Trypanosoma Antigens, Vaccine Compositions, and Related Methods

• US 20090324634A1
• Filed: 04/28/2008
• Published: 12/31/2009
• Est. Priority Date: 04/28/2007
• Status: Active Grant

• Current Assignee: iBio, Inc.
• Original Assignee: iBio, Inc.
• Inventors: Knapp, Elisabeth, Yusibov, Vidadi
• Granted Patent: US 8,778,348 B2
• Time in Patent Office: Days
• US Class Current: 424/191.1
• CPC Class Codes: A61K 2039/552 Veterinary vaccine
• CPC Class Codes: A61K 2039/6075 Viral proteins
• CPC Class Codes: A61K 39/005 Trypanosoma antigens
• CPC Class Codes: C07K 2319/35 containing a fusion for enhanced stability/folding during expression, e.g. fusions with chaperones or thioredoxin
• CPC Class Codes: C07K 2319/40 containing a tag for immunodetection, or an epitope for immunisation
• CPC Class Codes: C12N 15/8257 for the production of primary gene products, e.g. pharmaceutical products, interferon
• CPC Class Codes: C12N 15/8258 for the production of oral vaccines (antigens) or immunoglobulins

Abstract

The present invention relates to the intersection of the fields of immunology and protein engineering, and particularly to antigens and vaccines useful in prevention of infection by Trypanosoma protozoa. Provided are recombinant protein antigens, compositions, and methods for the production and use of such antigens and vaccine compositions.

Claims

1-143. -143. (canceled)

144. An isolated antigen comprising a component of a Trypanosoma protein fused to a thermostable protein or fused to alfalfa mosaic virus (AlMV) coat protein;

wherein the Trypanosoma protein component comprises at least one domain selected from the group consisting of alpha tubulin, beta tubulin, MAP15, and fragments thereof.

145. The isolated antigen of claim 144, wherein the Trypanosoma protein component is a polytope.

146. The isolated antigen of claim 144, wherein the Trypanosoma protein component comprises at least one domain selected from:

(a) alpha tubulin or an alpha tubulin fragment comprising an amino acid sequence as set forth in any of the sequences selected from the group consisting of SEQ ID NOs.;

1 and 4-11; and

(b) beta tubulin or a beta tubulin fragment comprising an amino acid sequence as set forth in any of the sequences selected from the group consisting of SEQ ID NOs.;

2 and 12-22.

147. The isolated antigen of claim 144, wherein the Trypanosoma protein component comprises an amino acid sequence comprising at least two sequences selected from the group consisting of any one of SEQ ID NOs.:

1-27.

148. The isolated antigen of claim 147, wherein the Trypanosoma protein component comprises a portion of alpha tubulin and a portion of beta tubulin, a plurality of portions of alpha tubulin, a plurality of portions of beta tubulin, or full length alpha tubulin and full length beta tubulin.

149. A vaccine composition comprising an antigen comprising a component of a Trypanosoma protein fused to a thermostable protein or fused to AlMV coat protein and a pharmaceutically acceptable carrier;

wherein the Trypanosoma protein component comprises at least one domain selected from the group consisting of a portion of alpha tubulin, a portion of beta tubulin, and a portion of MAP15; and

wherein the composition is capable of eliciting an immune response upon administration to a subject.

150. The vaccine composition of claim 149, wherein the Trypanosoma protein component comprises at least one domain selected from:

(a) alpha tubulin or alpha tubulin fragment comprising an amino acid sequence as set forth in any of the sequences selected from the group consisting of SEQ ID NOs.;

1 and 4-11.

(b) beta tubulin or beta tubulin fragment comprising an amino acid sequence as set forth in any of the sequences selected from the group consisting of SEQ ID NOs.;

2 and 12-22.

151. The vaccine composition of claim 149, wherein the Trypanosoma protein component comprises at least two domains comprising an amino acid sequence as set forth in any of the sequences selected from the group consisting of any one of SEQ ID NOs.:

1-27.

152. The vaccine composition of claim 151, wherein the Trypanosoma protein component comprises at least one portion of alpha tubulin and at least one portion of beta tubulin, a plurality of portions of alpha tubulin, a plurality of portions of beta tubulin, or full length alpha tubulin and full length beta tubulin.

