claims 1 to 18 , wherein the pmEVs are oxygen sparge.

20 . The pharmaceutical composition of claim 19 , wherein the pmEVs are ozygen sparged at about 0.1 vvm for at least two hours.

21 . The pharmaceutical composition of any one of claims 1 to 20 , wherein the dose of pmEVs is about 2×10 6 to about 2×10 16 particles.

22 . The pharmaceutical composition of any one of claims 1 to 21 , wherein the dose of pmEVs is about 5 mg to about 900 mg total protein.

23 . The pharmaceutical composition of any one of claims 1 to 22 , wherein the pharmaceutical composition is a solid dose form.

24 . The pharmaceutical composition of claim 23 , wherein the solid dose form comprises a tablet, a minitablet, a capsule, a pill, or a powder, or a combination of the foregoing.

25 . The pharmaceutical composition of claim 23 or 24 , wherein the solid dose form further comprises a pharmaceutically acceptable excipient.

26 . The pharmaceutical composition of any one of claims 23 to 25 , wherein the solid dose form comprises an enteric coating.

27 . The pharmaceutical composition of any one of claims 23 to 26 , wherein the solid dose form is formulated for oral administration.

28 . The pharmaceutical composition of any one of claims 1 to 22 , wherein the pharmaceutical composition is in the form of a suspension.

29 . The pharmaceutical composition of claim 28 , wherein the suspension is formulated for oral administration.

30 . The pharmaceutical composition of claim 29 , wherein the suspension comprises PBS, and optionally, sucrose or glucose.

31 . The pharmaceutical composition of claim 28 , wherein the suspension is formulated for intravenous, intraperitoneal, or intratumoral administration.

32 . The pharmaceutical composition of claim 31 , wherein the suspension comprises PBS.

33 . The pharmaceutical composition of any one of claims 28 to 32 , wherein the suspension further comprises a pharmaceutically acceptable excipient or a buffer.

34 . The pharmaceutical composition of any one of claims 1 to 33 , wherein the pmEvs are from Gram positive bacteria.

35 . The pharmaceutical composition of any one of claims 1 to 33 , wherein the pmEvs are from Gram negative bacteria.

36 . The pharmaceutical composition of claim 35 , wherein the Gram negative bacteria belongs to the class Negativicutes.

37 . The pharmaceutical composition of any one of claims 1 to 36 , wherein the pmEVs are from aerobic bacteria, anaerobic bacteria, acidophile bacteria, alkaliniphile bacteria, neutralophile bacteria, fastidious bacteria, nonfastidiouius bacteria, or a combination thereof.

38 . The pharmaceutical composition of any one of claims 1 to 37 , wherein the pmEVs are from one or more bacterial strain listed in Table 1, Table 2 or Table 3.

39 . The pharmaceutical composition of any one of claims 1 to 38 , wherein the composition further comprises one or more additional therapeutic agents.

40 . Use of a pharmaceutical composition of any one of claims 1 to 39 for the preparation of a medicament for the treatment of a disease.

41 . The use of claim 49 , wherein the disease is a cancer, an autoimmune disease, an inflammatory disease, a dysbiosis, and/or a metabolic disease.

42 . A method of treating a subject comprising administering to the subject a pharmaceutical composition of any one of claims 1 to 41 .

43 . The method of claim 42 , wherein the pmEVs are from bacteria that have been gamma irradiated, UV irradiated, heat inactivated, acid treated, oxygen sparged, or a combination thereof.

44 . The method of claim 42 , wherein the pmEVs are from live bacteria.

45 . The method of any one of claims 42 to 44 , wherein the composition activates or enhances of one or more immune responses in the subject.

46 . The method of claim 45 , wherein the one or more immune responses comprises a systemic immune response.

47 . The method of any one of claims 42 to 44 , wherein the composition suppresses an immune response in the subject.

48 . The method of any one of claims 42 to 44 , wherein the composition promotes immune activation in the subject.

49 . The method of any one of claims 42 to 48 , wherein the pharmaceutical composition comprising the pmEVs has comparable potency or increased potency compared to a pharmaceutical composition that contains whole microbes from the same bacterial strain from which the pmEVs were produced).

50 . The method of any one of claims 42 to 48 , wherein the pharmaceutical composition comprising the pmEVs has more therapeutically active microbial material compared to a pharmaceutical composition that contains whole microbesfrom which the pmEVs were obtained.

51 . The method of any one of claims 42 to 50 , wherein the subject is in need of treatment for a cancer.

52 . The method of any one of claims 42 to 50 , wherein the subject is in need of treatment for an autoimmune disease and/or an inflammatory disease.

53 . The method of any one of claims 42 to 50 , wherein the subject is in need of treatment for a dysbiosis.

54 . The method of any one of claims 42 to 50 , wherein the subject is in need of treatment for a metabolic disease.

55 . The method of any one of claims 42 to 50 , wherein the pharmaceutical composition is administered in combination with an additional therapeutic agent.

56 . The method of any one of claims 42 to 55 , wherein the composition comprises pmEVs from one strain of bacteria.

57 . The method of any one of claims 42 to 56 , wherein the pmEVs are lyophilized.

58 . The method of any one of claims 42 to 57 , wherein the pharmaceutical composition is orally administered.

59 . The method of any one of claims 42 to 57 , wherein the pharmaceutical composition is administered intravenously.

60 . The method of any one of claims 42 to 57 , wherein the pharmaceutical composition is administered intratumorally.

61 . The method of any one of claims 42 to 57 , wherein the pharmaceutical composition is administered subtumorally.

62 . The method of any one of claims 42 to 57 , wherein the pharmaceutical composition is administered by injection.

63 . A method for preparing a pharmaceutical composition comprising pmEVs in a suspension, the method comprising: combining pmEVs with a pharmaceutically acceptable buffer, thereby preparing the pharmaceutical composition.

64 . The method of claim 63 , wherein the pharmaceutically acceptable buffer comprises PBS.

65 . The method of claim 63 or 64 , wherein the suspension further comprises sucrose or glucose.

66 . The method of any one of claims 63 to 65 , wherein the pmEVs comprise about 2×10 6 to about 2×10 16 particles of pmEVs.

67 . The method of any one of claims 63 to 66 , wherein the pmEVs comprise about 5 mg to about 900 mg total protein.

68 . A pharmaceutical composition prepared by the method of any one of claims 62 to 67 .

69 . A method for preparing a solid dose form of pharmaceutical composition comprising pmEVs (e.g., a therapeutically effective amount thereof) in a solid dose form, the method comprising:

a) combining pmEVs with a pharmaceutically acceptable excipient; and

b) compressing the combined pmEVs and pharmaceutically acceptable excipient; thereby preparing a solid dose form of a pharmaceutical composition.

70 . The method of claim 69 , further comprising enterically coating the solid dose form.

71 . The method of claim 69 or 70 , wherein the solid dose form comprises a tablet or a minitablet.

72 . The method of any one of claims 69 to 71 , wherein the composition comprises pmEVs from one strain of bacteria.

73 . The method of any one of claims 69 to 72 , wherein the pmEVs are lyophilized.

74 . The method of any one of claims 69 to 73 , wherein the pmEVs comprise about 2×10 6 to about 2×10 16 particles.

75 . The method of any one of claims 69 to 74 , wherein the pmEVs comprise about 5 mg to about 900 mg total protein.

76 . A pharmaceutical composition prepared by the method of any one of claims 69 to 75 .