Compositions for and methods of identifying antigens
AssigneePresident and Fellows of Harvard College
InventorsDarren E. HIGGINS, Michael N. STARNBACH, Todd GIERAHN, Nadia R. ROAN
Life of this patent
Estimating the term…
Abstract
Replicable libraries having discrete members in defined locations for screening for antigens to a pathogenic organism are provided. Also provided are methods for using such libraries as well as a specific antigen, CT788, which induces T-cell activation during a Chlamydia infection.
Claims25
1-14. (canceled)
A vaccine comprising at least one epitope identified by a method of determining whether a library contains or encodes an immunogenic polypeptide, wherein said library comprises at least 20 discrete members in defined locations and said members each comprise a cell or virus comprising a pre-determined open reading frame of a polynucleotide encoding a polypeptide, or a portion thereof, differentially expressed in a neoplastic cell as compared to a corresponding normal cell, said pre-predetermined open reading frame operably linked to a promoter,
- said method comprising:
- (a) individually contacting a member of said library with a second cell capable of (i) endocytosing said cell or said virus and (ii) displaying a polypeptide encoded by said pre-determined open reading frame on its surface through the major histocompatibility complex (MHC class) I pathway;
- (b) individually contacting each member of step (a) with a cytotoxic T lymphocyte (CTL) cell derived from a mammal having, or having previously had, a neoplasm;
- (c) detecting whether said CTL cell is activated, wherein activation of said CTL cell indicates that said member of said library contains said pre-determined open reading frame which encodes at least a portion of said immunogenic polypeptide;
- and
- (d) identifying an epitope sufficient for CTL activation within said polypeptide, or portion thereof, determined to be immunogenic in step (c),
- further comprising a pharmaceutically acceptable carrier.
An epitope cocktail comprising at least 5 epitopes identified using a method of determining whether a library contains or encodes an immunogenic polypeptide, wherein said library comprises at least 20 discrete members in defined locations and said members each comprise a cell or virus comprising a pre-determined open reading frame of a polynucleotide encoding a polypeptide, or a portion thereof, differentially expressed in a neoplastic cell as compared to a corresponding normal cell, said pre-predetermined open reading frame operably linked to a promoter,
- said method comprising:
- (a) individually contacting a member of said library with a second cell capable of (i) endocytosing said cell or said virus and (ii) displaying a polypeptide encoded by said pre-determined open reading frame on its surface through the major histocompatibility complex (MHC class) I pathway;
- (b) individually contacting each member of step (a) with a cytotoxic T lymphocyte (CTL) cell derived from a mammal having, or having previously had, a neoplasm;
- (c) detecting whether said CTL cell is activated, wherein activation of said CTL cell indicates that said member of said library contains said pre-determined open reading frame which encodes at least a portion of said immunogenic polypeptide;
- and
- (d) identifying an epitope sufficient for CTL activation within said polypeptide, or portion thereof, determined to be immunogenic in step (c).
A composition comprising at least one immunogenic polypeptide, or a portion thereof, identified by a method of determining whether a library contains or encodes an immunogenic polypeptide, wherein said library comprises at least 20 discrete members in defined locations and said members each comprise a cell or virus comprising a pre-determined open reading frame of a polynucleotide encoding a polypeptide, or a portion thereof, differentially expressed in a neoplastic cell as compared to a corresponding normal cell, said pre-predetermined open reading frame operably linked to a promoter,
- said method comprising:
- (a) individually contacting a member of said library with a second cell capable of (i) endocytosing said cell or said virus and (ii) displaying a polypeptide encoded by said pre-determined open reading frame on its surface through the major histocompatibility complex (MHC class) I pathway;
- (b) individually contacting each member of step (a) with a cytotoxic T lymphocyte (CTL) cell derived from a mammal having, or having previously had, a neoplasm;
- and
- (c) detecting whether said CTL cell is activated, wherein activation of said CTL cell indicates that said member of said library contains said pre-determined open reading frame which encodes at least a portion of said immunogenic polypeptide.
A vaccine comprising at least one immunogenic polypeptide, or a portion thereof, identified by a method of determining whether a library contains or encodes an immunogenic polypeptide, wherein said library comprises at least 20 discrete members in defined locations and said members each comprise a cell or virus comprising a pre-determined open reading frame of a polynucleotide encoding a polypeptide, or a portion thereof, differentially expressed in a neoplastic cell as compared to a corresponding normal cell, said pre-predetermined open reading frame operably linked to a promoter,
- said method comprising:
- (a) individually contacting a member of said library with a second cell capable of (i) endocytosing said cell or said virus and (ii) displaying a polypeptide encoded by said pre-determined open reading frame on its surface through the major histocompatibility complex (MHC class) I pathway;
- (b) individually contacting each member of step (a) with a cytotoxic T lymphocyte (CTL) cell derived from a mammal having, or having previously had, a neoplasm;
- and
- (c) detecting whether said CTL cell is activated, wherein activation of said CTL cell indicates that said member of said library contains said pre-determined open reading frame which encodes at least a portion of said immunogenic polypeptide,
- further comprising a pharmaceutically acceptable carrier.
22-38. (canceled)
50-58. (canceled)
(canceled)
62-73. (canceled)
A pharmaceutical composition comprising polypeptide comprising a CT788 polypeptide or comprising an immunogenic fragment of said CT788 polypeptide, and a pharmaceutically acceptable carrier.
75-80. (canceled)
Dependencies are inferred from the claim text.