Vaccine containing a Shigella bacterium having an enhanced antigenic property
First Claim
1. A vaccine comprising a Shigella bacterium, or an immunogenic fragment or derivative thereof, said Shigella bacterium having an enhanced antigenic property, which is harvested from a culture of a Shigella species grown in vitro in a culture medium with a combination of conditions comprising:
- (a) about 0.05% to about 3% bile or about 0.025% to about 0.6% of one or more bile acids or salts thereof;
(b) at a temperature between about 30°
C. and about 42°
C.;
(c) in air or a gas mixture, wherein the gas mixture comprises (i) about 5% to about 20% CO2 with about 80% to about 95% air;
(ii) about 5% to about 20% CO2 with about 80% to about 95% N2 ;
or (iii) about 5% to about 10% O2 with about 10% to about 20% CO2 with about 70% to about 85% N2 ; and
(d) a divalent cation chelator selected from the group consisting of 0 to about 100 μ
M of 1,2-bis(2-aminophenoxy)ethane-N,N,N'"'"',N'"'"'-tetraacetic acid/acetoxymethyl ester, 0 to about 10 mM of ethylene-bis(oxyethylenenitrilo)-tetraacetic acid, and 0 to about 100 μ
M of ethylene-bis (oxyethylenenitrilo)-tetraacetic acid/acetoxymethyl ester,wherein said Shigella culture is at about early log phase, between early log phase and stationary phase, or at about stationary phase, and the enhanced antigenic property is a higher level of an immunogenic antigen or a new immunogenic antigen when compared to the antigenic property of bacteria from a culture of the Shigella species grown in brain heart infusion broth.
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Abstract
Methods using in vitro processes are disclosed for inducing or enhancing expression of enteric bacterial antigens or virulence factors. The methods, therefore, produce antigenically enhanced bacteria for use in vaccines. Also disclosed are methods for using the antigenically enhanced bacteria as well as the vaccines containing the enteric bacteria. Specifically, a whole enteric bacteria or components thereof, are provided by Shigella species. In addition to this microorganism there are other enteric bacteria, such as Campylobacter species and Helicobacter pylori, which are useful for inducing or enhancing expression of enteric bacterial antigens and/or virulence factors.
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Citations
16 Claims
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1. A vaccine comprising a Shigella bacterium, or an immunogenic fragment or derivative thereof, said Shigella bacterium having an enhanced antigenic property, which is harvested from a culture of a Shigella species grown in vitro in a culture medium with a combination of conditions comprising:
-
(a) about 0.05% to about 3% bile or about 0.025% to about 0.6% of one or more bile acids or salts thereof; (b) at a temperature between about 30°
C. and about 42°
C.;(c) in air or a gas mixture, wherein the gas mixture comprises (i) about 5% to about 20% CO2 with about 80% to about 95% air;
(ii) about 5% to about 20% CO2 with about 80% to about 95% N2 ;
or (iii) about 5% to about 10% O2 with about 10% to about 20% CO2 with about 70% to about 85% N2 ; and(d) a divalent cation chelator selected from the group consisting of 0 to about 100 μ
M of 1,2-bis(2-aminophenoxy)ethane-N,N,N'"'"',N'"'"'-tetraacetic acid/acetoxymethyl ester, 0 to about 10 mM of ethylene-bis(oxyethylenenitrilo)-tetraacetic acid, and 0 to about 100 μ
M of ethylene-bis (oxyethylenenitrilo)-tetraacetic acid/acetoxymethyl ester,wherein said Shigella culture is at about early log phase, between early log phase and stationary phase, or at about stationary phase, and the enhanced antigenic property is a higher level of an immunogenic antigen or a new immunogenic antigen when compared to the antigenic property of bacteria from a culture of the Shigella species grown in brain heart infusion broth. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. A vaccine comprising a Shigella bacterium, or an immunogenic fragment or derivative thereof, said Shigella bacterium having an enhanced antigenic property, which is harvested from a culture of a Shigella species grown in vitro in a culture medium with a combination of conditions comprising:
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(a) a divalent cation chelator selected from the group consisting of about 1.0 to about 25 μ
M of 1,2-bis(2-aminophenoxy)ethane-N,N,N'"'"',N'"'"'-tetraacetic acid/acetoxymethyl ester, about 0.5 to about 10 mM of ethylene-bis(oxyethylenenitrilo)-tetraacetic acid, and about 1 to about 100 μ
M of ethylene-bis(oxyethylenenitrilo)-tetraacetic acidlacetoxymethyl ester;(b) at a temperature between about 30°
C. and about 42°
C.; and(c) in air or a gas mixture, wherein the gas mixture comprises (i) about 5% to about 20% CO2 with about 80% to about 95% air;
(ii) about 5% to about 20% CO2 with about 80% to about 95% N2 ;
or (iii) about 5% to about 10% O2 with about 10% to about 20% CO2 with about 70% to about 85% N2,wherein said Shigella culture is at about early log phase, between early log phase and stationary phase, or at about stationary phase, and the enhanced antigenic property is a higher level of an immunogenic antigen or a new immunogenic antigen when compared to the antigenic property of bacteria from a culture of the Shigella species grown in brain heart infusion broth. - View Dependent Claims (11, 12, 13, 14, 15, 16)
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Specification