Leptospira with increased antigenic mass
The antigenic mass of Leptospira cultures can be significantly increased, independent from any increase in biomass, by supplementing Leptospira cultures with a specific type of fatty acid: a polyunsaturated C18 fatty acid. This provides advantages in the production Leptospira antigens. Also this enables the production of improved Leptospirosis vaccines, that are safer and more effective.
1. A vaccine against Leptospirosis comprising an inactivated Leptospira culture and a pharmaceutically acceptable carrier; wherein said inactivated Leptospira culture is prepared by a method for increasing the antigenic mass of a Leptospira culture comprising the steps of supplementing said Leptospira culture with at least 15 μg/ml of a polyunsaturated C18 fatty acid and inactivating said supplemented Leptospira culture; wherein the polyunsaturated C18 fatty acid is selected from the group consisting of linoleic acid, alpha-linolenic acid, gamma-linolenic acid, and combinations thereof; and wherein said inactivated supplemented Leptospira culture has an increased antigenic mass that exceeds the corresponding increase in biomass; wherein when the Leptospira is serogroup pomona the polyunsaturated C18 fatty acid is not alpha-linolenic acid.
2. A method for producing a vaccine against Leptospirosis comprising the steps of:
a. proliferating a Leptospira culture in an in vitro system,
b. supplementing said Leptospira culture with at least 15 μg/ml of a polyunsaturated C18 fatty acid, wherein the polyunsaturated C18 fatty acid is selected from the group consisting of linoleic acid, alpha-linolenic acid, gamma-linolenic acid, and combinations thereof, and wherein said supplemented Leptospira culture comprises an increased antigenic mass that exceeds the corresponding increase in biomass; wherein when the Leptospira is serogroup pomona the polyunsaturated C18 fatty acid is not alpha-linolenic acid,
c. inactivating and harvesting said supplemented Leptospira culture, and
d. admixing the inactivated Leptospira culture with a pharmaceutically acceptable carrier.
3. The method of claim 2 , wherein between steps a. and b. there is an additional step selected from the group consisting of harvesting, storage, and purification, of the Leptospira culture.
4. The method of claim 2 wherein the polyunsaturated C18 fatty acid is comprised by a vegetable oil.
5. A method for increasing the antigenic mass of a
Leptospira culture comprising the step of supplementing said Leptospira culture with a compound or composition that is relatively rich in a polyunsaturated C18 fatty acid; wherein the polyunsaturated C18 fatty acid is selected from the group consisting of linoleic acid, alpha-linolenic acid, gamma-linolenic acid, and combinations thereof; and wherein the compound or composition that is relatively rich in a polyunsaturated C18 fatty acid is a vegetable oil; wherein when the Leptospira is serogroup pomona the polyunsaturated C18 fatty acid is not alpha-linolenic acid.
6. The method of claim 2 , wherein the polyunsaturated C18 fatty acid comprises between 25 and 250 μg/ml linoleic acid.
7. The method of claim 2 , wherein the polyunsaturated C18 fatty acid comprises between 15 and 300 μg/ml linolenic acid.
8. The vaccine of claim 1 , wherein the polyunsaturated C18 fatty acid comprises between 15 and 300 μg/ml linolenic acid.
9. A vaccine against Leptospirosis comprising the inactivated Leptospira culture of claim 7 .
10. A vaccine against Leptospirosis comprising the inactivated Leptospira culture of claim 6 .
11. A vaccine against Leptospirosis comprising the inactivated Leptospira culture of claim 2 .
12. A vaccine against Leptospirosis comprising the inactivated Leptospira culture of claim 3 .
13. The vaccine of claim 2 , wherein said inactivating of the Leptospira culture to make the inactivated Leptospira culture is done by chemical inactivation.
14. The vaccine of claim 13 , wherein the chemical inactivation is done with formalin.
15. The method of claim 5 , further comprising the step of inactivating the supplemented Leptospira culture.
16. The vaccine of claim 1 , wherein the polyunsaturated C18 fatty acid comprises 25 and 250 μg/ml linoleic acid.
17. A vaccine against Leptospirosis comprising the inactivated Leptospira culture of claim 15 .