153. The vaccine composition of claim 149, further comprising a second antigen wherein the second antigen comprises a component of a Trypanosoma protein fused to a thermostable protein or fused to AlMV coat protein and a pharmaceutically acceptable carrier;

wherein the Trypanosoma protein component of the first antigen comprises at least one domain selected from the group consisting of a portion of alpha tubulin, a portion of beta tubulin, and a portion of MAP15, and the Trypanosoma protein component of the second antigen comprises at least one domain distinct from the first antigen selected from the group consisting of a portion of alpha tubulin, a portion of beta tubulin, and a portion of MAP15; and

wherein the composition is capable of eliciting an immune response upon administration to a subject.

154. The vaccine composition of claim 149, wherein the antigen is produced in a plant selected from a transgenic plant and a plant transiently expressing the antigen.

155. The vaccine composition of claim 149, wherein the composition comprises antigen which is purified, partially purified, or unpurified from plant cells, a plant, seeds, fruit, or an extract thereof.

156. A vaccine composition comprising at least two antigens, each of which comprises a component of a Trypanosoma protein, wherein the Trypanosoma protein components each comprise at least one domain selected from the group consisting of a portion of alpha tubulin, a portion of beta tubulin, and a portion of MAP15;

wherein at least one of the antigens is fused to a thermostable protein or fused to AlMV coat protein;

wherein the composition comprises a pharmaceutically acceptable carrier; and

wherein the composition is capable of eliciting an immune response upon administration to a subject.

157. The vaccine composition of claim 156, wherein at least one Trypanosoma protein component comprises at least one domain selected from:

(a) alpha tubulin or an alpha tubulin fragment comprising an amino acid sequence as set forth in any of the sequences selected from the group consisting of SEQ ID NOs.;

1 and 4-11; and

(b) beta tubulin or a beta tubulin fragment comprising an amino acid sequence as set forth in any of the sequences selected from the group consisting of SEQ ID NO.;

2 or any one of SEQ ID NOs.;

12-22.

158. A method for inducing a protective immune response against Trypanosoma infection in a subject comprising administering to a subject an effective amount of an anti-Trypanosoma vaccine composition, wherein the administration is sufficient to stimulate production of antigen specific antibodies or stimulate a cellular immune response by the subject;

thereby inducing a protective immune response;

wherein the vaccine composition comprises an antigen comprising a component of a Trypanosoma protein fused to a thermostable protein or fused to AlMV coat protein; and

wherein the Trypanosoma protein component comprises at least one domain selected from the group consisting of a portion of alpha tubulin, a portion of beta tubulin, and a portion of MAP15.

159. A method for producing an antigen protein comprising a component of a Trypanosoma protein fused to a thermostable protein or fused to AlMV coat protein, comprising:

preparing a nucleic acid construct encoding an antigen comprising a component of a Trypanosoma protein fused to a thermostable protein or fused to AlMV coat protein;

introducing the nucleic acid construct into a cell; and

incubating the cell under conditions favorable for expression of the antigen protein, thereby producing the antigen protein;

wherein the Trypanosoma protein component comprises at least one domain selected from the group consisting of a portion of alpha tubulin, a portion of beta tubulin, and a portion of MAP15.

160. The method of claim 159, wherein the Trypanosoma protein component comprises at least one domain selected from:

(a) alpha tubulin or alpha tubulin fragment comprising an amino acid sequence as set forth in any of the sequences selected from the group consisting of SEQ ID NOs.;

1 and 4-11; and

(b) beta tubulin or beta tubulin fragment comprising an amino acid sequence as set forth in any of the sequences selected from the group consisting of SEQ ID NOs.;

2 and 12-22.

161. An isolated nucleic acid construct comprising nucleic acid sequence encoding a component of a Trypanosoma protein fused to a thermostable protein or fused to AlMV coat protein;

and wherein the Trypanosoma protein component comprises at least one domain selected from the group consisting of a portion of alpha tubulin, a portion of beta tubulin, and a portion of MAP15.

162. The isolated nucleic acid construct of claim 161, wherein the Trypanosoma protein component comprises at least one domain selected from:

(a) alpha tubulin or alpha tubulin fragment comprising an amino acid sequence as set forth in any of the sequences selected from the group consisting of SEQ ID NOs.;

1 and 4-11; and

(b) beta tubulin or beta tubulin fragment comprising an amino acid sequence as set forth in any of the sequences selected from the group consisting of SEQ ID NOs.;

2 and 12-22.

163. A host cell comprising the nucleic acid construct of claim 161.

164. The host cell of claim 163, which is a plant cell, wherein the plant is selected from the Nicotiana genus.